Arylpiperazine derivatives, processes for their preparation and use

Abstract

The present application relates to the technical field of compound preparation, in particular to an arylpiperazine derivative, a preparation method and application. Compared with the prior art, the arylpiperazine derivative prepared by the present application is different from the known inhibitors, and the drug resistance problem of the prior art compound is avoided. Through preliminary pharmacological research, it is shown that the arylpiperazine derivative can significantly inhibit the invasion ability of lung cancer cells, and can be developed as a new antitumor drug. This discovery provides a new mechanism for lung cancer treatment and has clinical practical value.

Description

Technical Field

[0001] This invention relates to the field of compound preparation technology, specifically to an arylpiperazine derivative, its preparation method, and its application. Background Technology

[0002] Lung cancer is one of the leading causes of death and morbidity among malignant tumors worldwide. According to the World Health Organization, lung cancer causes millions of deaths annually, with non-small cell lung cancer (NSCLC) accounting for approximately 85% of these cases. The high mortality rate of lung cancer is closely related to its aggressive and metastatic nature. Current treatments include surgery, radiotherapy, chemotherapy, and targeted therapy; however, due to issues such as drug resistance and toxic side effects, there is still an urgent need in clinical practice to develop new and effective treatments.

[0003] Currently, some known small molecule inhibitors (such as EGFR inhibitors and ALK inhibitors) can delay lung cancer progression to some extent, but their effects on inhibiting cancer cell invasion and metastasis are limited, and drug resistance issues exist. Therefore, developing compounds with novel mechanisms of action and structures to inhibit the invasive ability of lung cancer cells has significant clinical implications and application prospects. Summary of the Invention

[0004] To overcome the above-mentioned technical defects, the present invention provides an arylpiperazine derivative, a preparation method and an application to inhibit the invasiveness and metastasis of lung cancer cells.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows: Firstly, arylpiperazine derivatives of Formula I or Formula II, or pharmaceutically acceptable salts thereof. Formula I, Formula II Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, naphthalene ring, anthracene ring, phenaene, tetraphenylene, pyrene, linear or angled pentaphenylene, hexaphenylene, indene or fluorene; R1 represents 1 to 2 substituents, which can be independently selected from hydrogen atoms, halogen atoms, C1-C5 haloalkyl groups, and hydroxyl groups; R2 is selected from ester group and C1-C5 alkoxy group.

[0006] Furthermore, the Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, and naphthalene ring; R1 can be independently selected from H, monosubstituted F, or monosubstituted C1-C3 haloalkyl; R2 can be independently selected from carboxylic acid ester group, sulfonate group, and C1-C3 alkoxy group.

[0007] Furthermore, the arylpiperazine derivative shown in Formula I is specifically: , , , , .

[0008] Furthermore, the arylpiperazine derivative shown in Formula II is specifically: , , .

[0009] Secondly, the use of the above-mentioned arylpiperazine derivatives or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating cancer is provided.

[0010] The following applications are also within the scope of protection of this invention: Furthermore, the tumor-related antigen or cell surface receptor is selected from one of MSG783, STEAP2, TrpM4, CRIPTO, CD21, HER2, EphB2R, and BAFF-R.

[0011] Further, the tumor is breast cancer, lung cancer, liver cancer, stomach cancer, colon cancer, rectal cancer, nasopharyngeal carcinoma, pancreatic cancer, thyroid cancer, prostate cancer, leukemia, lymphoma, kidney tumor, sarcoma, or blastoma; more preferably, the tumor is non-small cell lung cancer.

[0012] Furthermore, the drug uses the above-mentioned arylpiperazine derivative or its pharmaceutically acceptable salt as the active ingredient, and is formulated into a pharmaceutically acceptable dosage form with pharmaceutically acceptable excipients.

