Dehydrophenylastemizole compounds, methods of making and using the same

Abstract

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

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to dehydrophenylasiustin compounds, their preparation methods, and their applications. Background Technology

[0002] Plinabulin (NPI-2358) is a 2,5-diketopiperazine derivative, obtained through structural modification of the low-molecular-weight cyclic dipeptide phenylahistin, a metabolite produced by the marine fungus *Aspergillus* sp. It is a microtubule-binding agent. Plinabulin selectively acts near the colchicine binding site in endothelial tubules, inhibiting tubule polymerization and blocking microtubule assembly, thereby disrupting the endothelial cytoskeleton and inhibiting tumor blood flow. Experiments have shown that plinabulin acts on cells, causing cell arrest in early mitosis and inducing cell death. Recent studies have found that punabulin is a differentiation immune and stem cell modulator, as well as a guanine nucleotide exchange factor (GEF-H1) activator, which can target and alter the tumor microenvironment and disrupt tumor angiogenesis through multiple mechanisms of action. As an effective antigen-presenting cell (APC) inducer (by activating dendritic cell maturation), punabulin's durable anticancer effect is related to its activation of T cells (Cell Reports 2019, 28:13, 3367-3386).

[0003] This candidate drug, currently being developed by BeyondSpring Pharmaceuticals, is undergoing Phase III clinical trials. It is being used in combination with docetaxel for the treatment of non-small cell lung cancer and for the prevention of chemotherapy-induced neutropenia (CIN) in non-myeloid malignancies. An NDA has also been submitted for its whitening indication.

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

[0005]

[0006] Plinabulin has the molecular formula C 19 H 20 N4O2, with a molecular weight of 336.39 and CAS number 714272-27-2, exhibits good stability but has poor water solubility and significant toxic side effects. Further development of highly active and soluble microtubule inhibitors is still needed. Summary of the Invention

[0007] The technical problem to be solved by this invention is to address the deficiency of the relatively simple structure of existing microtubule inhibitors. This invention provides dehydrophenylastistin compounds, their preparation methods and applications. The compounds of this invention have novel structures and good activity.

[0008] To achieve the above-mentioned objectives, the present invention employs the following technical solution:

[0009] This invention provides compounds of formula (I), their tautomers, their stereoisomers, their pharmaceutically acceptable salts, or solvates thereof (referring to the aforementioned compounds of formula (I), their tautomers, their stereoisomers, or their pharmaceutically acceptable salts):

[0010]

[0011] in,

[0012] R 1 H, deuterium, halogen, C1-C8 alkyl, C1-C8 alkyl substituted with one or more halogens, C1-C8 alkoxy, C1-C8 alkoxy substituted with one or more halogens, C2-C8 alkenyl, benzoyl, benzoyl substituted with one or more halogens, phenoxy, or "phenoxy substituted with one or more halogens".

[0013] R 1a R 1b and R 1c It can be independently H, deuterium, halogen, or C1-C8 alkyl;

[0014] Or, R 1 R 1a Together with the carbon atoms they are attached to, they form C6-C. 10 The aryl group or "a 3-10 membered heteroaryl group selected from one or more of N, O and S, with 1-5 heteroatoms";

[0015] Or, R 1 R 1b Together with the carbon atoms they are attached to, they form C6-C. 10 The aryl group or "a 3-10 membered heteroaryl group selected from one or more of N, O and S, with 1-5 heteroatoms";

[0016] Or, R 1b R 1c Together with the carbon atoms they are attached to, they form C6-C. 10 The aryl group or "a 3-10 membered heteroaryl group whose heteroatoms are selected from one or more of N, O and S, and whose heteroatoms number 1-5";

[0017] The ring C is

[0018] R 2 It can be H, halogen, or benzyl;

[0019] R3 For one or more R 3-1 Substituted C1-C8 alkyl, C2-C8 alkynyl, or C2-C8 alkenyl;

[0020] R 3-1 Independently hydroxyl, C1-C8 alkoxy, -OTBS, NR 3-1-1 R 3-1-2 , or -OC(=O)-R 3 -1-3 ;

[0021] R 3-1-1 and R 3-1-2 Independently H, C1-C8 alkyl, or -C(=O)-OR 3-1-1-1 ;R 3-1-1-1 It is a C1-C8 alkyl or benzyl group;

[0022] R 3-1-3 It is a C1-C8 alkyl group or is composed of one or more R groups. 3-1-3-1 Replacement C6-C 10 Aryl; R 3-1-3-1 It can be independently OH or benzyloxy group;

[0023] R 4 It is a C1-C8 alkyl or C6-C 10 Aryl;

[0024] R 5 and R 6 The definition satisfies the following conditions: (1) R 5 For H, R 6 For C6-C 10 Aryl; or (2)R 5 For C6-C 10 Aryl, R 6 It is a C1-C8 alkyl or C6-C 10 Aryl;

[0025] R 7 and R 8 For H, or R 7 and R 8 Together with the carbon atoms they are attached to, they form C6-C. 10 aryl;

[0026] R 9 It is a C1-C8 alkyl, a C1-C8 alkyl substituted with one or more halogens, a C2-C8 alkenyl or benzyl;

[0027] R 10 R 11 R 12 and R 13Independently H, halogen, C1-C8 alkoxy group, or by one or more R groups 11-1 Substituted C1-C8 alkoxy or C1-C8 alkyl, R 10 R 11 R 12 and R 13 Not both H, or R 10 R 11 Together with the carbon atoms they are attached to, they form C6-C. 10 Aryl, or R 11 R 12 Together with the carbon atoms they are attached to, they form C6-C. 10 Aryl, or R 12 R 13 Together with the carbon atoms they are attached to, they form C6-C. 10 Aryl;

[0028] R 11-1 Independently for C3-C 10 cycloalkyl;

[0029] R 14 It is a C1-C8 alkyl, a C1-C8 alkyl substituted with one or more halogens, a C2-C8 alkenyl or benzyl;

[0030] R 15 R 16 and R 17 Independently H, halogen, C1-C8 alkoxy group, or by one or more R groups 11-1 Substituted C1-C8 alkoxy or C1-C8 alkyl.

[0031] In a preferred embodiment of the present invention, certain groups in the compound represented by formula (I), its tautomers, its stereoisomers, its pharmaceutically acceptable salts, or solvates of any of the foregoing are defined as follows, and unmentioned groups are as described in any embodiment of this application (hereinafter referred to as "in a certain embodiment of the present invention"), when R 1 When the halogen is halogen, the halogen is fluorine.

[0032] In one aspect of the present invention, when R 1 When it is a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0033] In one aspect of the present invention, when R 1 When the C1-C8 alkyl group is substituted with one or more halogens, the C1-C8 alkyl group substituted with one or more halogens is a C1-C4 alkyl group substituted with one or more halogens, such as trifluoromethyl.

[0034] In one aspect of the present invention, when R1 When it is a C1-C8 alkoxy group, the C1-C8 alkoxy group is a C1-C4 alkoxy group, such as a methoxy group.

[0035] In one aspect of the present invention, when R 1 When the C1-C8 alkoxy group is substituted with one or more halogens, the C1-C8 alkoxy group substituted with one or more halogens is a C1-C4 alkoxy group substituted with one or more halogens, such as a trifluoromethoxy group.

[0036] In one aspect of the present invention, when R 1 When the benzoyl group is substituted with one or more halogens, the halogen is fluorine. Preferably, the benzoyl group substituted with one or more halogens is...

[0037] In one aspect of the present invention, when R 1 When the phenoxy group is substituted with one or more halogens, the halogen is fluorine. Preferably, the phenoxy group substituted with one or more halogens is...

[0038] In one aspect of the present invention, when R 1a R 1b and R 1c When it is a halogen on its own, the halogen is fluorine.

[0039] In one aspect of the present invention, when R 1a R 1b and R 1c When independently a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0040] In one aspect of the present invention, when R 1 R 1a Together with the carbon atoms they are attached to, they form C6-C. 10 When the aryl group is present, the C6-C 10 The aryl group is phenyl.

[0041] In one aspect of the present invention, when R 1 R 1a When the carbon atoms connected to them form a "3-10 heteroaryl group with heteroatoms selected from one or more of N, O and S and having 1-5 heteroatoms", the "3-10 heteroaryl group with heteroatoms selected from one or more of N, O and S and having 1-5 heteroatoms" is "5-6 heteroaryl group with heteroatoms selected from one or more of N, O and S and having 1-2 heteroatoms"; for example, pyridyl.

[0042] In one aspect of the present invention, when R 2When the halogen is halogen, the halogen is chlorine, bromine or iodine.

[0043] In one aspect of the present invention, when R 3 For one or more R 3-1 When the substituted C1-C8 alkyl group is used, the C1-C8 alkyl group is a C1-C4 alkyl group, such as ethyl or n-propyl.

[0044] In one aspect of the present invention, when R 3 When it is a C2-C8 ynyl group, the C2-C8 ynyl group is a C2-C4 ynyl group, such as propynyl.

[0045] In one aspect of the present invention, when R 3 When it is a C2-C8 alkenyl group, the C2-C8 alkenyl group is a C2-C4 alkenyl group, such as allyl.

[0046] In one aspect of the present invention, when R 3-1 When independently a C1-C8 alkoxy group, the C1-C8 alkoxy group is a C1-C4 alkoxy group, such as a methoxy or ethoxy group.

[0047] In one aspect of the present invention, when R 3-1-1 and R 3-1-2 When independently a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0048] In one aspect of the present invention, when R 3-1-1-1 When it is a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0049] In one aspect of the present invention, when R 3-1-3 When it is a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as tert-butyl.

[0050] In one aspect of the present invention, when R 3-1-3 For one or more R 3-1-3-1 Replacement C6-C 10 In the aryl case, the C6-C 10 The aryl group is phenyl.

[0051] In one aspect of the present invention, when R 4 When it is a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as isopropyl or tert-butyl.

[0052] In one aspect of the present invention, when R 4 For C6-C 10 In the aryl case, the C6-C 10 The aryl group is phenyl.

[0053] In one aspect of the present invention, when R 7 and R 8 Together with the carbon atoms they are attached to, they form C6-C. 10 When the aryl group is present, the C6-C 10 The aryl group is phenyl.

[0054] In one aspect of the present invention, when R 9 When the alkyl group is C1-C8, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl, ethyl, n-propyl, isopropyl or n-butyl, or methyl, ethyl or isopropyl.

[0055] In one aspect of the present invention, when R 9 When the C1-C8 alkyl group is substituted with one or more halogens, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0056] In one aspect of the present invention, when R 9 When it is a C2-C8 alkenyl group, the C2-C8 alkenyl group is a C2-C4 alkenyl group, such as allyl.

[0057] In one aspect of the present invention, when R 10 R 11 R 12 and R 13 When it is a halogen on its own, the halogen is bromine.

[0058] In one aspect of the present invention, when R 10 R 11 R 12 and R 13 When independently a C1-C8 alkoxy group, the C1-C8 alkoxy group is a C1-C4 alkoxy group, such as methoxy, ethoxy, n-propoxy, isopropoxy, or n-butoxy.

[0059] In one aspect of the present invention, when R 10 R 11 R 12 and R 13 Independently for one or more R 11-1 When the C1-C8 alkoxy group is substituted, the C1-C8 alkoxy group is a C1-C4 alkoxy group, such as methoxy, ethoxy, or n-propoxy.

[0060] In one aspect of the present invention, when R 10 R 11 R 12 and R 13 When independently a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0061] In one aspect of the present invention, when R 10 R 11 Together with the carbon atoms they are attached to, they form C6-C. 10 In the aryl case, the C6-C 10 The aryl group is phenyl.

[0062] In one aspect of the present invention, when R 11 R 12 Together with the carbon atoms they are attached to, they form C6-C. 10 In the aryl case, the C6-C 10 The aryl group is phenyl.

[0063] In one aspect of the present invention, when R 12 R 13 Together with the carbon atoms they are attached to, they form C6-C. 10 In the aryl case, the C6-C 10 The aryl group is phenyl.

[0064] In one aspect of the present invention, when R 11-1 Independently for C3-C 10 In the case of cycloalkyl groups, the C3-C 10 The cycloalkyl group is a C5-C6 cycloalkyl group, such as a cyclohexyl group.

[0065] In one aspect of the present invention, when R 14 When it is a C1-C8 alkyl group, the C1-C8 alkyl group is a C1-C4 alkyl group, such as methyl.

[0066] In one aspect of the present invention, R 1 The phenoxy group is H, benzoyl, benzoyl group substituted with one or more halogens, phenoxy group, or "phenoxy group substituted with one or more halogens", preferably, R 1 It is benzoyl, benzoyl group substituted with one or more halogens, or phenoxy group substituted with one or more halogens.

[0067] In one aspect of the present invention, R 1a R 1b and R 1c H stands for H independently.

[0068] In one aspect of the present invention, R 2 It is H or halogen.

[0069] In one aspect of the present invention, R 3-1 Independently hydroxyl, C1-C8 alkoxy, NR 3-1-1 R 3-1-2 , or -OC(=O)- R 3-1-3.

[0070] In one aspect of the present invention, R 3-1-3 It is a C1-C8 alkyl group.

[0071] In one aspect of the present invention, R 3-1-1 and R 3-1-2 Independently H or -C (=O) -OR 3-1-1-1 .

[0072] In one aspect of the present invention, R 4 It is a C1-C8 alkyl group.

[0073] In one aspect of the present invention, R 7 and R 8 For H.

[0074] In one aspect of the present invention, R 9 It is a C1-C8 alkyl group.

[0075] In one aspect of the present invention, R 10 It is H or C1-C8 alkyl.

[0076] In one aspect of the present invention, R 11 It is H, C1-C8 alkoxy or C1-C8 alkyl, or R 11 R 12 Together with the carbon atoms they are attached to, they form C6-C. 10 Aryl.

[0077] In one aspect of the present invention, R 12 It is H, C1-C8 alkoxy or C1-C8 alkyl, or R 11 R 12 Together with the carbon atoms they are attached to, they form C6-C. 10 Aryl.

[0078] In one aspect of the present invention, R 13 It is H, methoxy, or C1-C8 alkyl.

[0079] In one aspect of the present invention, R 14 It is a C1-C8 alkyl group.

[0080] In one aspect of the present invention, R 15 R 16 and R 17 H stands for H independently.

[0081] In one aspect of the present invention, for

[0082]

[0083] In one aspect of the present invention, for In one aspect of the present invention, for

[0084] In one aspect of the present invention, for

[0085] In one aspect of the present invention, for

[0086] The present invention also provides the following compounds, their tautomers, their stereoisomers, their pharmaceutically acceptable salts, or solvates of any of the foregoing (referring to the aforementioned compounds, their tautomers, their stereoisomers, or their pharmaceutically acceptable salts):

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095] The pharmaceutically acceptable salt of the compound shown in formula (I) may be a salt prepared from the compound shown in formula (I) and a pharmaceutically acceptable acid, wherein the pharmaceutically acceptable acid may be an acid conventional in the art, such as an inorganic acid or an organic acid, wherein the inorganic acid is preferably hydrochloric acid, and the organic acid is preferably methanesulfonic acid. Preferably, the pharmaceutically acceptable acid is hydrochloric acid. More preferably, the pharmaceutically acceptable salt of the compound shown in formula (I) is a salt formed from the compound shown in formula (I) and hydrochloric acid in a molar ratio of 1:2.

[0096] The present invention also provides a method for preparing the compound shown in formula (I) above, which includes the following steps: the compound shown in formula (II) and the compound shown in formula (III) undergo a condensation reaction as shown below to obtain the compound shown in formula (I);

[0097]

[0098] Among them, rings C and R 1 R 1a R 1b and R 1c The definition is as stated in the previous item.

[0099] The present invention also provides a compound as shown in formula (II):

[0100]

[0101] The definition of ring C is as described in the previous item.

[0102] Preferably, the compound represented by formula (II) is any of the following compounds:

[0103]

[0104]

[0105] The present invention also provides a method for preparing the compound shown in formula (II) above, which includes the following steps: a condensation reaction is carried out between the 2,5-diketopiperazine derivative shown in formula (A) and the compound shown in formula (B) as shown below to obtain the compound shown in formula (II);

[0106]

[0107] The definition of ring C is as described in the previous item.

[0108] The present invention also provides a pharmaceutical composition comprising the above-described compounds, their tautomers, their stereoisomers, their pharmaceutically acceptable salts, or solvates of any of the foregoing, and pharmaceutical excipients.

[0109] In some formulations, the pharmaceutical excipient does not contain a cosolvent.

[0110] This invention also provides the use of the above-described compounds, their tautomers, their stereoisomers, their pharmaceutically acceptable salts, or solvates of any of the foregoing, or the above-described pharmaceutical compositions in the preparation of a medicament. Preferably, the medicament is used for the prevention and / or treatment of cancer. The cancer is preferably one or more of lung cancer, pancreatic cancer, and colon cancer. The compounds of this invention exhibit strong cytotoxicity against a variety of tumor cells; for example, all compounds of this invention exhibit proliferation-inhibiting activity against NCI-H460, BXPC-3, HT-29, and other cells, with an IC50 value of [missing information]. 50 Value <5μM.

[0111] The present invention also provides the use of the above-described compounds, their tautomers, their stereoisomers, their pharmaceutically acceptable salts, or solvates of any of the foregoing, or the above-described pharmaceutical compositions in the preparation of microtubule inhibitors.

[0112] In the described applications, the tubulin inhibitor can be used in mammalian organisms; it can also be used in vitro, primarily for experimental purposes, such as providing a standard or control sample for comparison, or preparing a kit according to conventional methods in the art to provide rapid detection of the effect of the tubulin inhibitor.

[0113] The present invention also provides a method for preventing and / or treating cancer, comprising administering to a patient a therapeutically effective dose of the aforementioned compound, its tautomer, its stereoisomer, its pharmaceutically acceptable salt, or a solvate of any of the foregoing, or the aforementioned pharmaceutical composition.

[0114] The compounds, their tautomers, their stereoisomers, their pharmaceutically acceptable salts, solvates of any of the foregoing, and pharmaceutical compositions of the present invention can be administered topically or systemically, for example, for enteral administration, such as rectal or oral administration, or for parenteral administration to mammals (especially humans). Exemplary combinations for rectal administration include suppositories that may contain, for example, suitable non-irritating excipients, such as cocoa butter, synthetic glycerides, or polyethylene glycol, which are solid at room temperature but melt and / or dissolve in the rectal lumen to release the drug. The compounds of the present invention can also be administered parenterally, for example, by inhalation, injection, or infusion, such as via intravenous, intra-arterial, intra-bone, intramuscular, intracerebral, extraventricular, intrasynovial, intrasternal, intrathecal, intralesional, intralesional, intracranial, intratumoral, intradermal, and subcutaneous injection or infusion.

[0115] The therapeutically effective amount of the active ingredient is as defined in the context and depends on the mammal species, weight, age, individual condition, individual pharmacokinetic parameters, the disease to be treated, and the route of administration. For enteral administration, such as oral administration, the compounds of the present invention can be formulated into a wide variety of dosage forms.

[0116] The effective amount of the compound, its tautomer, its stereoisomer, its pharmaceutically acceptable salt, its solvate or pharmaceutical composition thereof described in this invention can be easily determined by routine experiments. The most effective and convenient route of administration and the most appropriate formulation can also be determined by routine experiments.

[0117] The pharmaceutical excipients described herein may be those widely used in the pharmaceutical manufacturing field. Excipients primarily serve to provide a safe, stable, and functional pharmaceutical composition, and may also provide methods for enabling the active ingredient to dissolve at a desired rate after administration to a subject, or to promote the effective absorption of the active ingredient after administration to a subject. The pharmaceutical excipients may be inert fillers, or provide a function such as stabilizing the overall pH of the composition or preventing the degradation of the active ingredient. The pharmaceutical excipients may include one or more of the following: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0118] Substances that can be used as pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicates, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-blocking polymers, lanolin, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as carboxymethyl cellulose. Sodium, ethyl cellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salts; Ringer's solution; ethanol, phosphate buffer solution, and other non-toxic and suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coatings, sweeteners, flavorings and spices, preservatives and antioxidants.

[0119] The pharmaceutical compositions of the present invention can be prepared using any method known to those skilled in the art, based on the disclosure. For example, conventional mixing, dissolving, granulation, emulsification, grinding, encapsulation, embedding, or lyophilization processes.

[0120] Pharmaceutical dosage forms of the compounds of this invention can be provided in the form of immediate-release, controlled-release, sustained-release, or targeted drug delivery systems. Common dosage forms include solutions and suspensions, (micro)emulsions, ointments, gels and patches, liposomes, tablets, sugar-coated pills, soft-shell or hard-shell capsules, suppositories, ovules, implants, amorphous or crystalline powders, aerosols, and lyophilized formulations. Depending on the route of administration used, special devices may be required to administer or deliver the drug, such as syringes and needles, inhalers, pumps, injection pens, applicators, or special flasks. Pharmaceutical dosage forms often consist of the drug, excipients, and a container / sealing system. One or more excipients (also known as inactive ingredients) may be added to the compounds of this invention to improve or facilitate the manufacture, stability, administration, and safety of the drug, and to provide a method for obtaining the desired drug release profile. Therefore, the type of excipient added to the drug can be determined by various factors, such as the physical and chemical properties of the drug, the route of administration, and the preparation steps. Pharmaceutical excipients exist in this field and include those listed in various pharmacopoeias. (See the United States Pharmacopeia (USP), Japanese Pharmacopoeia (JP), European Pharmacopoeia (EP), and British Pharmacopoeia (BP); publications of the Center for Drug Evaluation and Research (CEDR) of the U.S. Food and Drug Administration (www.fda.gov), such as the Inactive Ingredient Guide (1996); and the Handbook of Pharmaceutical Additives (2002, Synapse Information Resources, Inc., Endicott NY, etc.).

[0121] The pharmaceutical dosage forms of the compounds of this invention can be manufactured by any method well known in the art, such as conventional mixing, sieving, dissolving, melting, granulation, manufacturing of sugar-coated pills, tableting, suspension, extrusion, spray drying, grinding, emulsification, (nano / micron) encapsulation, packaging, or lyophilization processes. As described above, the compositions of this invention may include one or more physiologically acceptable inactive ingredients that facilitate the processing of the active molecules into formulations for pharmaceutical use.

[0122] The pharmaceutical composition and dosage form may comprise one or more compounds of the present invention, one or more pharmaceutically acceptable salts thereof, or one or more solvates of any of the foregoing as the active ingredient. The pharmaceutically acceptable carrier may be solid or liquid. Solid formulations include powders, tablets, pills, lozenges, capsules, pods, suppositories, and dispersible granules. The solid carrier may also serve as one or more substances as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, preservative, tablet disintegrant, or encapsulating material. In powders, the carrier is typically a finely ground solid, which is a mixture with the finely ground active ingredient. In tablets, the active ingredient is typically mixed with a carrier having the necessary binding capacity in a suitable proportion and compacted to the desired shape and size. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, methylcellulose, sodium carboxymethyl cellulose, low-melting-point waxes, cocoa butter, etc. Formulations of the active compound may include encapsulating materials as carriers, providing capsules in which the active ingredient, with or without a carrier, is surrounded by a carrier bound to it.

[0123] Other forms suitable for oral administration include liquid formulations, including emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions, or solid formulations intended to be converted into a liquid form shortly before use. Emulsions can be prepared in solutions, such as aqueous solutions of propylene glycol, or may contain emulsifiers such as lecithin, sorbitan monooleate, or gum arabic. Aqueous solutions can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavorings, stabilizers, and thickeners. Aqueous suspensions can be prepared by dispersing finely particulate active ingredients in water using binders such as natural or synthetic gums, resins, methylcellulose, carboxymethylcellulose, and other commonly used suspending agents. Solid formulations include solutions, suspensions, and emulsions, which, in addition to the active ingredient, may contain colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0124] For parenteral administration, the pharmaceutical compositions of the present invention may be in the form of sterile injectable or infusionable formulations, for example, as sterile aqueous or oily suspensions. These suspensions may be formulated using suitable dispersants or wetting agents (e.g., Tween 80) and suspending agents according to techniques known in the art. Sterile injectable or infusionable formulations may also be sterile injectable or infusionable solutions or suspensions in non-toxic, parenteral-acceptable diluents or solvents. For example, the pharmaceutical composition may be a solution in 1,3-propanediol. Other examples of acceptable media and solvents that may be used in the pharmaceutical compositions of the present invention include, but are not limited to, mannitol, water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile non-volatile oils are commonly used as solvents or suspension media. Any mild non-volatile oil, including synthetic monoglycerides or diglycerides, may be used for this purpose. Fatty acids such as oleic acid and its glyceride derivatives may be used in the preparation of injections, as well as naturally occurring pharmaceutically acceptable oils such as olive oil or castor oil, particularly in their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants. Solutions intended for parenteral use may also include suitable stabilizers and, if necessary, buffering agents. Suitable stabilizers include antioxidants, such as sodium bisulfate, sodium sulfite, or ascorbic acid, citric acid, and their salts, and sodium EDTA, alone or in combination. Parenteral solutions may also contain preservatives, such as benzalkonium chloride, p-hydroxybenzoic acid, or propylparaben, and chlorobutanol.

[0125] The effective therapeutic dose can be estimated first using various methods well known in the art. The initial dose for animal studies can be based on the effective concentration established in cell culture assays. The appropriate dose range for individual humans can be determined, for example, using data obtained from animal studies and cell culture assays. In some embodiments, the compounds of the present invention can be prepared as oral formulations.

[0126] The effective amount or therapeutically effective dose of a drug (such as the compounds of this invention) refers to the amount of the drug or compound that causes improvement in individual symptoms or prolongs survival. The toxicity and therapeutic efficacy of the molecule can be determined in cell cultures or laboratory animals using standard pharmaceutical procedures, such as by measuring LD50. 50 (Dose that causes 50% lethality in the population) and ED 50 (The dose effective for 50% of the population). The dose ratio of toxicity to therapeutic effect is the therapeutic index, which can be expressed as LD50. 50 / ED 50 Drugs exhibiting a high therapeutic index are preferred.

[0127] An effective or therapeutically effective dose is the amount of a compound or pharmaceutical composition that will elicit a biological or medical response in a tissue, system, animal, or human that is being sought by researchers, veterinarians, physicians, or other clinicians. The dosage is preferably within the range of minimal or no toxicity. 50 The dosage may vary within the range of circulating concentrations. The dosage may vary within this range depending on the dosage form and / or route of administration. The appropriate formulation, route of administration, dosage, and dosing interval should be selected based on methods known in the art, taking into account the specific circumstances of the individual.

[0128] Dosage and intervals can be individually adjusted to provide a plasma level sufficient to achieve the desired effect; this is known as the minimum effective concentration (MEC). The MEC will vary from compound to compound but can be estimated, for example, from in vitro data and animal studies. The dose required to achieve the MEC will depend on individual characteristics and route of administration. In cases of local administration or selective uptake, the effective local concentration of the drug may be independent of plasma concentration.

[0129] The amount of medication or composition administered may vary depending on various factors, including the individual’s sex, age and weight, the severity of the illness, the method of administration and the prescribing physician’s judgment.

[0130] Unless otherwise specified, the terms used in this invention have the following meanings:

[0131] Those skilled in the art will understand that, according to conventions used in the art, the structural formulas of the groups described in this invention are... This refers to the fact that the corresponding group is connected to other fragments or groups in the compound through this site.

[0132] In this document, terms may be preceded and / or followed by a single dash "-" or a double dash "=" to indicate the bond order between the named substituent and the parent moiety; a single dash indicates a single bond, and a double dash indicates a double bond. In the absence of a single or double dash, a single bond is assumed to form between the substituent and its parent moiety. Furthermore, substituents are read "from left to right" unless otherwise indicated.

[0133] The term "multiple" refers to 2, 3, 4 or 5, preferably 2 or 3.

[0134] The term "pharmaceutically acceptable" means that the salts, solvents, excipients, etc., are generally non-toxic, safe, and suitable for patient use. The term "patient" preferably refers to a mammal, and more preferably a human.

[0135] The term "solvate" refers to a substance formed by the combination of the compound of this invention with a stoichiometric or non-stoichiometric solvent. Solvent molecules in a solvate can exist in an ordered or disordered arrangement. The solvents include, but are not limited to, water, methanol, and ethanol.

[0136] The terms "pharmaceutically acceptable salt" and "solvent" in the term "pharmaceuticalally acceptable salt solvate" refer, as described above, to substances formed by combining the compounds of the present invention with 1) a relatively non-toxic, pharmaceutically acceptable acid or base, or 2) a stoichiometric or non-stoichiometric solvent. The term "pharmaceuticalally acceptable salt solvate" includes, but is not limited to, hydrochloric acid monohydrate of the compounds of the present invention.

[0137] When any variable (e.g., R) 3-1 When a variable appears multiple times in the definition of a compound, the definition at each position is independent of the definitions at the other positions; their meanings are independent and do not affect each other. Therefore, if a group is surrounded by one, two, or three R... 3-1 Group substitution, meaning that the group can be replaced by up to 3 R groups. 3-1 Replace, the position R 3-1 Definition and other positions R 3-1 The definitions are independent of each other. Furthermore, combinations of substituents and / or variables are only permitted if the combination produces a stable compound.

[0138] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0139] The term "alkyl" refers to a saturated straight-chain or branched alkyl group having a specified number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and similar alkyl groups.

[0140] The term "alkoxy group" refers to the group -OR X , where R X It is an alkyl group as defined above.

