A methylpiperazine-chalcone derivative and its preparation method, application and pharmaceutical preparation

Abstract

The present invention belongs to the technical field of chalcone derivatives, and provides a methylpiperazine-chalcone derivative and its preparation method and application and pharmaceutical preparation. The preparation method of the methylpiperazine-chalcone derivative of the present invention comprises the following steps: vanillyl acetone, 3-chloro-3-methyl-1-butyne, hydrated copper chloride, anhydrous acetonitrile and 1,8-diazabicyclo[5.4.0]undec-7-ene are mixed and then subjected to condensation reaction to obtain a condensation compound; the condensation compound and pyridine are subjected to cyclization reaction to obtain an acetophenone intermediate; the acetophenone intermediate, 4-(4-methylpiperazine)benzaldehyde, an alcohol solvent and an alkali solution are mixed and then subjected to Claisen-Schmidt reaction to obtain a methylpiperazine-chalcone derivative.

Description

Technical Field

[0001] The invention relates to the technical field of chalcone derivatives, in particular to a methylpiperazine-chalcone derivative, a preparation method and application thereof, and a pharmaceutical preparation. Background Art

[0002] In view of the high incidence and mortality of cancer and the trend of younger patients, the research and development of new anti-tumor drugs has received widespread attention and is an important task in the current modern medical field. However, due to many problems such as tumor heterogeneity, drug resistance, immune escape, and drug side effects, the research and development of new anti-tumor drugs faces multiple challenges. Although traditional chemotherapy drugs are effective, they often have large toxic side effects on normal cells, and tumor cells develop drug resistance to them, reducing their efficacy. Although targeted therapy and immunotherapy can reduce damage to normal cells and improve efficacy to a certain extent, the therapeutic window is narrow and expensive. Therefore, the development of broad-spectrum, high-efficiency, low-toxic, and low-cost anti-tumor drugs is imminent.

[0003] In recent years, chalcone compounds have attracted much attention due to their wide range of biological activities, especially in the field of anti-tumor. Chalcone is a class of natural products with multiple pharmacological effects, which can affect the growth and survival of tumor cells through various mechanisms, such as inducing cell apoptosis, inhibiting tumor angiogenesis and regulating the cell cycle. Although chalcone and its derivatives have the advantages of multiple anti-tumor mechanisms, low price and high biocompatibility as anti-tumor drugs, due to their relatively simple structure, although they have anti-tumor activity, the activity is general and they are prone to off-target phenomena, so they have potential high toxic side effects and may also affect the patient's tolerance.

[0004] Therefore, it is of great significance to study and obtain a new methylpiperazine-chalcone derivative with enhanced anti-tumor activity and selectivity and reduced toxic side effects. Summary of the invention

[0005] The purpose of the present invention is to provide a methylpiperazine-chalcone derivative and a preparation method and application thereof as well as a pharmaceutical preparation in order to overcome the deficiencies of the prior art.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a methylpiperazine-chalcone derivative, the structural formula of the methylpiperazine-chalcone derivative is:

[0008] .

[0009] The present invention also provides a method for preparing the methylpiperazine-chalcone derivatives, comprising the following steps:

[0010] 1) vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, hydrated copper chloride, anhydrous acetonitrile and 1,8-diazabicyclo[5.4.0]undec-7-ene are mixed and subjected to a condensation reaction to obtain a condensation compound;

[0011] 2) subjecting the condensed compound to a cyclization reaction with pyridine to obtain an acetophenone intermediate;

[0012] 3) The acetophenone intermediate, 4-(4-methylpiperazine)benzaldehyde, alcohol solvent and alkali solution are mixed and subjected to Claisen-Schmidt reaction to obtain methylpiperazine-chalcone derivatives.

[0013] Preferably, in step 1), the molar ratio of vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, cupric chloride hydrate and 1,8-diazabicyclo[5.4.0]undec-7-ene is 17.85-18.25:26.88-27.28:0.05-0.06:26.88-27.28.