[0013] Thirdly, a method for preparing arylpiperazine derivatives is provided, and the reaction route is as follows:

[0014] Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, naphthalene ring, anthracene ring, phenaene, tetraphenylene, pyrene, linear or angled pentaphenylene, hexaphenylene, indene or fluorene; R1 represents 1 to 2 substituents, which can be independently selected from hydrogen atoms, halogen atoms, C1-C5 haloalkyl groups, and hydroxyl groups.

[0015] Fourthly, a pharmaceutical composition for targeted cancer therapy is provided, comprising the above-mentioned arylpiperazine derivative or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients.

[0016] Furthermore, the excipients include one or more of solvents, disintegrants, flavoring agents, preservatives, colorants, binders, lubricants, diluents, and drug carriers. Those skilled in the art will fully understand that the pharmaceutically acceptable carrier is generally recognized for this purpose and as an inactive ingredient in the pharmaceutical preparation.

[0017] The auxiliary agents include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, and release inhibitors. The diluent can be one or more of mannitol, sucrose, lactose, sorbitol, xylitol, polyethylene glycol, propylene glycol, vegetable oil, and mineral oil; the disintegrant can be one or more of croscarmellose sodium, colloidal silica, and citric acid; the binder can be one or more of starch paste, ethanol, water, and povidone alcohol solution; the preservative can be one or more of ethylparaben, propylparaben, sorbic acid, potassium sorbate, calcium propionate, sodium dehydroacetate, sodium diacetate, and sodium lactate; and the antioxidant can be ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetate, butylated hydroxytoluene, glycine, inositol, and other antioxidants. The flavoring agent may be one or more of the following: citric acid, sodium ascorbate, lecithin, malic acid, hydroquinone, citric acid, succinic acid, and sodium metabisulfite; the flavoring agent may be one or more of the following: aspartame, sucrose, xylitol, steviol glycosides, cyclamate, sorbitol, cocoa, pure vanilla, vanillin, ethyl vanillin, chocolate, malt, and mint; the suspending agent may be one or more of the following: xanthan gum, polyvinylpyrrolidone, sodium alginate, aluminum stearate, and hydrogenated vegetable oil; the emulsifier may be one or more of the following: alkyl sulfate, soap, dodecylbenzene sulfonate, lactate, sulfosuccinate, monoglyceride sulfonate, phosphate ester, siloxane, and taurine.

[0018] Furthermore, the dosage form of the pharmaceutical composition is a suspension, a dispersible oil suspension, a dry suspension, a wettable powder, an emulsifiable concentrate, or a water-dispersible granule.

[0019] The drug is administered via gastrointestinal and non-gastrointestinal routes.

[0020] Specifically, the non-gastrointestinal route of administration is selected from injection, respiratory administration, skin administration, mucosal administration, or cavity administration.

[0021] Non-gastrointestinal drug delivery preparations are selected from injections, sprays, aerosols, patches, etc.

[0022] Specifically, the gastrointestinal administration formulation is selected from tablets, capsules, powders, granules, pills, solutions, emulsions, or syrups.

[0023] The pharmaceutical composition of the present invention contains 0.1-90 wt% of the active ingredient.

[0024] Pharmaceutical compositions may be prepared according to methods known in the art. For this purpose, if desired, the active ingredient may be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to form a suitable administration or dosage form for human use.

[0025] In addition, colorants, preservatives, flavorings, tasters, sweeteners or other materials may be added to pharmaceutical preparations if necessary.

[0026] The dosage of the pharmaceutical composition of the present invention depends on many factors, such as the sex, age, weight, and individual response of the patient or animal, the route of administration, and the frequency of administration. The dosage can be administered as a single dose or in several doses, such as two, three, or four doses. The dosage level must be selected based on the specific route of administration, the severity of the condition being treated, and the patient's condition and medical history. However, it is the practice in the art to start with a dosage below the level required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0027] However, it should be recognized that the total daily dosage of the pharmaceutical compositions of the present invention must be determined by the attending physician within the bounds of reliable medical judgment. For any given patient, the specific therapeutically effective dosage level must be determined based on a variety of factors, including the disorder being treated and its severity; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate; the duration of treatment; any drugs used in combination or concurrently; and similar factors known in the medical field. For example, it is practiced in the art to start with a dose below the level required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved.

