Piperazine-substituted 3, 4-dihydro-1 (2H)-naphthalenone derivative, and preparation method and application thereof

Abstract

The invention discloses a piperazine-substituted 3, 4-dihydro-1 (2H)-naphthalenone derivative as well as a preparation method and an application of the piperazine-substituted 3, 4-dihydro-1 (2H)-naphthalenone derivative. The piperazine substituted 3, 4-dihydro-1 (2H)-naphthalenone derivative disclosed by the invention has a good tumor treatment effect. # imgabs0 #

Description

Technical Field

[0001] This application relates to the technical field of medicinal chemistry, and particularly to a piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, a preparation method thereof, and uses thereof. Background Art

[0002] In tumor cells, NF-κB is often abnormally activated, thereby regulating the expression of a series of genes related to tumor cell proliferation, survival, invasion, and metastasis. The continuous activation of the MAPK pathway is closely related to tumor progression, and promotes the proliferation and metastasis of tumor cells by affecting the cell cycle process, cell survival signals, and the expression of molecules related to cell migration. Nitrogen-containing heterocyclic compounds have shown great practicality in drug discovery and improvement due to their special chemical structures and various organic activities. They have become an important way to improve new anti-cancer drugs, and their pharmacological effects include antifungal, antimalarial, antimigraine, antidiabetic, antiproliferative, antihypertensive, anti-inflammatory, anti-obesity, anti-microbial, cardiovascular effects, and calcium antagonists. The prior art has regarded 3,4-dihydronaphthalen-1(2H)-one derivatives as potential drugs for new broad-spectrum antibacterial agents, antifungal agents, antimalarials, photosensitizers, improving enzyme inhibition characteristics, and treating cancer. Summary of the Invention

[0003] Object of the Invention: This application provides a piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, a preparation method thereof, and uses thereof. The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative of this application can be used to treat tumors.

[0004] Technical Solution: This application provides a piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, and its structural formula is as follows:

[0005]

[0006] Among them, R1 is N-methylpiperazine or N-ethylpiperazine, R2 is phenyl or pyridyl, and the phenyl or pyridyl is substituted by one or more R a groups, and the R a is selected from one or more of single or multiple fluoro groups, nitro groups, methyl groups, trifluoromethyl groups.

[0007] In some embodiments, the structural formula of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative is as follows:

[0008]

[0009] Among them, R1 is N-methylpiperazine or N-ethylpiperazine, and R a is selected from one or more of single or multiple fluoro groups, nitro groups, methyl groups, trifluoromethyl groups.

[0010] In some embodiments, Ra is selected from the following groups:

[0011] 2-F, 3-F, 4-F, 2-CF3, 3-CF3, 4-CF3, 3,5-CF3, 2,3-F, 2,4-F, 2,5-F, 2,6-F, 3,4-F, 3,5-F, 2,3,4-F, 2,3,5-F, 2,4,6-F, 2,4,5-F, 2-NO2, 3-NO2, 4-NO2.

[0012] The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives described in this application include the hydrochloride salts of piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives.

[0013] This application further provides a method for preparing piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives, comprising the following steps:

[0014] Add the first reactant in a solvent and the second reactant R2-CHO, and react in the presence of a catalyst to obtain a product. The reaction formula is:

[0015]

[0016] In some specific embodiments, the synthetic route of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives of this application is as follows:

[0017]

[0018] In some embodiments, the product is purified by washing with acetone to obtain the purified product.

[0019] In some embodiments, the solvent includes methanol.

[0020] In some embodiments, the catalyst is hydrochloric acid.

[0021] In some embodiments, the reaction temperature is 0°C ± 5°C.

[0022] In this application, the so-called room temperature can be 20°C ± 5°C.

[0023] In some embodiments, the molar ratio of the first reactant to the second reactant is (0.8 to 1.2):(1.6 to 2.4).

[0024] This application further provides a pharmaceutical composition, comprising the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives described above or the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives prepared by the above preparation method.

[0025] The present invention further provides the use of the 3,4-dihydro-1(2H)-naphthalenone derivative described above, or the 3,4-dihydro-1(2H)-naphthalenone derivative prepared by the preparation method described above, or the pharmaceutical composition in the preparation of a medicament for treating tumors.

[0026] In some embodiments, the tumors of the present application include liver cancer and / or primary liver cancer.

[0027] Term Explanation

[0028] Unless otherwise defined, all technical and scientific terms used in the present application have the same meaning as commonly understood by those skilled in the art. Unless otherwise specified, all patent documents and publicly disclosed materials referred to in the present application are incorporated herein by reference in their entirety. If there are multiple definitions for the same term in the present application, the definition in this section shall prevail.

[0029] "Substituted" means that a hydrogen atom is replaced by a substituent.

[0030] As used herein, the term "nitrogen-containing heterocycle" refers to N-methylpiperazine and N-ethylpiperazine.

[0031] As used herein, the term "pyridyl" is pyridin-3-yl.

[0032] Unless otherwise specified, "%" in the present application refers to mass percentage.