[0014] Preferably, the temperature of the condensation reaction in step 1) is 15-35° C., and the time of the condensation reaction is 7-9 hours.

[0015] Preferably, the temperature of the cyclization reaction in step 2) is 110-130° C., and the time of the cyclization reaction is 11-13 h.

[0016] Preferably, in step 3), the molar ratio of the acetophenone intermediate to 4-(4-methylpiperazine)benzaldehyde is 0.84-0.88:1.68-1.76; the alkali solution is a potassium hydroxide solution, and the mass concentration of the potassium hydroxide solution is 37-43%; and the alcohol solvent is anhydrous ethanol.

[0017] Preferably, the temperature of the Claisen-Schmidt reaction in step 3) is 15-35° C. and the time is 7-9 h.

[0018] The invention also provides the use of the methylpiperazine-chalcone derivatives in the preparation of anti-tumor drugs.

[0019] The present invention also provides a pharmaceutical preparation, which comprises an active ingredient and excipients, wherein the active ingredient comprises the methylpiperazine-chalcone derivative; the pharmaceutical preparation is an injection, tablet, capsule, aerosol, suppository, film, pill, ointment, controlled release agent, sustained release agent or nano preparation.

[0020] The beneficial effects of the present invention include the following:

[0021] 1) The methylpiperazine-chalcone derivatives of the present invention have a broad spectrum of anti-tumor activity, including but not limited to prostate cancer, colorectal cancer, ovarian cancer, lung cancer, liver cancer, etc.; and the anti-tumor activity is better than the current clinical first-line chemotherapy drug cisplatin, but the toxic side effects on normal cells are less than cisplatin.

[0022] 2) The methylpiperazine-chalcone derivatives of the present invention also have high anti-tumor activity and high biosafety in vivo.

[0023] 3) The methylpiperazine-chalcone derivatives of the present invention show good anti-tumor effects in both in vitro and in vivo models and have high biosafety. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a synthetic route for the methylpiperazine-chalcone derivatives of the present invention;

[0025] Figure 2 The methylpiperazine-chalcone derivative prepared in Example 1 1 H NMR nuclear magnetic resonance spectrum;

[0026] Figure 3 The methylpiperazine-chalcone derivative prepared in Example 1 13 C NMR nuclear magnetic resonance spectrum;

[0027] Figure 4 The in vivo antitumor activity of the methylpiperazine-chalcone derivatives prepared in Example 1;

[0028] Figure 5 The biosafety data of the methylpiperazine-chalcone derivatives prepared in Example 1, wherein a is the concentration of alanine aminotransferase, b is the concentration of aspartate aminotransferase, c is the concentration of creatinine, ns is the methylpiperazine-chalcone derivative, and * is cisplatin. DETAILED DESCRIPTION

[0029] The present invention provides a methylpiperazine-chalcone derivative, the structural formula of the methylpiperazine-chalcone derivative is:

[0030] .

[0031] The present invention also provides a method for preparing the methylpiperazine-chalcone derivatives, comprising the following steps:

[0032] 1) vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, hydrated copper chloride, anhydrous acetonitrile and 1,8-diazabicyclo[5.4.0]undec-7-ene are mixed and subjected to a condensation reaction to obtain a condensation compound;

[0033] 2) subjecting the condensed compound to a cyclization reaction with pyridine to obtain an acetophenone intermediate;

[0034] 3) The acetophenone intermediate, 4-(4-methylpiperazine)benzaldehyde, alcohol solvent and alkali solution are mixed and subjected to Claisen-Schmidt reaction to obtain methylpiperazine-chalcone derivatives.

[0035] In the present invention, the synthetic route of methylpiperazine-chalcone derivatives is as follows Figure 1 shown.