[0141] The term "alkenyl" refers to a straight-chain or branched olefin having a specific number of carbon atoms, containing one or more carbon-carbon double bonds and no carbon-carbon triple bonds. The one or more carbon-carbon double bonds can be internal or terminal. Examples of alkenes include vinyl, allyl, methyl vinyl, propenyl, butenyl, pentenyl, 1,1-dimethyl-2-propenyl, hexenyl, etc.

[0142] The term "alkynyl" refers to a straight-chain or branched hydrocarbon group (e.g., C2-C8 alkynyl, or C2-C4 alkynyl) having one or more triple bonds with a specific number of carbon atoms. These carbon-carbon triple bonds can be internal or terminal, such as in the propynyl group where the triple bond is internal. Or a propynyl group at the end of the triple bond wait.

[0143] The term "aryl" refers to C6-C 10 Aryl groups, such as phenyl or naphthyl.

[0144] The term "heteroaryl" refers to an aromatic group containing heteroatoms, preferably containing 1-5 independent 3-6 membered monocyclic or 9-10 membered bicyclic aromatic rings selected from nitrogen, oxygen, and sulfur. When it is a bicyclic ring, at least one ring is aromatic, such as furanyl, pyridinyl, pyridinyl, pyrimidinyl, pyrazinyl, thiophene, isozolyl, oxazolyl, diazolyl, imidazole, pyrroleyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, inzolyl, indololinyl, benzothiazolyl, benziisothiazolyl, benzozolyl, benziisozolyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, etc.

[0145] The term "cycloalkyl" refers to a saturated cycloalkyl group, such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0146] The term "heterocyclic alkyl" refers to a saturated cyclic group having heteroatoms, preferably a 3- to 10-membered saturated monocyclic or bicyclic group containing 1-5 independently selected cyclic heteroatoms chosen from N, O, and S. Examples of heterocyclic alkyl groups include: tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydropyridyl, tetrahydropyrroliyl, aza-butyl, thiazolyl, azoleyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazine, aza-heptyl, diaza-heptyl, oxazolidinyl, etc. Preferred heterocyclic groups are morpholin-4-yl, piperidin-1-yl, pyrrolidine-1-yl, thiomorpholin-4-yl, and 1,1-dioxo-thiomorpholin-4-yl.

[0147] The term “treatment” refers to a therapeutic approach. When a specific condition is involved, treatment means: (1) alleviating one or more biological manifestations of the disease or condition; (2) interfering with (a) one or more points in a biological cascade that causes or precipitates the condition or (b) one or more biological manifestations of the condition; (3) improving one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment; or (4) slowing the development of the condition or one or more biological manifestations of the condition.

[0148] The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0149] The term "patient" refers to any animal, preferably a mammal, that is about to receive or has already received administration of the compound or composition according to embodiments of the invention, with humans being the most preferred. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans, with humans being the most preferred.

[0150] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0151] 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 literature. Compared with Plinabulin, some compounds have comparable or even better antitumor activity, and have good development prospects. Attached Figure Description

[0152] Figure 1 The results are immunofluorescence results for the H460 cell line containing dehydrophenylastitin compounds. Detailed Implementation

[0153] The present invention will be further illustrated by way of embodiments below, but the present invention is not limited to the scope of the embodiments described herein.

[0154] Example 1: Preparation of (3Z,6Z)-3-((5-phenyl-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-1)

[0155]

[0156] 1) Synthesis of ethyl 2-chloro-3-oxo-3-phenylpropionate 6

[0157]

[0158] Take a 100 mL dry round-bottom flask, dissolve ethyl benzoyl chloride (10.0 g, 52.1 mmol) in 20 mL dry DCM, place in a cold well at 0 °C and stir for 15 min. Then slowly add sulfonyl chloride (7.72 g, 57.2 mmol) dropwise. After the addition is complete, move the reaction flask to room temperature and stir. When the temperature of the reaction solution reaches room temperature, transfer the round-bottom flask to an oil bath and reflux at 35 °C for 6 h. Monitor the reaction by LC-MS until complete. Add 200 mL of dichloromethane to the reaction solution and transfer the solution to a separatory funnel. Wash the organic phase successively with saturated sodium carbonate aqueous solution (30 mL), water (30 mL), and saturated brine (30 mL). Combine the aqueous phases, back-extract the aqueous phase three times with dichloromethane (100 mL * 3), combine the organic phases, and add anhydrous sodium sulfate to dry. Filter, concentrate the filtrate under reduced pressure to obtain 11.6 g of reddish-brown clear liquid, yield 99.0%. 1 H NMR (500MHz, DMSO-d6) δ8.03(d,J=7.3Hz,2H),7.74(t,J=7.4Hz,1H),7.60(t,J=7.9Hz,1H),6.63 (s,1H),4.20(q,J=7.1Hz,2H),1.14(t,J=7.1Hz,3H).MS(ESI)m / z:[M+Na] + Calculated value C 11 H 11 ClNaO3: 249.03, measured value: 248.85.

[0159] 2) Synthesis of ethyl 5-phenyl-1H-imidazolium-4-carboxylate 11b

[0160]

[0161] Take a 100 mL dry round-bottom flask and add 11.6 g (51.5 mmol) of ethyl 2-chloro-3-oxo-3-phenylpropionate, 23.2 g (515 mmol) of formamide, and 1.85 g (103 mmol) of water sequentially. Reflux the mixture in an oil bath at 180 °C for 14 h. After the reaction is complete, allow the reaction solution to cool to room temperature. Add 150 mL of water to the reaction solution, resulting in the formation of a brown solid. Extract the solution three times with 200 mL of ethyl acetate (200 mL x 3). Combine the organic phases, wash the organic phase with 200 mL of saturated brine, and back-extract the aqueous phase three times with 200 mL of ethyl acetate (200 mL x 3). Combine the organic phases and dry with anhydrous sodium sulfate. Filter under vacuum, concentrate the filtrate to a solid, add an appropriate amount of water to the crude product for purification, filter under vacuum, wash the filter cake with 100 mL of water (3 times), and dry the filter cake under vacuum at 50 °C to obtain 5.08 g of brown solid, with a yield of 45.6%. 1H NMR (400 MHz, DMSO-d6) δ7.85(s,1H),7.77(s,1H),7.39(dt,J=13.3,6.9Hz,4H),4.21(q,J=6.8Hz,2H),1.22(t, J=9.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C 12 H 13 N2O2: 217.10, measured value: 216.88.

[0162] 3) Synthesis of 5-phenyl-1H-imidazol-4-methanol 12b

[0163]

[0164] Ethyl 5-phenyl-1H-imidazolium-4-carboxylate 11b (5.08 g, 23.5 mmol) was used as the starting material for this reaction. Compound 12b was synthesized according to the preparation method of compound 12a, yielding 5.00 g of a yellowish-brown solid, with a yield of 99.9%. MS (ESI) m / z: [M+H] + Calculated value C 10 H 11 N2O: 175.09, measured value: 174.72.

[0165] 4) Synthesis of 5-phenyl-1H-imidazol-4-carboxaldehyde 13b

[0166]

[0167] 5-Phenylacetyl-1H-imidazol-4-methanol 12b (5.00 g, 28.7 mmol) was used as the starting material for this step of the reaction. Compound 13b was synthesized according to the preparation method of compound 13a, yielding 3.51 g of a yellowish-brown solid, with a yield of 71.1%. 1 H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 8.03 (s, 1H), 7.82 (d, J = 7.0Hz, 2H), 7.51-7.44 (m, 3H). MS (ESI) m / z: [M+H] + Calculated value C 10 H9N2O: 173.07, Measured value: 172.74.

[0168] Synthesis of (Z)-1-acetyl-3-((5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione 15b

[0169]

[0170] 5-Phenylacetyl-1H-imidazol-4-carboxaldehyde 13b (280 mg, 1.63 mmol) and 1,4-diacetylpiperazine-2,5-dione 14 (969 mg, 4.89 mmol) were used as starting materials for this step of the reaction. Compound 15b was synthesized according to the preparation method of compound 15a, yielding 240 mg of orange-red solid, with a yield of 47.5%. 1 H NMR(500MHz,DMSO-d6)δ13.14(s,1H),11.84(s,1H),8.12(s,1H),7.56(q,J=7.5Hz,4H), 7.46(t,J=6.8Hz,1H),6.90(s,1H),4.33(s,2H),2.49(d,J=8.2Hz,3H).MS(ESI)m / z:[M+ H] + Calculated value C 16 H 15 N4O3: 311.11, measured value: 310.81.

[0171] Synthesis of (6)(3Z,6Z)-3-((5-phenyl-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-1

[0172]

[0173] Take a 25 mL dry brown round-bottom flask, add (Z)-1-acetyl-3-((5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (80.0 mg, 0.26 mmol) and 3-(4-fluorobenzoyl)benzaldehyde 4 (88.2 mg, 0.39 mmol), DMF (4 mL), purge with nitrogen three times, add cesium carbonate (143.0 mg, 0.39 mmol) and anhydrous sodium sulfate (74.0 mg, 0.52 mmol), purge with nitrogen three times again, and stir the reaction at 45 °C for 24 h. After the reaction was complete as detected by LC-MS, the reaction solution was added dropwise to cold water, resulting in the precipitation of a large amount of pale yellow solid. The solid was filtered, and the filter cake was washed with water. The filter cake was then reconstituted with a mixed solvent of methanol and dichloromethane (VMeOH:VDCM = 1:3), filtered again, and the filtrate was concentrated under reduced pressure to a solid concentration. An appropriate amount of methanol was added, and the mixture was purified by ultrasonic slurrying. The purified product was then stored frozen, filtered again, and the filter cake was washed with methanol. The filter cake was then dried in a vacuum drying oven at 50°C to obtain 58.0 mg of an orange-red product (PLN-1-1), with a yield of 47.0%. mp: 248-250°C. 1H NMR (500MHz, DMSO-d6) δ13.10(s,1H),12.10(s,1H),10.41(s,1H),8.12(s,1H),7.91(dd,J=8.4, 5.7Hz,2H),7.83(s,1H),7.76(d,J=7.6Hz,1H),7.64(d,J=7.6Hz,1H),7.60-7.55(m ,5H),7.45(dd,J=7.3,4.7Hz,1H),7.40(t,J=8.7Hz,2H),6.84(s,1H),6.73(s,1H). 13 C NMR(125MHz,DMSO-d6)δ194.2,164.8,157.5,156.1,137.3,136.8,133.5*2,133.4,132.8*2,132.7,132.0,130.1 ,129.3,129.2*2,128.9,128.8,128.3,127.7*2,127.6,125.8,115.8,115.6,113.1,103.6.HRMS(ESI)m / z:[M+Na] + Calculated value C 28 H 19 FN4NaO3: 501.1333, measured value: 501.1336.

[0174] Example 2: Preparation of (3Z,6Z)-3-((5-tert-butyl-2-iodo-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-2)

[0175]

[0176] 1) Synthesis of 13g of 5-tert-butyl-2-iodo-1H-imidazol-4-carboxaldehyde

[0177]

[0178] Take a 100 mL dry brown round-bottom flask, and use 5-(tert-butyl)-1H-imidazolium-4-carboxaldehyde 13a (3.00 g, 19.7 mmol) and N-iodosuccinimide (4.44 g, 19.7 mmol) as the starting materials for this reaction step. Synthesize compound 13 g according to the preparation method of compound PLN-1-3, and obtain 4.74 g of white solid, with a yield of 86.5%. 1 H NMR(600MHz,DMSO-d6)δ13.47(s,1H),9.80(s,1H),1.35(s,9H).MS(ESI)m / z:[M+H]+ Calculated value C8H 12I N2O: 279.00, measured value: 278.71.

[0179] 2) Synthesis of 15g of (Z)-1-acetyl-3-((5-tert-butyl-2-iodo-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0180]

[0181] 13 g (1.00 g, 3.60 mmol) of 5-tert-butyl-2-iodo-1H-imidazol-4-carboxaldehyde and 14 g (1.43 g, 7.19 mmol) of 1,4-diacetylpiperazine-2,5-dione were used as starting materials for this reaction step. Compound 15 g was synthesized according to the preparation method of compound 15a, yielding 280 mg of yellow solid, with a yield of 18.7%. 1 H NMR (400MHz, DMSO-d6) δ11.44(s,1H),6.95(s,1H),4.30(s,2H),2.49(s,3H),1.36(s,9H).MS(ESI)m / z:[M+H] + Calculated value C 14 H 18 IN4O3: 417.04, measured value: 416.87.

[0182] Synthesis of (3Z,6Z)-3-((5-tert-butyl-2-iodo-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-2

[0183]

[0184] Using 15 g (140 mg, 0.34 mmol) of (Z)-1-acetyl-3-((5-tert-butyl-2-iodo-1H-imidazol-4-yl)methylene)piperazine-2,5-dione and 4-(4-fluorobenzoyl)benzaldehyde (115 mg, 0.50 mmol) as starting materials for this reaction, compound PLN-1-2 was synthesized according to the preparation method of compound PLN-1-1, yielding 75.0 mg of a pale yellow solid in 38.2% yield. mp: 171-173 °C. 1H NMR(600MHz, DMSO-d6)δ12.69(s,1H),11.67(s,1H),10.38(s,1H),7.91(dd,J=8.0,5.7Hz,2H),7.82(s,1H),7.76(d, J=7.5Hz,1H),7.64(d,J=7.6Hz,1H),7.59(t,J=7.6Hz,1H),7.40(t,J=8.6Hz,2H),6.81(s,1H),6.78(s,1H),1.36(s,9H). 13 C NMR (150MHz, DMSO-d6) δ194.2,164.8,157.6,156.2,145.1,137.3,134.4,133.5, 133.4,132.9,132.8,130.2,129.7,128.9,128.8,127.6,124.2,115.8,115.7,113.0,104.1,86.9,32.3,30.6*3.HRMS(ESI)m / z:[M+Na] + Calculated value C 26 H 22 FIN4NaO3: 607.0613, Measured value: 607.0620.

[0185] Example 3: Preparation of (3Z,6Z)-3-((2-bromo-5-tert-butyl-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-3)

[0186]

[0187] Take a 50 mL dry round-bottom flask and dissolve (3Z,6Z)-3-((5-tert-butyl-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzylmethyl)piperazine-2,5-dione 1 (70.0 mg, 0.15 mmol) in dry tetrahydrofuran (8 mL). Stir at room temperature for 2 min. Then add N-bromosuccinimide (27.2 mg, 0.15 mmol) to the round-bottom flask, heat to 70 °C, and reflux for 24 h. Monitor the reaction completion by LC-MS. After the reaction solution cools to room temperature, add saturated sodium thiosulfate aqueous solution dropwise until the reaction solution turns pale yellow. Then extract three times with ethyl acetate (100 mL * 3), combine the organic phases, wash the organic phase with saturated brine (100 mL), combine the aqueous phases, back-extract the aqueous phase three times with ethyl acetate (50 mL * 3), combine the organic phases, and dry with anhydrous sodium sulfate. The filtrate was filtered under reduced pressure and concentrated. It was then purified by reversed-phase column chromatography (methanol:water = 0:100, 5:95, 10:90, 15:85, 20:80, 25:75, 30:70, 35:65, 40:60, 45:55, 50:50, 55:45, 60:40) to give 29.1 mg of a pale yellow solid, yield 27.1%. mp: 156-158℃. 1 H NMR(500MHz,DMSO-d6)δ12.92 (s,1H),11.47(s,1H),10.37(s,1H),7.91(dd,J=8.8,5.6Hz,2H),7.82(s,1H),7.76(d,J=7.7Hz,1H),7. 64(d,J=7.6Hz,1H),7.58(t,J=7.6Hz,1H),7.40(t,J=8.8Hz,2H),6.81(s,1H),6.79(s,1H),1.36(s,9H). 13 C NMR(125MHz,DMSO-d6)δ194.2,164.8,157.5,156.1,137.3,133.5,133.4*2,132.8*2,132.3,130.1, 129.6,128.9,128.8,127.5,124.1,116.1,115.8,115.6,113.1,104.5,32.5,30.5*3.HRMS(ESI)m / z:[M+Na] + Calculated value C 26 H 22 BrFN4NaO3: 559.0752, measured value: 559.0751.

[0188] Example 4: Preparation of (3Z,6Z)-3-((5-tert-butyl-2-chloro-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-4)

[0189]

[0190] Synthesis of (Z)-1-acetyl-3-((5-tert-butyl-2-chloro-1H-imidazol-4-yl)methylene)piperazine-2,5-dione 15f

[0191]

[0192] (Z)-1-acetyl-3-((5-(tert-butyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione 15a (300 mg, 1.03 mmol) and N-chlorosuccinimide (138 mg, 1.03 mmol) were used as starting materials for this step of the reaction. Compound 15f was synthesized according to the preparation method of compound PLN-1-3, yielding 116 mg of a pale yellow solid, with a yield of 34.6%. 1 H NMR(400MHz,DMSO-d6)δ13.00(s,1H),11.11(s,1H),6.96(s,1H),4.30(s,2H),2.49(s,3H),1.37(s,9H).MS(ESI)m / z:[M+H] + Calculated value C 14 H 18 ClN4O3: 325.11, measured value: 324.85.

[0193] Synthesis of (2)(3Z,6Z)-3-((5-tert-butyl-2-chloro-1H-imidazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-4

[0194]

[0195] Using (Z)-1-acetyl-3-((5-(tert-butyl)-3-chloro-1H-imidazol-4-yl)methylene)piperazin-2,5-dione 15f (91.6 mg, 0.28 mmol) and 3-(4-fluorobenzoyl)benzaldehyde 4 (96.5 mg, 0.42 mmol) as starting materials for this reaction, compound PLN-1-4 was synthesized according to the preparation method of compound PLN-1-1 (reaction temperature adjusted to 70 °C), yielding 31.8 mg of an orange-yellow solid, with a yield of 22.9%. mp: 158–160 °C. 1H NMR (500MHz, DMSO-d6) δ11.41 (s, 1H), 10.37 (s, 1H), 7.91 (dd, J = 8.8, 5.5Hz, 2H), 7.82 (s,1H),7.75(d,J=7.7Hz,1H),7.64(d,J=7.7Hz,1H),7.58(t,J=7.6Hz,1H),7.40(t,J=8.8Hz,2H),6.81(s,1H),6.79(s,1H),1.37(s,9H). 13 C NMR(125MHz,DMSO-d6)δ194.2,165.8,163.8,157.5,156.1, 137.3,133.5*2,133.4*2,132.8*2,132.3,130.1,128.9,128.8,127.5,124.0,115.8,115.6,113.1,104.2,32.5,30.5*3.HRMS(ESI)m / z:[M+Na] + Calculated value C 26 H 22 ClFN4NaO3: 515.1257, Measured value: 515.1253.

[0196] Example 5: Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-3-benzyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-1-5)

[0197]

[0198] 1) Compound 1 (4.75 g, 30 mmol) was dissolved in acetic acid (19.4 mL) at 0°C. An aqueous solution of NaNO2 (5.18 g, 75 mmol) (16.5 mL) was added dropwise at 0°C. The mixture was stirred at 0°C for 2 h, then heated to room temperature and stirred for 2.5 h. The reaction endpoint was determined by LC-MS. The mixture was extracted three times with diethyl ether (50 mL x 3). The combined organic phases were washed with NaHCO3, dried over anhydrous sodium sulfate, filtered, and distilled under reduced pressure to give a pale yellow oil 2 (4.83 g, 25.82 mmol).

[0199] 2) Dry a 100 mL reaction flask, add compound 2 (4.83 g, 25.82 mmol), dissolve in anhydrous ethanol (20 mL), add 10% Pd / C (1.59 g), degas, add dropwise HCl-EtOH (2 M) solution (6.7 mL), degas, stir at room temperature for 24 h, filter and evaporate to dryness to obtain brownish-yellow paste 3 (3.5 g, 18.70 mmol).

[0200] 3) Dissolve raw material 3 (2.0 g, 9.54 mmol) in dry DCM, add triethylamine (3.98 mL, 28.62 mmol), stir, add phenylacetyl chloride (1.9 mL, 14.31 mmol) dropwise, stir overnight at room temperature, and detect the reaction endpoint by LC-MS to obtain compound 4 (2.22 g, 7.63 mmol).

[0201] 4) Dissolve starting material 4 (2.22 g, 7.63 mmol) in glacial acetic acid (30 mL), add ammonium acetate (2.94 g, 38.16 mmol), and reflux at 120°C overnight. After complete reaction, evaporate the solvent, neutralize with saturated sodium bicarbonate, and perform column chromatography with petroleum ether:ethyl acetate = 10:1 to give cyclized compound 5 (289 mg, 1.06 mmol).

[0202] 5) Dissolve LiAlH4 (122 mg, 3.22 mmol) in dry THF (5 mL) at 0°C. Dissolve starting material 5 (289 mg, 1.06 mmol) in dry THF (5 mL) and add it dropwise to the reaction system. Gradually raise the temperature to room temperature and react overnight. The reaction was confirmed to be complete by TLC. The reaction was quenched with water, extracted with EA, and separated by column chromatography to obtain compound 6 (223 mg, 0.97 mmol).

[0203] 6) Dissolve raw material 6 (223 mg, 0.97 mmol) in DCM (5 mL), add manganese dioxide (842 mg, 9.68 mmol), stir at room temperature for 48 h, detect the reaction by TLC, and separate compound 7 (15 mg, 0.066 mmol) by column chromatography.

[0204] 7) The second batch of compound 7 (716 mg, 3.14 mmol) was dissolved in dry DMF (20 mL), acetylated DKP (2.49 g, 12.56 mmol) was added, potassium carbonate (651 mg, 4.71 mmol) was added, the reaction was carried out at 50 degrees Celsius for 24 h, the reaction was detected by LC-MS, filtered, and compound 8 (166 mg, 0.45 mmol) was obtained by reverse column chromatography.

[0205] 8) Dry 400 mg of 4A molecular sieve, dissolve compound 8 (166 mg, 0.45 mmol) in dry DMF (4 mL), add p-fluorobenzoylbenzaldehyde (156 mg, 0.68 mmol), add cesium carbonate (222 mg, 0.68 mmol), react at 80 degrees Celsius in the dark for 48 h, and detect the final product B-1 by LC-MS. After recrystallization with ethanol, filter and dry to obtain yellow powder A (10 mg, 0.019 mmol).

[0206] Example 6: Preparation of (3Z,6Z)-3-((5-tert-butyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-6)

[0207]

[0208] 1) Synthesis of 5-tert-butyloxazole-4-methanol 12c

[0209]

[0210] A 1L dry round-bottom flask was placed in a -60℃ cold well. First, 250mL of dry tetrahydrofuran was added, followed by 20.2g (532mmol) of lithium aluminum hydride. A solution of 35.0g (178mmol) of ethyl 5-tert-butyl-1,3-oxazol-4-carboxylate 10a dissolved in 200mL of tetrahydrofuran was slowly added dropwise to the lithium aluminum hydride suspension. After the addition was complete, the temperature was gradually raised to -40℃, and the reaction was stirred for 4 hours. The reaction was monitored by LC-MS until complete. The reaction solution was then slowly quenched by adding it dropwise to ice water. The mixture was filtered, and the filter cake was washed twice with water (200mL*2) and twice with anhydrous ethanol (500mL*2). The filtrate was concentrated under reduced pressure to obtain 29.1g of a pale yellow oil, with a yield of 99.9%. MS (ESI) m / z: [M+ Na] + Calculated value C8H 12 NNaO2: 178.08, measured value: 177.77.

[0211] 2) Synthesis of 5-tert-butyloxazole-4-carboxaldehyde 13C

[0212]

[0213] 5-tert-butyloxazole-4-methanol 12c (29.1 g, 187 mmol) was used as the starting material for this reaction. Compound 13c was synthesized according to the preparation method of compound 13a, yielding 6.65 g of an orange-yellow oil, with a yield of 23.2%. 1 H NMR(400MHz, CDCl3)δ10.09(s,1H), 7.76(s,1H),1.46(s,9H).MS(ESI)m / z:[M+H] + Calculated value C8H 12 NO2: 154.09, Measured value: 153.68.

[0214] Synthesis of (Z)-1-acetyl-3-((5-tert-butyloxazol-4-yl)methylene)piperazine-2,5-dione 15c

[0215]

[0216] 5-tert-butyl-1,3-oxazol-4-carboxaldehyde 13c (1.00 g, 6.53 mmol) and 1,4-diacetylpiperazine-2,5-dione 14 (2.59 g, 13.1 mmol) were used as starting materials for this reaction. Compound 15c was synthesized according to the preparation method of compound 15a (the reaction temperature was adjusted to 45 °C) to give 850 mg of white solid, with a yield of 44.7%. 1 H NMR (400MHz, CDCl3) δ11.22(s,1H),7.83(s,1H),7.08 (s,1H),4.47(s,2H),2.64(s,3H),1.44(s,9H).MS(ESI)m / z:[M+Na] + Calculated value C 14 H 17 N3NaO4: 314.11, Measured value: 313.84.

[0217] Synthesis of (4)(3Z,6Z)-3-((5-tert-butyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-6

[0218]

[0219] (Z)-1-acetyl-3-((5-tert-butyloxazol-4-yl)methylene)piperazine-2,5-dione 15c (200 mg, 0.687 mmol) and 3-(4-fluorobenzoyl)benzaldehyde 4 (235 mg, 1.03 mmol) were used as starting materials for this reaction. Compound PLN-1-6 was synthesized according to the preparation method of compound PLN-1-1, yielding 244 mg of a pale yellow solid, with a yield of 77.4%. mp: 238-240℃. 1 H NMR (500MHz, DMSO- d6) δ11.12(s,1H),10.55(s,1H),8.60(s,1H),7.91(dd,J=8.8,5.5Hz,2H),7.82(s,1H),7.76(d,J=7.7Hz, 1H),7.64(s,1H),7.60(d,J=7.6Hz,1H),7.40(t,J=8.8Hz,2H),6.86(s,1H),6.72(s,1H),1.39(s,9H). 13C NMR (100MHz, DMSO-d6) δ194.2,164.8,158.0,157.0,156.3,150.2,137.3,133.5,133.4,133.3,132.9, 132.8,130.2,129.0,128.9,128.2,127.2,126.8,115.9,115.7,114.1,100.8,33.1,29.1*3.HRMS(ESI)m / z:[M+H] + Calculated value C 26 H 23 FN3O4: 482.1487, measured value: 482.1485.

[0220] Example 7: Preparation of (3Z,6Z)-3-((5-tert-butyl-2-phenyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-7)

[0221]

[0222] 1) Synthesis of ethyl 5-tert-butyl-2-phenyloxazol-4-carboxylate (10 h)

[0223]

[0224] Take a 100 mL dry round-bottom flask and add ethyl pivaloyl acetate (5.00 g, 29.0 mmol), benzylamine (7.78 g, 72.6 mmol), and sodium bicarbonate (18.4 g, 72.6 mmol) sequentially. After stirring, add iodine (18.4 g, 72.6 mmol) and water (50 mL). Heat to 80 °C and reflux for 2 h. Monitor the reaction completion by thin-layer chromatography. Stop heating and allow to cool at room temperature. Stir at room temperature and add saturated sodium thiosulfate until the solution is clear. Extract the reaction solution three times with ethyl acetate (150 mL * 3). Combine the organic phases, wash the organic phase with saturated brine, combine the aqueous phases, back-extract the aqueous phase twice with ethyl acetate (150 mL * 2), combine the organic phases, and dry with anhydrous sodium sulfate. The filtrate was filtered under reduced pressure and concentrated. It was then purified by column chromatography (PE:EA = 40:1, 30:1, 20:1, 15:1, 10:1) to obtain 5.29 g of a pale yellow oil, with a yield of 66.6%. 1 H NMR (400MHz, DMSO-d6) δ7.98-7.96(m,2H),7.60-7.53(m,3H),4.32(q,J=7.1Hz,2H),1.47(s,9H), 1.32(t,J=7.1Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C16 H 20 NO3: 274.14, Measured value: 273.85.

[0225] 2) Synthesis of 5-tert-butyl-2-phenyloxazol-4-methanol for 12 hours

[0226]

[0227] Ethyl 5-tert-butyl-2-phenyloxazol-4-carboxylate 10h (3.59 g, 13.1 mmol) was used as the starting material for this reaction. Compound 12h was synthesized according to the preparation method of compound 12c, yielding 3.43 g of a pale yellow oil, with a yield of 99.9%. MS (ESI) m / z: [M+H] + Calculated value C 14 H 18 NO2: 232.13, measured value: 231.88.

[0228] 3) Synthesis of 5-tert-butyl-2-phenyloxazol-4-carboxaldehyde (13h)

[0229]

[0230] 5-tert-butyl-2-phenyloxazol-4-methanol 12h (3.43 g, 14.8 mmol) was used as the starting material for this step of the reaction. Compound 13h was synthesized according to the preparation method of compound 13a, yielding 1.37 g of orange-yellow solid, with a yield of 40.3%. 1 H NMR(400MHz,DMSO-d6)δ10.09 (s,1H),8.02-8.00(m,2H),7.58-7.56(m,3H),1.49(s,9H).MS(ESI)m / z:[M+Na] + Calculated value C 14 H 15 NNaO2: 252.10, measured value: 251.85.

[0231] Synthesis of (Z)-1-acetyl-3-((5-tert-butyl-2-phenyloxazol-4-yl)methylene)piperazine-2,5-dione for 15 hours

[0232]

[0233] 5-tert-butyl-2-phenyloxazol-4-carboxaldehyde 13h (500 mg, 2.18 mmol) and 1,4-diacetylpiperazine-2,5-dione 14 (865 mg, 4.36 mmol) were used as starting materials for this step of the reaction. Compound 15h was synthesized according to the preparation method of compound 15a (the reaction temperature was adjusted to 45 °C) to give 591 mg of brown solid, with a yield of 73.8%. 1 H NMR (400MHz, CDCl3) δ11.42(s,1H),8.05-8.03(m, 2H),7.51-7.49(m,3H),7.12(s,1H),4.51(s,1H),2.66(s,3H),1.51(s,9H).MS(ESI)m / z:[M+Na] + Calculated value C 20 H 21 N3NaO4: 390.14, measured value: 390.03.