[0033] Advantageous Effects: The present application provides a piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, a preparation method and uses thereof. The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative of the present application exhibits good anti-tumor activity and can be used for treating tumors. Description of the Drawings

[0034] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the drawings in the following description are only some embodiments of the present application. For those skilled in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0035] Figure 1This is the test result of the apoptosis of HepG2 cells induced by the compound DHN 28 of this application. Among them, Figure A is the detection result by flow cytometry after incubation with DHN 28. From left to right in Figure A are the results after incubation with the DMSO group, the results after incubation with the compound DHN 28 at a concentration of 0.46 μM, the results after incubation with the compound DHN 28 at a concentration of 0.92 μM, and the results after incubation with the compound DHN 28 at a concentration of 1.84 μM. Figure B is the result of the apoptosis distribution percentage of the compound DHN 28 on HepG2 cells, *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001;

[0036] Figure 2 This is the detection result by flow cytometry after the compound DHN 28 of this application is incubated with the cells HepG2. Among them, from left to right are the results after incubation with the DMSO group, the results after incubation with the compound DHN 28 at a concentration of 0.46 μM, the results after incubation with the compound DHN 28 at a concentration of 0.92 μM, and the results after incubation with the compound DHN 28 at a concentration of 1.84 μM;

[0037] Figure 3 This is the real-time fluorescence PCR test result of the relative expression levels of apoptosis-related genes BAX, BCL-2, and C-caspase-3 induced by the compound DHN 28 of this application. Among them, the left figure is the relative expression level of the BAX gene after incubation with DHN 28, the middle figure is the relative expression level of the BCL-2 gene after incubation with DHN 28, and the right figure is the relative expression level of the C-caspase-3 gene after incubation with DHN 28, *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001;

[0038] Figure 4 This is the effect of the compound DHN 28 of this application on the nuclear translocation of p65 protein in TNF-α-induced HepG2 cells;

[0039] Figure 5 This is the scratch test result of HepG2 cells treated with the compound DHN 28 of this application;

[0040] Figure 6 This is the result of the effect of the compound DHN 28 of this application on the migration ability of HepG2 cells. Among them, Figure A is the crystal violet staining result, and Figure B is the absorbance result measured after decolorization with a 33% acetic acid solution;

[0041] Figure 7This is the result of the inhibition of the activation and expression of NF-κB and MAPK-related proteins by the compound DHN 28 of the present application. Among them, Figure A shows the effect of the compound DHN 28 on the expression levels of NF-κB and MAPK-related proteins, and Figures B-E show the expression results of P-p65 and p65, P-IκBα and IκBα, ERK and P-ERK, and P-p38 and p38. Compared with the control group: *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001. Detailed implementation manners

[0042] The following will clearly and completely describe the technical solutions in the embodiments of the present application with reference to the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present application. It should be understood that the foregoing general description and the following detailed description are merely exemplary and explanatory, and have no limitation on any claim. It should be noted that in the specification and the appended claims, unless otherwise stated in the text, the singular forms such as "a", "an", and "this" include plural referents. It should also be noted that unless otherwise stated, "or" represents "and / or". In addition, similar terms such as "comprising" and "including" are not restrictive, and the term "including" means "including but not limited to". The terms first, second, third, etc. are only used as labels and do not impose numerical requirements or establish an order. The various embodiments of the present application may exist in a range form; it should be understood that the description in a range form is only for convenience and brevity, and should not be construed as a rigid limitation on the scope of the present application; therefore, it should be considered that the described range description has specifically disclosed all possible sub-ranges and single values within that range. For example, it should be considered that the range description from 1 to 6 has specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers within the counted range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. Additionally, whenever a numerical range is indicated herein, it means including any cited number (fraction or integer) within the indicated range.

[0043] I. Test raw materials and test methods

[0044] 1.1 Chemical reagents were purchased from Leyan Technology Co., Ltd. (Beijing) and Bide Pharmaceutical Technology Co., Ltd. (Shanghai).

[0045] The first reactant in the present application can be obtained through the prior art or synthesized in the laboratory. Taking 6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one as an example, the synthesis method is as follows:

[0046]

[0047] Weigh N-methylpiperazine (16.00 g, 0.16 mol) and K2CO3 (27.64 g, 0.20 mol) and place them in a 100 mL three-necked flask. Add 10 mL of DMF and stir at 40 °C for 12 hours. 6-Fluoro-3,4-dihydro-2H-naphthalen-1-one (3.28 g, 0.02 mol) is added dropwise to the reaction system, and the temperature rises to 110 °C for reaction. After refluxing for 8 hours, the reaction ends. The reaction system is filtered, and the filtrate is washed with distilled water and saturated brine. The combined organic extracts are dried (Na2SO4), and the solvent is distilled off under reduced pressure. The residue is eluted with dichloromethane:methanol (20:1, v / v) by column chromatography to obtain the dark brown target product 6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one with a yield of 87.2%. In this application, the reactants used for synthesizing compounds 1-35 can be obtained by the same synthesis method as that of reactant 1, except that the reaction raw materials are replaced with the reaction raw materials corresponding to the groups of this compound.

[0048] 1.2 Test methods

[0049] (1) In dimethyl sulfoxide (DMSO-d6) solvent with tetramethylsilane (TMS) as the internal standard, use 1 1H NMR, Bruker Avance 400 MHz or 600 MHz spectrometer and 13 13C-NMR, Bruker Avance 100 MHz or 150 MHz spectrometer to record spectra. Chemical shift (δ) is in ppm, and coupling constant (J) is in Hz. All melting factors are measured on a digital melting factor meter.

[0050] (2) Anti-hepatocellular carcinoma activity experiment: Mouse HepG2 cells are provided by Procell (China Wuhan, China) and cultured in a complete medium of MEM medium + 10% fetal bovine serum and 1% streptomycin / penicillin antibiotics. Take cells in the logarithmic growth phase for the following tests. According to the manufacturer's instructions, use CCK-8 to detect anti-hepatocellular carcinoma activity. Cells are grown and treated in a 96-well plate and incubated with CCK-8 reagent at 37 °C for 1 hour. Absorbance is measured at 450 nm.