[0036] In the present invention, the molar ratio of the vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, cupric chloride hydrate and 1,8-diazabicyclo[5.4.0]undec-7-ene in step 1) is preferably 17.85-18.25:26.88-27.28:0.05-0.06:26.88-27.28, further preferably 17.95-18.15:26.98-27.18:0.053-0.057:26.98-27.18, and more preferably 18.05:27.08:0.05416-0.055:27.08.

[0037] In the present invention, the temperature of the condensation reaction in step 1) is preferably 15-35°C, more preferably 20-30°C, and more preferably 25°C; the time of the condensation reaction is preferably 7-9h, more preferably 7.5-8.5h, and more preferably 8h.

[0038] In the present invention, the condensation compound is 1-(3-methoxy-4-((2-methylbut-3-en-2-yl)oxy)phenyl)ethan-1-one.

[0039] In the present invention, the mixing in step 1) is preferably performed by adding vanillyl acetonitrile, 3-chloro-3-methyl-1-butyne and hydrated copper chloride to anhydrous acetonitrile, and then adding 1,8-diazabicyclo[5.4.0]undec-7-ene, and stirring during the condensation reaction; after the condensation reaction is completed, the solvent anhydrous acetonitrile is removed by vacuum distillation, and the obtained condensation compound can be directly used in the next cyclization reaction without further treatment.

[0040] In the present invention, the temperature of the cyclization reaction in step 2) is preferably 110-130°C, more preferably 115-125°C, and more preferably 120°C; the time of the cyclization reaction is preferably 11-13h, more preferably 11.5-12.5h, and more preferably 12h.

[0041] In the present invention, the cyclization reaction in step 2) is preferably: adding solvent pyridine to the condensation compound, and carrying out the cyclization reaction under stirring and reflux conditions; after the cyclization reaction is completed, the obtained product is sequentially subjected to quenching reaction, vacuum distillation, and extraction, and the extracted organic layer is sequentially dried, vacuum distilled again, and purified by silica gel column chromatography to obtain an acetophenone intermediate; the reagent for quenching the reaction is preferably water, the purpose of vacuum distillation is to remove the solvent pyridine, the residue obtained by vacuum distillation is dissolved in dichloromethane and distilled water for extraction, the number of extractions is preferably 2 to 4 times, more preferably 3 times, the reagent used for drying is preferably anhydrous magnesium sulfate, the purpose of vacuum distillation again is to remove dichloromethane, and the residue obtained after removing dichloromethane is purified by silica gel column chromatography, the reagents used for silica gel column chromatography are preferably petroleum ether and ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is preferably 19 to 21:1, more preferably 20:1.

[0042] The present invention has no particular limitation on the specific amounts of reagents used for quenching reaction, extraction and silica gel column chromatography purification after the reaction is completed, and can be adjusted according to actual needs.

[0043] In the present invention, the acetophenone intermediate is 6-acetyl-8-methoxy-2,2-dimethyl-2H-1-benzopyran.

[0044] In the present invention, the molar ratio of the acetophenone intermediate and 4-(4-methylpiperazine)benzaldehyde in step 3) is preferably 0.84-0.88:1.68-1.76, more preferably 0.85-0.87:1.7-1.74, and more preferably 0.861-0.865:1.72-1.73; the alkali solution is preferably a potassium hydroxide solution, and the mass concentration of the potassium hydroxide solution is preferably 37-43%, more preferably 39-41%, and more preferably 40%; the alcohol solvent is preferably anhydrous ethanol.

[0045] The present invention has no particular limitation on the amount of the alkali solution and the alcohol solvent, and the amount can be adjusted according to actual needs to ensure that the reaction proceeds smoothly.

[0046] In the present invention, the temperature of the Claisen-Schmidt reaction in step 3) is preferably 15-35°C, more preferably 20-30°C, and more preferably 25°C; the time is preferably 7-9h, more preferably 7.5-8.5h, and more preferably 8h.