[0028] Generally speaking, the dosage of the pharmaceutical composition of the present invention for mammals, especially humans, calculated based on the active ingredient, can be between 1 and 1000 mg / kg body weight / day, for example between 1 and 500 mg / kg body weight / day, for example between 50 and 500 mg / kg body weight / day, or 100 and 500 mg / kg, or 150 and 500 mg / kg, or 200 and 500 mg / kg, or 250 and 500 mg / kg.

[0029] The salts of the arylpiperazine derivatives described in this invention mainly include one of the following: hydrobromide, hydroiodide, hydrochloride, perchlorate, sulfate, maleate, fumarate, malate, tartrate, citrate, benzoate, amygdalinate, methanesulfonate, ethanesulfonate, benzenesulfonate, oxalate, phosphate, succinate, oxalate, lactate.

[0030] The pharmaceutical salts of the arylpiperazine derivatives described in this invention are prepared by the following method: the arylpiperazine derivative is dissolved in an organic solvent, then an acid is added, and the mixture is stirred at room temperature. The precipitated solid is an inorganic or organic acid salt of the arylpiperazine derivative.

[0031] Compared with existing technologies, the arylpiperazine derivative prepared in this invention, unlike existing known inhibitors, avoids the drug resistance problem of existing compounds. Preliminary pharmacological studies have shown that it can significantly inhibit the invasive ability of lung cancer cells and can be developed into a novel anti-tumor drug. This discovery will provide a new mechanism of action for the treatment of lung cancer and has clinical practical value. Attached Figure Description

[0032] Figure 1 The product obtained in Example 1 1 H NMR spectrum; Figure 2 The product obtained in Example 1 13 C NMR spectrum; Figure 3 The product obtained in Example 2 1 H NMR spectrum; Figure 4 The product obtained in Example 2 13 C NMR spectrum; Figure 5 The product obtained in Example 3 1 H NMR spectrum; Figure 6 The product obtained in Example 3 13 C NMR spectrum; Figure 7 The product obtained in Example 4 1 H NMR spectrum; Figure 8 The product obtained in Example 4 13 C NMR spectrum; Figure 9 The product obtained in Example 5 1 H NMR spectrum; Figure 10 The product obtained in Example 5 13 C NMR spectrum;

[0033] Figure 11The results show the cell migration ability of arylpiperazine derivatives. Figure 12 The antitumor activity of arylpiperazine derivatives in mice; in, Figure 12 a is a graph showing the changes in mouse body weight; Figure 12 b shows the changes in tumor volume in mice; Figure 13 The effect of arylpiperazine derivatives on organ tumor metastasis in mice; Figure 14 The effect of arylpiperazine derivatives on mouse survival time; Figure 15 The effects of arylpiperazine derivatives on serum biochemical parameters in mice; Figure 16 The effects of arylpiperazine derivatives on organ tissues in mice. Detailed Implementation

[0034] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be described in detail below with reference to specific embodiments. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise specified, the test materials used in the following embodiments were purchased from conventional biochemical reagent stores. Unless otherwise stated, percentages and parts are by weight. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those familiar with the art. Furthermore, any methods and materials similar to or equivalent to those described herein can be applied to the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0035] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0036] Since tumor metastasis is the core mechanism of cancer death by damaging the function of key organs, inducing systemic metabolic disorders and immune collapse, the inventors conducted multi-omics sequencing on collected patient samples and discovered an important target, stem2. They also conducted extensive functional verification of stem2 and found that inhibiting stem2 can inhibit tumor metastasis. Therefore, based on the target stem2, they developed a novel small molecule inhibitor, arylpiperazine derivative, to inhibit the invasive ability of lung cancer cells.