[0051] (3) Western blotting experiment: HepG2 cells (2.0×10 per well in a 6-well plate) 6Cells were exposed to the drug for 2 hours and then to TNF-α (10 ng / mL) for 24 hours. Lysis was performed using a potent RIPA lysis buffer containing phosphatase inhibitors (RIPA lysis buffer: phosphatase inhibitor = 99:1). After sonication lysis was completed on ice and then centrifuged. After determining the protein concentration, the protein solution was boiled. The protein sample was added to a 10% polyacrylamide gel for electrophoresis. The protein was transferred to a PVDF membrane and incubated with 5% non-fat milk for 2 hours. Subsequently, it was incubated overnight at 4 °C with antibodies against IκB-α, P-IκB-α, p65, P-p65, p38, P-p38, ERK, P-ERK, GAPDH (antibodies purchased from Cell Signaling Technoligy, USA). It was incubated with goat anti-rabbit IgG / HRP antibody (Beijing Solarbio Science & Technology Co., Ltd.) for 1.0 h at room temperature. Detection was performed after using an ECL developing solution (Shanghai Noren Biomedical Technology Co., Ltd.). Protein intensity was detected using Image Lab software and analyzed using ImageJ software.

[0052] (5) Cell apoptosis assay: HepG2 cells were seeded in 12-well plates at a density of 2×10 5 cells / well. Then they were treated with compound concentrations of 0, 0.5 μM, 1 μM, and 2 μM for 24 hours. The cells were collected in centrifuge tubes and washed twice with pre-cooled PBS. The PBS was discarded after centrifugation. The cells were resuspended in 200 μL of binding buffer. Then 5 μL of Annexin V-FITC and 5 μL of propidium iodide (Wuhan Aidy Anti-Bio Technology Co., Ltd.) were added to the solution in sequence. Cell apoptosis was detected using a flow cytometer (BD FACS Calibur). Finally, the cells were analyzed using a flow cytometer.

[0053] (6) Cell cycle assay: First, HepG2 cells were collected, and a control group and experimental groups were set up. They were seeded in 6-well plates at a density of 3×10 5 cells / mL, and the medium containing the compound at final concentrations of 0, 0.5 μM, 1 μM, and 2 μM was added. After 2 h, TNF-α (10 ng / mL) was added to treat the cells. After incubation for 22 h, according to the manufacturer's instructions, a DNA cycle kit (Wuhan Aidy Anti-Bio Technology Co., Ltd.) was used to detect the proportion of each cell cycle. 400 μL of PI staining solution was added and mixed well, and the cells were incubated in the dark at 4 °C for 30 min. Detection was performed using a flow cytometer (BD FACS Calibur), and the red fluorescence at an excitation wavelength of 488 nm was recorded.

[0054] (7) Real-time fluorescence quantitative PCR assay: Using 6×10 5Inoculate into a six-well plate at a density of cells / mL, and add media with final concentrations of the compound being 0, 0.5 μM, 1 μM, and 2 μM respectively. After 2 h, add TNF-α with a final concentration of 10 ng / mL. After 22 h, extract the RNA of each component according to the instructions of the SteadyPure Quick RNA Extraction Kit (Hunan Akribio Biotechnology Co., Ltd.). After measuring the RNA concentration, reverse transcribe it into cDNA, and then use a fluorescence quantitative PCR instrument to detect the gene expression level. Select GAPDH (glyceraldehyde-3-phosphate dehydrogenase) as the internal reference, and use the 2 -ΔΔCt method to calculate the relative expression levels of each gene.

[0055] (7) Immunofluorescence experiment: Seed HepG2 cells at a density of 2×10 5 cells in a confocal culture dish, and incubate them in a constant temperature incubator with compound concentrations of 1.5 μM, 3.0 μM, and 6.0 μM for 2 h, and then act with the inducer TNF-α (10 ng / mL) for 2 h. Wash the cells, fix them with 4% paraformaldehyde at 37 °C for 20 minutes, and permeabilize them by adding 500 μL of 0.3% Triton-X 100 for 30 minutes. Then block the cells with 5% BSA at room temperature for 2 h, and incubate them with the primary antibody NF-κB / p65 (1:200) overnight at 4 °C. Wash the cells with PBS, then incubate them with the secondary antibody (1:1000) for 30 minutes, wash away the unbound secondary antibody, and then incubate them in DAPI at room temperature in the dark for 30 minutes, and image the cells using a Leica confocal microscope (LEICA Stellaris 5).

[0056] (7) Cell scratch and migration experiments: For cells in good growth state, prepare a single-cell suspension, seed 2 mL of cells containing 6×10 5 cells / mL into a six-well plate, use a 10 μL pipette tip to draw a straight line on the plate, and wash away the floating cells that have been scratched with PBS. Add the compound, and take pictures with an inverted microscope at 0 and 24 h respectively.

[0057] Evaluate cell migration using a Transwell chamber with a pore size of 8 μm (Corning, NY, product number #3422). Add cells (5×10 4 cells) suspended in 200 μL of medium to the upper chamber, and add 600 μL of medium containing 10% FBS to the lower chamber. After incubating for 24 h, wipe off the cells inside the upper chamber with a cotton swab, fix them with 4% paraformaldehyde at room temperature for 30 minutes, and stain them with 0.1% crystal violet for 20 minutes. Take pictures under an inverted microscope. Decolorize with 33% acetic acid solution for one hour, and measure the absorbance at a wavelength of 580 nm.

[0058] II. Compound synthesis

[0059] Example 1: Synthesis of Compounds 1 - 17

[0060] Compound 1 (DHN 1):

[0061] (E)-2-(2-Fluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0062] Dissolve 2-fluorobenzaldehyde (2.22 g, 0.02 mol) and 6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one (2.44 g, 0.01 mol) in 1 mL of methanol, stir in an ice-water bath, then introduce HCl gas for 1 h, and stir the reaction at 0 °C for 2 - 3 days, monitoring by TLC during the period. After the reaction is completed, filter the suspension, and wash and purify the filter residue with acetone. Compound 1 is obtained as a pink powder with a yield of 39.6%.