[0047] In the present invention, the mixing in step 4) is preferably: adding the acetophenone intermediate and 4-(4-methylpiperazine)benzaldehyde to anhydrous ethanol, and then adding alkali solution; stirring during the Claisen-Schmidt reaction, after the Claisen-Schmidt reaction is completed, the reaction product is sequentially quenched, vacuum distilled, and extracted, and the organic layer obtained by extraction is sequentially dried, vacuum distilled again, and recrystallized to obtain methylpiperazine-chalcone derivatives; the reagent used for quenching is preferably a saturated ammonium chloride solution; the purpose of vacuum distillation is to remove the solvent anhydrous ethanol, and the residue of vacuum distillation is dissolved in dichloromethane and distilled water for extraction, the number of extractions is preferably 2 to 4 times, and more preferably 3 times, the reagent used for drying is preferably anhydrous magnesium sulfate, the purpose of vacuum distillation again is to remove dichloromethane, and a small amount of methanol is added to the residue of vacuum distillation again for recrystallization.

[0048] The invention also provides the use of the methylpiperazine-chalcone derivatives in the preparation of anti-tumor drugs.

[0049] In the present invention, the tumor preferably includes prostate cancer, ovarian cancer, lung cancer, liver cancer and bile duct cancer.

[0050] The present invention also provides a pharmaceutical preparation, which comprises an active ingredient and excipients, wherein the active ingredient comprises the methylpiperazine-chalcone derivative; the pharmaceutical preparation is preferably an injection, tablet, capsule, aerosol, suppository, film, pill, ointment, controlled release agent, sustained release agent or nano preparation.

[0051] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0052] Example 1

[0053] 18.05mmol of vanillyl ethyl ketone, 27.08mmol of 3-chloro-3-methyl-1-butyne and 54.16μmol of hydrated copper chloride were added to 12mL of anhydrous acetonitrile, and then 27.08mmol of 1,8-diazabicyclo[5.4.0]undec-7-ene was added. The mixture was stirred at 800rpm for 8h at 25°C for condensation reaction. After the condensation reaction, the solvent anhydrous acetonitrile was removed by distillation under reduced pressure to obtain a condensation compound: 1-(3-methoxy-4-((2-methylbut-3-en-2-yl)oxy)phenyl)ethane-1-one.

[0054] 15 mL of solvent pyridine was added to the condensation compound, and the mixture was stirred and refluxed at 120° C. and 800 rpm for 12 h to carry out a cyclization reaction. After the cyclization reaction was completed, water was added to the obtained product to quench the reaction, and the solvent pyridine was removed by vacuum distillation. The residue obtained by vacuum distillation was dissolved in a mixed solution of 30 mL of dichloromethane and 30 mL of distilled water and extracted three times. The organic layer obtained by extraction was dried over anhydrous magnesium sulfate overnight, and dichloromethane was removed by vacuum distillation. The obtained residue was purified by silica gel column chromatography. The silica gel column chromatography purification (rate of 5 drops / s) used a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 20:1. After purification, an acetophenone intermediate: 6-acetyl-8-methoxy-2,2-dimethyl-2H-1-benzopyran was obtained. The acetophenone intermediate was an orange-brown solid with a mass of 3.1 g and a yield of 73.99%.

[0055] 861.03 μmol of acetophenone intermediate and 1.72 mmol of 4-(4-methylpiperazine)benzaldehyde were added to 8 mL of anhydrous ethanol, and then a potassium hydroxide solution with a mass concentration of 40% was added dropwise, and the resulting reaction solution was stirred at 25°C for 8 h to perform a Claisen-Schmidt reaction; after the reaction was completed, a saturated ammonium chloride solution was added to quench, and the solvent anhydrous ethanol was removed by distillation under reduced pressure. The residue was dissolved in a mixed solution of 30 mL of dichloromethane and 30 mL of distilled water and extracted three times. The organic layer obtained by extraction was dried over anhydrous magnesium sulfate, and dichloromethane was removed by distillation under reduced pressure. 5 drops of methanol were added to the residue for recrystallization to obtain a methylpiperazine-chalcone derivative, which was a yellow solid with a mass of 120 mg and a yield of 44.4%.