[0037] In some specific embodiments, arylpiperazine derivatives or pharmaceutically acceptable salts thereof are provided, having the structural formula shown in Formula I: Formula I; Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, naphthalene ring, anthracene ring, phenaene, tetraphenylene, pyrene, linear or angled pentaphenylene, hexaphenylene, indene or fluorene; R1 represents 1 to 2 substituents, which can be independently selected from hydrogen atoms, halogen atoms, and C1-C5 haloalkyl groups; In some specific embodiments, the Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, and naphthalene ring; R1 can be independently selected from H, monosubstituted F, or monosubstituted C1-C3 haloalkyl; In some preferred embodiments, the arylpiperazine derivative represented by Formula I is specifically: , , , , .

[0038] In some specific embodiments, arylpiperazine derivatives or pharmaceutically acceptable salts thereof are provided, having the structural formula shown in Formula II: Formula II Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, naphthalene ring, anthracene ring, phenaene, tetraphenylene, pyrene, linear or angled pentaphenylene, hexaphenylene, indene or fluorene; R1 represents 1 to 2 substituents, which can be independently selected from hydrogen atoms, halogen atoms, and C1-C5 haloalkyl groups; R2 is selected from ester group and C1-C5 alkoxy group.

[0039] In some specific embodiments, the Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, and naphthalene ring; R1 can be independently selected from H, monosubstituted F, or monosubstituted C1-C3 haloalkyl; R2 can be independently selected from carboxylic acid ester group, sulfonate group, and C1-C3 alkoxy group.

[0040] In some preferred embodiments, the arylpiperazine derivative represented by Formula II is specifically: , , .

[0041] In another specific embodiment, a method for preparing arylpiperazine derivatives is provided, and the reaction route is as follows:

[0042] Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, naphthalene ring, anthracene ring, phenaene, tetraphenylene, pyrene, linear or angled pentaphenylene, hexaphenylene, indene or fluorene; R1 represents 1 to 2 substituents, which can be independently selected from hydrogen atoms, halogen atoms, and C1-C5 haloalkyl groups.

[0043] In some embodiments of this implementation, It is obtained using the following route:

[0044] In some embodiments of this implementation, the Ar is selected from benzene ring, thiophene ring, furan ring, pyrrole, pyridine, benzothiophene, benzofuran, benzopyrrole, benzopyridine, and naphthalene ring; R1 can be independently selected from H, monosubstituted F, or monosubstituted C1-C3 haloalkyl.

[0045] The technical solution of the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings. It should be understood that the following embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0046] Example 1: Synthesis of arylpiperazine derivatives (1) Add 1 eq of 4-trifluoromethylbenzoic acid and 1–2 drops of anhydrous DMF to a dry round-bottom flask, cool to 0°C, and slowly add 1.2–1.5 eq of thionyl chloride; slowly heat to 70–80°C and stir the reaction for 1–2 h; after the raw material disappears, evaporate excess SOCl2 under reduced pressure and concentrate under reduced pressure to obtain 4-trifluoromethylbenzoyl chloride;

[0047] (2) Dissolve 3 mmol of 4-amino-N-piperazinylnitrobenzene in 15 mL of anhydrous DCM, place it in a dry three-necked flask, add triethylamine, cool to 0°C in an ice-water bath, and stir continuously for 10 min. Separately, dissolve 1 eq of 4-trifluoromethylbenzoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution, maintaining 0°C to prevent side reactions. After the addition, continue stirring in an ice bath for 30 min, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete, wash the organic layer with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate. The crude product is purified by silica gel column chromatography with petroleum ether / ethyl acetate = 3:1 to obtain the nitrated product.