[0234] Preparation of 5)(3Z,6Z)-3-((5-tert-butyl-2-phenyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-7

[0235]

[0236] A 25 mL dry round-bottom flask was used as the starting material for this reaction. (Z)-1-acetyl-3-((5-tert-butyl-2-phenyloxazol-4-yl)methylene)piperazine-2,5-dione 15h (100 mg, 0.27 mmol) and 3-(4-fluorobenzoyl)benzaldehyde 4 (93.1 mg, 0.41 mmol) were added. Following the preparation method of compound PLN-1-1 (reaction temperature adjusted to 70 °C), compound PLN-1-7 was synthesized, yielding 118 mg of a pale yellow solid, with a yield of 81.2%. mp: 259-261 °C. 1 H NMR(500MHz,DMSO-d6)δ11.21(s,1H),10.57(s,1H),8.00 (dd,J=7.4,2.1Hz,2H),7.92(dd,J=8.7,5.6Hz,1H),7.84(s,1H),7.77(d,J=7.7Hz,1H),7.66(d ,J=7.7Hz,1H),7.62-7.58(m,4H),7.41(t,J=8.8Hz,2H),6.88(s,1H),6.75(s,1H),1.46(s,9H). 13C NMR(125 MHz, DMSO-d6)δ194.1,164.8,158.6,158.0,157.0,156.2,137.3,133.5,133.4,133.3,132.8*2,131.3,130.2,130.1 ,129.4*2,128.9*2,127.2,126.9,126.0*2,125.5,115.8,115.6,114.0,100.8,33.3,29.2*3.HRMS(ESI)m / z:[M+Na] + Calculated value C 32 H 26 FN3NaO4: 558.1800, measured value: 558.1797.

[0237] Example 8: Preparation of (3Z,6Z)-3-((5-phenyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-8)

[0238]

[0239] 1) Synthesis of ethyl 5-phenyloxazol-4-carboxylate 10d

[0240]

[0241] Ethyl isocyanate 7 (10.0 g, 88.4 mmol) and benzoic anhydride 8 (24.0 g, 106 mmol) were used as the starting materials for this reaction. Compound 16d was synthesized according to the preparation method of compound 10a, yielding 7.83 g of a yellow oily substance with a yield of 40.8%. 1 HNMR(500MHz, DMSO-d6)δ8.57(s,1H),7.95(dd,J=6.8,3.0Hz,2H),7.54-7.48(m,3H),4.29(q,J=7.1Hz,2H),1.26(t, J=7.1Hz,3H).MS(ESI)m / z:[M+Na] + Calculated value C 12 H 11 NNaO3: 240.06, measured value: 239.85.

[0242] 2) Synthesis of 5-phenyloxazole-4-methanol 12d

[0243]

[0244] Ethyl 5-phenyl-1,3-oxazol-4-carboxylate 10d (7.20 g, 33.2 mmol) was used as the starting material for this reaction. Compound 12d was synthesized according to the preparation method of compound 12c, yielding 6.57 g of a yellowish-brown solid, with a yield of 99.9%. MS (ESI) m / z: [M+Na] + Calculated value C 10 H9NNaO2: 198.05, measured value: 197.78.

[0245] 3) Synthesis of 5-phenyloxazole-4-carboxaldehyde 13d

[0246]

[0247] 5-Phenylexaazole-4-methanol 12d (6.57 g, 37.5 mmol) was used as the starting material for this reaction. Compound 13d was synthesized according to the preparation method of compound 12a, yielding 1.36 g of orange-yellow solid, with a yield of 21.0%. 1 H NMR(400MHz,DMSO-d6)δ10.04(s,1H), 8.65(s,1H),8.08-8.05(m,2H),7.61-7.58(m,3H).MS(ESI)m / z:[M+H] + Calculated value C 10 H8NO2: 174.05, measured value: 173.71.

[0248] 4)(Z)-1-acetyl-3-((5-phenyloxazol-4-yl)methylene)piperazine-2,5-dione 15d

[0249]

[0250] 5-Phenyrazol-4-carboxaldehyde 13d (500 mg, 2.89 mmol) and 1,4-diacetylpiperazine-2,5-dione 14 (1.14 g, 5.77 mmol) were used as starting materials for this reaction. Compound 15d was synthesized according to the preparation method of compound 15a (the reaction temperature was adjusted to 45 °C), yielding 689 mg of brown solid with a yield of 76.6%. 1 H NMR(400MHz,DMSO-d6)δ10.84(s,1H),8.83(s,1H),7.71(dd, J=7.3,1.3Hz,3H),7.62(m,3H),7.54(d,J=7.4Hz,1H),4.36(s,2H),2.51(s,5H).MS(ESI)m / z:[M+Na] + Calculated value C 16 H 13N3NaO4: 334.08, measured value: 333.85.

[0251] Synthesis of (5)(3Z,6Z)-3-((5-phenyloxazol-4-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-8

[0252]

[0253] Using (Z)-1-acetyl-3-((5-phenyloxazol-4-yl)methylene)piperazine-2,5-dione 15d (100 mg, 0.32 mmol) and 3-(4-fluorobenzoyl)benzaldehyde 4 (110 mg, 0.48 mmol) as starting materials for this reaction, compound PLN-1-8 was synthesized according to the preparation method of compound PLN-1-1, yielding 69.4 mg of a pale yellow solid in a yield of 45.1%. mp: 228-230℃. 1 H NMR(500MHz,DMSO-d6) δ11.06(s,1H),10.63(s,1H),8.82(s,1H),7.91(dd,J=8.7,5.6Hz,2H),7.84(s,1H),7.77(d,J=7.6Hz ,1H),7.71(d,J=7.6Hz,2H),7.66(d,J=7.7Hz,1H),7.60(m,3H),7.51(t,J=7.4Hz,1H),7.40(t,J=8.8 Hz,1H),6.89(s,1H),6.75(s,1H). 13 C NMR(125MHz,DMSO-d6)δ194.1,164.8,156.8,156.2,151.8,148.3, 137.3,133.5,133.4,133.2,132.8*2,130.3,130.2,129.7,129.5*2,129.0,128.9 ,128.6,127.1,126.9,126.4*2,115.8,115.6,114.4,99.6.HRMS(ESI)m / z:[M+Na] + Calculated value C 28 H 18 FN3NaO4: 502.1174, measured value: 502.1174.

[0254] Example 9: Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-methylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-9)

[0255]

[0256] Take a 10 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-methylimidazolium-2-carboxaldehyde (46.6 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), anhydrous sodium sulfate (80.2 mg, 0.56 mmol), and DMF (3 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 45 °C oil bath, and stir for 22 h. Monitor the reaction by LC-MS; the starting material spot disappears. After the reaction is complete, add the reaction solution dropwise to 30 mL of cold water at 4 °C. No solid precipitates. Extract with DCM (100 mL * 3) and EA (100 mL). Combine the organic phases, dry to anhydrous sodium sulfate, filter, concentrate under reduced pressure, and dry. Methanol was ultrasonically homogenized, placed in a 0°C refrigerator to stand, filtered, the filter cake was washed with cold methanol, and dried in a 50°C vacuum drying oven to obtain 78.2 mg of yellow solid, with a yield of 69%.

[0257] 1 H NMR (500MHz, DMSO-d6) δ12.23(s,1H),10.44(s,1H),7.42(t,J=7.9Hz,1H),7.36(d,J=0.7 Hz,1H),7.28(d,J=7.7Hz,1H),7.24(dd,J=11.6,4.8Hz,3H),7.19(s,1H),7.14– 7.10(m,2H),6.92(dd,J=8.1,2.2Hz,1H),6.80(s,1H),6.59(s,1H),3.77(s,3H). 13 C NMR((125MHz,DMSO-d6)δ159.14, 157.24,157.05,156.72,156.66,152.56,152.54,143.49,134.91,130.28,128.46,128.28,126.82,124 .59,123.00,120.60,120.53,119.02,117.98,116.63,116.44,114.55,95.92,32.49.MS(ESI)m / z:[M+H] + Calculated value C 22 H 18 FN4O3: 405.1357, measured value: 405.1352. Melting point: 238-240℃.

[0258] Example 10: Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-ethylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-10)

[0259]

[0260] 1) Preparation of 1-ethylimidazol-2-carboxaldehyde (14o)

[0261]

[0262] 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] + Calculated value of C6H9N2O: 125.0709, measured value: 124.69.

[0263] 2) Synthesis of the final product PLN-1-10

[0264]

[0265] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-ethylimidazolium-2-carboxaldehyde (52.5 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 45 °C with stirring for 22 h. A yellow solid of 101.3 mg was obtained, with a yield of 86%.

[0266] 1H NMR (500MHz, DMSO-d6) δ12.29(s,1H),10.44(s,1H),7.44–7.39(m,2H),7.28(d,J=7.8Hz, 1H),7.23(dt,J=6.1,4.4Hz,3H),7.20(s,1H),7.16–7.09(m,2H),6.92(dd,J=8.1, 2.1Hz,1H),6.80(s,1H),6.59(s,1H),4.17(q,J=7.2Hz,2H),1.32(t,J=7.2Hz,3H). 13 C NMR(125MHz,DMSO-d6)δ159.14, 157.23,157.03,156.76,156.68,152.57,152.55,142.57,134.93,130.28,128.68,128.55,126.82,124. 60,121.30,120.59,120.52,119.03,117.98,116.63,116.44,114.56,95.69,16.38.MS(ESI)m / z:[M+Na] + Calculated value C 23 H 19 FN4O3 Na: 441.1339, measured value: 441.1332. Melting point: 214-216℃.

[0267] Example 11 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-propylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-11)

[0268]

[0269] 1) Synthesis of 1-propylimidazol-2-carboxaldehyde (14p)

[0270]

[0271] The compound 14o 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%. 1H 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] + Calculated value C7H 11 N2O: 139.0866, measured value: 138.64.

[0272] 2) Synthesis of the final product PLN-1-11

[0273]

[0274] The compound PLN-1-9 was prepared using the same synthetic method. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-propylimidazolium-2-carboxaldehyde (58.5 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 50 °C with stirring for 25 h. A yellow solid of 59.5 mg was obtained, with a yield of 49%.

[0275] 1 H NMR (500MHz, DMSO-d6) δ12.32(s,1H),10.44(s,1H),7.44–7.38(m,2H),7.28(d,J=7.8Hz, 1H),7.26–7.22(m,3H),7.19(s,1H),7.14–7.09(m,2H),6.92(dd,J=8.1,2.1Hz,1H),6. 80(s,1H),6.59(s,1H),4.11(t,J=7.1Hz,2H),1.75–1.64(m,2H),0.84(t,J=7.4Hz,3H). 13 C NMR (125MHz, DMSO- d6) δ159.14,157.23,157.03,156.76,156.68,152.57,152.55,142.93,134.93,130.27,128.68,128.42,126.82, 124.60,121.99,120.58,120.51,119.02,117.97,116.62,116.44,114.54,95.77,46.77,24.02,10.80.MS(ESI)m / z:[M+Na]+ Calculated value C 24 H 21 FN4O3 Na: 455.1495, measured value: 455.1507. Melting point: 168-170℃.

[0276] Example 12 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-butylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-1-12)

[0277]

[0278] 1) Synthesis of 1-Butylimidazol-2-carboxaldehyde (14q)

[0279]

[0280] The compound 14o 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=15.0,11.2,7.5Hz,2H),1.30(dd,J=15.2,7.5Hz,2H),0.90(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C8H 13 N2O: 153.1022, measured value: 152.73.

[0281] 2) Synthesis of the final product PLN-1-12

[0282]

[0283] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-butylimidazolium-2-carboxaldehyde (64.4 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 50 °C with stirring for 24 h. A yellow solid of 77.3 mg was obtained, with a yield of 61%.

[0284] 1 H NMR (500MHz, DMSO-d6) δ12.31(s,1H),10.44(s,1H),7.45–7.39(m,2H),7.28(d,J=7.8Hz, 1H),7.26–7.21(m,3H),7.19(s,1H),7.14–7.10(m,2H),6.92(dd,J=8.1,2.2Hz,1H),6.80(s,1H),6.59 (s,1H),4.14(t,J=7.1Hz,2H),1.73–1.59(m,2H),1.25(dq,J=14.7,7.3Hz,2H),0.88(t,J=7.4Hz,3H). 13 C NMR (125MHz, DMSO-d6) δ159.14,157.23,157.03,156.76,156.68,152.55,142.88,134.93,130.28, 128.68,128.43,126.82,124.60,121.96,120.58,120.51,119.03,117.97,11 6.62,116.44,114.55,95.74,45.05,32.75,19.17,13.43.MS(ESI)m / z:[M+H] + Calculated value C 25 H 24 FN4O3: 447.1827, measured value: 447.1819. Melting point: 158-160℃.

[0285] Example 13 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-isopropylimidazol)-2-methylene)piperazine-2,5-dione (PLN-1-13)

[0286]

[0287] Synthesis of 1-isopropylimidazol-2-carboxaldehyde (14r)

[0288]

[0289] The compound 14o was prepared according to the synthetic method described for compound 14o. 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, the mixture was reacted in DMF (10 mL) 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] + Calculated value C7H 11 N2O: 139.0866, Measured value: 139.04.

[0290] 2) Synthesis of the final product PLN-1-13

[0291]

[0292] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-isopropylimidazolium-2-carboxaldehyde (58.5 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 50 °C with stirring for 22 h. A yellow solid of 78.8 mg was obtained, with a yield of 65%. Melting point: 198-200 °C. 1 H NMR(500MHz,DMSO-d6)δ12.36(s,1H),10.44 (s,1H),7.54(d,J=1.0Hz,1H),7.42(t,J=7.9Hz,1H),7.28(d,J=7.7Hz,2H),7.26–7.21(m,2H),7.20( s,1H),7.14–7.09(m,2H),6.92(dd,J=8.1,2.1Hz,1H),6.80(s,1H),6.64(s,1H),4.75(dt,J=13.2,6.6 Hz,1H),1.40(d,J=6.6Hz,6H). 13C NMR(125MHz,DMSO-d6)δ159.14,157.24,157.04,156.81,156.68,152.55,142.22,134.93,130.28,128.83,128.77,1 26.82,124.60,120.59,120.52,119.02,117.98,116.63,116.44,114.54,95.76,47.09,23.15*2.MS(ESI)m / z:[M+H] + Calculated value C 24 H 22 FN4O3: 433.1670, measured value: 433.1675.

[0293] Example 14 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((1-isopropylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-14)

[0294]

[0295] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorobenzoylphenyl)methylene)piperazine-2,5-dione (100 mg, 0.27 mmol), 1-isopropylimidazolium-2-carboxaldehyde (56.6 mg, 0.41 mmol), cesium carbonate (133.4 mg, 0.41 mmol), and anhydrous sodium sulfate (77.5 mg, 0.55 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 45 °C with stirring for 23 h. A yellow solid of 76.6 mg was obtained, with a yield of 63%.

[0296] 1 H NMR(500MHz,DMSO-d6)δ12.38(s,1H),10.63(s,1H),7.95–7.90(m,2H),7.85(s,1H),7.77 (d,J=7.7Hz,1H),7.66(d,J=7.7Hz,1H),7.60(t,J=7.7Hz,1H),7.55(d,J=1.1Hz,1H),7.41(t,J=8.8 Hz,2H),7.28(d,J=0.7Hz,1H),6.89(s,1H),6.64(s,1H),4.76(dt,J=13.2,6.6Hz,1H),1.40(d,J=6.6Hz,6H). 13C NMR (125MHz, DMSO-d6) δ194.14,165.75,163.75,156.96,156.62,142.23,137.28,133.50, 133.44,133.29,132.83,132.75,130.26,128.95,128.87,128.83,128.79,127. 26,118.01,115.79,115.62,114.15,95.80,47.08,23.16*2.MS(ESI)m / z:[M+H] + Calculated value C 25 H 22 FN4O3: 445.1670, measured value: 455.1669. Melting point: 222-224℃.

[0297] Example 15 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-(2-fluoro)ethylimidazolium)-2-methylene)piperazine-2,5-dione (PLN-1-15)

[0298]

[0299] 1) Synthesis of 1-(2-fluoro)ethylimidazolium-2-carboxaldehyde (14t)

[0300]

[0301] Take a 25 mL dry round-bottom flask and add imidazole-2-carboxaldehyde (200 mg, 2.08 mmol), DMF (5 mL), potassium carbonate (345.2 mg, 2.50 mmol), and 1-bromo-2-fluoroethane (317.1 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 213.0 mg of an oily liquid, yield 72%. 1 H NMR(500MHz, CDCl3)δ9.84(s,1H),7.32(s,1H),7.25(s,1H),4.75(s,2H),4.72– 4.68(m,1H),4.67–4.61(m,1H).MS(ESI)m / z:[M+H] + Calculated value of C6H8FN2O: 143.0615, measured value: 142.62.

[0302] 2) Synthesis of the final product PLN-1-15

[0303]

[0304] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-(2-fluoro)ethylimidazolium-2-carboxaldehyde (60.2 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 50 °C with stirring for 25 h. A yellow solid of 69.5 mg was obtained, with a yield of 56%.

[0305] 1 H NMR(500MHz,DMSO-d6)δ12.29(s,1H),10.44(s,1H),7.43–7.39(m,2H),7.31–7.27(m,2H), 7.26–7.21(m,2H),7.20(s,1H),7.12(ddd,J=10.5,5.3,3.2Hz,2H),6.92(dd,J=8.1,2.0Hz,1H),6.80( s,1H),6.65(s,1H),4.73(t,J=4.5Hz,1H),4.64(t,J=4.5Hz,1H),4.55(t,J=4.5Hz,1H),4.49(t,J=4.5 Hz,1H). 13 C NMR (125MHz, DMSO-d6) δ159.14,157.24,157.04,156.74,156.70,152.55,143.59,134.92, 130.28,128.86,128.65,126.80,124.60,122.04,120.59,120.53,119.03,117.9 8,116.63,116.44,114.57,95.95,83.64,82.30,45.90,45.75.MS(ESI)m / z:[M+H] + Calculated value C 23 H 19 F2N4O3: 437.1420, measured value: 437.1424. Melting point: 245-247℃.

[0306] Example 16 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-propenylimidazol)-2-methylene)piperazine-2,5-dione (PLN-1-16)

[0307]

[0308] 1) Synthesis of 1-propenylimidazol-2-carboxaldehyde (14u)

[0309]

[0310] The compound 14t was prepared according to the synthetic method described for compound 14t. 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=10.2,0.8H z,1H),5.10(dd,J=17.0,0.7Hz,1H),5.02(d,J=5.8Hz,2H).MS(ESI)m / z:[M+H] + Calculated value of C7H9N2O: 137.0709, measured value: 136.65.

[0311] 2) Synthesis of the final product PLN-1-16

[0312]

[0313] The compound PLN-1-9 was prepared using the same synthetic method. The reaction mixture was prepared in an oil bath at 50°C with (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-propenylimidazolium-2-carboxaldehyde (57.6 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol), and DMF (3 mL). The mixture was stirred for 22 h. A yellow solid of 68.6 mg was obtained, with a yield of 56%.

[0314] 1H NMR (500MHz, DMSO-d6) δ12.26(s,1H),10.44(s,1H),7.42(t,J=7.9Hz,1H),7.39(d,J=0.8 Hz,1H),7.28(d,J=4.9Hz,2H),7.26–7.21(m,2H),7.19(s,1H),7.16–7.09(m,2H),6.92(dd,J=8.1,2.2H z,1H),6.80(s,1H),6.54(s,1H),6.01(ddd,J=22.1,10.2,5.0Hz,1H),5.20(dd,J=10.3,1.2Hz,1H),4.90 (dd,J=17.1,1.3Hz,1H),4.85(d,J=4.9Hz,2H). 13 C NMR(125MHz,DMSO-d6)δ159.14,157.23,157.03, 156.69, 152.56, 143.18, 134.92, 134.03, 130.27, 128.76, 128.53, 126.80, 124.59, 122.13, 120. 58,120.52,119.03,117.98,116.76,116.62,116.44,114.59,95.89,47.56.MS(ESI)m / z:[M+Na] + Calculated value C 24 H 19 FN4O3Na: 453.1339, measured value: 453.1335. Melting point: 206-208℃.

[0315] Example 17 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-benzylimidazol)-2-methylene)piperazine-2,5-dione (PLN-1-17)

[0316]

[0317] 1) Synthesis of 1-benzylimidazol-2-carboxaldehyde (14v)

[0318]

[0319] The compound 14t was prepared according to the synthetic method described for compound 14t. 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 reacted in DMF (10 mL) in an oil bath at 50 °C with stirring for 6 h. Thin-layer chromatography yielded 166.5 mg of an oily liquid, with a yield of 43%. 1H 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] + Calculated value C 11 H 11 N2O: 187.0866, measured value: 186.72.

[0320] 2) Synthesis of the final product PLN-1-17

[0321]

[0322] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-benzylimidazolium-2-carboxaldehyde (78.8 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 45 °C with stirring for 24 h. A yellow solid of 123.0 mg was obtained, with a yield of 91%.

[0323] 1 H NMR (500MHz, DMSO-d6) δ12.26(s,1H),10.41(s,1H),7.53(d,J=0.8Hz,1H),7.41(t,J=7.9 Hz,1H),7.36(t,J=7.5Hz,2H),7.32–7.21(m,5H),7.18(s,1H),7.16–7.09( m,4H),6.91(dd,J=8.2,2.1Hz,1H),6.78(s,1H),6.59(s,1H),5.46(s,2H). 13 C NMR(125MHz,DMSO-d6)δ159.13,157.22,157.01, 156.69,156.62,152.56,152.54,143.28,137.20,134.92,130.26,128.96,128.80,128.73,127.71,126.78,1 26.64,124.59,122.53,120.57,120.50,119.03,117.98,116.62,116.43,114.60,95.73,48.52.MS(ESI)m / z: [M+H] + Calculated value C 28 H22 FN4O3: 481.1670, measured value: 481.1660. Melting point: 206-208℃.

[0324] Example 18 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((1-isopropylbenzimidazole)-2-methylene)piperazine-2,5-dione (PLN-1-18)

[0325]

[0326] 1) Synthesis of 1-isopropyl-2-carboxybenzimidazole (14y)

[0327]

[0328] The compound 14o was prepared according to the synthetic method described above. Starting materials included 2-formylbenzimidazole (100 mg, 0.68 mmol), potassium carbonate (114.6 mg, 0.82 mmol), and 2-iodopropane (139.6 mg, 0.82 mmol), and DMF (3 mL). The mixture was stirred in an oil bath at 50 °C for 6 h. 86.3 mg of an oily liquid was obtained, with a yield of 67%. 1 H NMR (400MHz, CDCl3) δ10.11 (s, 1H), 7.93 (d, J = 7.7Hz, 1H),7.67(d,J=8.3Hz,1H),7.47–7.41(m,1H),7.37(dd,J=11.0,4.0Hz,1H),5.88(dt,J= 13.9,7.0Hz,1H),5.88(dt,J=13.9,7.0Hz,1H),1.67(d,J=7.0Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C 11 H 13 N2O: 189.1022, measured value: 189.24.

[0329] 2) Synthesis of the final product (PLN-1-18)

[0330]

[0331] The compound PLN-1-9 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), 1-isopropyl-2-carboxybenzimidazole (79.7 mg, 0.42 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 50 °C with stirring for 24 h. A yellow solid of 126.3 mg was obtained, with a yield of 93%.

[0332] 1 H NMR(500MHz,DMSO-d6)δ12.61(s,1H),10.64(s,1H),7.84–7.80(m,1H),7.78(dd,J=6.3, 2.8Hz,1H),7.43(t,J=7.9Hz,1H),7.33–7.28(m,3H),7.25(dd,J=12.2,5.4Hz,3H),7.16–7.09(m,2H), 6.94(dd,J=8.1,1.9Hz,1H),6.87(s,1H),6.83(s,1H),5.11(dq,J=13.6,6.8Hz,1H),1.62(d,J=6.9Hz, 6H). 13 C NMR (125MHz, DMSO-d6) δ159.62,157.71,157.49,157.43,156.89,153.04,148.71,142.66, 135.32,133.68,132.77,130.75,125.20,123.75,123.22,121.06,120.99,119.60,119.44, 118.60,117.11,116.92,115.91,112.84,97.75,95.17,47.87,21.73*2.MS(ESI)m / z:[M+H] + Calculated value C 28 H 24 FN4O3: 483.1827, measured value: 483.1820. Melting point: 263-265℃.

[0333] Example 19 Preparation of (3Z,6Z)-3-(pyridin-2-ylmethylene)-6-(3-(4-fluorobenzoyl)benzylmethylene)piperazine-2,5-dione (PLN-1-19)

[0334]

[0335]

[0336] Take a 25 mL dry brown round-bottom flask and add (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (100 mg, 0.273 mmol), pyridine-2-carboxaldehyde 25a (35.1 mg, 0.328 mmol), cesium carbonate (134 mg, 0.410 mmol), anhydrous sodium sulfate (77.6 mg, 0.546 mmol), and DMF (2.5 mL) sequentially. Purge with nitrogen three times and stir at 45 °C for 24 h. Monitor the reaction completion by LC-MS. After the reaction is complete, add the reaction mixture dropwise to cold water; a large amount of solid precipitates. Filter the mixture, and wash the filter cake with water. Redissolve the filter cake in a mixed solvent of methanol and dichloromethane (V... 甲醇 V 二氯甲烷 =1:3), filter, concentrate the filtrate under reduced pressure to a solid, add an appropriate amount of methanol and ultrasonically beat, freeze overnight, filter, wash the filter cake with methanol, and dry the filter cake in a vacuum drying oven at 50℃ to obtain 80.0 mg of pale yellow solid, with a yield of 70.9%. mp: 259-261℃. 1 H NMR(500MHz,DMSO-d6)δ12.60(s,1H),10.63(s, 1H),8.73(d,J=4.1Hz,1H),7.93-7.89(m,3H),7.85(s,1H),7.78(d,J=7.7Hz,1H),7.67(dd,J=7.8,5 .2Hz,2H),7.60(t,J=7.6Hz,1H),7.42-7.36(m,3H),6.90(s,1H),6.73(s,1H).HRMS(ESI)m / z:[M+Na] + Calculated value C 24 H 16 FN3NaO3: 436.1068, measured value: 436.1062.

[0337] Example 20: Preparation of (3Z,6Z)-3-((6-methoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-20)

[0338]

[0339] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (50.0 mg, 0.14 mmol) and 6-methoxypyridine-2-carboxaldehyde (22.5 mg, 0.16 mmol) as starting materials for this reaction, compound PLN-1-20 was synthesized according to the preparation method of compound PLN-1-19, yielding 39.0 mg of a pale yellow solid in a yield of 64.4%. mp: 216-218 °C. 1 H NMR(500MHz,DMSO-d6) δ12.32(s,1H),10.61(s,1H),7.92(dd,J=8.8,5.5Hz,2H),7.85(s,1H),7.82-7.77(m,2H),7.66(d,J=7 .7Hz,1H),7.60(t,J=7.7Hz,1H),7.40(t,J=8.8Hz,2H),7.27(d,J=7.3Hz,1H),6.87(s,1H),6.84(d,J= 8.2Hz,1H),6.66(s,1H),3.99(s,3H). 13 C NMR(100MHz,DMSO-d6)δ194.2,164.8,162.5,157.0,156.4, 151.9,140.5,137.3,133.6,133.5,133.3,132.9,132.8,130.5,130.4,129.1,128 .9,127.3,119.8,115.9,115.7,114.3,110.0,108.1,53.7.HRMS(ESI)m / z:[M+Na] + Calculated value C 25 H 18 FN3NaO4: 466.1174, Measured value: 466.1170.

[0340] Example 21 Preparation of (3Z,6Z)-3-((5-methoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-21)

[0341]

[0342] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (50.0 mg, 0.14 mmol) and 5-methoxypyridine-2-carboxaldehyde (22.5 mg, 0.16 mmol) as starting materials for this reaction, compound PLN-1-21 was synthesized according to the preparation method of compound PLN-1-19, yielding 42.3 mg of a pale yellow solid in a yield of 69.9%. mp: 237-239 °C. 1 H NMR(500MHz,DMSO-d6)δ 12.34(s,1H),10.54(s,1H),8.48(d,J=3.0Hz,1H),7.92(dd,J=8.8,5.5Hz,2H),7.85(s,1H),7.78(d,J =7.7Hz,1H),7.66(dd,J=8.2,5.0Hz,2H),7.59(t,J=7.7Hz,1H),7.53(dd,J=8.8,3.0Hz,1H),7.40(t,J =8.8Hz,2H),6.87(s,1H),6.72(s,1H),3.89(s,3H). 13 C NMR(100MHz,DMSO-d6)δ194.2,164.8,157.2, 156.6,154.5,147.1,137.3,136.7,133.6,133.5,133.4,132.9,132.8,130.4,129.0,128 .9,128.7,127.7,127.3,122.1,115.9,115.7,114.0,108.2,55.9.HRMS(ESI)m / z:[M+Na] + Calculated value C 25 H 18 FN3NaO4: 466.1174, Measured value: 466.1167.