[0056] The methylpiperazine-chalcone derivatives prepared in Example 1 were characterized by NMR. 1 H NMR nuclear magnetic resonance spectrum Figure 2 As shown, 13 C NMR nuclear magnetic resonance spectrum Figure 3 As shown by Figure 2 and Figure 3 The data obtained are:

[0057] 1H-NMR (400 MHz, Chloroform-d) δ 7.76 (d, J = 15.5 Hz, 1H), 7.56 (d,J = 8.6 Hz, 2H), 7.52 (d, J = 1.9 Hz, 1H), 7.41 – 7.33 (m, 2H), 6.90 (d, J =8.8 Hz, 2H), 6.38 (d, J = 9.9 Hz, 1H), 5.68 (d, J = 9.8 Hz, 1H), 3.94 (s,3H), 3.35 (t, J = 5.0 Hz, 4H), 2.60 (t, J = 5.0 Hz, 4H), 2.38 (s, 3H), 1.52(s, 6H). 13C-NMR (101 MHz, Chloroform-d) δ 188.72, 152.64, 148.55, 146.48,131.34, 131.12, 130.14, 125.69, 122.07, 121.02, 120.28, 118.15, 114.98,112.15, 56.45, 54.90, 47.86, 46.21, 28.33.

[0058] Figure 2 and Figure 3 It shows that the methylpiperazine-chalcone derivative prepared by the present invention is: (E)-1-(8-methoxy-2,2-dimethyl-2H-benzopyran-6-yl)-3-(4-(4-methylpiperazine-1-yl)phenyl)prop-2-en-1-one.

[0059] Example 2

[0060] 17.95mmol of vanillyl ethyl ketone, 26.98mmol of 3-chloro-3-methyl-1-butyne and 53μmol of hydrated copper chloride were added to 11mL of anhydrous acetonitrile, and then 26.98mmol of 1,8-diazabicyclo[5.4.0]undec-7-ene was added. The mixture was stirred at 800rpm for 7.5h at 20°C for condensation reaction. After the condensation reaction, the solvent anhydrous acetonitrile was removed by distillation under reduced pressure to obtain a condensation compound: 1-(3-methoxy-4-((2-methylbut-3-en-2-yl)oxy)phenyl)ethane-1-one.

[0061] 14 mL of solvent pyridine was added to the condensation compound, and the mixture was stirred and refluxed at 115° C. and 800 rpm for 13 h to carry out a cyclization reaction. After the cyclization reaction was completed, water was added to the obtained product to quench the reaction, and the solvent pyridine was removed by vacuum distillation. The residue obtained by vacuum distillation was dissolved in a mixed solution of 30 mL of dichloromethane and 30 mL of distilled water and extracted three times. The organic layer obtained by extraction was dried over anhydrous magnesium sulfate overnight, and dichloromethane was removed by vacuum distillation. The obtained residue was purified by silica gel column chromatography. The silica gel column chromatography purification (rate of 5 drops / s) used a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 19:1. After purification, an acetophenone intermediate: 6-acetyl-8-methoxy-2,2-dimethyl-2H-1-benzopyran was obtained. The acetophenone intermediate was an orange-brown solid with a yield of 78%.

[0062] 850 μmol of acetophenone intermediate and 1.7 mmol of 4-(4-methylpiperazine)benzaldehyde were added to 7.5 mL of anhydrous ethanol, and then a potassium hydroxide solution with a mass concentration of 39% was added dropwise, and the resulting reaction solution was stirred at 20°C for 8.5 h to perform a Claisen-Schmidt reaction; after the reaction was completed, a saturated ammonium chloride solution was added to quench, and the solvent anhydrous ethanol was removed by vacuum distillation, and the residue was dissolved in a mixed solution of 30 mL of dichloromethane and 30 mL of distilled water, and extracted three times, and the organic layer obtained by extraction was dried over anhydrous magnesium sulfate, and dichloromethane was removed by vacuum distillation, and 5 drops of methanol were added to the residue for recrystallization to obtain a methylpiperazine-chalcone derivative, which was a yellow solid with a yield of 44%.