[0063] Mp: 174 - 183 °C;

[0064] 1 H NMR (600 MHz, DMSO-d6) δ 11.22 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.59 (s, 1H), 7.52 (td, J = 7.7, 1.7 Hz, 1H), 7.46 (tdd, J = 7.6, 5.3, 1.7 Hz, 1H), 7.34–7.27 (m, 2H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.5 Hz, 1H), 4.13 (d, J = 13.9 Hz, 2H), 3.47 (d, J = 12.4 Hz, 3H), 3.37–3.30 (m, 2H), 3.14–3.02 (m, 2H), 2.94–2.91 (m, 2H), 2.89 (dd, J = 8.0, 5.5 Hz, 2H), 2.80 (d, J = 4.0 Hz, 3H).

[0065] 1313C NMR(151MHz, DMSO-d6) δ 184.95, 160.55 (d, J = 247.3Hz), 153.18, 146.09, 138.75, 131.31 (d, J = 2.7Hz), 131.21 (d, J = 8.4Hz), 130.11, 126.88 (d, J = 3.2Hz), 124.92 (d, J = 3.4Hz), 124.33, 123.68 (d, J = 13.8Hz), 116.18 (d, J = 21.6Hz), 113.80, 112.81, 52.11, 44.25, 42.33, 29.02, 27.54.

[0066] Compound 2 (DHN 2):

[0067] (E)-2-(3-Fluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one

[0068] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 3-fluorobenzaldehyde. The obtained product, Compound 2, was a white powder with a yield of 62.7%.

[0069] Mp: 181 - 182 °C;

[0070] 1 1H NMR(600MHz, Chloroform-d) δ 8.05 (d, J = 8.8Hz, 1H), 7.73 (s, 1H), 7.39–7.30 (m, 1H), 7.18 (d, J = 7.7Hz, 1H), 7.10 (d, J = 9.8Hz, 1H), 7.02 (td, J = 8.5, 2.5Hz, 1H), 6.84 (dd, J = 8.9, 2.5Hz, 1H), 6.61 (d, J = 2.5Hz, 1H), 3.42 (t, J = 5.1Hz, 4H), 3.06 (t, J = 6.5Hz, 2H), 2.88 (t, J = 6.5Hz, 2H), 2.57 (t, J = 5.1Hz, 4H), 2.36 (d, J = 1.1Hz, 3H).

[0071] 1313C NMR (151 MHz, DMSO-d6) δ 184.95, 162.56 (d, J = 243.7 Hz), 154.36, 145.79, 138.54 (d, J = 7.9 Hz), 137.86, 132.93 (d, J = 2.3 Hz), 130.92 (d, J = 8.6 Hz), 130.04, 126.36 (d, J = 2.7 Hz), 123.42, 116.63 (d, J = 21.5 Hz), 115.57 (d, J = 21.1 Hz), 113.10, 111.86, 54.72, 46.71, 46.09, 29.02, 27.26.

[0072] Compound 3 (DHN 3):

[0073] (E)-2-(4-Fluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0074] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 4-fluorobenzaldehyde. The obtained product, Compound 3, was a yellow powder with a yield of 74.3%.

[0075] Mp: 182 - 183 °C;

[0076] 1 1H NMR (600 MHz, DMSO-d6) δ 11.28 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.62 (s, 1H), 7.57 (dd, J = 8.4, 5.6 Hz, 2H), 7.29 (t, J = 8.6 Hz, 2H), 7.02 (d, J = 8.9 Hz, 1H), 6.89 (s, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.47 (d, J = 12.3 Hz, 2H), 3.33 (t, J = 13.1 Hz, 2H), 3.10 (q, J = 11.9, 11.5 Hz, 2H), 3.01 (d, J = 6.5 Hz, 2H), 2.88 (d, J = 6.6 Hz, 2H), 2.79 (d, J = 4.2 Hz, 3H).

[0077] 1313C NMR (151 MHz, DMSO-d6) δ 185.29, 162.37 (d, J = 246.8 Hz), 153.09, 145.79, 136.31, 133.67, 132.52 (d, J = 8.3 Hz), 132.46, 130.01, 124.58, 115.98 (d, J = 21.3 Hz), 113.77, 112.82, 52.13, 44.30, 42.34, 28.96, 27.14.

[0078] Compound 4 (DHN 4):

[0079] (E)-2-(2-Trifluoromethylbenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0080] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 2-trifluoromethylbenzaldehyde. The obtained product, Compound 4, was a yellow powder with a yield of 30.3%.

[0081] Mp: 185 - 188 °C;

[0082] 1 1H NMR (600 MHz, DMSO-d6) δ 11.24 (s, 1H), 7.89 (d, J = 8.9 Hz, 1H), 7.84 (d, J = 8.0 Hz, 1H), 7.75 (t, J = 6 Hz, 1H), 7.72 (s, 1H), 7.61 (t, J = 6 Hz, 1H), 7.55 (d, J = 7.6 Hz, 1H), 7.04 (d, J = 8.9 Hz, 1H), 6.90 (s, 1H), 4.13 (d, J = 14.1 Hz, 2H), 3.48 (d, J = 12.3 Hz, 2H), 3.34 (t, J = 13.2 Hz, 2H), 3.10 (q, J = 11.3 Hz, 2H), 2.85 (t, J = 6 Hz, 2H), 2.81 (s, 2H), 2.80 (s, 3H).

[0083] 13 13C NMR (151 MHz, DMSO-d6) δ 185.01, 153.28, 146.14, 139.18, 134.66, 132.96, 131.32, 130.33, 130.12, 129.11, 127.91 (q, J = 29.8 Hz), 126.50 (q, J = 5.5 Hz), 124.62 (q, J = 273.7 Hz), 124.17, 113.83, 112.84, 52.07, 44.20, 42.30, 29.18, 27.25.