[0048] (3) Add the nitro compound (2 mmol) to the reaction flask, dissolve it in 30 ml of dichloromethane / methanol (1:1), add the catalytic amount of palladium on carbon, stir overnight at room temperature under hydrogen, filter, and obtain the appropriate amount of product;

[0049] (4) Add 1 eq of 1-naphthoic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0°C, and slowly add 1.2–1.5 eq of thionyl chloride. Slowly heat to 70–80°C and stir the reaction for 1–2 hours; after the starting material disappears, evaporate excess SOCl2 under reduced pressure and concentrate under reduced pressure to obtain 1-naphthoyl chloride;

[0050] (5) Dissolve amine (2 mmol) in 10 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 1-naphthoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate to obtain ZD94. The crude product is purified by silica gel column chromatography with petroleum ether / ethyl acetate = 2:1, with a yield of 79%. Perform purity and mass spectrometry analysis on the product. Figure 1-2 ); .

[0051] Example 2 Synthesis of arylpiperazine derivatives (1) Add 1 eq of 4-fluorobenzoic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0°C, slowly add 1.2–1.5 eq of thionyl chloride, slowly heat to 70–80°C, stir the reaction for 1–2 hours, after the starting material disappears, evaporate excess SOCl2 under reduced pressure, and concentrate under reduced pressure to obtain 4-fluorobenzoyl chloride;

[0052] (2) Dissolve 3 mmol of 4-amino-N-piperazinylnitrobenzene in 15 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 4-fluorobenzoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution while maintaining 0°C. After the addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 h. After the reaction is complete, wash the organic layer with saturated NaHCO3, water, and saturated NaCl, respectively, and dry, filter, and concentrate. Purify the crude product by silica gel column chromatography with petroleum ether / ethyl acetate = 3:1 to obtain the nitrated product.

[0053] (3) Add the nitro compound (2 mmol) to the reaction flask, dissolve it in 30 ml of dichloromethane / methanol (1:1), add the catalytic amount of palladium on carbon, stir overnight at room temperature under hydrogen, filter, and obtain the appropriate amount of product;

[0054] (4) Add 1 eq of 1-naphthoic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0°C, and slowly add 1.2–1.5 eq of thionyl chloride. Slowly heat to 70–80°C and stir the reaction for 1–2 hours; after the starting material disappears, evaporate excess SOCl2 under reduced pressure and concentrate under reduced pressure to obtain 1-naphthoyl chloride;

[0055] (5) Dissolve amine (2 mmol) in 10 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 1-naphthoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add ice water to terminate the reaction. Wash the organic layer with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate to obtain ZD95. The crude product is purified by silica gel column chromatography with petroleum ether / ethyl acetate = 2:1, with a yield of 75%. Its purity and mass spectrometry are analyzed. Figure 3-4 ); .

[0056] Example 3 Synthesis of arylpiperazine derivatives (1) Add 1 eq of 4-pyridinecarboxylic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0 °C, and slowly add 1.2–1.5 eq of thionyl chloride. Slowly heat to 70–80 °C, stir the reaction for 1–2 hours, and after the starting material disappears, evaporate excess SOCl2 under reduced pressure and concentrate under reduced pressure to obtain 4-pyridinecarboxylic acid chloride;

[0057] (2) Dissolve 3 mmol of 4-amino-N-piperazinylnitrobenzene in 15 mL of anhydrous DCM, place it in a dry three-necked flask, add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 4-pyridinecarboxyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours; after the reaction is complete, wash the organic layer with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate; purify the crude product by silica gel column chromatography with petroleum ether / ethyl acetate = 3:1 to obtain the nitrated product;

[0058] (3) Add the nitro compound (2 mmol) to the reaction flask, dissolve it in 30 ml of dichloromethane / methanol (1:1), add the catalytic amount of palladium on carbon, and stir overnight at room temperature under hydrogen. Filter to remove the palladium on carbon and obtain the appropriate amount of product;

[0059] (4) Add 1 eq of 1-naphthoic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0°C, slowly add 1.2–1.5 eq of thionyl chloride, slowly heat to 70–80°C, stir the reaction for 1–2 hours, after the starting material disappears, evaporate excess SOCl2 under reduced pressure, and concentrate under reduced pressure to obtain 1-naphthoyl chloride;

[0060] (5) Dissolve amine (2 mmol) in 10 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 1-naphthoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate to obtain ZD96. The crude product is purified by silica gel column chromatography with petroleum ether / ethyl acetate = 2:1, with a yield of 73%. Perform purity and mass spectrometry analysis on the crude product. Figure 5-6 ); .