[0343] Example 22 Preparation of (3Z,6Z)-3-((4-methoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-22)

[0344]

[0345] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (50.0 mg, 0.14 mmol) and 4-methoxypyridin-2-carboxaldehyde (22.5 mg, 0.16 mmol) as starting materials for this reaction, compound PLN-1-22 was synthesized according to the preparation method of compound PLN-1-19, yielding 41.4 mg of a pale yellow solid in a yield of 68.4%. mp: 253-255 °C. 1 H NMR (500MHz, CDCl3) δ13.06 (s,1H),8.47(d,J=5.8Hz,1H),8.25(s,1H),7.86(dd,J=8.9,5.4Hz,2H),7.77-7.75(m,2H),7 .63-7.58(m,2H),7.19(t,J=8.6Hz,2H),7.09(s,1H),6.87(s,1H),6.68(s,1H),3.88(s,3H). 13 C NMR(125MHz, Pyridine-d5)δ194.5,167.0,165.0,158.4,157.8,157.5,138.8,135.5,135.2,134.4,133.8,133.5,133.4,132.8, 131.6,130.0,129.8,129.2,116.4,116.2,115.3,112.3,109.6,109.3,55.8.HRMS(ESI)m / z:[M+Na] + Calculated value C 25 H 18 FN3NaO4: 466.1174, Measured value: 466.1165.

[0346] Example 23 Preparation of (3Z,6Z)-3-((3-methoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-23)

[0347]

[0348] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (50.0 mg, 0.14 mmol) and 3-methoxypyridine-2-carboxaldehyde (22.5 mg, 0.16 mmol) as the starting materials for this reaction, compound PLN-1-23 was synthesized according to the preparation method of compound PLN-1-19, yielding 35.6 mg of a pale yellow solid in a yield of 58.8%. mp: 274-276 °C. 1H NMR(500MHz,DMSO-d6)δ 12.57(s,1H),10.62(s,1H),8.32(d,J=4.8Hz,1H),7.92(dd,J=8.8,5.5Hz,2H),7.85(s,1H),7.78(d,J= 7.6Hz,1H),7.66(d,J=7.7Hz,1H),7.61-7.58(m,2H),7.43-7.39(m,3H),7.00(s,1H),6.90(s,1H),3.92 (s,3H).HRMS(ESI)m / z:[M+Na] + Calculated value C 25 H 18 FN3NaO4: 466.1174, measured value: 466.1171.

[0349] Example 24: Preparation of (3Z,6Z)-3-((5-hydroxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-24)

[0350]

[0351] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (100 mg, 0.27 mmol) and 5-hydroxypyridine-2-carboxaldehyde 25k (40.3 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-24 was synthesized according to the preparation method of compound PLN-1-19, yielding 40.1 mg of a pale yellow solid in a yield of 34.2%. mp: 243-245℃. 1 H NMR(500MHz,DMSO-d6) δ12.35(s,1H),8.28(d,J=2.7Hz,1H),7.92(dd,J=8.7,5.6Hz,2H),7.84(s,1H),7.78(d,J=7.7Hz,1H) ,7.65(d,J=7.7Hz,1H),7.59(t,J=7.7Hz,1H),7.52(d,J=8.6Hz,1H),7.40(t,J=8.8Hz,2H),7.26(dd, J=8.5,2.8Hz,1H),6.86(s,1H),6.66(s,1H). 13C NMR(100MHz,DMSO-d6)δ194.3,164.8,157.3,156.5, 140.1,138.5,137.3*2,133.6,133.5*2,132.9,132.8,130.4,128.9,128.1,1 27.9,127.5,127.4,123.7,115.9,115.7,113.7,108.8.HRMS(ESI)m / z:[M+H] + Calculated value C 24 H 17 FN3O4: 430.1198, measured value: 430.1195.

[0352] Example 25: Preparation of (3Z,6Z)-3-((3-hydroxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-25)

[0353]

[0354]

[0355] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (100 mg, 0.27 mmol) and 3-hydroxypyridine-2-carboxaldehyde 25 (40.3 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-25 was synthesized according to the preparation method of compound PLN-1-19, yielding 47.7 mg of a pale yellow solid in a yield of 40.8%. mp: 260-262℃. 1 H NMR(500MHz,DMSO-d6) δ12.61(s,1H),10.73(s,1H),10.56(s,1H),8.21(d,J=3.4Hz,1H),7.92(dd,J=8.8,5.5Hz,2H),7.85( s,1H),7.78(d,J=7.7Hz,1H),7.66(d,J=7.7Hz,1H),7.60(t,J=7.6Hz,1H),7.41(t,J=8.8Hz,2H),7.34 (d,J=7.0Hz,1H),7.25(dd,J=8.3,4.6Hz,1H),7.02(s,1H),6.88(s,1H). 13C NMR(100MHz,DMSO-d6) δ194.2,164.8,157.2,156.6,152.8,142.8,139.3,137.3,133.6,133.5,133.4,132.9,132.8,13 0.4,129.6,129.0,128.9,127.3,124.0,123.1,115.9,115.7,114.1,102.4.HRMS(ESI)m / z:[M+H] + Calculated value C 24 H 17 FN3O4: 430.1198, measured value: 430.1196.

[0356] Example 26 Preparation of (3Z,6Z)-3-(3-(p-fluorophenoxy)benzene)methylene-6-((N-methylpyrrole)-2-methylene)piperazine-2,5-dione (PLN-1-26)

[0357]

[0358] Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), N-methyl-2-pyrrolecarboxaldehyde (46.19 mg, 0.34 mmol), cesium carbonate (137.9 mg, 0.42 mmol), anhydrous sodium sulfate (80.2 mg, 0.56 mmol), and DMF (3 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection, and in an oil bath at 45 °C. Stir the reaction for 24 h. Monitor the reaction by LC-MS; the starting material spot disappears. After the reaction was complete, the reaction solution was added dropwise to 30 ml of cold water at 4 °C, filtered, the filter cake was washed with cold water, dried in a vacuum drying oven at 50 °C, ultrasonically homogenized with methanol, placed in a refrigerator at 0 °C to stand, filtered, the filter cake was washed with cold methanol, subjected to plate chromatography (MeOH:DCM = 1:200), passed through a short column (MeOH:DCM = 1:10), concentrated under reduced pressure, ultrasonically homogenized with methanol, placed in a refrigerator at 0 °C to stand, filtered, the filter cake was washed with cold methanol, and dried in a vacuum drying oven at 50 °C to obtain 4.5 mg of yellow solid, with a yield of 3.95%. Using Z)-1-acetyl-3-((3-p-fluorophenoxyphenyl)methylene)piperazine-2,5-dione (100 mg, 0.28 mmol), N-methyl-2-pyrrolecarboxaldehyde (46.2 mg, 0.34 mmol), cesium carbonate (137.9 mg, 0.42 mmol), and anhydrous sodium sulfate (80.2 mg, 0.56 mmol) as starting materials, the reaction was carried out in DMF (3 mL) in an oil bath at 45 °C with stirring for 24 h. The reaction was monitored by LC-MS, and the starting material spot disappeared. After the reaction was complete, the reaction solution was added dropwise to 30 mL of cold water at 4°C, filtered, the filter cake was washed with cold water, dried in a vacuum drying oven at 50°C, ultrasonically homogenized with methanol, placed in a refrigerator at 0°C to stand, filtered, the filter cake was washed with cold methanol, subjected to plate chromatography (MeOH:DCM = 1:200), passed through a short column (MeOH:DCM = 1:10), concentrated under reduced pressure, ultrasonically homogenized with methanol, placed in a refrigerator at 0°C to stand, filtered, the filter cake was washed with cold methanol, and dried in a vacuum drying oven at 50°C to obtain 4.5 mg of yellow solid, yield 4%.

[0359] 1 H NMR (500MHz, DMSO-d6) δ10.27(s,1H),9.56(s,1H),7.41(t,J=8.0Hz,1H),7.28(d,J=7.7 Hz,1H),7.23(d,J=8.6Hz,2H),7.19(s,1H),7.12(dd,J=9.0,4.5Hz,2H),6.99(s,1H),6.91(dd,J= 8.1,1.9Hz,1H),6.80(d,J=3.4Hz,1H),6.75(s,1H),6.72(s,1H),6.20–6.13(m,1H),3.66(s,3H).13 C NMR (125MHz, DMSO-d6) δ157.03, 135.18, 130.26, 127.24, 126.05, 124.46, 120.57, 120.50, 118 .88,117.79,116.63,116.44,113.48,112.18,108.86,104.73,33.75.MS(ESI)m / z:[M+Na] + Calculated value C 23 H 18 FN3O3Na: 426.1224, measured value: 426.1219. Melting point: 211-213℃.

[0360] Example 27 Preparation of (3Z,6Z)-3-((3-methylpyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-27)

[0361]

[0362] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (80.0 mg, 0.22 mmol) and 3-methylpyridin-2-carboxaldehyde 25f (31.7 mg, 0.26 mmol) as starting materials for this reaction, compound PLN-1-27 was synthesized according to the preparation method of compound PLN-1-19, yielding 80.6 mg of a pale yellow solid in a yield of 86.4%. mp: 244-247 °C. 1 H NMR(400MHz,DMSO-d6) δ10.69(s,1H),8.56(s,1H),7.92(t,J=5.7Hz,2H),7.85(s,1H),7.77(t,J=9.2Hz,2H),7.66(d,J=7.5H z,1H),7.60(t,J=7.6Hz,1H),7.41(t,J=8.7Hz,2H),7.29(dd,J=7.5,5.0Hz,1H),6.90(s,1H),6.76(s, 1H),2.42(s,3H). 13CNMR(100MHz,DMSO-d6)δ194.3,163.6,157.1,156.8,153.0,146.0,139.0,137.3, 133.7,133.6,133.5*2,133.3,132.9,132.8,131.2,130.4,129.1,128.9,122.5,115.9,115.7,114.5,103.7,18.4.HRMS(ESI)m / z:[M+H] + Calculated value C 25 H 19 FN3O3: 428.1405, measured value: 428.1402.

[0363] Example 28 Preparation of (3Z,6Z)-3-((4-methylpyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-28)

[0364]

[0365] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (100 mg, 0.27 mmol) and 4-methylpyridin-2-carboxaldehyde 25 g (39.7 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-28 was synthesized according to the preparation method of compound PLN-1-19, yielding 87.2 mg of a pale yellow solid in a yield of 74.7%. mp: 241-243 °C. 1 H NMR(400MHz,DMSO-d6) δ10.61(s,1H),8.57(d,J=5.1Hz,1H),7.91(dd,J=8.9,5.5Hz,2H),7.84(s,1H),7.77(d,J=7.6Hz,1H ),7.66(d,J=7.8Hz,1H),7.59(t,J=7.6Hz,1H),7.50(s,1H),7.40(t,J=8.9Hz,2H),7.21(d,J=5.1Hz, 1H),6.89(s,1H),6.65(s,1H),2.35(s,3H). 13C NMR(100MHz,DMSO-d6)δ194.2,164.8,157.0,156.6, 154.4,148.7,148.2,137.3,133.6,133.5,133.3,132.9,132.8,131.1,130.4,129 .0,128.9,127.2*2,123.5,115.9,115.7,114.4,107.9,20.6.HRMS(ESI)m / z:[M+H] + Calculated value C 25 H 19 FN3O3: 428.1405, measured value: 428.1403.

[0366] Example 29: Preparation of (3Z,6Z)-3-((5-methylpyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-29)

[0367]

[0368] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (100 mg, 0.27 mmol) and 5-methylpyridin-2-carboxaldehyde 25 h (39.7 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-29 was synthesized according to the preparation method of compound PLN-1-19, yielding 99.1 mg of a pale yellow solid in a yield of 84.9%. mp: 255-257 °C. 1 H NMR(400MHz,DMSO-d6) δ12.53(s,1H),10.62(s,1H),8.57(s,1H),7.92(t,J=8.2Hz,2H),7.85(s,1H),7.75(dd,J=17.4,7.7Hz, 2H),7.66(d,J=7.6Hz,1H),7.59(t,J=9.3Hz,2H),7.41(t,J=8.7Hz,2H),6.88(s,1H),6.70(s,1H),2.35 (s,3H). 13C NMR(100MHz,DMSO-d6)δ194.2,163.6,157.1,156.6,152.0,148.8,138.2,137.3,133.6,133.5,133.3, 132.9,132.8,132.4,130.4,130.3,129.0,128.9,127.3,126.2,115.9,115.7,114.2,108.0,18.0.HRMS (ESI)m / z:[M+H] + Calculated value C 25 H 19 FN3O3: 428.1405, measured value: 428.1401.

[0369] Example 30: Preparation of (3Z,6Z)-3-((6-methylpyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-30)

[0370]

[0371] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (50.0 mg, 0.14 mmol) and 6-methylpyridin-2-carboxaldehyde 25 (19.8 mg, 0.16 mmol) as the starting materials for this reaction, compound PLN-1-30 was synthesized according to the preparation method of compound PLN-1-19, yielding 37.0 mg of a pale yellow solid in a yield of 63.4%. mp: 220-223 °C. 1 H NMR (500MHz, DMSO-d6) δ12.80 (s, 1H), 10.61 (s, 1H), 7.92 (dd, J=8.8, 5.6Hz, 2H), 7.85 (s, 1H), 7.80 (t, J=7.8Hz, 2H), 7.66 (d, J =7.7Hz,1H),7.60(t,J=7.7Hz,1H),7.47(d,J=7.8Hz,1H),7.41(t,J=8.8Hz,2H),7.25(d,J=7.7Hz,1H),6.89(s,1H),6.68(s,1H),2.58(s,3H). 13C NMR(100MHz,DMSO-d6)δ194.2,163.6,157.1,157.0,156.6,154.0,138.1,137.3,133.6,133.5,133.3,132.9,13 2.8,130.9,130.4,129.0,128.9,127.3,123.8,122.2,115.9,115.7,114.2,107.8,24.1.HRMS(ESI)m / z:[M+Na] + Calculated value C 25 H 18 FN3NaO3: 450.1224, measured value: 450.1222.

[0372] Example 31: Preparation of (3Z,6Z)-3-((5-bromopyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-31)

[0373]

[0374] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (100 mg, 0.27 mmol) and 5-bromopyridin-2-carboxaldehyde 25 L (61.0 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-31 was synthesized according to the preparation method of compound PLN-1-19, yielding 65.3 mg of a brownish-yellow solid in a yield of 48.6%. mp: 256-259 °C. 1 H NMR(400MHz,DMSO-d6)δ 12.11(s,1H),10.69(s,1H),8.87(s,1H),8.15(d,J=8.4Hz,1H),7.91(t,J=7.9Hz,2H),7.84(s,1H),7 .77(d,J=7.5Hz,1H),7.65(d,J=8.3Hz,2H),7.60(t,J=7.6Hz,1H),7.40(t,J=8.6Hz,2H),6.90(s,1H), 6.73(s,1H). 13C NMR(125MHz,Pyridine-d5)δ194.5,166.0,159.2,158.1,157.9,154.3,140.4,138.8,135.1,134.4,133.8,133 .5,133.4,133.0,131.6,130.1,129.8,129.0,127.9,119.2,116.4116.2,115.64,107.8.HRMS(ESI)m / z:[M+H] + Calculated value C 24 H 16 BrFN3O3: 492.0354, measured value: 492.0351.

[0375] Example 32: Preparation of (3Z,6Z)-3-(quinoline-2-ylmethylene)-6-(3-(4-fluorobenzoyl)benzylmethylene)piperazine-2,5-dione (PLN-1-32)

[0376]

[0377] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (100 mg, 0.27 mmol) and quinoline-2-carboxaldehyde 25 mg (51.6 mg, 0.33 mmol) as starting materials for this reaction, compound PLN-1-32 was synthesized according to the preparation method of compound PLN-1-19, yielding 75.5 mg of a brownish-yellow solid in a yield of 59.7%. mp: 241-243 °C. 1 H NMR(400MHz,DMSO-d6)δ10.74 (s,1H),8.44(d,J=8.5Hz,1H),8.00(d,J=8.4Hz,2H),7.92(t,J=8.2Hz,2H),7.86(d,J=10.9Hz,2H), 7.80(m,J=13.0,2H),7.63(dt,J=20.8,7.8Hz,3H),7.41(t,J=8.7Hz,2H),6.94(s,1H),6.86(s,1H). 13C NMR(100MHz,Pyridine-d5)δ194.5,166.0,158.2*2,156.3,147.4,138.7,137.7,135.1,134.4,134.1,133.9, 133.5,133.4,131.7,131.0,130.1,129.8,129.1,129.1,128.5,127.6,127.4,124.7,116.4,116.2,115.7,108.4.HRMS(ESI)m / z:[M+H] + Calculated value C 28 H 19 FN3O3: 464.1405, measured value: 464.1403.

[0378] Example 33: Preparation of (3Z,6Z)-3-(isoquinoline-3-ylmethylene)-6-(3-(4-fluorobenzoyl)benzylmethylene)piperazine-2,5-dione (PLN-1-33)

[0379]

[0380] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (70.0 mg, 0.19 mmol) and isoquinoline-3-carboxaldehyde 25n (36.0 mg, 0.23 mmol) as starting materials for this reaction, compound PLN-1-33 was synthesized according to the preparation method of compound PLN-1-19, yielding 64.0 mg of a pale yellow solid in a yield of 72.3%. mp: 278-280℃. 1 H NMR(400MHz,DMSO-d6)δ12.52 (s,1H),10.63(s,1H),9.48(s,1H),8.20(d,J=8.2Hz,1H),8.13(s,1H),7.98(d,J=8.0Hz,1H),7.93(dd,J=8.9,5.5Hz,2H),7.86(t,J=7.0Hz,2 H),7.80(d,J=7.6Hz,1H),7.74(t,J=7.5Hz,1H),7.66(d,J=7.7Hz,1H),7.60(t,J=7.6Hz,1H),7.41(t,J=8.9Hz,2H),6.89(s,1H),6.85(s,1H). 13C NMR(100MHz, DMSO-d6)δ194.3,163.6,157.2,156.5,152.1,148.1,137.3,135.8,133.6,133.5*2,133.0,132.9,131.9, 130.4*2,129.9*2,129.0,128.5,128.4*2,128.3,127.0,126.8,123.2,115.9,115.7.HRMS(ESI)m / z:[M+H] + Calculated value C 28 H 19 FN3O3: 464.1405, measured value: 464.1402.

[0381] Example 34: Preparation of (3Z,6Z)-3-(isoquinoline-1-ylmethylene)-6-(3-(4-fluorobenzoyl)benzylmethylene)piperazine-2,5-dione (PLN-1-34)

[0382]

[0383]

[0384] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (70.0 mg, 0.19 mmol) and isoquinoline-1-carboxaldehyde 25 (36.0 mg, 0.23 mmol) as the starting materials for this reaction, compound PLN-1-34 was synthesized according to the preparation method of compound PLN-1-19, yielding 67.2 mg of a pale yellow solid, with a yield of 75.9%. mp: 274-276℃. HRMS (ESI) m / z: [M+H] + Calculated value C 28 H 19 FN3O3: 464.1405, measured value: 464.1403.

[0385] Example 35: Preparation of (3Z,6Z)-3-((5-ethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-35)

[0386]

[0387] 1) Synthesis of 5-ethoxypyridine-2-aldehyde 25p

[0388]

[0389] Take a 25 mL dry round-bottom flask, and add 20 mg (25.0 mg, 0.20 mmol) of 5-hydroxypyridine-2-carboxaldehyde, 22.1 mg (0.20 mmol) of bromoethane, 28.1 mg (0.20 mmol) of potassium carbonate, and 1.5 mL of DMF in sequence. Purge the mixture three times under nitrogen protection, and stir the mixture in a 100 °C oil bath for 1 h. Monitor the reaction completion by LC-MS. After the reaction solution cools to room temperature, add excess water (10 mL), extract three times with ethyl acetate (50 mL * 3), combine the organic phases, wash the organic phase with saturated brine (50 mL), combine the aqueous phases, back-extract the aqueous phase three times with ethyl acetate (50 mL * 3), combine the organic phases, and dry with anhydrous sodium sulfate. Filter under vacuum, concentrate the filtrate under reduced pressure, and purify by column chromatography (PE:EA = 1:1) to obtain 14.3 mg of a pale yellow oil, yield 53.1%. 1 H NMR (400MHz, CDCl3) δ9.98 (s, 1H), 8.41 (s, 1H), 7.95 (d, J = 8.6Hz, 1H), 7.27 (dd, J = 2.7, 2.8Hz, 1H), 4.30 (t, J = 6.7Hz, 2H), 4.13 (t, J = 6.6 Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C8H 10 NO2: 152.07, measured value: 151.57.

[0390] 2. Synthesis of (3Z,6Z)-3-((5-ethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-35

[0391]

[0392] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (56.4 mg, 0.15 mmol) and 5-ethoxypyridine-2-aldehyde 25p (28.0 mg, 0.19 mmol) as the starting materials for this step, compound PLN-1-35 was synthesized according to the preparation method of compound PLN-1-19, yielding 47.3 mg of a pale yellow solid in a yield of 67.1%. mp: 229-231 °C. 1H NMR(500MHz,DMSO-d6) δ12.33(s,1H),10.53(s,1H),8.46(d,J=3.0Hz,1H),7.92(dd,J=8.8,5.5 Hz, 2H), 7.84 (s, 1H), 7.77 (d, J = 7.7Hz, 1H), 7.65 (dd, J = 8.2, 3.4Hz, 2H), 7.59 (t, J=7.7Hz, 1H), 7.51 (dd, J=8.8, 3.0Hz, 1H), 7.40 (t, J=8.8Hz, 2H) ,6.87(s,1H),6.72(s,1H),4.17(q,J=7.0Hz,2H),1.37(t,J=7.0Hz,3H). 13 C NMR (100 MHz, DMSO-d6) δ194.2,163.6,157.2,156.6,153.8,146.9,137.3,136.9,133.6,133.5,133.4,132.9,132.8, 130.4,129.0,128.9,128.7,127.8,127.3,122.5,115.9,115.7,113.9,108.3,64.1,14.5.HRMS(ESI)m / z:[M+H] + Calculated value C 26 H 21 FN3O4: 458.1511, measured value: 458.1507.

[0393] Example 36: Preparation of (3Z,6Z)-3-((5-propoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-36)

[0394]

[0395] 1) Synthesis of 5-propoxypyridine-2-carboxaldehyde 25q

[0396]

[0397] 5-hydroxypyridine-2-carboxaldehyde 20 (50.0 mg, 0.41 mmol) and n-iodopropane (69.0 mg, 0.41 mmol) were used as the starting materials for this step of the reaction. Compound 25q was synthesized according to the preparation method of compound 25p, yielding 48.1 mg of a pale yellow oil with a yield of 71.7%. 1H NMR (400MHz, CDCl3) δ9.99 (s, 1H), 8.43 (s, 1H), 7.95 (d, J = 8.6Hz, 1H), 7.29 (d, J = 2.8Hz, 1H), 4.06 (t,J=6.5Hz,2H),1.88(h,J=7.0Hz,2H),1.07(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C9H 12 NO2: 166.09, measured value: 165.67.

[0398] 2. Synthesis of (3Z,6Z)-3-((5-ethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-36

[0399]

[0400] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (79.1 mg, 0.22 mmol) and 5-propoxypyridine-2-carboxaldehyde 25q (42.7 mg, 0.26 mmol) as starting materials for this reaction, compound PLN-1-36 was synthesized according to the preparation method of compound PLN-1-19, yielding 65.8 mg of a pale yellow solid in a yield of 64.6%. mp: 223-225℃. 1 H NMR (500MHz, DMSO- d6) δ12.33(s,1H),10.53(s,1H),8.47(d,J=2.8Hz,1H),7.92(dd,J=8.5,5.6Hz,2H),7.84(s,1H),7.77 (d,J=7.7Hz,1H),7.65(dd,J=8.0,5.4Hz,2H),7.59(t,J=7.6Hz,1H),7.52(dd,J=8.7,2.9Hz,1H),7.40(t ,J=8.7Hz,2H),6.87(s,1H),6.71(s,1H),4.08(t,J=6.5Hz,2H),1.80-1.73(m,2H),0.99(t,J=7.4Hz,3H). 13C NMR (100MHz, DMSO-d6) δ194.2,164.8,157.2,156.6,153.9,146.9,137.3,136.9,133.6,133.5, 133.4,132.9,132.8,130.4,129.0,128.9,128.7,127.7,127.3,122.5,115.9,115.7,113.9,108.3,69.8,21.9,10.3.HRMS(ESI)m / z:[M+H] + Calculated value C 27 H 23 FN3O4: 472.1667, measured value: 472.1664.

[0401] Example 37 Preparation of (3Z,6Z)-3-((5-butoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-37)

[0402]

[0403] 1) Synthesis of 5-butoxypyridine-2-carboxaldehyde 25r

[0404]

[0405] 5-hydroxypyridine-2-carboxaldehyde 20 (50.0 mg, 0.41 mmol) and iodobutane (74.7 mg, 0.41 mmol) were used as the starting materials for this reaction step. Compound 25r was synthesized according to the preparation method of compound 25p, yielding 43.5 mg of a colorless transparent oil with a yield of 59.8%. 1 H NMR (400MHz, CDCl3) δ9.99 (s, 1H), 8.42 (d, J = 2.8Hz, 1H), 7.95 (d, J = 8.1Hz, 1H), 7.28 (dd, J = 8.7, 2.8Hz,1H),4.10(t,J=6.4Hz,2H),1.86-1.79(m,2H),1.57-1.47(m,2H),1.00(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C 10 H 14 NO2: 180.10, measured value: 179.68.

[0406] 2. Synthesis of (3Z,6Z)-3-((5-butoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-37

[0407]

[0408] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (64.9 mg, 0.18 mmol) and 5-butoxypyridine-2-carboxaldehyde 25r (38.1 mg, 0.21 mmol) as starting materials for this reaction, compound PLN-1-37 was synthesized according to the preparation method of compound PLN-1-19, yielding 67.0 mg of a pale yellow solid in a yield of 77.9%. mp: 224-226 °C. 1 H NMR (400MHz, DMSO- d6) δ12.33(s,1H),10.56(s,1H),8.46(s,1H),7.91(t,J=8.7Hz,2H),7.84(s,1H),7.77(d,J=7.5Hz,1H), 7.65(dd,J=7.9,4.8Hz,2H),7.59(t,J=7.6Hz,1H),7.51(d,J=8.7Hz,1H),7.40(t,J=8.7Hz,2H),6.8 7(s,1H),6.71(s,1H),4.10(t,J=6.4Hz,2H),1.72(p,J=6.5Hz,2H),1.44(h,J=7.3Hz,2H),0.94(t,J= 7.4Hz, 3H). 13 C NMR(100MHz,DMSO-d6)δ194.2,163.6,157.2,156.5,153.9,146.9,137.3,136.9,133.5 *2,133.4 132.9,132.8,130.4,129.0,128.9,128.6,127.7,127.3,122.5,115.9,115.7,113.9,108.3,68.0,30.6,18.7,13.7.HRMS(ESI)m / z:[M+H] + Calculated value C 28 H 25 FN3O4: 486.1824, measured value: 486.1819.

[0409] Example 38 Preparation of (3Z,6Z)-3-((5-isopropoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-38)

[0410]

[0411] 1) Synthesis of 5-hydroxy-N-methoxy-N-methylpyridine amide 22

[0412]

[0413] Take a 100 mL dry round-bottom flask, dissolve 5-hydroxypyridinecarboxylic acid 21 (1.00 g, 7.19 mmol) in DMF, stir at room temperature for 1 min, then add N-methyl-N-methoxyamine hydrochloride (1.05 g, 10.8 mmol), EDCI (2.07 g, 10.8 mmol), HOBT (1.65 g, 12.2 mmol), and triethylamine (2.91 g, 28.8 mmol) sequentially, and stir the reaction at room temperature for 6.5 h. After the reaction is complete as monitored by LC-MS, add an appropriate amount of saturated ammonium chloride aqueous solution to the reaction solution until the solution becomes clear. Extract three times with ethyl acetate (100 mL * 3), combine the organic phases, wash the organic phase with saturated brine (100 mL), combine the aqueous phases, back-extract the aqueous phase three times with ethyl acetate (80 mL * 3), combine the organic phases, and dry with anhydrous sodium sulfate. The filtrate was filtered under reduced pressure and concentrated. It was then purified by column chromatography (PE:EA = 1:1) to obtain 860 mg of a brownish-yellow oily substance, with a yield of 65.7%. 1 H NMR (400MHz, CDCl3) δ8.16 (s, 1H), 7.60 (d, J = 8.6Hz, 1H), 7.19 (dd, J = 8.6, 2.7Hz, 1H), 3.72 (s, 3H), 3.44 (s, 3H). MS (ESI) m / z: [M+H] + Calculated value C8H 11 FN2O3: 183.08, measured value: 182.71.

[0414] 2) Synthesis of 5-isopropoxy-N-methoxy-N-methylpyridine amide 24S

[0415]

[0416] Take a 100 mL dry round-bottom flask, dissolve 300 mg (1.65 mmol) of 5-hydroxy-N-methoxy-N-methylpyridine amide 22 in 10 mL of dry DCM, stir at room temperature for 1 min, then add isopropanol (149 mg, 2.47 mmol), triphenylphosphine (562 mg, 2.14 mmol), and diisopropyl azodicarbonate (433 mg, 2.14 mmol) sequentially, and stir at room temperature for 30 min. After the reaction is complete as monitored by LC-MS, add 50 mL of water to the reaction solution, extract three times with DCM (100 mL * 3), combine the organic phases, wash the organic phase with 100 mL of saturated brine, combine the aqueous phases, back-extract the aqueous phase three times with DCM (80 mL * 3), combine the organic phases, and dry with anhydrous sodium sulfate. The filtrate was filtered, concentrated under pressure, and purified by column chromatography (PE:EA = 10:1, 8:1, 5:1, 2:1) to obtain 223 mg of a pale yellow oil, with a yield of 45.4%. 1 H NMR (400MHz, CDCl3) δ8.23(s,1H),7.70(d,J=8.6Hz,1H),7.20(dd,J=8.7,2.9Hz,1H),4.62(hept, J=6.0Hz,1H),3.76(s,3H),3.41(s,3H),1.36(d,J=8.1Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C 11 H 17 FN2O3: 225.12, measured value: 224.83.