[0063] Example 3

[0064] 18.15mmol of vanillyl ethyl ketone, 27.18mmol of 3-chloro-3-methyl-1-butyne and 55μmol of hydrated copper chloride were added to 13mL of anhydrous acetonitrile, and then 27.18mmol of 1,8-diazabicyclo[5.4.0]undec-7-ene was added. The mixture was stirred at 800rpm for 8.5h at 30°C for condensation reaction. After the condensation reaction, the solvent anhydrous acetonitrile was removed by distillation under reduced pressure to obtain a condensation compound: 1-(3-methoxy-4-((2-methylbut-3-en-2-yl)oxy)phenyl)ethane-1-one.

[0065] 16 mL of solvent pyridine was added to the condensation compound, and the mixture was stirred and refluxed at 125° C. and 800 rpm for 11 h to carry out a cyclization reaction. After the cyclization reaction was completed, water was added to the obtained product to quench the reaction, and the solvent pyridine was removed by vacuum distillation. The residue obtained by vacuum distillation was dissolved in a mixed solution of 32 mL of dichloromethane and 32 mL of distilled water and extracted three times. The organic layer obtained by extraction was dried over anhydrous magnesium sulfate overnight, and dichloromethane was removed by vacuum distillation. The obtained residue was purified by silica gel column chromatography. The silica gel column chromatography purification (rate of 5 drops / s) used a mixed solution of petroleum ether and ethyl acetate with a volume ratio of 21:1. After purification, an acetophenone intermediate: 6-acetyl-8-methoxy-2,2-dimethyl-2H-1-benzopyran was obtained. The acetophenone intermediate was an orange-brown solid with a yield of 85%.

[0066] 870 μmol of acetophenone intermediate and 1.74 mmol of 4-(4-methylpiperazine)benzaldehyde were added to 8.5 mL of anhydrous ethanol, and then a potassium hydroxide solution with a mass concentration of 41% was added dropwise, and the resulting reaction solution was stirred at 30°C for 7.5 h to perform a Claisen-Schmidt reaction; after the reaction was completed, a saturated ammonium chloride solution was added to quench, and the solvent anhydrous ethanol was removed by vacuum distillation, and the residue was dissolved in a mixed solution of 32 mL of dichloromethane and 32 mL of distilled water and extracted three times, and the organic layer obtained by extraction was dried over anhydrous magnesium sulfate, and dichloromethane was removed by vacuum distillation, and 5 drops of methanol were added to the residue for recrystallization to obtain a methylpiperazine-chalcone derivative, which was a yellow solid with a yield of 44%.

[0067] Application Example 1 Determination of the in vitro antitumor activity of methylpiperazine-chalcone derivatives by diphenyltetrazolium bromide (MTT) method