[0084] Compound 5 (DHN 5):

[0085] (E)-2-(3,5-Bis(trifluoromethyl)benzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0086] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 3,5-trifluoromethylbenzaldehyde. The obtained product, Compound 5, was a yellow powder with a yield of 25.7%.

[0087] Mp: 192 - 194 °C;

[0088] 1 H NMR (600 MHz, DMSO-d6) δ 11.51 (s, 1H), 8.15 (s, 2H), 8.09 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.72 (s, 1H), 7.03 (dd, J = 9.0, 2.5 Hz, 1H), 6.89 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.46 (d, J = 12.2 Hz, 2H), 3.39–3.32 (m, 2H), 3.14–3.05 (m, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.88 (t, J = 6.5 Hz, 2H), 2.78 (d, J = 4.6 Hz, 3H).

[0089] 13 C NMR (151 MHz, DMSO-d6) δ 184.37, 152.85, 145.54, 139.25, 138.39, 130.97, 130.45 (q, J = 65.0, 32.1 Hz), 130.12, 129.70, 123.70, 123.27 (q, J = 273.0 Hz), 121.68–121.41 (m), 113.36, 112.27, 51.60, 43.73, 41.81, 28.36, 26.58.

[0090] Compound 6 (DHN 6):

[0091] (E)-2-(2,3-Difluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0092] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 2,3-difluorobenzaldehyde. The resulting product, compound 6, was a yellow powder with a yield of 61.6%.

[0093] Mp: 168 - 172 °C;

[0094] 1 1H NMR (600 MHz, DMSO-d6) δ 11.25 (s, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.55 (s, 1H), 7.48 (dtd, J = 10.0, 7.9, 1.8 Hz, 1H), 7.36–7.27 (m, 2H), 7.04 (dd, J = 8.9, 2.6 Hz, 1H), 6.91 (d, J = 2.5 Hz, 1H), 4.13 (d, J = 11.6 Hz, 2H), 3.47 (d, J = 12.0 Hz, 2H), 3.32 (s, 2H), 3.10 (s, 2H), 2.90 (s, 4H), 2.80 (s, 3H).

[0095] 13 13C NMR (151 MHz, DMSO-d6) δ 163.29, 131.83, 128.98 (dd, J = 245.6, 12.4 Hz), 126.71 (dd, J = 248.2, 12.6 Hz), 124.75, 118.60, 108.75, 105.10 (d, J = 3.4 Hz), 104.68 (d, J = 10.7 Hz), 104.15 (t, J = 2.8 Hz), 103.87 (dd, J = 7.3, 4.5 Hz), 102.73, 96.54 (d, J = 16.9 Hz), 92.39, 91.36, 30.68, 22.80, 20.91, 7.54, 6.20.

[0096] Compound 7 (DHN 7):

[0097] (E)-2-(2,4-Difluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0098] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 2,4-difluorobenzaldehyde. The resulting product, compound 7, was a yellow powder with a yield of 58.9%.

[0099] Mp: 166 - 169 °C;

[0100] 11H NMR (600 MHz, DMSO-d6) δ 11.17 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.58 (td, J = 8.6, 6.5 Hz, 1H), 7.52 (s, 1H), 7.37 (ddd, J = 10.5, 9.3, 2.6 Hz, 1H), 7.19 (td, J = 8.5, 2.6 Hz, 1H), 7.03 (dd, J = 9.0, 2.6 Hz, 1H), 6.89 (d, J = 2.6 Hz, 1H), 4.12 (d, J = 13.9 Hz, 2H), 3.47 (d, J = 12.3 Hz, 2H), 3.31 (d, J = 12.9 Hz, 2H), 3.09 (q, J = 11.0 Hz, 2H), 2.89 (d, J = 5.7 Hz, 4H), 2.79 (s, 3H).

[0101] 13 13C NMR (151 MHz, DMSO-d6) δ 184.84, 162.77 (dd, J = 250.0, 12.3 Hz), 160.75 (dd, J = 250.0, 12.3 Hz), 153.19, 146.07, 138.78, 132.56 (dd, J = 9.9, 4.2 Hz), 130.11, 125.97 (d, J = 2.5 Hz), 124.28, 120.30 (dd, J = 14.1, 3.8 Hz), 113.80, 112.80, 112.21 (dd, J = 21.3, 3.4 Hz), 104.84 (t, J = 26.1 Hz), 52.11, 44.24, 42.33, 28.96, 27.50.

[0102] Compound 8 (DHN 8):

[0103] (E)-2-(2,6-Difluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0104] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 2,6-difluorobenzaldehyde. The obtained product, Compound 8, was a purple powder with a yield of 37.9%.

[0105] Mp: 182 - 186 °C;

[0106] 11H NMR (600 MHz, DMSO-d6) δ 10.64 (s, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.53 (p, J = 7.5 Hz, 1H), 7.30 (s, 1H), 7.22 (t, J = 8.1 Hz, 2H), 7.04 (d, J = 9.0 Hz, 1H), 6.90 (s, 1H), 4.15 (d, J = 14.0 Hz, 2H), 3.50 (d, J = 12.1 Hz, 2H), 3.27 (t, J = 13.1 Hz, 2H), 3.12 (t, J = 11.4 Hz, 2H), 2.89 (t, J = 6.5 Hz, 2H), 2.82 (d, J = 4.5 Hz, 3H), 2.67 (d, J = 6.4 Hz, 2H).

[0107] 13 13C NMR (151 MHz, DMSO-d6) δ 184.41, 160.85 (dd, J = 247.64 Hz, 7.55 H), 153.25, 146.43, 141.74, 131.54 (t, J = 10.2 Hz), 130.19, 124.06, 120.82, 113.86, 113.21 (d, J = 19.3 Hz), 112.86, 112.46 dd, J = 24.16 Hz, J = 4.53 Hz), 52.24, 44.26, 42.44, 29.03, 28.48.