[0417] 3) Synthesis of 5-isopropoxypyridine-2-carboxaldehyde 25s

[0418]

[0419] Take a 50 mL dry, sealed round-bottom flask, purge with nitrogen three times, dissolve diisobutylaluminum hydride (199 mg, 1.40 mmol) in THF (5 mL), stir at -78 °C for 10 min, then slowly add a 2 mL THF solution of 5-isopropoxy-N-methoxy-N-methylpyridine amide 24s (209 mg, 0.93 mmol). After the addition is complete, stir the reaction at -78 °C for 1.5 h. Monitor the reaction completion by LC-MS, add 20 mL of saturated sodium potassium tartrate aqueous solution to the reaction mixture, and move the reaction flask to room temperature and stir for 1 h. Extract three times with ethyl acetate (50 mL * 3), combine the organic phases, wash the organic phase with saturated brine (50 mL), combine the aqueous phases, back-extract the aqueous phase three times with ethyl acetate (40 mL * 3), combine the organic phases, and dry with anhydrous sodium sulfate. The filtrate was filtered under reduced pressure and concentrated. It was then purified by column chromatography (PE:EA = 10:1, 8:1) to obtain 109.4 mg of a pale yellow oil, with a yield of 71.0%. 1 H NMR(400MHz, CDCl3)δ9.97(s,1H),8.39(s,1H), 7.94(d,J=8.7Hz,1H),7.27-7.25(m,1H),4.71(hept,J=5.8Hz,1H),1.40(d,J=6.1Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C9H 12 NO2: 166.09, Measured value: 165.65.

[0420] Synthesis of (4)(3Z,6Z)-3-((5-isopropoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-38

[0421]

[0422] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (121 mg, 0.33 mmol) and 5-isopropoxypyridine-2-carboxaldehyde 25s (65.2 mg, 0.40 mmol) as the starting materials for this reaction, compound PLN-1-38 was synthesized according to the preparation method of compound PLN-1-19, yielding 131 mg of a pale yellow solid in 84.3% yield. mp: 234-236℃. 1H NMR (500MHz, DMSO- d6) δ12.33(s,1H),10.53(s,1H),8.44(d,J=2.9Hz,1H),7.92(dd,J=8.7,5.6Hz,2H),7.84(s,1H),7.77 (d,J=7.7Hz,1H),7.62(dt,J=15.3,8.0Hz,3H),7.51(dd,J=8.8,2.9Hz,1H),7.40(t ,J=8.8Hz,2H),6.87(s,1H),6.71(s,1H),4.83-4.75(m,1H),1.31(d,J=6.0Hz,6H). 13 C NMR(100MHz,DMSO-d6)δ 194.2,164.8,157.2,156.6,152.8,146.7,137.8,137.3,133.6,133.5,133.4,132.9,132.8,130.4,1 29.0*2,128.6,127.8,127.4,123.4,115.9,115.7,113.9,108.3,70.4,21.6*2.HRMS(ESI)m / z:[M+H] + Calculated value C 27 H 23 FN3O4: 472.1667, measured value: 472.1663.

[0423] Example 39 Preparation of (3Z,6Z)-3-((5-(2-cyclohexylethoxy)pyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-39)

[0424]

[0425] 1) Synthesis of 5-(2-cyclohexylethoxy)pyridine-2-aldehyde 25t

[0426]

[0427] 5-hydroxypyridine-2-carboxaldehyde 20 (50.0 mg, 0.41 mmol) and 2-bromoethylcyclohexane (77.6 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25t was synthesized according to the preparation method of compound 25p, yielding 74.5 mg of a pale yellow oil with a yield of 78.6%. 1H NMR (400MHz, CDCl3) δ9.99(s,1H),8.42(s,1H),7.95(dd,J=8.6,0.5Hz,1H),7.28(dd,J=5.2,4.2 Hz,1H),4.32-4.28(m,10H),4.21-4.15(m,3H),3.74-3.69(m,2H).MS(ESI)m / z:[M+H] + Calculated value C 14 H 20 NO2: 234.15, measured value: 233.84.

[0428] Synthesis of (2)(3Z,6Z)-3-((5-(2-cyclohexylethoxy)pyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-39

[0429]

[0430] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (87.0 mg, 0.24 mmol) and 5-(2-cyclohexylethoxy)pyridine-2-carboxaldehyde 25 t (66.5 mg, 0.29 mmol) as starting materials for this reaction, compound PLN-1-39 was synthesized according to the preparation method of compound PLN-1-19, yielding 115 mg of a pale yellow solid in 89.6% yield. mp: 217-219℃. 1 H NMR(500MHz, DMSO-d6)δ12.62(s,1H),8.34(s,1H),8.25(s,1H),7.86(dd,J=8.5,5.5Hz,2H),7.75(t,J=7.4Hz,2H), 7.63-7.57(m,2H),7.30(d,J=8.7Hz,1H),7.24-7.17(m,3H),7.07(s,1H),6.72(s,1H),4. 09(t,J=6.6Hz,2H),1.78-1.63(m,7H),1.31-1.17(m,4H),0.99(dd,J=22.7,10.8Hz,2H). 13C NMR (100MHz, Pyridine- d5) δ194.8,166.0,158.6,157.9,154.8,148.2,138.7,137.3,135.3,134.4,133.8,133.5,133.4,131.6,130.3, 129.9,129.8,129.4,127.9,122.9,116.4,116.2,114.8,109.4,67.3,36.9,35.0,33.6*2,27.0,26.8*2.HRMS(ESI)m / z:[M+H] + Calculated value C 32 H 31 FN3O4: 540.2293, measured value: 540.2291.

[0431] Example 40: Preparation of (3Z,6Z)-3-((5-benzyloxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-40)

[0432]

[0433] 1) Synthesis of 5-benzyloxy-N-methoxy-N-methylpyridine amide 24u

[0434]

[0435] 5-hydroxy-N-methoxy-N-methylpyridine amide 22 (300 mg, 1.65 mmol) and benzyl alcohol (267 mg, 2.47 mmol) were used as the starting materials for this step of the reaction. Compound 24u was synthesized according to the preparation method of compound 24s, yielding 169 mg of a pale yellow oil with a yield of 37.8%. 1 H NMR (400MHz, DMSO-d6) δ8.36 (s, 1H), 7.62-7.54 (m, 2H), 7.48 (d, J = 7.2Hz, 2H), 7.39 (dt, J = 15.8, 7.1Hz,3H),5.24(s,2H),3.67(s,3H),3.27(s,3H).MS(ESI)m / z:[M+H] + Calculated value C 15 H 17 N2O3: 273.12, Measured value: 272.79.

[0436] 2) Synthesis of 5-benzyloxypyridine-2-carboxaldehyde 25u

[0437]

[0438] 5-Benzyloxy-N-methoxy-N-methylpyridine amide 24u (169 mg, 0.62 mmol) was used as the starting material for this step of the reaction. Compound 25u was synthesized according to the preparation method of compound 25s, yielding 109 mg of a pale yellow oil with a yield of 82%. 1 H NMR (400MHz, CDCl3) δ9.99(s,1H),8.51(d,J=2.3Hz,1H),7.95(d,J=8.7Hz,1H),7.43-7.42(m,3H),7.40-7.35(m,3H),5.21(s,2H).MS(ESI)m / z:[M+H] + Calculated value C 13 H 12 NO2: 214.09, measured value: 213.79.

[0439] Synthesis of (3Z,6Z)-3-((5-benzyloxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-40

[0440]

[0441] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (102 mg, 0.28 mmol) and 5-benzyloxypyridine-2-carboxaldehyde 25 u (71.0 mg, 0.33 mmol) as the starting materials for this reaction, compound PLN-1-40 was synthesized according to the preparation method of compound PLN-1-19, yielding 121 mg of a yellowish-white solid, with a yield of 84.1%. mp: 220-223℃. 1 H NMR(500MHz,DMSO-d6) δ12.32(s,1H),10.54(s,1H),8.54(d,J=2.8Hz,1H),7.92(dd,J=8.6,5.6Hz,2H),7.84(s,1H),7.77(d ,J=7.6Hz,1H),7.66(t,J=8.6Hz,2H),7.60(t,J=8.6Hz,2H),7.49(d,J=7.3Hz,2H),7.43-7.35(m,5H), 6.88(s,1H),6.71(s,1H),5.26(s,2H). 13C NMR(100MHz,DMSO-d6)δ194.3,164.9,157.2,156.6,153.6, 147.3,137.3,137.2,136.2,133.6,133.5,133.4,133.0,132.9,130.4,129.1,129.0,128.9,128. 7*2,128.3,128.1*2,127.7,127.4,123.1,115.9,115.7,114.0,108.2,70.0.HRMS(ESI)m / z:[M+H] + Calculated value C 31 H 23 FN3O4: 520.1667, measured value: 520.1663.

[0442] Example 41: Preparation of (3Z,6Z)-3-((5-phenylethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)phenylmethylene)piperazine-2,5-dione (PLN-1-41)

[0443]

[0444] 1) Synthesis of 5-phenylethoxy-N-methoxy-N-methylpyridine amide 24v

[0445]

[0446] 5-hydroxy-N-methoxy-N-methylpyridine amide 22 (300 mg, 1.65 mmol) and phenylethanol (302 mg, 2.47 mmol) were used as the starting materials for this step of the reaction. Compound 24v was synthesized according to the preparation method of compound 24s, yielding 291 mg of a pale yellow oil, with a yield of 46.2%. 1 H NMR (400MHz, CDCl3) δ8.24 (s, 1H), 7.67 (d, J = 8.5Hz, 1H), 7.32-7.18 (m, 6H), 4.23 (t, J = 7.0Hz, 2H),3.73(s,3H),3.38(s,3H),3.10(t,J=7.0Hz,2H).MS(ESI)m / z:[M+H] + Calculated value C 16 H 19 N2O3: 287.14, Measured value: 286.80.

[0447] 2) Synthesis of 5-phenylethoxypyridine-2-carboxaldehyde 25v

[0448]

[0449] 5-Phenylacetyl-N-methoxy-N-methylpyridine amide 24v (281 mg, 0.98 mmol) was used as the starting material for this step of the reaction. Compound 25v was synthesized according to the preparation method of compound 25s, yielding 178 mg of a colorless, transparent oil with a yield of 80.1%. 1 H NMR (400MHz, CDCl3) δ9.98 (s, 1H), 8.42 (d, J = 2.8Hz, 1H), 7.94 (d, J = 8.7Hz, 1H), 7.36-7.26 (m, 6H), 4.31 (t, J = 6.9 Hz,2H),3.16(t,J=6.9Hz,2H).MS(ESI)m / z:[M+H] + Calculated value C 14 H 14 NO2: 228.10, measured value: 227.77.

[0450] Synthesis of (3Z,6Z)-3-((5-phenylethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)phenylmethylene)piperazine-2,5-dione 26v

[0451]

[0452] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (79.5 mg, 0.22 mmol) and 5-phenylethoxypyridine-2-carboxaldehyde 25v (59.0 mg, 0.26 mmol) as the starting materials for this reaction, compound PLN-1-41 was synthesized according to the preparation method of compound PLN-1-19, yielding 98.7 mg of a pale yellow solid in 85.2% yield. mp: 175-178℃. 1 H NMR (500MHz, DMSO- d6) δ12.32(s,1H),10.53(s,1H),8.47(d,J=2.9Hz,1H),7.91(dd,J=8.7,5.6Hz,2H),7.84(s,1H),7.77 (d,J=7.7Hz,1H),7.62(dt,J=15.3,8.2Hz,3H),7.53(dd,J=8.8,2.9Hz,1H),7.40(t,J=8.8Hz,2H),7. 35-7.30(m,4H),7.23(t,J=6.8Hz,1H),6.87(s,1H),6.71(s,1H),4.35(t,J=6.9Hz,2H),3.08(t,J=6.9 Hz, 2H). 13C NMR (100MHz, DMSO-d6) δ194.2,163.6,157.2,156.6,153.7,147.1,137.9,137.3,136.9, 133.6,133.5*2,133.4,132.9,132.8,130.4,129.1,129.0*2,128.9,128.7,128.4,127 .7,127.4,126.5,122.7,115.9,115.7,113.9,108.2,68.9,34.7.HRMS(ESI)m / z:[M+H] + Calculated value C 32 H 25 FN3O4: 534.1824, measured value: 534.1823.

[0453] Example 42 Preparation of (3Z,6Z)-3-((3-ethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-42)

[0454]

[0455] 1) Synthesis of 3-ethoxypyridine-2-carboxaldehyde 25w

[0456]

[0457] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and bromoethane (44.2 mg, 0.41 mmol) were used as starting materials for this reaction. Compound 25w was synthesized according to the preparation method of compound 25p, yielding 32.8 mg of a pale yellow solid with a yield of 53.4%. 1 H NMR (400MHz, DMSO-d6) δ10.21(s,1H),8.34(d,J=4.3Hz,1H),7.73(d,J=8.6Hz,1H),7.63(dd,J=8.6,4.3 Hz,1H),4.21(q,J=7.0Hz,2H),1.38(t,J=7.0Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C8H 10 NO2: 152.07, measured value: 151.65.

[0458] Synthesis of (2)(3Z,6Z)-3-((3-ethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26w

[0459]

[0460] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (56.1 mg, 0.15 mmol) and 3-ethoxypyridine-2-carboxaldehyde 25w (27.6 mg, 0.18 mmol) as the starting materials for this step, compound PLN-1-42 was synthesized according to the preparation method of compound PLN-1-19, yielding 33.6 mg of a pale yellow solid in a yield of 48.0%. mp: 225-227℃. 1 H NMR (500MHz, DMSO- d6) δ12.63(s,1H),10.61(s,1H),8.30(d,J=4.7Hz,1H),7.92(dd,J=8.7,5.5Hz,2H),7.84(s,1H),7.78 (d,J=7.6Hz,1H),7.66(d,J=7.7Hz,1H),7.61-7.56(m,2H),7.39(dt,J=8.5,6.8 Hz,3H),7.01(s,1H),6.89(s,1H),4.18(q,J=6.9Hz,2H),1.41(t,J=6.9Hz,3H). 13 C NMR(100MHz,DMSO-d6)δ194.2,157.1, 156.7,153.1,144.0,139.8,137.3,133.6,133.5,133.4,133.3,132.9,132.8,130.5,130.3,12 9.0,128.9,127.2,123.8,119.9,115.9,115.6,114.3,101.4,64.2,14.5.HRMS(ESI)m / z:[M+H] + Calculated value C 26 H 21 FN3O4: 458.1511, measured value: 458.1505.

[0461] Example 43: Preparation of (3Z,6Z)-3-((3-propoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-43)

[0462]

[0463] 1) Synthesis of 3-propoxypyridine-2-carboxaldehyde 25x

[0464]

[0465] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and n-iodopropane (69.0 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25x was synthesized according to the preparation method of compound 25p, yielding 67.1 mg of a pale yellow oil with a yield of 70.6%. 1 H NMR (400MHz, CDCl3) δ10.43(s,1H),8.38(d,J=4.3Hz,1H),7.45(dd,J=8.6,4.3Hz,1H),7.39(d,J= 8.6Hz,1H),4.07(t,J=6.1Hz,2H),1.91(h,J=7.4Hz,2H),1.09(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C9H 12 NO2: 166.09, measured value: 165.67.

[0466] 2. Synthesis of (3Z,6Z)-3-((3-propoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26x

[0467]

[0468] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (38.1 mg, 0.10 mmol) and 3-propoxypyridine-2-carboxaldehyde 25x (20.5 mg, 0.12 mmol) as starting materials for this reaction, compound PLN-1-43 was synthesized according to the preparation method of compound PLN-1-19, yielding 41.2 mg of a pale yellow solid in 83.4% yield. mp: 188-191℃. 1 H NMR (500MHz, DMSO- d6) δ (s, 1H), 10.61 (s, 1H), 8.30 (d, J = 4.6Hz, 1H), 7.92 (dd, J = 8.7, 5.6Hz, 2H), 7.85 (s, 1H), 7.78 (d, J = 7.6Hz,1H),7.66(d,J=7.7Hz,1H),7.59(dd,J=15.6,7.9Hz,2H),7.42-7.36(m,3H), 7.02(s,1H),6.89(s,1H),4.09(t,J=6.5Hz,2H),1.81(m,2H),1.02(t,J=7.4Hz,3H). 13C NMR(100MHz,DMSO-d6)δ194.6, 163.6,157.1,156.8,153.3,144.2,139.9,137.4,133.7,133.6,133.4,132.9,132.9,130.7,130.5,1 29.1,129.0,127.4,123.9,120.0,116.0,115.8,114.4,101.3,68.2,19.0,13.8.HRMS(ESI)m / z:[M+H] + Calculated value C 27 H 23 FN3O4: 472.1667, Measured value: 472.1662.

[0469] Example 44 Preparation of (3Z,6Z)-3-((3-butoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-44)

[0470]

[0471] 1) Synthesis of 3-butoxypyridine-2-carboxaldehyde 25y

[0472]

[0473] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and iodobutane (74.7 mg, 0.41 mmol) were used as the starting materials for this step of the reaction. Compound 25y was synthesized according to the preparation method of compound 25p, yielding 54.9 mg of an orange-yellow oil with a yield of 75.4%. 1 H NMR (400MHz, CDCl3) δ10.42(s,1H),8.38(d,J=2.9Hz,1H),7.45(dd,J=8.6,4.3Hz,1H),7.40(d,J= 7.3Hz,1H),4.11t,J=6.5Hz,2H),1.90-1.83(m,2H),1.59-1.49(m,2H),1.00(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C 10 H 14 NO2: 180.10, measured value: 179.69.

[0474] 2. Synthesis of (3Z,6Z)-3-((3-butoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione PLN-1-44

[0475]

[0476] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (84.6 mg, 0.23 mmol) and 3-butoxypyridine-2-carboxaldehyde 25y (49.6 mg, 0.28 mmol) as the starting materials for this reaction, compound PLN-1-44 was synthesized according to the preparation method of compound PLN-1-19, yielding 71.0 mg of a pale yellow solid in a yield of 63.3%. mp: 188-191℃. 1 H NMR (400MHz, DMSO- d6) δ10.63(s,1H),8.28(s,1H),7.91(t,J=5.64Hz 2H),7.84(s,1H),7.78(d,J=7.5Hz,1H),7.66(d,J= 7.6Hz,1H),7.59(dd,J=12.4,8.0Hz,2H),7.43-7.35(m,3H),7.00(s,1H),6.88(s,1H),4 .11(t,J=6.4Hz,2H),1.77(p,J=6.6Hz,2H),1.47(h,J=7.4Hz,2H),0.95(t,J=7.4Hz,3H). 13 C NMR (100MHz, DMSO- d6) δ194.2,164.8,157.1,156.7,153.2,144.1,139.9,137.3,133.6,133.5,133.3,132.9,132.8,130.6,130.4, 129.0,128.9,127.3,123.8,120.0,115.9,115.7,114.3,101.3,68.1,30.5,18.8,13.7.HRMS(ESI)m / z:[M+H] + Calculated value C 28 H 25 FN3O4: 486.1824, measured value: 486.1822.

[0477] Example 45: Preparation of (3Z,6Z)-3-((3-isopropoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-45)

[0478]

[0479] 1) Synthesis of 3-isopropoxypyridine-2-carboxaldehyde 25z

[0480]

[0481] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and 2-bromopropane (49.9 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25z was synthesized according to the preparation method of compound 25p, yielding 21.6 mg of a pale yellow oil with a yield of 32.2%. 1 H NMR (400MHz, CDCl3) δ10.44(s,1H),8.38(s,1H),7.71(dd,J=5.8,3.3Hz,1H),7.52(dd,J=5.8,3.3Hz,1H), 4.68(hept,J=6.0Hz,1H),1.43(d,J=6.0Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C9H 12 NO2: 166.09, Measured value: 165.65.

[0482] Synthesis of (2)(3Z,6Z)-3-((3-isopropoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26Z

[0483]

[0484] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (39.9 mg, 0.11 mmol) and 3-isopropoxypyridin-2-aldehyde 25z (21.6 mg, 0.13 mmol) as starting materials for this step, compound PLN-1-45 was synthesized according to the preparation method of compound PLN-1-19, yielding 42.7 mg of a pale yellow solid in 83.2% yield. mp: 179-181℃. 1 H NMR (400MHz, DMSO- d6) δ10.64(s,1H),8.28(d,J=3.6Hz,1H),7.91(dd,J=8.9,5.5Hz,2H),7.84(s,1H),7.77(d,J=7.6Hz, 2H),7.66(d,J=7.8Hz,2H),7.59(t,J=7.7Hz,1H),7.41(t,J=8.9Hz,1H),7.36(dd,J=8.5 ,4.7Hz,1H),6.99(s,1H),6.89(s,1H),4.77(hept,J=5.9Hz,1H),1.33(d,J=6.0Hz,6H). 13C NMR (100MHz, DMSO- d6) δ194.2,164.8,157.1,156.7,152.2,144.7,139.8,137.3,133.6,133.5,133.3,132.9,132.8,130.5,130.4, 129.0,128.9,127.2,123.8,121.2,115.9,115.7,114.3,101.6,70.7,21.6*2.HRMS(ESI)m / z:[M+H] + Calculated value C 27 H 23 FN3O4: 472.1667, measured value: 472.1663.

[0485] Example 46: Preparation of (3Z,6Z)-3-((3-cyclohexylmethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzylmethylene)piperazine-2,5-dione (PLN-1-46)

[0486]

[0487] 1) Synthesis of 25aa of 3-cyclohexylmethoxypyridine-2-carboxaldehyde

[0488]

[0489] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and bromomethylcyclohexane (71.9 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25aa was synthesized according to the preparation method of compound 25p, yielding 46.2 mg of a pale yellow oil with a yield of 51.9%. 1 H NMR (400MHz, DMSO-d6) δ10.20(s,1H),8.33(dd,J=4.4,1.2Hz,1H),7.73(d,J=7.6Hz,1H),7.62 (dd,J=8.6,4.4Hz,1H),3.95(d,J=6.2Hz,2H),1.85-1.64(m,6H),1.31-1.03(m,4H).MS(ESI)m / z:[M+H] + Calculated value C 13 H 18 NO2: 220.13, measured value: 219.80.

[0490] 2. Synthesis of (3Z,6Z)-3-((3-cyclohexylmethoxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26aa

[0491]

[0492] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazin-2,5-dione 17 (56.4 mg, 0.15 mmol) and 3-cyclohexylmethoxypyridine-2-carboxaldehyde 25aa (40.4 mg, 0.18 mmol) as the starting materials for this reaction, compound PLN-1-46 was synthesized according to the preparation method of compound PLN-1-19, yielding 65.5 mg of a pale yellow solid in 80.9% of the product. mp: 171-173 °C. 1 H NMR (400MHz, DMSO-d6) δ10.64(s,1H),8.29(d,J=3.6Hz,1H),7.91(dd,J=8.9,5.5Hz,2H),7.84(s,1H),7.78(d,J= 7.6Hz, 1H), 7.66 (d, J = 7.8Hz, 1H), 7.59 (dd, J = 15.1, 7.5Hz, 2H), 7.41 (t, J = 8.9Hz, 2H), 7.36 (dd, J = 8. 5,4.7Hz,1H),7.01(s,1H),6.88(s,4H),3.92(d,J=6.1Hz,2H),1.83-1.65(m,6H),1.32-1.03(m,5H). 13 C NMR (100MHz, DMSO-d6) δ194.2,164.8,157.0,156.7,153.3,144.1,139.9,137.3,133.6,133.5,133.4, 132.9,132.8,130.7,130.4,129.0,128.9,127.3,123.8,120.0,115.9,11 5.7,114.3,101.2,73.4,36.8,29.2*2,26.1,25.2*2.HRMS(ESI)m / z:[M+H] + Calculated value C 31 H 29 FN3O4: 526.2137, measured value: 526.2132.

[0493] Example 47 Preparation of (3Z,6Z)-3-((3-(2-cyclohexylethoxy)pyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-47)

[0494]

[0495] 1) Synthesis of 3-cyclohexylmethoxypyridine-2-carboxaldehyde 25ab

[0496]

[0497] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and 2-bromoethylcyclohexane (77.6 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25ab was synthesized according to the preparation method of compound 25p, yielding 69.1 mg of a pale yellow oil with a yield of 72.9%. 1 H NMR (400MHz, CDCl3) δ10.42(s,1H),8.38(dd,J=4.2,1.2Hz,2H),7.46-7.39(m,2H),4.14(t,J=6.7 Hz,2H),1.78-1.65(m,7H),1.31-1.15(m,4H),1.04-0.94(m,3H).MS(ESI)m / z:[M+H] + Calculated value C 14 H 20 NO2: 234.15, measured value: 233.84.

[0498] Synthesis of (2)(3Z,6Z)-3-((3-(2-cyclohexylethoxy)pyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26ab

[0499]

[0500] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (87.0 mg, 0.24 mmol) and 3-cyclohexylmethoxypyridine-2-carboxaldehyde 25ab (66.5 mg, 0.29 mmol) as starting materials for this reaction, compound PLN-1-47 was synthesized according to the preparation method of compound PLN-1-19, yielding 115 mg of a pale yellow solid in 89.6% yield. mp: 195-197℃. 1H NMR(500MHz, DMSO-d6)δ12.59(s,1H),10.59(s,1H),8.29(d,J=4.7Hz,1H),7.92(dd,J=8.7,5.6Hz,2H),7.86(s,1H), 7.79(d,J=7.6Hz,1H),7.66(d,J=7.7Hz,1H),7.59(t,J=7.8Hz,2H),7.42-7.35(m,3H),7.02(s,1H),6. 88(s,1H),4.15(t,J=6.7Hz,2H),1.74-1.60(m,7H),1.26-1.13(m,4H),0.98(dd,J=23.9,12.1Hz,2H). 13 C NMR(100MHz,DMSO-d6)δ194.2,157.0,156.7,153.2,144.1,139.9,137.3,133.5*2,133.4,133.3, 132.9,132.8,130.5,130.4,129.0,128.9,127.2,123.8,120.1,115.9,115.6 ,114.3,101.4,66.7,35.8,34.3,32.7*2,26.0,25.8*2.HRMS(ESI)m / z:[M+H] + Calculated value C 32 H 31 FN3O4: 540.2293, measured value: 540.2288.

[0501] Example 48 Preparation of (3Z,6Z)-3-((3-benzyloxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione (PLN-1-48)

[0502]

[0503] 1) Synthesis of 3-benzyloxypyridine-2-carboxaldehyde 25ac

[0504]

[0505] 3-hydroxypyridine-2-carboxaldehyde 18 (50.0 mg, 0.41 mmol) and benzyl bromide (69.4 mg, 0.41 mmol) were used as starting materials for this step of the reaction. Compound 25ac was synthesized according to the preparation method of compound 25p, yielding 48.2 mg of a pale yellow oil with a yield of 55.7%. 1H NMR (400MHz, DMSO-d6) δ10.22(s,1H),8.37(d,J=5.4Hz,1H),7.82(d,J=8.6Hz,1H),7.65(dd,J=8.6,4.4 Hz,1H),7.52(d,J=7.4Hz,2H),7.41(t,J=7.4Hz,2H),7.34(t,J=7.2Hz,1H),5.32(s,2H).MS(ESI)m / z:[M+H] + Calculated value C 13 H 12 NO2: 214.09, measured value: 213.79.

[0506] 2. Synthesis of (3Z,6Z)-3-((3-benzyloxypyridin-2-yl)methylene)-6-(3-(4-fluorobenzoyl)benzyl)piperazine-2,5-dione 26ac

[0507]

[0508] Using (Z)-1-acetyl-3-(3-(4-fluorobenzoyl)methylene)piperazine-2,5-dione 17 (62.7 mg, 0.17 mmol) and 3-benzyloxypyridine-2-carboxaldehyde 25ac (43.8 mg, 0.21 mmol) as starting materials for this reaction, compound PLN-1-48 was synthesized according to the preparation method of compound PLN-1-19, yielding 59.2 mg of a pale yellow solid in a yield of 66.6%. mp: 192-194 °C. 1 H NMR (500MHz, DMSO- d6) δ12.62(s,1H),10.62(s,1H),8.32(d,J=4.6Hz,1H),7.91(dd,J=8.6,5.6Hz,2H),7.84(s,1H),7.77 (d,J=7.6Hz,1H),7.66(dd,J=8.2,3.1Hz,2H),7.59(t,J=7.6Hz,1H),7.48( d,J=7.3Hz,2H),7.44-7.37(m,6H),7.03(s,1H),6.89(s,1H),5.28(s,2H). 13C NMR(100MHz,DMSO-d6)δ194.2,164.8,157.0,156.7, 152.8,144.3,140.2,137.3,136.2,133.6,133.5,133.3,132.9,132.8,130.8,130.4,129.1,128.9,1 28.7*2,128.3,128.0*2,127.2,123.7,120.5,115.9,115.7,114.4,101.2,69.9.HRMS(ESI)m / z:[M+H] + Calculated value C 31 H 23 FN3O4: 520.1667, measured value: 520.1664.