[0068] The cells in the logarithmic growth phase (prostate cancer cell DU145, colorectal cancer cell HCT116, ovarian cancer cell SKOV3, lung cancer cell A549, liver cancer cell Huh-7, gastric cancer cell SGC, human normal liver cell MIHA) were digested with 0.25% trypsin, centrifuged, and counted. Each cell was transferred to a 50 mL centrifuge tube and mixed evenly. The cells were plated into a 96-well plate at a cell density of 6000 cells / well using a spray gun, 160 μL / well. After the cell morphology is restored, 40 μL of culture medium containing corresponding concentrations of the methylpiperazine-chalcone derivatives of Example 1 (using the half-dilution method, the concentrations of the methylpiperazine-chalcone derivatives are 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.5625 μM, and 0.78125 μM in sequence) is added to each well, so that the final volume of each well is 200 μL. A blank control group (without compound) and a blank well (to eliminate background interference) are set up, and the 96-well plate with the drug added is returned to the incubator (37°C) for 48 hours. Then, add 20 μL of MTT working solution (5 mg / mL) to each well in the dark, continue to incubate in the incubator for 4 hours, use a vacuum pump to remove the supernatant to ensure that there is no liquid residue in each well, add DMSO (100 μL / well) with a spray gun, and keep it in the dark at room temperature and shake it horizontally for 10 minutes to fully dissolve the formazan. Finally, measure the absorbance value of each well at a wavelength of 490 nm on the microplate reader, mark the corresponding wells, and save the experimental results. Use the formula to calculate the absorbance of each group IC 50 value. IC 50 The calculation formula is as follows: IC 50 =X m -I[P-(3-P m -P n ) / 4], where X m =lg maximum dose (i.e., the logarithm of the maximum dose); I = lg (maximum dose / adjacent dose), i.e., the logarithm of the maximum dose divided by the adjacent dose; P is the sum of the positive reaction rates; P m is the maximum positive reaction rate; P n The minimum positive reaction rate.

[0069] The in vitro antitumor activity results of the methylpiperazine-chalcone derivatives and cisplatin prepared in Example 1 are shown in Table 1. The methylpiperazine-chalcone derivatives have good antitumor activity against prostate cancer cells DU145, colorectal cancer cells HCT116, ovarian cancer cells SKOV3, lung cancer cells A549, liver cancer cells Huh-7, and gastric cancer cells SGC. IC 50The values ​​were all less than 10 μM; however, the toxicity of MIHA to normal human liver cells was weak. IC 50 The value is greater than 50 μM; the experimental results show that the methylpiperazine-chalcone derivatives of the present invention have good selectivity for tumor cells. In addition, the methylpiperazine-chalcone derivatives have good selectivity for the above tumor cells. IC 50 The values ​​were lower than those of cisplatin, the most widely used first-line chemotherapy drug in clinical practice, indicating that its anti-tumor activity is better than cisplatin; especially its effect on MIHA of normal human hepatocytes IC 50 The value is 4 times higher than that of cisplatin, indicating that it causes less damage to normal cells than cisplatin.

[0070] Table 1 In vitro antitumor activity of derivatives and cisplatin determined by MTT assay

[0071]

[0072] Application Example 2 Evaluation of the in vivo antitumor activity of methylpiperazine-chalcone derivatives

[0073] The in vivo antitumor activity of methylpiperazine-chalcone derivatives was evaluated in a nude mouse xenograft tumor model. Six-week-old BALB / c nude mice were subcutaneously inoculated with DU145 cells suspended in PBS. 3 The tumor-bearing mice were randomly divided into 2 groups, 5 mice in each group. Subsequently, one group was injected with 5 mg / kg of the methylpiperazine-chalcone derivative of Example 1 through the tail vein, and the other group was injected with normal saline through the tail vein (as the control group), once every two days for a total of 14 days. The mice were euthanized, and the tumor tissues were taken to observe the changes in tumor size.

[0074] The results of the in vivo antitumor activity test of the methylpiperazine-chalcone derivatives and normal saline in Example 1 are as follows: Figure 4 As shown by Figure 4 It can be seen that compared with the control group, the administration of methylpiperazine-chalcone derivatives can significantly inhibit the growth of DU145 tumors, indicating that methylpiperazine-chalcone derivatives have excellent in vivo anti-tumor activity.

[0075] Application Example 3 Biosafety Assessment of Methylpiperazine-Chalcone Derivatives

[0076] In view of the serious toxic side effects of current chemotherapy drugs on the liver and kidneys, the effects of the methylpiperazine-chalcone derivatives of Example 1 on the liver and kidneys were further evaluated. After the sudden death of the mice in Application Example 2, blood was collected and centrifuged at 3000 rpm for 5 min. The supernatant was collected and the concentrations of ALT, AST and creatinine in the serum were measured using alanine aminotransferase (ALT / GPT), aspartate aminotransferase (AST / GOT) and creatinine (Cr) assay kits, respectively.