[0061] Example 4 Synthesis of arylpiperazine derivatives (1) In a dry round-bottom flask, add 1 eq of 3-thiophenecarboxylic acid, 1–2 drops of anhydrous DMF (catalyst), cool to 0°C, and slowly add 1.2–1.5 eq of thionyl chloride. Slowly heat to 70–80°C and stir the reaction for 1–2 hours. After the starting material disappears, remove excess SOCl2 under reduced pressure and concentrate under reduced pressure to obtain 3-thiophenecarboxylic acid chloride;

[0062] (2) Dissolve 3 mmol of 4-amino-N-piperazinylnitrobenzene in 15 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 3-thiopheneformyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer (Na2SO4), filter, and concentrate. Purify the crude product by silica gel column chromatography with petroleum ether / ethyl acetate = 3:1 to obtain 723 mg of product (yield 76%).

[0063] (3) Add the nitro compound (2 mmol) to the reaction flask, dissolve it in 30 ml of dichloromethane / methanol (1:1), add the catalytic amount of palladium on carbon, and stir overnight at room temperature under hydrogen. Filter to remove the palladium on carbon and obtain the appropriate amount of product;

[0064] (4) Add 1 eq of 1-naphthoic acid and 1–2 drops of anhydrous DMF (catalyst) to a dry round-bottom flask, cool to 0°C, slowly add 1.2–1.5 eq of thionyl chloride, slowly heat to 70–80°C, stir the reaction for 1–2 hours, after the raw material disappears, evaporate excess SOCl2 under reduced pressure, and concentrate under reduced pressure to obtain 1-naphthoyl chloride;

[0065] (5) Dissolve amine (2 mmol) in 10 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 1-naphthoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate to obtain ZD97. The crude product is purified by silica gel column chromatography with petroleum ether / ethyl acetate = 2:1, and its purity and mass spectrometry are analyzed. Figure 7-8 ); .

[0066] Example 5 Synthesis of arylpiperazine derivatives (1) In a dry round-bottom flask, add 1 eq of 2-fluorobenzoic acid and 1–2 drops of anhydrous DMF (catalyst), cool to 0°C, slowly add 1.2–1.5 eq of thionyl chloride, slowly heat to 70–80°C, and stir the reaction for 1–2 hours. After the starting material disappears, remove excess SOCl2 by vacuum distillation, and concentrate under vacuum to obtain 2-fluorobenzoyl chloride;

[0067] (2) Dissolve 3 mmol of 4-amino-N-piperazinylnitrobenzene in 15 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 2-fluorobenzoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, and concentrate. Purify the crude product by silica gel column chromatography with petroleum ether / ethyl acetate = 3:1 to obtain 720 mg of product (yield 73%).

[0068] (3) Add the nitro compound (2 mmol) to the reaction flask, dissolve it in 30 ml of dichloromethane / methanol (1:1), add the catalytic amount of palladium on carbon, stir overnight at room temperature under hydrogen, filter to remove palladium on carbon, and obtain the appropriate amount of product;

[0069] (4) Add 1 eq of 1-naphthoic acid and 1–2 drops of anhydrous DMF to a dry round-bottom flask, cool to 0°C, and slowly add 1.2–1.5 eq of thionyl chloride; slowly heat to 70–80°C and stir the reaction for 1–2 hours. After the starting material disappears, remove excess SOCl2 by vacuum distillation and concentrate under vacuum to obtain 1-naphthoyl chloride;