[0509] Example 49 Synthesis of (3Z,6Z)-3-(3-p-fluorobenzoxy)benzylmethyl-6-((1-methyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-1)

[0510]

[0511] Synthesis of (1)(Z)-1-acetyl-3-((1-methyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (18b)

[0512]

[0513] Take a 25 mL dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.36 mmol), DMF (2.5 mL), and cesium carbonate (353.8 mg, 1.09 mmol) sequentially. Place the flask in a -30 °C cold trap for 20 min, then add iodomethane (513.7 mg, 3.62 mmol) dropwise. Continue stirring at room temperature for 2 h. Monitor the reaction by LC-MS; the starting material has reacted completely. Add the reaction solution dropwise to 4 °C cold water, stir for 20 min, filter, and wash the filter cake with water. Dry in a vacuum drying oven at 50 °C to obtain 60.0 mg of a light yellow solid, yield 57%. 1 H NMR(500MHz, CDCl3)δ11.98(s,1H),7.39(s,1H),7.02(s,1H),4.46(s,2H), 3.67(s,3H),3.31–3.16(m,1H),2.64(s,3H),1.39(d,J=7.2Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C14 H 19 N4O3: 291.1452, measured value: 290.75.

[0514] 2) Synthesis of the final product (PLN-5-1)

[0515]

[0516] The compound PLN-1-1 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((1-methyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (30 mg, 0.10 mmol), p-fluorobenzoxybenzaldehyde (27 mg, 0.12 mmol), cesium carbonate (51 mg, 0.16 mmol), and anhydrous sodium sulfate (30 mg, 0.21 mmol). The mixture was reacted in DMF (3 mL) in an oil bath at 55 °C with stirring for 24 h. 17 mg of a pale yellow solid was obtained, with a yield of 37%. Melting point: 218-219 °C. 1 H NMR(500MHz,DMSO-d6)δ11.98(s,1H),10.12 (s,1H),7.84(s,1H),7.41(t,J=7.9Hz,1H),7.29-7.20(m,3H),7.17(s,1H),7.12(dd,J=8.9,4.5Hz,2H ),6.90(d,J=6.6Hz,1H),6.70(d,J=13.7Hz,2H),3.68(s,3H),3.31-3.23(m,1H),1.32(d,J=7.1Hz,6H). 13 C NMR(125MHz,DMSO-d6)δ159.1,157.2,157.0,156.8(J C-F =155.7Hz),152.6,138.6,138.1,135.2, 132.2,130.3,127.3,124.4,123.8,120.5*2(J C-F =8.4Hz),118.9,117.7,116.6*2(J C-F =23.2Hz),113.1,104.1, 32.2,23.8,21.9*2.MS(ESI):m / z 447.1825[M+H] + MPa: 218-219℃.

[0517] Example 50: Preparation of (3Z,6Z)-3-(3-p-fluorobenzoxy)benzylmethyl-6-((1-ethyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-2)

[0518]

[0519] Synthesis of (Z)-1-acetyl-3-((1-ethyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (18d)

[0520]

[0521] The compound 18b was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.54 mmol), cesium carbonate (530.7 mg, 1.63 mmol), and iodoethane (846.8 mg, 5.43 mmol). The mixture was stirred in DMF (4 mL) at 0 °C for 1.5 h. A light yellow solid of 111.8 mg was obtained, with a yield of 68%. 1 H NMR (500MHz, CDCl3) δ12.02(s,1H),7.45(s,1H),7.05(s,1H),4.46(s,2H),3.99(q,J=7.3Hz,2H),3.16(dt,J=14.4, 7.2Hz,1H),2.64(s,3H),1.45(t,J=7.3Hz,3H),1.41(d,J=7.2Hz,6H).MS(ESI)m / z:[M+H] + Calculated value C 15 H 21 N4O3: 305.1608, measured value: 304.82.

[0522] 2) Synthesis of the final product (PLN-5-2)

[0523]

[0524] The compound PLN-1-1 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((1-ethyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (152.2 mg, 0.50 mmol), p-fluorobenzoxybenzaldehyde (130 mg, 0.60 mmol), and cesium carbonate (244 mg, 0.75 mmol). The mixture was reacted in DMF (2 mL) in an oil bath at 55 °C with stirring for 24 h. 70.0 mg of a pale yellow solid was obtained, with a yield of 30%. Melting point: 195-198 °C. 1H NMR(500MHz,DMSO-d6)δ12.02(s,1H),10.13(s,1H),7.92(s,1H),7.42(t,J=7.9Hz,1H), 7.31-7.22(m,3H),7.18(s,1H),7.15-7.11(m,2H),6.92(dd,J=8.1,2.1Hz,1H),6.72(d, J=9.5Hz,2H),4.07(q,J=7.2Hz,2H),3.32-3.22(m,1H),1.34(d,J=7.0Hz,9H). 13 C NMR(125MHz, DMSO-d6)δ159.1,157.2,157.0,156.8(J C-F =154.2Hz),152.6,138.3,136.7,135.2,132.1,130.3,127.2,124.4,123.7,120.5*2(J C-F =8.4Hz),118.9,117.7,116.5*2(J C-F =23.2Hz),113.1,104.2,39.7,23.7,22.5*2, 16.5.MS(ESI):m / z 461.1973[M+H] + MPa: 195-198℃.

[0525] Example 51 Synthesis of (3Z,6Z)-3-(3-p-fluorobenzoxy)benzylmethyl-6-((1-n-propyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-3)

[0526]

[0527] Synthesis of (Z)-1-acetyl-3-((1-n-propyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (18f)

[0528]

[0529] The compound 18b was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.54 mmol), cesium carbonate (530.7 mg, 1.63 mmol), and iodopropane (922.9 mg, 5.43 mmol), with 4 mL of DMF, and stirred at room temperature for 2 h. A light yellow solid was obtained in 74 mg, yielding 43%. 1H NMR(500MHz, CDCl3)δ12.02(s,1H),7.42(s,1H),7.06(s,1H),4.46(s,2H),4.01–3.80(m,2H),3.24–2.99(m,1H),2.64 (s,3H),1.79(dd,J=14.8,7.4Hz,2H),1.41(d,J=7.2Hz,6H),0.98(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C 16 H 23 N4O3: 319.1765, measured value: 319.23.

[0530] 2) Synthesis of the final product (PLN-5-3)

[0531]

[0532] The compound PLN-1-1 was prepared according to the synthetic method. Starting materials included (Z)-1-acetyl-3-((1-n-propyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (75 mg, 0.24 mmol), p-fluorobenzoxybenzaldehyde (62 mg, 0.28 mmol), and cesium carbonate (116 mg, 0.35 mmol). The reaction mixture was stirred in an oil bath at 55 °C for 24 h in DMF (6 mL). 75 mg of a pale yellow solid was obtained, with a yield of 67%. Melting point: 201-203 °C. 1 H NMR (500MHz, DMSO-d6) δ12.00(s,1H),10.12(s,1H),7.89(s,1H),7.41(t,J= 7.9Hz,1H),7.30-7.20(m,3H),7.17(s,1H),7.12(dd,J=9.0,4.5Hz,2H),6.94-6.87(m,1H),6.71(d,J= 8.0Hz,2H),3.97(t,J=7.3Hz,2H),3.31-3.18(m,1H),1.71-1.67(m,2H),1.32(d,J=7.1Hz,6H),0.87(t, J=7.3Hz,3H). 13 C NMR(125MHz,DMSO-d6)δ159.1,157.2,157.0,156.8(J C-F =152.6Hz),152.6,138.5,137.2,135.2,132.1,130.3,127.2,124.4,123.7,120.5*2(J C-F=8.5Hz),118.9,117.7,116.5*2(J C-F =23.2 Hz),113.1,104.2,46.08,24.0,23.6,22.6*2,10.7.MS(ESI):m / z475.2133[M+H] + MPa: 201-203℃.

[0533] Example 52 Synthesis of (3Z,6Z)-3-(3-p-fluorophenoxy)benzylmethyl-6-((1-n-butyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-4)

[0534]

[0535] Synthesis of (Z)-1-acetyl-3-((1-n-butyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (18g)

[0536]

[0537] The compound 18b was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), cesium carbonate (1.1 g, 3.26 mmol), and n-butane iodide (2.0 g, 10.86 mmol). The reaction was carried out in 5 mL of DMF solution with stirring at room temperature for 4 h. 148.0 mg of a brownish-brown solid was obtained, with a yield of 41%. 1 H NMR (400MHz, CDCl3) δ12.03(s,1H),7.42(s,1H),7.05(s,1H),4.45(s,2H),3.90(t,J=7.4Hz,2H),3.13(dt,J=14.3,7.1 Hz,1H),2.64(s,3H),1.80–1.67(m,2H),1.46–1.25(m,8H),0.97(t,J=7.4Hz,3H).MS(ESI)m / z:[M+H] + Calculated value C 17 H 25 N4O3: 333.1921, measured value: 332.86.

[0538] 2) Synthesis of the final product (PLN-5-4)

[0539]

[0540] The compound PLN-1-1 was prepared according to the synthetic method described above. Starting materials included (Z)-1-acetyl-3-((1-n-butyl-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.31 mmol), p-fluorophenoxybenzaldehyde (78.1 mg, 0.36 mmol), cesium carbonate (147.0 mg, 0.45 mmol), and anhydrous sodium sulfate (85.5 mg, 0.60 mmol). The reaction mixture was stirred in DMF (3.5 mL) in an oil bath at 50 °C for 25 h. A pale yellow solid of 90.3 mg was obtained, with a yield of 61%. Melting point: 203-204 °C. 1 H NMR(500MHz,DMSO-d6)δ12.01 (s,1H),10.16(s,1H),7.90(s,1H),7.41(t,J=7.9Hz,1H),7.24(ddd,J=8.6,7.3,5.4Hz,3H),7.17(s,1H ),7.14–7.09(m,2H),6.90(dd,J=8.1,2.0Hz,1H),6.71(s,1H),6.69(s,1H),4.00(t,J=7.4Hz,2H),3.21 (dt,J=14.2,7.1Hz,1H),1.69–1.58(m,2H),1.34–1.24(m,8H),0.90(t,J=7.4Hz,3H). 13 C NMR (125 MHz, DMSO-d6) δ159.13,157.38,157.23,157.04,156.16,152.59,138.44,137.17,135.20,132.09,130.28, 127.23,124.42,123.68,120.57,120.50,118.89,117.74,116.63,116.44, 113.16,104.23,44.38,32.71,23.67,22.63*2,19.18,13.47.MS(ESI):m / z 489.01[M+H] + MPa: 203-204℃.

[0541] Example 53 Synthesis of (3Z,6Z)-3-(3-(benzoyl)benzene)methylene-6-((5-isopropyl-1-ethylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-5)

[0542]

[0543] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (150 mg, 0.54 mmol), DMF (4 ml), and cesium carbonate (530.65 mg, 1.63 mmol) sequentially. Place the flask in a 0 °C cold trap for 20 min, then add iodoethane (846.75 mg, 5.43 mmol) dropwise and continue stirring for 1.5 h. LC-MS analysis showed the reaction was complete. The reaction solution was added dropwise to 0 °C cold water, stirred for 10 min, filtered, and the filter cake was washed with water. The solution was dried under vacuum at 50 °C to obtain 101 mg of a light yellow solid, yield 61.12%.

[0544] 2) Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-ethylimidazol-4-yl)methylene)piperazine-2,5-dione (95 mg, 0.31 mmol), DMF (2 ml), 1-2 (85.49 mg, 0.37 mmol), cesium carbonate (152.55 mg, 0.47 mmol), and anhydrous sodium sulfate (88.67 mg, 0.62 mmol) sequentially. Exhaust the gas, place under nitrogen protection, and in an oil bath at 55 °C. Stir and react for 24 h. After the reaction is complete, dropwise add the reaction solution to 50 ml of cold water at 4 °C, filter, dissolve in methanol and dichloromethane (50 ml: 20 ml), filter, concentrate under reduced pressure, and slurry with EA. Place at -30 °C overnight, filter, wash the filter cake with cold EA, and dry under vacuum at 50 °C to obtain 82 mg of a pale yellow solid, yield 55.60%.

[0545] 1 H NMR(500MHz,DMSO-d6)δ12.02(s,1H),10.35(s,1H),7.91(s,1H),7.84-7.80(m,3H),7.75(d, J=7.6Hz,1H),7.69(t,J=7.4Hz,1H),7.63(d,J=7.7Hz,1H),7.58(t,J=7.7Hz,3H),6.80(s,1H), 6.69(s,1H),4.05(q,J=7.2Hz,2H),3.27-3.17(m,1H),1.33(dd,J=7.0,5.5Hz,9H).MS(ESI):m / z 455.14 [M+H] + MPa: 214-216℃.

[0546] Example 54 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-allylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-6)

[0547]

[0548] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.73 mmol), potassium carbonate (300.1 mg, 2.17 mmol), and dropwise add DMF (5 ml) containing bromopropene (175.2 mg, 1.45 mmol). Stir the reaction mixture at 45 °C for 2 h. The reaction mixture is then added dropwise to cold water at 4 °C, resulting in the precipitation of a solid. The solid is filtered, the filter cake is washed with water, dried under vacuum at 50 °C, slurried with methanol, and allowed to stand in a -30 °C refrigerator for at least 2 h. The mixture is then filtered, washed with cold methanol, and dried to obtain 135.5 mg of a brownish-yellow solid, with a yield of 59.17%.

[0549] 2) Take a 10ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(propenylimidazol-4-yl)methylene)piperazine-2,5-dione (100mg, 0.32mmol), 3-p-fluorobenzoylbenzaldehyde (118.95mg, 0.52mmol), cesium carbonate (154.67mg, 0.47mmol), anhydrous sodium sulfate (89.90mg, 0.63mmol), and DMF ( 4 ml of the mixture was vented, and under nitrogen protection, placed in an oil bath at 45°C and stirred for 20 h. The reaction was monitored by LC-MS until the starting material spot disappeared. After the reaction was complete, the reaction solution was added dropwise to 40 ml of cold water at 4°C, filtered, and the filter cake was washed with cold water. The filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. The methanol was ultrasonically slurried, placed in a -30°C refrigerator to stand, filtered, and the filter cake was washed with cold methanol and dried under vacuum at 50°C to give 114.5 mg of a yellow solid, with a yield of 74.67%.

[0550] 1H NMR(500MHz,DMSO-d6)δ11.99(s,1H),10.36(s,1H),7.94-7.89(m,2H),7.88(s,1H),7.82(s, 1H),7.75(d,J=7.52Hz,1H),7.64(d,J=7.63Hz,1H),7.59(t,J=7.63Hz,1H),7.40(t,J=8.74Hz,2H),6 .81(s,1H),6.71(s,1H),6.01(ddd,J=4.97,10.23,15.29Hz,1H),5.22(d,J=10.32Hz,1H),4.94(d,J= 17.16Hz,1H),4.73(s,2H),3.15(dt,J=7.05,14.20Hz,1H),1.30(d,J=7.07Hz,6H).MS(ESI):m / z485.14[M+H] + MPa: 200-202℃.

[0551] Example 55 Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-propynylimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-7)

[0552]

[0553] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), potassium carbonate (450.16 mg, 3.26 mmol), and dropwise add DMF (6 ml) of bromopropyne (258.33 mg, 2.17 mmol). Stir the reaction mixture at 45 °C for 2 h. The reaction mixture is then added dropwise to cold water at 4 °C, resulting in the precipitation of a solid. The solid is filtered, the filter cake is washed with water, dried under vacuum at 50 °C, slurried with methanol, and allowed to stand in a -30 °C refrigerator for at least 2 h. The mixture is then filtered, washed with cold methanol, and dried to obtain 232.0 mg of a brownish-yellow solid, with a yield of 67.97%.

[0554] 2) Take a 25ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(propenylimidazol-4-yl)methylene)piperazine-2,5-dione (150mg, 0.48mmol), 3-p-fluorobenzoylbenzaldehyde (130.82mg, 0.57mmol), cesium carbonate (233.48mg, 0.72mmol), anhydrous sodium sulfate (135.71mg, 0.96mmol), D MF (5 ml), under venting and nitrogen protection, was placed in an oil bath at 45 °C and stirred for 17 h. The reaction was monitored by LC-MS. After the reaction was complete, the reaction solution was added dropwise to 40 ml of cold water at 4 °C, filtered, and the filter cake was washed with cold water. The filter cake was dissolved in a mixture of methanol and dichloromethane (1:3), filtered, and concentrated under reduced pressure. The methanol was ultrasonically slurried, placed in a -30 °C refrigerator to stand, filtered, and the filter cake was washed with cold methanol and dried under vacuum at 50 °C to give 166.0 mg of yellow solid, with a yield of 72.02%.

[0555] 1 H NMR (500MHz, DMSO-d6) δ11.90 (s, 1H), 10.38 (s, 1H), 7.92 (dd, J = 9.14, 17.20Hz, 3H), 7.82 (s, 1H),7.76(d,J=7.43Hz,1H),7.64(d,J=7.54Hz,1H),7.59(t,J=7.58Hz,1H),7.40(t,J=8.70Hz,2H),6. 82(s,1H),6.69(s,1H),5.02(s,2H),3.58(s,1H),3.37-3.25(m,1H),1.35(d,J=7.03Hz,6H).MS(ESI): m / z 483.10[M+H] + MPa: 150-152℃.

[0556] Example 56 Preparation of (3Z,6Z)-3-(4-p-fluorobenzoyl)benzylmethyl-6-((1-(2-methoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-8)

[0557]

[0558] 1. (Z)-1-acetyl-3-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0559]

[0560] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-bromo-2-methoxyethane (300 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. The solution was transferred to a 100ml single-necked flask, rinsed with ethanol, and concentrated under reduced pressure to obtain an orange oily substance. The solution was then redissolved with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filtered, and the filter cake was washed with the mixed solvent (V ethanol: V dichloromethane = 1:5). The solution was concentrated under reduced pressure to obtain 400mg of a brown oily mixture, which was not purified and proceeded directly to the next step.

[0561] 2. (3Z,6Z)-3-(4-p-fluorobenzoyl)benzylmethyl-6-((1-(2-methoxyethyl)-5-isopropyl-1H-

[0562] Imidazol-4-yl)methylene)piperazine-2,5-dione

[0563]

[0564] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.90 mmol), 3-(4-fluorobenzoyl)benzaldehyde (246 mg, 1.08 mmol), cesium carbonate (438 mg, 1.35 mmol), anhydrous sodium sulfate (255 mg, 1.80 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the mixture in an oil bath at 45 °C and stir for 20 h. Transfer the mixture to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, then redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurried, filtered at -20°C, and dried to obtain 160 mg of pale yellow solid, with a yield of 29.52%. 1H NMR(500MHz,DMSO- d6)δ12.00(s,1H),10.34(s,1H),7.93–7.89(m,2H),7.85(s,1H),7.82(s,1H),7.75(d,J=7.5Hz,1H),7.6 4(d,J=7.5Hz,2H),7.59(t,J=7.6Hz,1H),7.40(t,J=8.5Hz,1H),6.81(s,1H),6.71(s,1H),4.20(t,J=4.8 Hz,2H),3.58(t,J=4.9Hz,2H),3.29–3.17(m,4H),1.31(d,J=7.0Hz,6H).MS(ESI):m / z 503.17 [M+H] + MPa: 209-211℃.

[0565] Example 57 Preparation of (3Z,6Z)-3-(4-p-fluorobenzoyl)benzylmethyl-6-((1-(2-ethoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-9)

[0566]

[0567] 1. (Z)-1-acetyl-3-((1-(2-ethoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0568]

[0569] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-bromo-2-ethoxyethane (330 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. The solution was transferred to a 100ml single-necked flask, rinsed with ethanol, and concentrated under reduced pressure to obtain an orange oily substance. The solution was then redissolved with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filtered, and the filter cake was washed with the mixed solvent (V ethanol: V dichloromethane = 1:5). The solution was concentrated under reduced pressure to obtain 400mg of a brown oily mixture, which was not purified and proceeded directly to the next step.

[0570] 2. (3Z,6Z)-3-(4-p-fluorobenzoyl)benzylmethyl-6-((1-(2-ethoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0571]

[0572] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((1-(2-ethoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.86 mmol), 3-(4-fluorobenzoyl)benzaldehyde (224 mg, 1.03 mmol), cesium carbonate (419 mg, 1.29 mmol), anhydrous sodium sulfate (244 mg, 1.72 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the flask in an oil bath at 45 °C and stir for 20 h. Transfer the solution to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurried, filtered at -20°C, and dried to obtain 100 mg of pale yellow solid, with a yield of 17.95%. 1 H NMR(500MHz,DMSO- d6)δ12.01(s,1H),10.34(s,1H),7.96–7.89(m,2H),7.85(s,1H),7.82(s,1H),7.75(d,J=7.5Hz,1H), 7.64(d,J=7.6Hz,1H),7.58(t,J=7.7Hz,1H),7.40(t,J=8.2Hz,2H),6.81(s,1H),6.71(s,1H),4.18(d, J=4.3Hz,2H),3.61(t,J=4.5Hz,2H),3.42(q,J=6.9Hz,2H),3.25(dt,J=13.4,6.7Hz,1H),1.32(d,J=7.1Hz,6H),1.08(t,J=6.9Hz,3H).MS(ESI):m / z 517.15[M+H] + MPa: 205-207℃.

[0573] Example 58 Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-10)

[0574] 1)(Z)-1-acetyl-3-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0575]

[0576] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-bromo-2-methoxyethane (300 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. Transfer to a 100ml single-necked flask, rinse with ethanol, concentrate under reduced pressure to obtain an orange oily substance, then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filter, wash the filter cake with the mixed solvent (V ethanol: V dichloromethane = 1:5), concentrate under reduced pressure to obtain 400mg of brown oily mixture, unpurified, proceed directly to the next step.

[0577] 2)(3Z,6Z)-3-(-3-(4-fluorophenoxy)benzylmethyl)-6-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0578]

[0579] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((5-isopropyl-1-(2-methoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.90 mmol), 3-(4-fluorophenoxy)benzaldehyde (233 mg, 1.08 mmol), cesium carbonate (438 mg, 1.35 mmol), anhydrous sodium sulfate (255 mg, 1.80 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the mixture in an oil bath at 45 °C and stir for 20 h. Transfer the mixture to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, and then redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3). Filter the mixture, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurryed, placed at -20℃ for 2 hours, filtered, and dried to obtain 145 mg of pale yellow solid, with a yield of 27.25%.

[0580] 1H NMR(500MHz,dmso)δ11.78(s,1H),10.14(s,1H),7.77(s,1H),7.59(s,1H),7.44(d,J=7.5Hz, 1H),7.33(t,J=7.9Hz,1H),7.21(t,J=8.7Hz,2H),7.13-7.02(m,2H),6.78(d,J=7.9Hz,1H),6.56(s,1H ),6.51(s,1H),4.17(t,J=4.9Hz,2H),3.57(t,J=5.0Hz,2H),3.25(s,3H),3.24-3.16(m,1H),1.31(d,J =7.1Hz,6H).MS(ESI):m / z 491.06 [M+H] + MPa: 159-162℃.

[0581] Example 59 Preparation of (3Z,6Z)-3-(-3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(2-ethoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-11)

[0582] 1)(Z)-1-acetyl-3-((1-(2-ethoxyethyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0583]

[0584] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-bromo-2-ethoxyethane (330 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. Transfer to a 100ml single-necked flask, rinse with ethanol, concentrate under reduced pressure to obtain an orange oily substance, then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filter, wash the filter cake with the mixed solvent (V ethanol: V dichloromethane = 1:5), concentrate under reduced pressure to obtain 400mg of brown oily mixture, unpurified, proceed directly to the next step.

[0585] 2)(3Z,6Z)-3-(-3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(2-ethoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0586]

[0587] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((5-isopropyl-1-(2-ethoxyethyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.86 mmol), 3-(4-fluorophenoxy)benzaldehyde (222 mg, 1.03 mmol), cesium carbonate (419 mg, 1.29 mmol), anhydrous sodium sulfate (244 mg, 1.72 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the mixture in an oil bath at 45 °C and stir for 20 h. Transfer the mixture to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, then redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurryed, placed at -20℃ for 2 hours, filtered, and dried to obtain 95 mg of pale yellow solid, with a yield of 17.33%.

[0588] 1 H NMR (500MHz, dmso) δ11.99 (s, 1H), 10.15 (s, 1H), 7.85 (s, 1H), 7.41 (t, J = 7.7Hz, 1H), 7.26 (d, J=10.1Hz,1H),7.23(d,J=8.3Hz,2H),7.17(s,1H),7.12(dd,J=6.9,4.4Hz,2H),6.90(d,J=8.1Hz,1H), 6.72(s,1H),6.71(s,1H),4.19(s,2H),3.61(s,2H),3.42(q,J=6.7Hz,2H),3.28-3.22(m,1H),1.31(d, J=6.8Hz,6H),1.07(t,J=6.8Hz,3H).MS(ESI):m / z 505.26[M+H] + MPa: 177-180℃.

[0589] Example 60: Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(3-methoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-12)

[0590] 1)(Z)-1-acetyl-3-((5-isopropyl-1-(3-methoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0591]

[0592] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 1-bromo-3-methoxypropane (330 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. The solution was transferred to a 100ml single-necked flask, rinsed with ethanol, and concentrated under reduced pressure to obtain an orange oily substance. The solution was then redissolved with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filtered, and the filter cake was washed with the mixed solvent (V ethanol: V dichloromethane = 1:5). The solution was concentrated under reduced pressure to obtain 400mg of a brown oily mixture, which was not purified and proceeded directly to the next step.

[0593] 2)(3Z,6Z)-3-(3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(3-methoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0594]

[0595] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((5-isopropyl-1-(3-methoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.86 mmol), 3-(4-fluorophenoxy)benzaldehyde (222 mg, 1.03 mmol), cesium carbonate (419 mg, 1.29 mmol), anhydrous sodium sulfate (244 mg, 1.72 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the mixture in an oil bath at 45 °C and stir for 20 h. Transfer the mixture to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurryed, placed at -20℃ for 2 hours, filtered, and dried to obtain 90 mg of pale yellow solid, with a yield of 16.42%.

[0596] 1H NMR(500MHz,dmso)δ11.99(s,1H),10.12(s,1H),7.87(s,1H),7.41(t,J=7.9Hz,1H),7.27(d, J=7.7Hz,1H),7.23(t,J=8.6Hz,2H),7.17(s,1H),7.15-7.08(m,2H),6.90(d,J=8.1Hz,1H),6.71(s,1H ),6.70(s,1H),4.06(t,J=7.2Hz,2H),3.35-3.28(m,2H),3.25(s,2H),3.23-3.18(m,1H),2.00–1.81(m, 3H),1.32(d,J=7.1Hz,6H).MS(ESI):m / z 505.12[M+H] + MPa: 194-196℃.

[0597] Example 61: Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(3,3-dimethoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-13)

[0598] 1)(Z)-1-acetyl-3-((1-(3,3-dimethoxypropyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0599]

[0600] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 3-bromo-1,1-dimethoxypropane (395 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. Transfer to a 100ml single-necked flask, rinse with ethanol, concentrate under reduced pressure to obtain an orange oily substance, then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filter, wash the filter cake with the mixed solvent (V ethanol: V dichloromethane = 1:5), concentrate under reduced pressure to obtain 400mg of brown oily mixture, unpurified, proceed directly to the next step.

[0601] 2. Preparation of (3Z,6Z)-3-(3-(4-fluorophenoxy)benzene)methylene)-6-((5-isopropyl-1-(3,3-dimethoxypropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0602]

[0603] Take a 25 mL dry reaction flask, (Z)-1-acetyl-3-((1-(3,3-dimethoxypropyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.79 mmol), 3-(4-fluorophenoxy)benzaldehyde (205 mg, 0.95 mmol), cesium carbonate (385 mg, 1.18 mmol), anhydrous sodium sulfate (224 mg, 1.58 mmol), and DMF (6 mL). Under nitrogen protection and protected from light, place the mixture in an oil bath at 45 °C and stir for 20 h. Transfer the mixture to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. The methanol was slurryed, placed at -20℃ for 2 hours, filtered, and dried to obtain 80 mg of pale yellow solid, with a yield of 13.79%.

[0604] 1 H NMR (500MHz, DMSO-d6) δ11.99(s,1H),10.08(s,1H),7.90(s,1H),7.41(t,J=7.9Hz,1H),7.27 (d,J=7.8Hz,1H),7.23(t,J=8.7Hz,2H),7.17(s,1H),7.14-7.09(m,2H),6.90(d,J=8.1Hz,1H),6.72(s, 1H),6.70(s,1H),4.39(t,J=5.3Hz,1H),4.04(t,J=7.3Hz,2H),3.26(s,6H),3.23-3.17(m,1H),1.97(dd, J=13.0,6.6Hz,2H),1.33(d,J=7.0Hz,6H).MS(ESI):m / z 535.00[M+H] + MPa: 176-178℃.

[0605] Example 62 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(hydroxyethyl ethyl acetate)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-14)

[0606] 1)(Z)-2-(4-((4-acetyl-3,6-dioxopiperazin-2-ylidene)methyl)-5-isopropyl-1H-imidazolyl)ethyl acetate

[0607]

[0608] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), ethyl 2-bromo-ethyl (360 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Under nitrogen protection, place the flask in an oil bath at 70 °C and stir for 24 h. Transfer to a 100ml single-necked flask, rinse with ethanol, concentrate under reduced pressure to obtain an orange oily substance, then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filter, wash the filter cake with the mixed solvent (V ethanol: V dichloromethane = 1:5), concentrate under reduced pressure to obtain 400mg of brown oily mixture, unpurified, proceed directly to the next step.