[0108] Compound 9 (DHN 9):

[0109] (E)-2-(3,4-Difluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0110] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 3,4-difluorobenzaldehyde. The obtained product, Compound 9, was a yellow powder with a yield of 29.4%.

[0111] Mp: 187 - 191 °C;

[0112] 111H NMR (600 MHz, DMSO-d6) δ 11.43 (s, 1H), 7.86 (d, J = 8.8 Hz, 1H), 7.65–7.59 (m, 1H), 7.57 (s, 1H), 7.52 (q, J = 9.2 Hz, 1H), 7.38 (t, J = 6.0 Hz, 1H), 7.02 (dd, J = 9.0, 2.5 Hz, 1H), 6.90 (s, 1H), 4.12 (d, J = 13.9 Hz, 2H), 3.47 (d, J = 12.2 Hz, 2H), 3.35 (t, J = 13.1 Hz, 2H), 3.11 (t, J = 12.0 Hz, 2H), 3.01 (d, J = 6.6 Hz, 2H), 2.88 (t, J = 6.5 Hz, 2H), 2.79 (d, J = 4.5 Hz, 3H).

[0113] 13 13C NMR (151 MHz, DMSO-d6) δ 184.64, 152.73, 150.38–149.91 (m), 148.68–148.25 (m), 145.44, 137.01, 133.23 (dd, J = 6.5, 3.9 Hz), 132.11, 129.62, 127.05 (dd, J = 6.3, 3.1 Hz), 123.94, 118.69 (d, J = 17.1 Hz), 117.67 (d, J = 16.8 Hz), 113.32, 112.33, 51.59, 43.78, 41.83, 28.42, 26.64.

[0114] Compound 10 (DHN 10):

[0115] (E)-2-(3,5-Difluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0116] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 3,5-difluorobenzaldehyde. The obtained product, Compound 10, was a yellow powder with a yield of 48.5%.

[0117] Mp: 186 - 189 °C;

[0118] 11H NMR (600 MHz, DMSO-d6) δ 11.25 (s, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.57 (s, 1H), 7.28 (dt, J = 9.2, 2.3 Hz, 1H), 7.25 (d, J = 6.6 Hz, 2H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.47 (d, J = 12.1 Hz, 2H), 3.32 (d, J = 11.8 Hz, 2H), 3.10 (d, J = 11.1 Hz, 2H), 3.02 (t, J = 5.7 Hz, 2H), 2.89 (t, J = 6.5 Hz, 2H), 2.79 (s, 3H). 13 13C NMR (151 MHz, DMSO-d6) δ 184.96, 162.78 (dd, J = 245.8, 13.3 Hz), 153.22, 146.01, 139.60 (d, J = 9.8 Hz), 138.69, 132.27, 130.12, 124.29, 113.79, 113.18 (dd, J = 20.2, 5.4 Hz), 112.75, 104.23 (t, J = 26.0 Hz), 52.11, 44.25, 42.34, 28.82, 27.18.

[0119] Compound 11 (DHN 11):

[0120] (E)-2-(2,3,5-Trifluorobenzylidene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0121] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 2,3,5-trifluorobenzaldehyde. The obtained product, Compound 11, was a yellow powder with a yield of 23.5%.

[0122] Mp: 170 - 175 °C;

[0123] 11H NMR (600 MHz, DMSO-d6) δ 11.21 (s, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.61–7.57 (m, 1H), 7.48 (s, 1H), 7.27 (dt, J = 8.4, 3.5 Hz, 1H), 7.03 (dd, J = 8.9, 2.6 Hz, 1H), 6.90 (d, J = 2.7 Hz, 1H), 4.14 (d, J = 14.1 Hz, 2H), 3.47 (d, J = 12.1 Hz, 2H), 3.34 (s, 2H), 3.10 (s, 2H), 2.89 (s, 4H), 2.79 (s, 3H).

[0124] 13 13C NMR (151 MHz, DMSO-d6) δ 184.45, 157.37 (dd, J = 243.1, 10.8 Hz), 153.31, 151.56–148.63 (m), 146.26, 145.00 (dd, J = 245.0, 13.5 Hz), 141.00, 130.21, 126.87–126.49 (m), 124.71, 124.00, 113.81, 112.90 (d, J = 3.02 Hz), 112.74, 106.39 (dd, J = 28.6, 21.3 Hz), 52.07, 44.17, 42.30, 28.83, 27.56.

[0125] Compound 12 (DHN 12):

[0126] (E)-6-(4-Methylpiperazin-1-yl)-2-(2,4,6-trimethylbenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0127] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced with 2,4,5-trifluorobenzaldehyde. The obtained product, Compound 12, was a purple powder with a yield of 54.4%.

[0128] Mp: 163-165 °C;

[0129] 11H NMR (600 MHz, DMSO-d6) δ 11.49 (s, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.33 (t, J = 8.8 Hz, 2H), 7.23 (s, 1H), 7.04 (d, J = 8.8 Hz, 1H), 6.91 (s, 1H), 4.14 (d, J = 13.8 Hz, 2H), 3.48 (d, J = 12.1 Hz, 2H), 3.42–3.35 (m, 4H), 2.89 (t, J = 6.3 Hz, 2H), 2.80 (s, 3H), 2.66 (t, J = 6.4 Hz, 2H).

[0130] 13 13C NMR (151 MHz, DMSO-d6) δ 183.84, 162.22 (dt, J = 248.2, 16.1 Hz), 159.77 (ddd, J = 248.5, 15.4, 10.4 Hz), 152.82, 145.94, 141.47, 129.73, 123.48, 119.51, 113.37, 112.35, 109.58 (td, J = 20.3, 4.5 Hz), 101.42–100.77 (m), 51.60, 43.70, 41.83, 28.54, 27.97.