[0070] (5) Dissolve amine (2 mmol) in 10 mL of anhydrous DCM and place it in a dry three-necked flask. Add triethylamine (TEA, 2 eq), cool to 0°C in an ice-water bath, and stir continuously for 10 minutes. Separately, dissolve 1 eq of 1-naphthoyl chloride in a small amount of DCM and slowly add it dropwise to the above solution. Maintain 0°C for dropwise addition to prevent side reactions. After dropwise addition, continue stirring in an ice bath for 30 minutes, then warm to room temperature and continue stirring for 6–12 hours. After the reaction is complete as detected by TLC, add 10 mL of ice water to terminate the reaction. Wash the organic layer in a separatory funnel with saturated NaHCO3, water, and saturated NaCl, respectively. Dry the organic layer, filter, concentrate, and obtain crude product ZD93. Purify by silica gel column chromatography with petroleum ether / ethyl acetate = 2:1, yield 74%. Perform purity and mass spectrometry analysis on the product. Figure 1-2 ); .

[0071] Example 6: Investigating the effect of arylpiperazine derivatives on tumor cell migration ability The effect of the compounds of this invention on the migration ability of tumor cells. A549 cells were seeded in 6-well plates. After the cells grew to 90–100% confluence, a "wound" was gently made along a straight line in the center of the well using a sterile 200 μL pipette tip, followed by slow rinsing three times with PBS to remove detached cells. The experiment was divided into 6 groups: DMSO control group, Example 1 (ZD94 group), Example 2 (ZD95 group), Example 3 (ZD96 group), Example 4 (ZD97 group), and Example 5 (ZD93 group), with multiple concentration gradients of each compound. The medium was replaced with serum-free medium containing 0.5% FBS, and the corresponding concentrations of the compound were added. The cells were cultured at 37°C and 5% CO2 for 24 hours. Images of the same field of view of the scratched area were taken at 0 hours and 24 hours, and the scratch closure area was measured and calculated using ImageJ software to characterize cell migration ability. Each experiment was repeated at least 3 times, and the data are expressed as mean ± standard deviation (mean ± SD).

[0072] from Figure 11 As can be seen, compared with the DMSO control group, the compounds in each example inhibited cell migration to the scratch region to varying degrees within 24 hours. ZD93 performed best, achieving a migration rate of approximately 50% of that of DMSO at around 40 nm. The other compounds generally achieved a migration rate of approximately 50% of that of DMSO at around 80 nm.

[0073] Example 7: Investigation of the in vivo antitumor activity of arylpiperazine derivatives Laboratory animals: 6–8-week-old BALB / c nude mice.

[0074] Animal grouping and administration: Mice were acclimatized for one week before the experiment. Logarithmic growth phase A549 cells were digested with trypsin, resuspended in PBS, and counted. Each mouse was subcutaneously injected with 1 × 10⁻⁶ cells into the right axilla. 6 A549 cells were seeded at a volume of 100 μL. Drug intervention began on day 5 post-seeding. Mice were randomly assigned to five groups: Veh group (vector control, DMSO), ZD93_1 group (ZD93, 10 mg / kg), ZD93_2 group (ZD93, 20 mg / kg), PTX group (paclitaxel, 5 mg / kg), and PTX + ZD93 group (paclitaxel 5 mg / kg + ZD93 10 mg / kg), with six or eight mice in each group. All drugs were administered intraperitoneally every other day. The observation period lasted up to day 30.

[0075] (1) Effects of drug administration on mouse body weight and tumor size Starting from the onset of drug treatment, mouse body weight and tumor volume were measured every 2–3 days, and the results were as follows: Figure 12 As shown.