[0609] 2)(3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(hydroxyethyl ethyl acetate)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0610]

[0611] Take a 25 mL dry reaction flask, add (Z)-2-(4-((4-acetyl-3,6-dioxopiperazin-2-ylidene)methyl)-5-isopropyl-1H-imidazol-1-yl)ethyl acetate (300 mg, 0.83 mmol), 3-(4-fluorophenoxy)benzaldehyde (215 mg, 0.99 mmol), cesium carbonate (405 mg, 1.24 mmol), anhydrous sodium sulfate (236 mg, 1.66 mmol), and DMF (6 mL), and place under nitrogen protection, protected from light, in an oil bath at 45 °C with stirring for 20 h. Transfer to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, then redissolve with a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. Flash purification was performed with methanol: 76% and water: 24% to obtain the product. The product was concentrated under reduced pressure to give 90 mg of a pale yellow solid, with a yield of 16.10%.

[0612] 1H NMR(500MHz,dmso)δ11.96(s,1H),10.12(s,1H),7.91(s,1H),7.41(t,J=7.9Hz,1H),7.27(d, J=7.7Hz,1H),7.23(t,J=8.6Hz,2H),7.17(s,1H),7.16–7.08(m,2H),6.91(d,J=8.1Hz,1H),6.72(s, 1H),6.70(s,1H),4.30(d,J=4.4Hz,2H),4.27(d,J=4.1Hz,2H),3.25–3.18(m,1H),2.01(s,3H),1.33 (d,J=7.0Hz,6H).MS(ESI):m / z 519.19[M+H] + MPa: 185-187℃.

[0613] Example 63 Synthesis of (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-(N-tert-butoxycarbonyl)ethylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-15)

[0614]

[0615] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-methyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.21 mmol), N-tert-butoxycarbonyl-3-bromoethylamine (541.65 mg, 2.42 mmol), cesium carbonate (787.50 mg, 2.42 mmol), sodium iodide (181.14 mg, 1.21 mmol), 4A molecular sieve (500 mg), and DMF (5 ml) in sequence. Exhaust the gas, place the flask under nitrogen protection in a 70°C oil bath, and stir for 4 h. After the reaction is complete, concentrate under reduced pressure, dry, then dissolve in methanol and dichloromethane (1:3), filter, concentrate under reduced pressure, and dry to obtain 378.43 mg of light brown crude solid, yield 80.0%. The product was not purified and directly added to the next step.

[0616] 2) Take a 25 mL dry brown round-bottom flask and add Z)-1-acetyl-3-((5-methyl-1-(N-tert-butoxycarbonyl)ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (378.43 mg, 0.97 mmol), 2-p-fluorobenzoylbenzaldehyde (264.75 mg, 1.16 mmol), cesium carbonate (472.50 mg, 1.45 mmol), anhydrous sodium sulfate (274.65 mg, 1.93 mmol), and DMF (6 mL) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 55 °C oil bath, and stir for 24 h. Monitor the reaction by LC-MS. After the reaction is complete, concentrate under reduced pressure, dry, then dissolve in methanol and dichloromethane (1:3), filter, concentrate under reduced pressure, dry, elute with a gradient, concentrate under reduced pressure, and give 52.8 mg of a pale yellow solid, yield 7.81%.

[0617] 1 H NMR (500MHz, DMSO-d6) δ = 11.84 (s, 1H), 10.34 (s, 1H), 7.95-7.88 (m, 3H), 7.85 (d, J = 6.0, 1H), 7.82(s,1H),7.57(dt,J=15.1,7.6,2H),7.40(t,J=8.8,2H),7.00(t,J=5.7,1H),6.74(s,1H) ,6.53(s,1H),4.01(t,J=5.8,2H),3.22(d,J=5.9,2H),2.29(s,3H),1.37(s,9H).MS(ESI):m / z 560.16[M+H] + MPa: 189-191 ℃.

[0618] Example 64 Synthesis of (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-16)

[0619]

[0620] Take a 100 ml dry round-bottom flask and add (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-(N-tert-butyloxycarbonyl)ethylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (35.1 mg, 0.063 mmol), methanol (5 ml), and hydrochloric acid (0.5 ml) sequentially. Place the flask in an oil bath at 40 °C and stir for 12 h. Monitor the reaction by LC-MS. After the reaction is complete, concentrate under reduced pressure, dry, add 0.5 ml of methanol and dropwise add acetone (2 ml). A solid precipitates out. Place the flask in a -20 °C refrigerator and let it stand for more than 2 h. Filter the flask, wash the filter cake with acetone, and dry it under vacuum at 50 °C to give 14 mg of brownish-yellow solid, yield 44.96%.

[0621] 1 H NMR (500MHz, DMSO-d6) δ = 11.70 (s, 1H), 10.38 (s, 1H), 8.36 (s, 3H), 8.17 (s, 1H), 7.94-7.88 (m, 2H),7.82(s,1H),7.75(d,J=7.4,1H),7.64(d,J=7.5,1H),7.59(t,J=7.6,1H),7.40(t,J=8.6,2H),6.82(s,1H), 6.58(s,1H),4.32(t,J=6.3,2H),3.20(d,J=5.6,2H),2.34(s,3H).MS(ESI):m / z 460.15[M+H] + MPa: 180-182℃.

[0622] Example 65 Synthesis of (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-(N-tert-butoxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-17)

[0623]

[0624] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-methyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.21 mmol), N-tert-butoxycarbonyl-3-bromopropylamine (575.54 mg, 2.42 mmol), cesium carbonate (787.50 mg, 2.42 mmol), sodium iodide (181.14 mg, 1.21 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 70 °C oil bath, and stir for 3 h. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution, wash the filter cake with water, dissolve it in methanol and dichloromethane, filter, concentrate under reduced pressure, and dry to obtain 367 mg of a light brown solid (yield 74.90%). The solid is not purified and directly added to the next step.

[0625] 2) Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-methyl-1-(N-tert-butyloxycarbonyl)propylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (367 mg, 0.90 mmol), 2-p-fluorobenzoylbenzaldehyde (247.89 mg, 1.09 mmol), cesium carbonate (442.37 mg, 1.36 mmol), anhydrous sodium sulfate (257.13 mg, 1.81 mmol), and DMF (5 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 60 °C oil bath, and stir for 21 h. Monitor the reaction by LC-MS. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution, wash the filter cake with cold water, dissolve it in methanol and dichloromethane, filter, and concentrate under reduced pressure. Extract the aqueous phase with EA until clear. Combine the organic phases and concentrate under reduced pressure. The filter cake and organic phase sample were combined and eluted with a gradient to give 260 mg of pale yellow solid, with a yield of 50.10%.

[0626] 1 H NMR (500MHz, DMSO-d6) δ = 11.87 (s, 1H), 10.37 (s, 1H), 7.97-7.88 (m, 3H), 7.86 (s, 1H), 7.82 (d,J=7.2,1H),7.62(d,J=7.5,1H),7.57(t,J=7.6,1H),7.40(t,J=8.7,2H),6.95(s,1H),6.77(s,1H),6.56 (s,1H),3.97(t,J=6.8,2H),2.93(d,J=5.8,2H),2.30(s,3H),1.90-1.70(m,2H),1.39(s,9H).MS(ESI):m / z 574.23[M+H] + MPa: 206-208℃.

[0627] Example 66 Synthesis of (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-18)

[0628]

[0629] Take a 100 ml dry round-bottom flask and add (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((5-methyl-1-(N-tert-butyloxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (100 mg, 0.21 mmol), methanol (15 ml), water (1 ml), and hydrochloric acid (2 ml, 12 mmol / ml) in sequence. Place the flask in an oil bath at 30 °C and stir for 12 h. After the reaction is complete as monitored by LC-MS, allow it to cool naturally to room temperature. A white solid precipitates, which is filtered, the filter cake is washed with methanol, and dried to give 82 mg of white solid, with a yield of 84.50%.

[0630] 1 H NMR (500MHz, DMSO-d6) δ = 11.65 (s, 1H), 10.39 (s, 1H), 8.27 (s, 1H), 8.20 (s, 3H), 7.91 (dd, J=8.7,5.6,2H),7.82(s,1H),7.75(d,J=7.7,1H),7.64(d,J=7.7,1H),7.59(t,J=7.6,1H),7.40(t,J=8.8,2H) ,6.82(s,1H),6.57(s,1H),4.16(t,J=7.0,2H),2.80(dd,J=12.9,6.6,2H),2.32(s,3H),2.08-1.98(m,2H).MS (ESI):m / z 474.16 [M+H] + MPa: 247-248℃.

[0631] Example 67 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-isopropyl-1-(3-(dimethylamino)propyl)methylene)piperazine-2,5-dione (PLN-5-19)

[0632] 1. (Z)-1-acetyl-3-((1-(3-(dimethylamino)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0633]

[0634] Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.08 mmol), 3-chloro-N,N-dimethylpropyl-1-amine (261 mg, 2.16 mmol), cesium carbonate (704 mg, 2.16 mmol), potassium iodide (180 mg, 1.08 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) in sequence. Exhaust the gas, protect with nitrogen, place in an oil bath at 70 °C, and stir to react for 24 h. The solution was transferred to a 100ml single-necked flask, rinsed with ethanol, and concentrated under reduced pressure to obtain an orange oily substance. The solution was then redissolved with a mixed solvent of ethanol and dichloromethane (V ethanol: V dichloromethane = 1:5), filtered, and the filter cake was washed with the mixed solvent (V ethanol: V dichloromethane = 1:5). The solution was concentrated under reduced pressure to obtain 400mg of a brown oily mixture, which was not purified and proceeded directly to the next step.

[0635] 2. (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-isopropyl-1-(3-(dimethylamino)propyl)methylene)piperazine-2,5-dione

[0636]

[0637] Take a 25 mL dry reaction flask, add (Z)-1-acetyl-3-((1-(3-(dimethylamino)propyl)-5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.83 mmol), 3-(4-fluorophenoxy)benzaldehyde (180 mg, 0.83 mmol), cesium carbonate (405 mg, 1.24 mmol), anhydrous sodium sulfate (239 mg, 1.66 mmol), and DMF (6 mL). Under nitrogen purging and protection from light, place the flask in an oil bath at 45 °C and stir for 20 h. Transfer the solution to a 100 mL single-necked flask, rinse with ethanol, concentrate under reduced pressure, redissolve in a mixed solvent of ethanol and dichloromethane (V ethanol:V dichloromethane = 1:3), filter, wash the filter cake with the mixed solvent (V ethanol:V dichloromethane = 1:3), and concentrate under reduced pressure. Flash purification was performed with methanol: 60%, 0.5% formic acid, and water: 40% to obtain the product. The product was concentrated under reduced pressure to give 138 mg of a pale yellow solid, with a yield of 32.16%.

[0638] 1H NMR(500MHz,DMSO-d6)δ12.00(s,1H),10.13(s,1H),7.88(s,1H),7.40(t,J=7.6Hz,1H), 7.28-7.21(m,3H),7.17(s,1H),7.12(s,2H),6.90(d,J=7.4Hz,1H),6.71(s,2H),4.02(t,J=6.4Hz,2H ),3.27–3.14(m,1H),2.14(s,6H),2.00(t,J=8.0Hz,2H),1.94–1.72(m,2H),1.33(d,J=5.9Hz,6H).MS (ESI):m / z 518.22[M+H] + MPa: 164-166℃.

[0639] Example 68 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxoethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-20)

[0640]

[0641] Preparation of (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0642]

[0643] A dry 250 mL eggplant-shaped reaction flask was used to dissolve 5-isopropyl-1H-imidazol-4-carboxaldehyde (5.0 g, 36.19 mmol) in 120 mL of dry DMF. 1,4-diacetyl-piperazine-2,5-dione (14.34 g, 72.38 mmol) and cesium carbonate (17.69 g, 54.29 mmol) were added. The mixture was degassed and reacted at 25 °C in the dark for 24 h. The reaction was monitored by LC-MS. The mixture was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, filtered, and evaporated to dryness. The mixture was then slurried with ethyl acetate, filtered, and dried to give compound (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (4.39 g, 15.89 mmol), with a yield of 43.9%.

[0644] Preparation of (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxoethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0645]

[0646] Take a dry 25 mL eggplant-shaped reaction flask, dissolve (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (50 mg, 0.18 mmol) in 4 mL of dry DMF, add 2-iodotert-butyldimethylsilylethanol (156 mg, 0.54 mmol), add cesium carbonate (177 mg, 0.54 mmol), degas, react at 50 °C in the dark for 24 h, and detect the reaction by LC-MS. The sample was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, filtered, and evaporated to dryness. The mixture was then separated by methanol / water gradient chromatography on a C18 Flash column to give compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (34.2 mg, 0.08 mmol), in a yield of 42.3%, as a yellow solid.

[0647] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxoethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0648]

[0649] Take a dry 25 mL eggplant-shaped reaction flask, dissolve (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (108 mg, 0.25 mmol) in 8 mL of dry DMF, add 3-(4-fluorophenoxy)-benzaldehyde (65 mg, 0.30 mmol), add cesium carbonate (122 mg, 0.37 mmol), and degas at 50 °C in the dark for 24 h. Detect the reaction by LC-MS. Wash with water, extract with ethyl acetate, collect the organic phase, dry, filter and evaporate to dryness, and separate by methanol / water gradient chromatography on a C18 Flash column to give the compound dissolved in (3Z,6Z)-3-(3-p-fluorophenoxy)benzylmethyl-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (80 mg, 0.14 mmol), yield 54.5%, as a yellow solid.

[0650] Example 69 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene)-6-((5-(isopropyl)-1-(hydroxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-21)

[0651]

[0652] Dissolve (3Z,6Z)-3-(3-p-fluorophenoxy)benzylmethyl-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (57 mg, 0.10 mmol) in 3 mL of dry THF and 1 M THF solution of TBAF (1.11 mL, 0.11 mmol). React at 25 °C in the dark for 5 h. The reaction was confirmed to be complete by LC-MS. The mixture was filtered, evaporated to dryness, washed with water, extracted with ethyl acetate, evaporated to dryness, and slurried with methanol to give compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzylmethyl-6-((5-(isopropyl)-1-(hydroxyethyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (14 mg, 0.04 mmol), yield 17.1%, in a yellow papery form.

[0653] 1 H NMR (500MHz, Pyridine-d5) δ11.35(s,1H),7.95(s,1H),7.44(d,J=10.5Hz,2H),7.34(d,J= 15.1Hz,3H),7.17(t,J=8.2Hz,2H),7.04(s,2H),6.95(d,J=7.1Hz,2H),4.14(s,2H),3.99(s,2H),3.29-3.14(m,1H),1.30(d,J=6.9Hz,6H).

[0654] Example 70: Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(hydroxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-22)

[0655]

[0656] Preparation of (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxane)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0657]

[0658] Take a dry 25 mL eggplant-shaped reaction flask, dissolve (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol) in 4 mL of dry DMF, add 3-iodotert-butyldimethylsilylethanol (649 mg, 2.16 mmol), add cesium carbonate (704 mg, 2.16 mmol), degas, react at 50 °C in the dark for 24 h, and detect the reaction by LC-MS. The sample was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, filtered, and evaporated to dryness. The mixture was then separated by methanol / water gradient chromatography on a C18 Flash column to give compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (38 mg, 0.08 mmol), in a yield of 11.7%, as a yellow solid.

[0659] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxane)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0660]

[0661] Take a dry 25 mL eggplant-shaped reaction flask, dissolve the intermediate (Z)-1-acetyl-3-((5-(isopropyl)-1-(tert-butyldimethylsiloxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (30 mg, 0.07 mmol) in 4 mL of dry DMF, add 3-(4-fluorophenoxy)-benzaldehyde (18 mg, 0.08 mmol), add cesium carbonate (33 mg, 0.10 mmol), degas, react at 50 °C in the dark for 12 h, and detect the reaction by LC-MS. The sample was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, and slurried with methanol to give compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzylmethyl-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (30 mg, 0.05 mmol), in 74.1% yield, as a yellow solid.

[0662] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(hydroxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0663]

[0664] Dissolve (3Z,6Z)-3-(3-p-fluorophenoxy)benzyl)methylene-6-((5-(isopropyl)-1-(tert-butyldimethylsiloxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (120 mg, 0.20 mmol) in 3 mL of dry THF and 1 M THF solution of TBAF (1.11 mL, 0.11 mmol). React at 25 °C in the dark for 5 h. The reaction was confirmed to be complete by LC-MS. The solution was filtered, evaporated to dryness, washed with water, extracted with ethyl acetate, evaporated to dryness, and slurried with methanol to give (3Z,6Z)-3-(3-p-fluorophenoxy)benzyl)methylene-6-((5-(isopropyl)-1-(hydroxypropyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (86 mg, 0.18 mmol), yield 87.7%, as a yellow solid.

[0665] 1 H NMR(500MHz,Pyridine-d5)δ11.35(s,2H),7.87(s,1H),7.44(dd,J=18.7,11.0Hz,2H),7.37- 7.30(m,3H),7.17(t,J=8.6Hz,2H),7.04(dd,J=8.9,4.4Hz,2H),6.98-6.92(m,1H),4.19(t,J= 7.1Hz,2H),3.80(t,J=5.7Hz,2H),3.19-3.16(m,1H),2.09-1.94(m,2H),1.32(d,J=7.1Hz,6H). 13 C NMR(125 MHz, Pyridine-d5)δ160.7,159.0,158.5,153.5,139.3,137.8,137.2,134.2,131.3,129.5,125. 7,124.8,121.8*2,119.6,118.4,117.3*2,117.2,114.1,105.9,58.5,43.0,35.0,25.1,23.1*2.

[0666] Example 71: Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(2-hydroxy-benzoate propionyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-23)

[0667]

[0668] 2-Benzyloxybenzoic acid-(3-iodo)-propyl ester

[0669]

[0670] Take a dry 50 mL eggplant-shaped reaction flask, dissolve 3-iodopropanol (1 g, 5.38 mmol) in 20 mL of dry DCM, add 2-benzyloxybenzoic acid (1.47 g, 6.45 mmol), add DPTS (3.16 g, 10.75 mmol), add DIC (1.67 mL, 10.75 mmol), and react at 25 °C in the dark for 12 h. The reaction was confirmed to be complete by LC-MS. Wash with water, extract with ethyl acetate, collect the organic phase, dry to anhydrous sodium sulfate, filter and evaporate to dryness, and perform column chromatography (P / E = 20 / 1) to give compound 2-benzyloxybenzoic acid-(3-iodo)-propyl ester (2.05 g, 5.17 mmol), yield 96.4%, colorless paste.

[0671] Preparation of (Z)-1-acetyl-3-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0672]

[0673] Take a dry 50 mL eggplant-shaped reaction flask, dissolve (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (561 mg, 2.03 mmol) in 25 mL of dry DMF, add 2-benzyloxybenzoic acid-(3-iodo)-propyl ester (2.41 g, 6.08 mmol), add cesium carbonate (1.98 g, 6.08 mmol), degas, react at 50 °C in the dark for 24 h, and detect the reaction by LC-MS. The sample was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, filtered, and evaporated to dryness. The mixture was then separated by methanol / water gradient chromatography on a C18 Flash column to give compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (505 mg, 0.93 mmol), in a yield of 45.7%, as a yellow solid.

[0674] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-2-2-73)

[0675]

[0676] Compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (1.3 g, 2.39 mmol) was dissolved in 30 mL of dry DMF, 3-(4-fluorophenoxy)-benzaldehyde (620 mg, 2.86 mmol) and cesium carbonate (1.17 g, 3.58 mmol) were added, the mixture was degassed, and the reaction was carried out at 50 °C in the dark for 24 h. The reaction was confirmed to be complete by LC-MS. The sample was filtered, evaporated to dryness, washed with water, extracted with ethyl acetate, and separated by a methanol / water gradient using a C18 Flash column chromatography. The methanol slurry yielded (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (540 mg, 0.77 mmol), in 32.3% yield, as a yellow solid.

[0677] 1 H NMR (500MHz, CDCl3) δ12.19 (s, 1H), 8.05 (s, 1H), 7.82 (dd, J = 7.7, 1.7Hz, 1H), 7.53-7.44 (m, 3H),7.41-7.27(m,5H),7.14-6.99(m,7H),6.95(s,1H),6.93-6.89(m,2H),6.85(s,1H),5.16(s,2H),4. 28(t,J=5.7Hz,2H),3.78(t,J=7.1Hz,2H),3.05-3.01(m,1H),2.06-2.03(m,2H),1.34(d,J=7.2Hz,6H). 13 C NMR (125MHz, CDCl3) δ166.7,159.0,158.3,157.2,153.5,138.7,136.4,136.4,135.3,134.0,133.4,132.2,130. 9,128.8*3,128.4,127.9*3,127.4,123.8,122.6,121.3*2,120.9,120.3,117.9,117.8,116.7*2,113.9,113.5, 106.4,71.0,61.1,42.0,30.3,24.7,22.8*2.

[0678] Example 72 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(2-hydroxy-benzoate propionyl)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-24)

[0679]

[0680] The compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzyl)methylene-6-((5-(isopropyl)-1-(2-benzyloxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 0.43 mmol) was dissolved in 30 mL of methanol and 30 mL of ethyl acetate. 200 mg of Pd / C was added, and the mixture was degassed. The reaction was carried out at 25 °C in the dark for 3 h under hydrogen atmosphere. The reaction was detected by LC-MS. The solution was filtered and evaporated to dryness. C18 Flash column chromatography with a methanol / water gradient was used to obtain the compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzyl)methylene-6-((5-(isopropyl)-1-(2-hydroxy-propylbenzoate)-imidazol-4-yl)methylene)piperazine-2,5-dione (82 mg, 0.13 mmol), in a yield of 31.2%, as a yellow powder.

[0681] 1 H NMR (500MHz, CDCl3) δ12.15(s,1H),10.61(s,1H),8.06(s,1H),7.76(d,J=7.5Hz,1H),7.56- 7.45(m,2H),7.39(t,J=8.0Hz,1H),7.13-6.98(m,6H),6.98-6.83(m,5H),4.41(t,J=5.8Hz,2 H),4.13(t,J=7.2Hz,2H),3.19-3.13(m,1H),2.28-2.23(m,2H),1.40(dd,J=13.3,7.3Hz,6H).

[0682] Example 73 Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(propylphosphite)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-25)

[0683]

[0684] Preparation of dibutyl-3-iodopropyl phosphate

[0685]

[0686] 3-Iodopropanol (300 mg, 0.43 mmol) was dissolved in 10 mL of dry dichloromethane. Pyridine (0.95 mL, 11.75 mmol) was added at 0 °C, followed by DMAP (18 mg, 0.15 mmol). Di-tert-butyl phosphate chloride (1.99 g, 3.72 mmol) was added dropwise, and the reaction was carried out at 25 °C for 3 h. The reaction was monitored by LC-MS. The solution was filtered and evaporated to dryness. The solution was separated by column chromatography from petroleum ether:ethyl acetate = 15:1 to 5:1 to give dibutyl-3-iodopropane phosphate (659 mg, 1.74 mmol), with a yield of 59.9%.

[0687] Preparation of (Z)-1-acetyl-3-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0688]

[0689] Take a dry 50 mL eggplant-shaped reaction flask, dissolve (Z)-1-acetyl-3-((5-(isopropyl)-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (500 mg, 1.81 mmol) in 25 mL of dry DMF, add dibutyl-3-iodopropyl phosphate (2.05 g, 5.43 mmol), add cesium carbonate (1.77 g, 5.43 mmol), degas, react at 50 °C in the dark for 24 h, and detect the reaction by LC-MS. The sample was washed with water, extracted with ethyl acetate, and the organic phase was collected, dried, filtered, and evaporated to dryness. The mixture was then separated by methanol / water gradient chromatography on a C18 Flash column to give compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione (324 mg, 0.62 mmol), in a yield of 34.0%, as a yellow solid.

[0690] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione

[0691]

[0692] Compound (Z)-1-acetyl-3-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione (289 mg, 0.55 mmol) was dissolved in 10 mL of dry DMF, 3-(4-fluorophenoxy)-benzaldehyde (143 mg, 0.66 mmol) and cesium carbonate (269 mg, 0.82 mmol) were added, the mixture was degassed, and the reaction was carried out at 50 °C for 24 h. The reaction was detected by LC-MS. The mixture was filtered and evaporated to dryness, then separated by methanol / water gradient chromatography using a C18 Flash column. The methanol slurry was then used to give the compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione (226 mg, 0.33 mmol), with a yield of 60.2%.

[0693] 1 H NMR (500MHz, CDCl3) δ12.17(s,1H),8.04(s,1H),7.50(s,1H),7.38(t,J=7.9Hz,1H),7.15- 6.98(m,5H),6.91(dd,J=19.4,15.4Hz,4H),4.11-4.05(m,9H),3.16-3.11(m,1H),2 .20-2.01(m,3H),1.70-1.67(m,4H),1.43-1.39(m,8H),0.97-0.93(t,J=7.4Hz,6H).

[0694] Preparation of (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(propylphosphite)-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-2-2-21)

[0695]

[0696] The compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione (50 mg, 0.07 mmol) was dissolved in 3 mL of dry DMF, and TMSBr (0.48 mL, 3.66 mmol) was added. The mixture was degassed and reacted at 45 °C for 4 h. The reaction was detected by LC-MS. The solution was filtered and evaporated to dryness, and then separated by a methanol / water gradient using a C18 Flash column to give the compound (3Z,6Z)-3-(3-p-fluorophenoxy)benzene)methylene-6-((5-(isopropyl)-1-(dibutylpropyl phosphate)-imidazol-4-yl)methylene)piperazine-2,5-dione (20 mg, 0.04 mmol), with a yield of 49.2%.

[0697] 1 H NMR(500MHz,DMSO-d6)δ11.66(s,1H),10.25(s,1H),8.22(s,1H),7.41(t,J=7.9Hz,1H), 7.30-7.20(m,3H),7.17(s,1H),7.15-7.09(m,2H),6.91(dd,J=8.2,1.7Hz,1H),6.74(s,1H),6.64(s,1H ),4.18(t,J=7.2Hz,2H),3.56(t,J=6.4Hz,2H),3.23-3.20(m,1H),2.28-2.24(m,2H),1.32(d,J=7.1Hz, 6H). 13 C NMR (125MHz, DMSO-d6) δ158.2,157.1,157.0,156.5,152.6,138.1,136.7,135.1,130.3,127.1, 124.5,120.6*2,118.9,117.8,116.6*2,116.5,113.8,109.5,102.9,43.6,33.3,30.9,23.6,22.2*2.

[0698] Example 74 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-26)

[0699]

[0700] 1) Take a 25 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), N-tert-butoxycarbonyl-3-bromoethylamine (324.43 mg, 1.45 mmol), cesium carbonate (471.69 mg, 1.45 mmol), potassium iodide (120.17 mg, 0.72 mmol), 4A molecular sieve (400 mg), and DMF (4 ml) in sequence. Exhaust the gas, place the flask under nitrogen protection in a 70°C oil bath, and stir for 3.5 h. After the reaction is complete, concentrate under reduced pressure, dry, then dissolve in methanol and dichloromethane (1:3), filter, concentrate under reduced pressure, and dry to obtain 242.80 mg of light brown crude solid, yield 80.00%. The product was not purified and directly added to the next step.

[0701] 2) Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(N-tert-butyloxycarbonyl)ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (242 mg, 0.58 mmol), 2-p-fluorobenzoylbenzaldehyde (158.00 mg, 0.69 mmol), cesium carbonate (281.96 mg, 0.87 mmol), anhydrous sodium sulfate (163.89 mg, 1.15 mmol), and DMF (4 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 55 °C oil bath, and stir for 24 h. Monitor the reaction by LC-MS. After the reaction is complete, concentrate under reduced pressure, dry, then dissolve in methanol and dichloromethane (1:3), filter, concentrate under reduced pressure, and dry. Elute using a gradient, concentrate under reduced pressure, and give 64.4 mg of a pale yellow solid, yield 19.00%.

[0702] 1 H NMR (500MHz, DMSO-d6) δ = 12.00 (s, 1H), 10.35 (s, 1H), 7.91 (dt, J = 8.4, 4.2, 2H), 7.83 (s, 1H), 7.77(d,J=11.6,2H),7.63(d,J=7.7,1H),7.58(t,J=7.6,1H),7.40(t,J=8.8,2H),7.03(t,J=5.7,1H ),6.79(s,1H),6.70(s,1H),4.05(t,J=5.8,2H),3.25–3.15(m,3H),1.38–1.27(m,15H).MS(ESI):m / z 588.25[M+H] + MPa: 204-206℃.

[0703] Example 75 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-27)

[0704]

[0705] Take a 25 ml dry round-bottom flask and add (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)ethylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (51.8 mg, 0.088 mmol), methanol (5 ml), and hydrochloric acid (1 ml, 12 mmol / ml) sequentially. Place the flask in an oil bath at 40 °C and stir for 4 h. Monitor the reaction by LC-MS. After the reaction is complete, concentrate under reduced pressure, dry, add 0.5 ml of methanol, and add acetone (2 ml) dropwise. A solid precipitates out. Place the flask in a -20 °C refrigerator and let it stand for more than 2 h. Filter the flask, wash the filter cake with acetone, and dry it under vacuum at 50 °C to give 24 mg of brownish-yellow solid, yield 76.92%.