[0131] Compound 13 (DHN 13):

[0132] (E)-6-(4-Methylpiperazin-1-yl)-2-(2,4,5-trifluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0133] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 2,4,6-trifluorobenzaldehyde. The obtained product, Compound 13, was a yellow powder with a yield of 43.1%.

[0134] Mp: 187 - 191 °C;

[0135] 1 1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.68 (dt, J = 10.6, 5.8 Hz, 2H), 7.46 (s, 1H), 7.03 (d, J = 8.9 Hz, 1H), 6.90 (s, 1H), 4.13 (d, J = 13.3 Hz, 2H), 3.47 (s, 2H), 3.10 (s, 2H), 2.89 (s, 4H), 2.80 (s, 3H).

[0136] 1313C NMR (151 MHz, DMSO-d6) δ 184.15, 155.49 (dd, J = 246.8, 9.8 Hz), 152.79, 150.93–148.12 (m), 146.70, 145.70, 139.33, 129.70, 124.57, 123.67, 120.62–119.68 (m), 118.52 (dd, J = 19.7, 4.2 Hz), 113.35, 112.31, 106.42 (dd, J = 29.1, 21.5 Hz), 51.68, 43.78, 41.90, 28.37, 26.96.

[0137] Compound 14 (DHN 14):

[0138] (E)-6-(4-Methylpiperazin-1-yl)-2-(2-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0139] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 2-nitrobenzaldehyde. The obtained product, Compound 14, was a yellow powder with a yield of 64.5%.

[0140] Mp: 182 - 187 °C;

[0141] 1 1H NMR (600 MHz, DMSO-d6) δ 11.01 (s, 1H), 8.18 (d, J = 8.2 Hz, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.82 (t, J = 7.5 Hz, 1H), 7.75 (s, 1H), 7.67 (t, J = 7.8 Hz, 1H), 7.58 (d, J = 7.6 Hz, 1H), 7.04 (dd, J = 9.0, 2.5 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.47 (d, J = 12.0 Hz, 2H), 3.31 (s, 2H), 3.10 (s, 2H), 2.87 (t, J = 6.4 Hz, 2H), 2.79 (d, J = 8.9 Hz, 5H).

[0142] 1313C NMR (151 MHz, DMSO-d6) δ 185.00, 153.23, 148.52, 146.18, 137.91, 134.24, 131.94, 131.51, 131.01, 130.12, 130.02, 125.27, 124.26, 113.83, 112.85, 52.11, 44.23, 42.33, 29.17, 27.16.

[0143] Compound 15 (DHN 15):

[0144] (E)-6-(4-Methylpiperazin-1-yl)-2-(3-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0145] The synthesis method is the same as that of Example 1, except that 2-fluorobenzaldehyde is replaced by 3-nitrobenzaldehyde. The obtained product, Compound 15, is a yellow powder with a yield of 64.5%.

[0146] Mp: 180 - 187 °C;

[0147] 1 1H NMR (600 MHz, DMSO-d6) δ 8.28 (s, 1H), 8.21 (d, J = 8.2 Hz, 1H), 7.94 (d, J = 7.7 Hz, 1H), 7.84 (d, J = 8.8 Hz, 1H), 7.74 (t, J = 7.9 Hz, 1H), 7.67 (s, 1H), 6.95 (d, J = 9.0 Hz, 1H), 6.79 (s, 1H), 3.38 (t, J = 5.2 Hz, 4H), 3.02 (t, J = 6.5 Hz, 2H), 2.87 (d, J = 6.6 Hz, 2H), 2.43 (t, J = 5.0 Hz, 4H), 2.23 (s, 3H).

[0148] 13 13C NMR (151 MHz, DMSO-d6) δ 184.73, 154.42, 148.40, 145.81, 139.04, 137.83, 136.53, 131.84, 130.51, 130.10, 124.39, 123.30, 123.27, 113.12, 111.83, 54.73, 46.70, 46.11, 28.99, 27.19.

[0149] Compound 16 (DHN 16):

[0150] (E)-6-(4-Methylpiperazin-1-yl)-2-(4-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0151] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 4-nitrobenzaldehyde. The obtained product, Compound 16, was a yellow powder with a yield of 86.7%.

[0152] Mp: 190 - 195 °C;

[0153] 1 H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.28 (d, J = 8.3 Hz, 2H), 7.88 (d, J = 8.9 Hz, 1H), 7.77 (d, J = 8.4 Hz, 2H), 7.67 (s, 1H), 7.03 (dd, J = 9.0, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.17–4.09 (m, 2H), 3.47 (d, J = 11.9 Hz, 2H), 3.30 (s, 2H), 3.15–3.06 (m, 2H), 3.04 (t, J = 6.6 Hz, 2H), 2.90 (t, J = 6.5 Hz, 2H), 2.79 (s, 3H).

[0154] 13 C NMR (151 MHz, DMSO-d6) δ 184.93, 153.26, 147.19, 146.01, 142.96, 139.67, 132.29, 131.34, 130.18, 124.26, 124.05, 113.83, 112.75, 52.13, 44.25, 42.35, 28.90, 27.33.

[0155] Compound 17 (DHN 17):

[0156] (E)-2-((5-Fluoropyridin-2-yl)methylene)-6-(4-methylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one

[0157] The synthesis method was the same as that of Example 1, except that 2-fluorobenzaldehyde was replaced by 5-fluoropyridine-2-carbaldehyde. The obtained product, Compound 17, was a yellow powder with a yield of 56.4%.