[0076] from Figure 12 As can be seen, during continuous administration, the overall body weight of mice in each group remained stable without a significant decreasing trend, indicating that neither ZD93 monotherapy nor the combination of ZD93 and paclitaxel (PTX) treatment caused significant systemic toxicity. Compared with the PTX group, the ZD93 group showed more stable body weight, suggesting good tolerability. Figure 12 As shown in b, compared with the Veh control group, ZD93 treatment did not significantly inhibit tumor growth. The PTX group showed some tumor-suppressing effect, while the PTX + ZD93 combination group showed the most significant tumor-suppressing effect, suggesting that ZD93 cannot inhibit tumor growth.

[0077] (2) Effects of drug administration on organ metastasis in mice After the experiment, the mice in each group were dissected, and major organs such as the lungs, liver, and spleen were removed. The presence of distant metastases was observed visually and statistically analyzed. The organ status of each mouse was marked with red (with metastasis) or gray (without metastasis).

[0078] from Figure 13 As can be seen, distant metastasis cases were detected in the Veh group, ZD93_1 group and PTX group, while no metastasis was found in the ZD93_2 group and PTX + ZD93 group, indicating that ZD93 can effectively inhibit tumor metastasis at a certain dose.

[0079] (3) Effect of drug administration on mouse survival time After drug administration, survival was observed, and natural survival time was monitored until the experimental endpoint. Statistical analysis was performed using Kaplan–Meier survival curves, and differences between groups were assessed using the log-rank test.

[0080] from Figure 14 As can be seen, compared with the control group, both the ZD93_1 group and the PTX + ZD93 group can prolong the survival time of mice, and the PTX + ZD93 group has the longest survival time, indicating that ZD93 can improve the prognosis of lung cancer.

[0081] (4) Effects of drug administration on mouse organs Orbital blood was collected from mice after the last administration of the drug, and serum was obtained by centrifugation. Routine biochemical indicators such as ALT, AST, BUN, CREA, GLU, and ALB were measured to evaluate liver and kidney toxicity and metabolic effects.

[0082] from Figure 15As can be seen, there were no significant increases in ALT and AST levels in mice in both the Veh and ZD93_1 groups, indicating that ZD93 did not cause liver function damage. Total protein (TP) levels were normal in both groups. CREA and BUN levels were also not elevated in mice in both groups, indicating negligible nephrotoxicity. Overall, this indicates that ZD93 did not cause significant liver and kidney toxicity at effective doses, demonstrating good systemic safety.

[0083] Mouse heart, liver, spleen, lungs, kidneys, and other major organs were collected, fixed in 4% paraformaldehyde, routinely embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E). Images were captured using an Olympus CX43 microscope and CellSens Entry software (see [link to image]). Figure 16 ).

[0084] HE staining results showed that the major organs of mice in the ZD93-1 treatment group, such as the heart, liver, spleen, lungs, and kidneys, had intact structures without obvious inflammation, necrosis, or structural disorder, which was comparable to that of the Veh group.

[0085] Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Those skilled in the art, under the guidance of the present invention, can make various similar representations without departing from the spirit and claims of the present invention, and such modifications all fall within the protection scope of the present invention.

Claims

1. The use of an arylpiperazine derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for inhibiting the migration or metastasis of lung cancer cells, wherein the arylpiperazine derivative is specifically: 、 、 、 、 。 2. Use according to claim 1, characterized in that, The relevant antigens or cell surface receptors of the lung cancer cells are selected from one of MSG783, STEAP2, TrpM4, CRIPTO, CD21, HER2, EphB2R, and BAFF-R.

3. The application according to claim 1, characterized in that, The drug uses an arylpiperazine derivative or a pharmaceutically acceptable salt thereof as its active ingredient and is formulated into a pharmaceutically acceptable dosage form with pharmaceutically acceptable excipients.

4. A pharmaceutical composition for targeted cancer therapy, comprising the arylpiperazine derivative of claim 1 or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients.

5. The pharmaceutical composition according to claim 4, characterized in that, The excipients include one or more of the following: solvents, disintegrants, flavoring agents, preservatives, colorants, binders, lubricants, diluents, and drug carriers.

Images