[0706] 1 H NMR (500MHz, DMSO-d6) δ = 11.89 (s, 1H), 10.40 (s, 1H), 8.31 (s, 3H), 8.06 (s, 1H), 7.91 (dd, J=8.7,5.6,2H),7.82(s,1H),7.75(d,J=7.7,1H),7.64(d,J=7.7,1H),7.59(t,J=7.6,1H),7.41(t,J=8.8,2H) ,6.82(s,1H),6.70(s,1H),4.34(t,J=6.7,2H),3.26(dt,J=14.2,7.1,1H),3.15(dd,J=12.0,6.0,2H),1.33(d, J=7.1,6H).MS(ESI):m / z 488.21[M+H] + MPa: 190-192℃.

[0707] Example 76 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-28)

[0708]

[0709] 1) Take a 50 ml dry round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (300 mg, 1.09 mmol), N-tert-butoxycarbonyl-3-bromopropylamine (517.09 mg, 2.17 mmol), cesium carbonate (707.55 mg, 2.17 mmol), sodium iodide (162.75 mg, 1.09 mmol), 4A molecular sieve (500 mg), and DMF (6 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in an 80 °C oil bath, and stir for 4 h. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution, wash the filter cake with water, dissolve it in methanol and dichloromethane, filter, concentrate under reduced pressure, and dry to obtain 393 mg of a light brown solid, with a yield of 83.49%. The solid is not purified and is directly added to the next step.

[0710] 2) Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (393 mg, 0.91 mmol), 3-p-fluorobenzoylbenzaldehyde (310.33 mg, 1.36 mmol), cesium carbonate (443.05 mg, 1.36 mmol), anhydrous sodium sulfate (257.53 mg, 1.81 mmol), and DMF (5 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 60 °C oil bath, and stir for 21 h. Monitor the reaction by LC-MS. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution; the solid is viscous. Wash the filter cake with cold water, then dissolve it in methanol and dichloromethane, filter, and concentrate under reduced pressure. Extract the aqueous phase with EA until clear. Combine the organic phases and concentrate under reduced pressure. The filter cake and organic phase sample were combined and eluted with a gradient to give 200 mg of pale yellow solid, with a yield of 43.98%.

[0711] 1 H NMR (500MHz, DMSO-d6) δ = 12.01 (s, 1H), 10.35 (s, 1H), 7.91 (s, 3H), 7.82 (s, 1H), 7.75 (d, J = 6.5, 1H),7.66-7.56(m,2H),7.40(s,2H),6.98(s,1H),6.81(s,1H),6.69(s,1H),4.02 (s,2H),3.20(s,1H),2.95(s,2H),1.79(s,2H),1.47-1.24(m,15H).MS(ESI):m / z 602.25[M+H] + MPa: 179-180℃.

[0712] Example 77 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-29)

[0713]

[0714] Take a 50 mL dry round-bottom flask and add (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (475.8 mg, 0.79 mmol), methanol (30 mL), and hydrochloric acid (3.30 mL, 12 mmol / mL) sequentially. Place the flask in an oil bath at 40 °C and stir for 11 h. Monitor the reaction by LC-MS. After the reaction is complete, concentrate under reduced pressure, adjust the pH to neutral with saturated sodium bicarbonate aqueous solution, concentrate again under reduced pressure, redissolve in ethanol:dichloromethane (1:3), remove salt by filtration, and perform C18 reverse-phase column chromatography with gradient elution. Concentrate under reduced pressure to give 310 mg of yellow solid, yield 78.07%.

[0715] 1 H NMR (500MHz, DMSO-d6) δ = 11.82 (s, 1H), 10.41 (s, 1H), 8.17 (s, 3H), 7.93–7.86 (m, 2H), 7.81 (s,1H),7.75(d,J=7.7,1H),7.64(d,J=7.7,1H),7.58(t,J=7.6,1H),7.43–7.37(m,2H),6.81(s,1H),6.67(d, J=3.6,1H),4.19(dt,J=14.5,7.4,2H),3.28–3.19(m,1H),2.81(dd,J=13.3,6.5,2H),2.04–1.95(m,2H),1.32 (dd,J=7.0,2.4,6H).MS(ESI):m / z 502.21[M+H] + MPa: 199-201℃.

[0716] Example 78 Synthesis of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-(3-N-benzyloxycarbonylpiperazinyl)propylamino-imidazol-4-yl)methylene)piperazin-2,5-dione (PLN-5-30)

[0717]

[0718] Take a 25 mL dry round-bottom flask and add (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((5-isopropyl-1-propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (60 mg, 0.12 mmol), pyridine (28.39 mg, 0.36 mmol), and dichloromethane (1.5 mL) dropwise. Place the flask in a 0 °C cold trap and stir for 10 min. Add a solution of benzyloxychloride (30.60 mg, 0.18 mmol) in dichloromethane (1.5 mL) dropwise. After the addition is complete, transfer the flask to room temperature and stir for 1 h. Monitor the reaction by LC-MS. Once the reaction is complete, quench with methanol, concentrate under reduced pressure, load onto a C18 column, and perform reverse-phase column chromatography with gradient elution. Concentrate under reduced pressure to give 42 mg of a yellow solid, yield 55.24%.

[0719] 1 H NMR (500MHz, DMSO-d6) δ12.01(s,1H),10.35(s,1H),7.90(s,3H),7.82(s,1H),7.75(d,J=6.08 Hz,1H),7.69-7.55(m,2H),7.47-7.26(m,7H),6.81(s,1H),6.69(s,1H),5.03(s,2H),4.03(s,2H),3.20(s, 1H),3.04(s,2H),1.83(s,2H),1.31(d,J=4.75Hz,6H).MS(ESI):m / z 636.33[M+H] + MPa: 175-178℃.

[0720] Example 79 Synthesis of (3Z,6Z)-3-benzylmethyl-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-31)

[0721]

[0722] 1) Take a 50 ml dry two-necked round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (200 mg, 0.72 mmol), N-tert-butoxycarbonyl-3-bromopropylamine (344.8 mg, 1.45 mmol), cesium carbonate (471.80 mg, 1.45 mmol), sodium iodide (108.50 mg, 0.72 mmol), 4A molecular sieve (300 mg), and DMF (4 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 60 °C oil bath, and stir for 10 h. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution, wash the filter cake with water, dissolve it in methanol and dichloromethane, filter, concentrate under reduced pressure, and dry to obtain 256 mg of a light brown solid, yield 81.57%. The solid is not purified and directly added to the next step.

[0723] 2) Take a 25 ml dry brown round-bottom flask and add (Z)-1-acetyl-3-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylaminoimidazol-4-yl)methylene)piperazine-2,5-dione (256 mg, 0.59 mmol), cesium carbonate (288.61 mg, 0.89 mmol), anhydrous sodium sulfate (167.76 mg, 1.18 mmol), benzaldehyde (94.00 mg, 0.89 mmol), and DMF (4 ml) sequentially. Exhaust the gas, place the flask under nitrogen protection in a 60 °C oil bath, and stir for 18 h. Monitor the reaction by LC-MS. After the reaction is complete, drop the reaction solution into 4 °C cold water. A solid precipitates out. Filter the solution, wash the filter cake with cold water, dissolve it in methanol and dichloromethane, filter, and concentrate under reduced pressure. Extract the aqueous phase with EA until clear. Combine the organic phases and concentrate under reduced pressure. The filter cake and organic phase sample were combined and subjected to reverse column chromatography with gradient elution to give 99 mg of pale yellow solid, with a yield of 34.96%.

[0724] 1 H NMR (500MHz, DMSO-d6) δ = 11.98 (s, 1H), 10.04 (s, 1H), 7.91 (s, 1H), 7.59–7.25 (m, 5H), 6.98 (s,1H),6.73(d,J=29.1,2H),4.02(s,2H),3.21(s,1H),2.95(s,2H),1.80(s,2H),1.36(15H).MS(ESI):m / z480.23[M+H] + MPa: 210-213℃.

[0725] Example 80: Synthesis of (3Z,6Z)-3-benzylmethyl-6-((5-isopropyl-1-propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-32)

[0726]

[0727] In a dry round-bottom flask, (3Z,6Z)-3-benzylmethyl-6-((5-isopropyl-1-(N-tert-butyloxycarbonyl)propylamino-imidazol-4-yl)methylene)piperazine-2,5-dione (60 mg, 0.12 mmol), methanol (10 ml), water (1 ml), and hydrochloric acid (2 ml, 12 mmol / ml) were added sequentially. The mixture was placed in an oil bath at 30 °C and stirred for 12 h. The reaction was monitored by LC-MS. After the reaction was complete, the pH was adjusted to 8 with saturated sodium hydroxide aqueous solution, and the mixture was concentrated under reduced pressure. The solution was redissolved in methanol and dichloromethane, filtered, and concentrated under reduced pressure. The mixture was then extracted with ethyl acetate and water, and the combined organic phases were concentrated under reduced pressure to give 30 mg of a yellow solid, with a yield of 63.20%.

[0728] 1 H NMR (500MHz, DMSO-d6) δ = 7.88 (s, 1H), 7.54 (d, J = 7.2, 2H), 7.41 (t, J = 7.2, 2H), 7.32 (d, J = 7.1, 1H),6.74(s,1H),6.69(s,1H),4.06(s,2H),3.24(m,2H),2.54(s,2H),1.73(s,2H),1.33(d,J=6.8,6H).MS(ESI):m / z 380.09[M+H] + MPa: 207-210℃.

[0729] Example 81: Preparation of (3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((4-(3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (PLN-5-33)

[0730] 1)(3Z)-1-acetyl-3-((4-(N-Boc-3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0731]

[0732] Take a 25 ml dry brown round-bottom flask and add (3Z)-1-acetyl-3-((5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (40.00 mg, 0.13 mmol), N-Boc-3-aminopropyl bromide (61.43 mg, 0.26 mmol), cesium carbonate (84.06 mg, 0.26 mmol), potassium iodide (21.41 mg, 0.13 mmol), 4A molecular sieve (40 mg), and DMF (2.5 ml) sequentially. Purge the mixture three times under nitrogen protection. Place the flask in a 75°C oil bath and stir for 4 h. After the reaction is complete, add the reaction solution dropwise to cold water. A reddish-brown solid precipitates. Filter the solution, wash the filter cake with water, redissolve the filter cake in methanol and dichloromethane, filter again, and concentrate the filtrate under reduced pressure to obtain 41.8 mg of an orange-yellow solid, with a yield of 69.37%.

[0733] 2)(3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((4-(N-Boc-3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0734]

[0735] Take a 25 ml dry brown round flask and add (3Z)-1-acetyl-3-((4-(N-Boc-3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (41.8 mg, 0.090 mmol), 3-(4-fluorobenzoyl)benzaldehyde (30.55 mg, 0.13 mmol), cesium carbonate (43.66 mg, 0.13 mmol), anhydrous sodium sulfate (25.43 mg, 0.18 mmol), and DMF (2 ml) in sequence. Place the flask in a 50 °C oil bath and stir for 21 h. After the reaction is complete, add the reaction solution dropwise to 20 ml of cold water at 4 °C. A pale yellow solid precipitates out. Filter the solution, wash the filter cake with cold water, dissolve the filter cake in a 1:3 mixture of methanol and dichloromethane, filter, and concentrate under reduced pressure. Methanol was ultrasonically homogenized, placed in a -30°C refrigerator to stand, filtered, the filter cake was washed with cold methanol, and dried under vacuum at 50°C to obtain 34.5 mg of orange-yellow solid, with a yield of 60.71%.

[0736] 3)(3Z,6Z)-3-(3-(p-fluorobenzoyl)benzene)methylene-6-((4-(3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione

[0737]

[0738] Take a 25 ml dry round-bottom brown bottle and add (3Z,6Z)-3-(2-(p-fluorobenzoyl)benzene)methylene-6-((4-(N-Boc-3-aminopropyl)-5-phenyl-1H-imidazol-4-yl)methylene)piperazine-2,5-dione (27 mg, 0.042 mmol), concentrated hydrochloric acid (0.18 ml), and methanol (3 ml) sequentially. The mixture is purged with nitrogen three times and placed in an oil bath at 30 °C with stirring for 12 h. After the reaction is complete, concentrate the solution under reduced pressure to a solid state, redissolve it with 0.5 ml of methanol, and add it dropwise to a round-bottom flask containing 10 ml of ethyl acetate. A pale yellow solid precipitates. After the addition is complete, place the flask in a cold well at 0 °C and stir for 1 h. Filter the solution, wash the filter cake with ethyl acetate, and dry it under vacuum at 50 °C to obtain 10.6 mg of a pale yellow solid. Yield: 48.18%.

[0739] 1 H NMR(500MHz,DMSO-d6)δ11.83(s,1H),10.39(s,1H),8.25(s,1H),7.99(s,3H),7.90(m,2H), 7.80(s,1H),7.74(d,J=7.5Hz,1H),7.60(m,5H),7.47(d,J=7.0Hz,2H),7.39(t,J=8.7Hz,2H),6 .84(s,1H),6.32(s,1H),4.13(t,J=6.6Hz,2H),2.65(d,J=5.6Hz,2H),1.83(m,2H).MS(ESI):m / z 536.10[M +H] + MPa: 263-265℃.

[0740] Example 82: Preparation and hydrochloric acid content test of dehydrophenyl acetylene hydrochloride.

[0741] 1) General methods for salt formation of compounds:

[0742] Take 1.0 eq of the compound and place it in a dry round-bottom flask. Dissolve the compound in an appropriate amount of methanol and add concentrated hydrochloric acid (10.0 eq) dropwise. Stir the mixture at room temperature in the dark for 2 h. After the reaction is complete, concentrate the mixture under reduced pressure until no solvent remains. Dissolve the solid in a small amount of methanol and add it dropwise to ethyl acetate or acetone. A solid precipitates out. Place the mixture in a 0°C cold trap and stir for 2 h. Filter the mixture and wash the filter cake with cold ethyl acetate or acetone. Dry the filter cake under vacuum at 50°C to obtain a white solid. Form a salt with hydrochloric acid in a quantitative molar ratio (compound:hydrochloric acid = 1:2).

[0743] 2) Test and analysis of the pure hydrochloric acid content of dehydrophenylastistin hydrochloride compounds

[0744] 1. Reagents and reagents

[0745] Sodium hydroxide (analytical grade), potassium hydrogen phthalate (standard); phenolphthalein indicator solution: 1g phenolphthalein is added to 100ml ethanol.

[0746] 2. Configuration

[0747] Take an appropriate amount of sodium hydroxide, add water and shake to dissolve it into a saturated solution. After cooling, place it in a polyethylene plastic bottle, let it stand for several days, and use it after it has clarified.

[0748] Sodium hydroxide titrant (0.1 mol / L): Take 5.6 ml of clear saturated sodium hydroxide solution, add freshly boiled and cooled water to make 1000 ml, and shake well.

[0749] 3. Calibration

[0750] 3.1 Operating Procedures

[0751] Sodium hydroxide titrant (0.1 mol / L): Accurately weigh approximately 0.6 g of potassium hydrogen phthalate (dried to constant weight at 105 °C), add 50 ml of freshly boiled and cooled water, shake to dissolve as much as possible; add 2 drops of phenolphthalein indicator, and titrate with this solution; near the endpoint, ensure the potassium hydrogen phthalate is completely dissolved, titrating until the solution turns pink. Each 1 ml of sodium hydroxide titrant (0.1 mol / L) is equivalent to 20.42 mg of potassium hydrogen phthalate.

[0752] 3.2 Calculation Formula

[0753] The concentration C (mol / L) of the sodium hydroxide titrant is calculated using the following formula:

[0754] C(mol / L)=(m*1.000) / (V*204.2)

[0755] In the formula: m is the amount of potassium hydrogen phthalate weighed (mg);

[0756] V represents the volume (ml) of this titrant consumed;

[0757] 204.2 represents the number of milligrams of potassium hydrogen phthalate equivalent to 1 ml of sodium hydroxide solution (1.000 mol / L).

[0758] 4. Titration

[0759] Accurately weigh the test reagent (approximately 10 mg), add 50 ml of freshly boiled and cooled water, shake to dissolve; add 2 drops of phenolphthalein indicator, titrate with standardized sodium hydroxide solution until the solution turns pink at the endpoint, and record the volume of sodium hydroxide solution consumed, V. 消耗 .

[0760] 5. Results Analysis

[0761] n (氢氧化钠)=C(mol / L)*V 消耗

[0762] n (试药) =m 试药 / M 试药

[0763] The salt formation ratio is: n (氢氧化钠) / n (试药)

[0764] Experimental results show that, according to the calculation, the salt formation ratio of the punabulin morpholine derivative of this invention to hydrochloric acid is 1:2, that is, one molecule of the derivative binds to two molecules of hydrochloric acid.

[0765] Effect Example 1: Inhibition test of cell proliferation by compound or its hydrochloride

[0766] Tumor cells (NCI-H460, BxPC-3, HT-29) in logarithmic growth phase were digested, centrifuged, and re-vortexed with fresh culture medium. Cell counting was performed using a cell counting chamber, with 2000-6000 cells per well. 100 μL of culture medium (containing cells) was added to each well of a 96-well plate, which was then incubated at 37°C, 5% CO2. After complete cell attachment, the test sample and Plinabulin were diluted to different concentrations using fresh culture medium, with four wells for each concentration. After 72 hours of drug treatment, 20 μL of 5 mg / L MTT was added to each well, and the plate was incubated for 3-4 hours. The culture medium and MTT were then discarded, and 100 μL of DMSO was added to each well. The 96-well plate was then placed on a 96-well plate shaker to completely dissolve the purple formazan. The absorbance (OD value) was then measured at 490 nm using a microplate reader. This experiment also requires the setting of zeroing wells (blank culture medium, MTT, DMSO). Cell inhibition rate = 1 - (OD value of drug-treated group - OD value of zeroing group) / (OD value of blank group - OD value of zeroing group) × 100%.

[0767] The results of the cell proliferation inhibition test of the compounds of the present invention or their hydrochlorides are shown in Table 1.

[0768] Table 1. Inhibition of cell proliferation by the compounds of the present invention.

[0769]

[0770]

[0771]

[0772] Note: NCI-H460 is a human non-small cell lung cancer cell line, BXPC-3 is a human pancreatic cancer cell line, and HT-29 is a human colon cancer cell line. Punabulin is a positive control. The control group is DMSO without the sample.

[0773] Example 2: Compound Solubility Experiment

[0774] 1) Establishment of the standard curve

[0775] Weigh approximately 1 mg of the compound and dissolve it in DMSO to prepare a 1 mg / mL stock solution. Dilute the stock solution with methanol 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, 50,000, 10,000, 5,000, 1,000, 500, and 250 ng / mL.

[0776] 2) Method for determining sample solubility

[0777] Weigh approximately 1 mg of each compound into a 1.5 mL brown EP tube and add 1 mL of ultrapure water. 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℃) and shake at 100 rpm for 24 h to allow complete dissolution equilibrium to be reached. After 24 h, quickly filter the supernatant through a 0.45 μm microporous membrane, discard the initial filtrate, and take 200 μL of the subsequent filtrate, diluting it with 200 μL of methanol. Repeat the determination at least three times. Inject the sample into LC-MS under the chromatographic conditions, determine the peak area, and calculate the equilibrium solubility of each compound in pure water.

[0778] 3) Calculation and Results

[0779] The results of the solubility experiments of the compounds are shown in Table 2 below:

[0780] Table 2

[0781] Compound Name solubility Plinabulin <250(ng / mL) PLN-1-13-1 <250(ng / mL) PLN-1-14-1 <250(ng / mL) PLN-5-29 833.10±49.65 PLN-5-6 <250 PLN-5-4 <250 PLN-5-8 <250 PLN-5-9 <250 PLN-5-10 <250

[0782] Example 3: Solubility test of dehydrophenyl acetylene hydrochloride.

[0783] 1) Experimental methods

[0784] Weigh approximately 2 mg of dehydrophenylastusine hydrochloride into a 0.5 mL brown EP tube and add 0.2 mL of ultrapure water. Vortex and place in an incubator shaker, maintaining the temperature at (37±1℃) and shaking at 100 rpm for 24 h to allow complete dissolution equilibrium. Observe the dissolution process.

[0785] 2) Experimental Results

[0786] The solubility results of dehydrophenylastitin hydrochloride compounds are shown in Table 3 below:

[0787] Table 3

[0788]

[0789]

[0790] Example 4: Immunofluorescence Experiment

[0791] 1) Experimental methods

[0792] 1. Preparation of cell slides: In a clean bench, place sterile round coverslips flat in a 12-well plate. Digest NPI-H460 cells in the logarithmic growth phase with trypsin, collect the cells by centrifugation, and resuspend them in fresh culture medium. Prepare 1×10⁻⁶ cells per well. 5 One cell was added to each well containing a sterile coverslip. After the cells adhered (about 24 hours), Plinabulin (10 nM), PLN-5-6 (2 nM), PLN-5-8 (2 nM) and PLN-5-9 (2 nM) were added and the cells were cultured for another 24 hours.

[0793] 2. Fixation and blocking of cell slides: Wash cell slides three times with PBS for 3 min each time; fix the slides with 4% paraformaldehyde for 15 min, wash the slides three times with PBS for 3 min each time; add 0.5% Triton X-100 prepared with PBS and let stand for 20 min, wash the slides three times with PBS for 3 min each time; discard the PBS, add 5% ordinary goat serum to the slides and cover the slides, and block at room temperature.

[0794] 3. Add primary antibody: Absorb the blocking solution with absorbent paper, add sufficient primary antibody (prepared with PBS in a certain ratio) to the slide, and incubate overnight at 4°C in a humidified chamber.

[0795] 4. Add fluorescent secondary antibody: Wash the slide three times with PBS for 3 minutes each time; discard the PBS, add the diluted fluorescent secondary antibody, incubate in a humidified chamber at 37°C for 1 hour, and then wash the slide three times with PBS for 3 minutes each time. (From the time you add the fluorescent secondary antibody onwards, each step must be performed in the dark.)

[0796] 5. Counterstain cell nuclei: Add DAPI staining solution and incubate at room temperature in the dark for 10 min.

[0797] 6. Mounting: Place the slides in PBS and wash three times on a decolorizing shaker for 5 minutes each time. After blotting the slides with absorbent paper, mount them with anti-fluorescence quenching mounting medium.

[0798] 7. Microscopic examination and photography: Observe and acquire images under a fluorescence microscope after the slides have been prepared. (DAPI is excited by ultraviolet light at a wavelength of 330-380 nm and emits blue light at a wavelength of 420 nm; FITC is excited by ultraviolet light at a wavelength of 465-495 nm and emits green light at a wavelength of 515-555 nm; CY3 is excited by ultraviolet light at a wavelength of 510-560 nm and emits red light at a wavelength of 590 nm).

[0799] 2) Experimental Results

[0800] The IOD value was calculated using Image Pro Plus 6.0 software, and the results are shown in Table 4 below. Figure 1 As shown:

[0801] Table 4

[0802]

[0803] Experiments showed that the cells in the untreated blank control group had normal morphology and intact microtubule structures, exhibiting a regular spindle shape. After treatment with the compound for 24 hours, microtubules atrophied and even broke down. Immunofluorescence results and β-tubulin fluorescence values ​​indicated that compound PLN-5-6 was more effective than Plinabulin in inhibiting microtubule polymerization.

[0804] Example 5: Western Blot Experiment

[0805] 1) Experimental methods

[0806] (1) Extraction of total cellular protein

[0807] NCI-H460 cells in logarithmic growth phase were injected with 1×10⁻⁶ cells. 6 Cells were seeded at a density of cells / well in 6-well plates. After 24 h of adherent culture, plinabulin (10 nM), PLN-5-4 (5 nM), PLN-5-4 (10 nM), PLN-5-6 (5 nM), and PLN-5-6 (10 nM) were added to the plates for further incubation for 24 h. The culture medium was then aspirated, and the cells were washed twice with PBS. The cells were lysed on ice with RIPA lysis buffer containing protease and phosphatase inhibitors. After 30 min on ice, the lysate was collected in 1.5 ml EP tubes and centrifuged at 12000 rpm for 10 min at 4 °C. The supernatant was then collected as total cellular protein.

[0808] (2) BCA method for determining protein concentration

[0809] A stock solution of BSA with a stock concentration of 2 mg / ml was diluted with distilled water to concentrations of 2, 1, 0.5, 0.25, 0.125, and 0.0625 mg / ml. A blank control (distilled water) was also included. The protein solution after lysis and centrifugation was diluted with distilled water at a ratio of 1:8. The protein content in a series of BSA solutions and sample protein solutions of different concentrations was determined using the following methods:

[0810] First, add 25 μl of solution A (prepared by mixing solution S and solution A at a volume ratio of 1:50) to each sample tube. Then, add 5 μl of BSA solution and protein solution of different concentrations. After gently shaking to mix, quickly add 200 μl of solution B and incubate at room temperature in the dark for 15 min. Read the absorbance value at 562 nm using a spectrophotometer. Plot a regression curve using the absorbance values ​​of the diluted BSA solutions from the standards. Calculate the protein concentration of the test sample using the OD value of the test sample.

[0811] (3) Glue pouring

[0812] ① Rinse the glued glass plate with distilled water and let it air dry vertically.

[0813] ② Prepare 10 ml of 15% separating gel according to the above method, add 10 μl of TEMED and 100 μl of 10% ammonium persulfate, mix well and immediately pour the gel up to 2-3 mm below the lower edge of the comb. Seal the liquid surface with isopropanol to remove air bubbles and isolate air. Let stand at room temperature for 45 minutes until the separating gel is completely solidified.

[0814] ③ After the separating gel has completely solidified, pour off the isopropanol on top, wash three times with deionized water, and then blot dry with filter paper.

[0815] ④ Prepare 5ml of 5% stacking gel, add 5μl of TEMED and 50μl of 10% APS, mix well and immediately pour the gel to the top, then vertically insert the Teflon comb and let it stand at room temperature for 20 minutes to allow the stacking gel to solidify.

[0816] ⑤ After the stacking gel has completely solidified, remove the comb, place the gel in the electrophoresis tank, add electrophoresis buffer, and rinse the sample wells with electrophoresis buffer to remove air bubbles.

[0817] (4). Electrophoresis

[0818] ① Take the protein extract from each treatment group, adjust the protein concentration, and mix it with an equal volume of 1× loading buffer to obtain the loading solution.

[0819] ② Boil the sample solution in 100℃ boiling water for 5 minutes to denature the protein, cool it quickly on ice, and centrifuge at 3000 rpm for 1 minute.

[0820] ③ Add 50 μg of sample solution to each well, and leave one well for 5 μl of pre-stained marker. Fill with electrophoresis buffer, cover the tank, and turn on the power. First, use 80V constant voltage electrophoresis for about 15 minutes. After the indicator bromofrancolene enters the separating gel, switch to 120V constant voltage electrophoresis. When the indicator reaches about 0.5 cm from the bottom of the gel, turn off the power and remove the gel plate.

[0821] (5) Transfer protein and immunoassay

[0822] ① Just before the electrophoresis is about to end, soak the PVDF membrane in methanol for 15 seconds, then rinse it with transfer buffer for 2 minutes, let it stand for 5 minutes, and then start the subsequent operations.

[0823] ② Pry the gel in water, trim the gel, and then immerse the gel in transfer buffer for 15 minutes to equilibrate.

[0824] ③ Prepare the transfer membrane "sandwich" in the following order: black side (negative electrode) → sponge → filter paper → gel → PVDF membrane → filter paper → sponge → red side (positive electrode). After each layer is laid, remove air bubbles before laying the next layer. Preparing the sandwich in the transfer buffer can avoid the formation of air bubbles.

[0825] ④ Connect the positive and negative electrodes, place the transfer box into the electroporator with the membrane facing the positive electrode, and add the transfer buffer.

[0826] ⑤ Place the electroporator in ice water and transfer the membrane at a constant current of 100mA for 90 minutes.

[0827] ⑥ After the transfer is complete, quickly remove the PVDF membrane and seal it in 5% BSA at room temperature for 2 hours.

[0828] ⑦ Remove the membrane and wash it with TBST for 5 minutes × 3 times on a shaker.

[0829] ⑧ Add TBST diluted caspase-3 (antibody diluted according to the brand instructions) and GAPDH (antibody diluted according to the brand instructions) to the incubation bag and incubate overnight at 4°C.

[0830] ⑨ Wash the membrane with TBST for 5 min × 3 times, and incubate at room temperature for 2 h with horseradish peroxidase (HRP)-labeled goat anti-rabbit secondary antibody (Beyotime brand, 1:7000) or horseradish peroxidase (HRP)-labeled goat anti-mouse secondary antibody (Beyotime brand, 1:7000).

[0831] ⑩ Wash the membrane with TBST for 5 min × 3 times. React the membrane with chemiluminescence detection reagent (reagent A: reagent B = 1:1) for 2 min, and then expose and develop it in a chemiluminescence developer.

[0832] 2) Experimental Results

[0833] The results are shown in Table 5 below:

[0834] Table 5

[0835]

[0836] The experimental results showed that after 24 hours of treatment, comparing the relative gray values ​​of Caspase-3 / GAPDH proteins, PLN-5-4 and PLN-5-6 were more effective than Plinabulin in promoting apoptosis in the NCI-H460 cell line.

Claims

1. The following compounds or their pharmaceutically acceptable salts: 。 2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and pharmaceutical excipients.

3. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a medicament for the prevention and / or treatment of cancer.

4. The application as described in claim 3, characterized in that, The cancer is one or more of the following: lung cancer, pancreatic cancer, colon cancer, and liver cancer.

5. The use of the compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a microtubule inhibitor.

Images