[0077] The biosafety data of methylpiperazine-chalcone derivatives and cisplatin prepared in Example 1 are as follows: Figure 5 As shown, wherein, a is the concentration of alanine aminotransferase, b is the concentration of aspartate aminotransferase, c is the concentration of creatinine, ns is a methylpiperazine-chalcone derivative, * is cisplatin, and Figure 5 It can be seen that compared with the control group, cisplatin treatment significantly affected the levels of ALT / AST and Cr in serum, indicating that it has huge toxic side effects on the liver and kidneys; however, methylpiperazine-chalcone derivatives treatment did not change the levels of ALT / AST and Cr in serum, indicating that it has basically no toxic side effects on the liver and kidneys and has high biosafety.

[0078] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A methylpiperazine-chalcone derivative, characterized in that: The structural formula of the methylpiperazine-chalcone derivative is: 。 2. The method for preparing the methylpiperazine-chalcone derivatives according to claim 1, characterized in that: The following steps are included: 1) vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, hydrated copper chloride, anhydrous acetonitrile and 1,8-diazabicyclo[5.4.0]undec-7-ene are mixed and subjected to a condensation reaction to obtain a condensation compound; 2) subjecting the condensed compound to a cyclization reaction with pyridine to obtain an acetophenone intermediate; 3) The acetophenone intermediate, 4-(4-methylpiperazine)benzaldehyde, alcohol solvent and alkali solution are mixed and subjected to Claisen-Schmidt reaction to obtain methylpiperazine-chalcone derivatives.

3. The method for preparing the methylpiperazine-chalcone derivatives according to claim 2, characterized in that: In step 1), the molar ratio of vanillyl ethyl ketone, 3-chloro-3-methyl-1-butyne, cupric chloride hydrate and 1,8-diazabicyclo[5.4.0]undec-7-ene is 17.85-18.25: 26.88-27.28: 0.05-0.06: 26.88-27.

28.

4. The method for preparing the methylpiperazine-chalcone derivatives according to claim 2 or 3, characterized in that: Step 1) The condensation reaction temperature is 15-35°C, and the condensation reaction time is 7-9h.

5. The method for preparing methylpiperazine-chalcone derivatives according to claim 4, characterized in that: Step 2) The temperature of the cyclization reaction is 110-130° C., and the time of the cyclization reaction is 11-13 hours.

6. The method for preparing methylpiperazine-chalcone derivatives according to claim 5, characterized in that: Step 3) The molar ratio of the acetophenone intermediate to 4-(4-methylpiperazine)benzaldehyde is 0.84-0.88:1.68-1.76; the alkali solution is a potassium hydroxide solution, and the mass concentration of the potassium hydroxide solution is 37-43%; the alcohol solvent is anhydrous ethanol.

7. The method for preparing the methylpiperazine-chalcone derivatives according to claim 5 or 6, characterized in that: Step 3) The temperature of the Claisen-Schmidt reaction is 15-35° C. and the time is 7-9 h.

8. Use of the methylpiperazine-chalcone derivatives according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor cells are prostate cancer cell DU145, colorectal cancer cell HCT116, ovarian cancer cell SKOV3, lung cancer cell A549, liver cancer cell Huh-7, and gastric cancer cell SGC.

9. A pharmaceutical preparation, characterized in that The pharmaceutical preparation comprises an active ingredient and excipients, wherein the active ingredient comprises the methylpiperazine-chalcone derivative according to claim 1; the pharmaceutical preparation is an injection, tablet, capsule, aerosol, suppository, film, pill, ointment, controlled release agent, sustained release agent or nano preparation.

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