[0158] Mp: 213 - 218 °C;

[0159] 11H NMR (600 MHz, Deuterium Oxide) δ 8.79 (s, 1H), 8.28 (td, J = 8.3, 7.5, 2.7 Hz, 1H), 8.03 (dd, J = 9.1, 4.8 Hz, 1H), 7.95 (d, J = 8.9 Hz, 1H), 7.56 (s, 1H), 7.01 (d, J = 8.0 Hz, 1H), 6.86 (s, 1H), 4.18 (d, J = 14.3 Hz, 2H), 3.65 (d, J = 12.4 Hz, 2H), 3.32 (t, J = 13.6 Hz, 2H), 3.22 (t, J = 12 Hz, 2H), 3.09 (t, J = 6.5 Hz, 2H), 2.97 (s, 3H), 2.94 (t, J = 6.6 Hz, 2H).

[0160] 13 13C NMR (151 MHz, DMSO-d6) δ 185.63, 158.44 (d, J = 256.7 Hz), 153.13, 151.91 (d, J = 4.3 Hz), 146.37, 139.63, 138.07 (d, J = 23.9 Hz), 130.92, 130.15, 129.13 (d, J = 4.8 Hz), 124.38 (d, J = 2.6 Hz), 124.25, 113.76, 112.77, 52.06, 44.25, 42.30, 28.90, 26.85.

[0161] Example 2: Synthesis of Compounds 18 - 35

[0162] Compound 18 (DHN 18):

[0163] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2-fluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0164] Dissolve 2-fluorobenzaldehyde (2.22 g, 0.02 mol) and 6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one (2.58 g, 0.01 mol) in 1 mL of methanol, stir in an ice-water bath, then introduce HCl gas for 1 h, and stir the reaction at 0 °C for 2 - 3 days, monitoring by TLC during the period. After the reaction is completed, filter the suspension, and wash and purify the filter residue with acetone. Obtain Compound 20 as a yellow powder with a yield of 52.6%.

[0165] Mp: 172 - 174 °C;

[0166] 11H NMR (600 MHz, DMSO-d6) δ 11.13 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.58 (s, 1H), 7.52 (td, J = 7.7, 1.8 Hz, 1H), 7.45 (tdd, J = 7.5, 5.3, 1.7 Hz, 1H), 7.33–7.26 (m, 2H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.89 (d, J = 2.6 Hz, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.2 Hz, 2H), 3.34 (dd, J = 13.1, 10.4 Hz, 2H), 3.18–3.11 (m, 2H), 3.04 (dt, J = 12.4, 9.2 Hz, 2H), 2.94–2.79 (m, 4H), 1.28 (t, J = 7.3 Hz, 3H).

[0167] 13 13C NMR (151 MHz, DMSO-d6) δ 184.95, 160.55 (d, J = 247.2 Hz), 153.22, 146.09, 138.76, 131.31 (d, J = 2.7 Hz), 131.21 (d, J = 8.5 Hz), 130.11, 126.88 (d, J = 3.3 Hz), 124.92 (d, J = 3.3 Hz), 124.34, 123.68 (d, J = 14.0 Hz), 116.18 (d, J = 21.6 Hz), 113.79, 112.78, 51.00, 50.10, 44.29, 29.02, 27.55, 9.24.

[0168] Compound 19 (DHN 19):

[0169] (E)-6-(4-Ethylpiperazin-1-yl)-2-(3-fluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0170] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced with 3-fluorobenzaldehyde. The obtained product, Compound 19, was a yellow powder with a yield of 60.6%.

[0171] Mp: 123-128 °C;

[0172] 11H NMR (600 MHz, DMSO-d6) δ 11.17 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.61 (d, J = 2.5 Hz, 1H), 7.50 (td, J = 8.1, 6.2 Hz, 1H), 7.37–7.32 (m, 2H), 7.23 (td, J = 8.6, 2.5 Hz, 1H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.54 (d, J = 12.2 Hz, 2H), 3.37 (d, J = 11.9 Hz, 2H), 3.15 (p, J = 7.2 Hz, 2H), 3.09–3.00 (m, 4H), 2.89 (t, J = 6.5 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0173] 13 13C NMR (151 MHz, DMSO-d6) δ 185.19, 162.55 (d, J = 243.6 Hz), 153.19, 145.92, 138.43 (d, J = 7.8 Hz), 137.63, 133.33 (d, J = 2.2 Hz), 130.96 (d, J = 8.6 Hz), 130.07, 126.41 (d, J = 2.6 Hz), 124.46, 116.67 (d, J = 21.6 Hz), 115.69 (d, J = 21.0 Hz), 113.76, 112.75, 51.00, 50.11, 44.22, 28.93, 27.22, 9.24.

[0174] Compound 20 (DHN 20):

[0175] (E)-6-(4-Ethylpiperazin-1-yl)-2-(4-fluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0176] The synthesis method is the same as that of Example 2, except that 2-fluorobenzaldehyde is replaced by 4-fluorobenzaldehyde. The obtained product, Compound 20, is a yellow powder with a yield of 23.9%.

[0177] Mp: 165 - 166 °C;

[0178] 11H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 7.87 (d, J = 8.7 Hz, 1H), 7.62 (s, 1H), 7.57 (t, J = 6.5 Hz, 2H), 7.29 (t, J = 8.7 Hz, 2H), 7.03 (d, J = 8.8 Hz, 1H), 6.90 (s, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.54 (d, J = 12.2 Hz, 2H), 3.38 (s, 2H), 3.17–3.14 (m, 2H), 3.10–2.98 (m, 4H), 2.88 (t, J = 6.4 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0179] 13 13C NMR (151 MHz, DMSO-d6) δ 185.29, 162.36 (d, J = 246.9 Hz), 153.13, 145.79, 136.31, 133.67, 132.52 (d, J = 8.4 Hz), 132.46, 130.01, 124.58, 115.98 (d, J = 21.3 Hz), 113.76, 112.78, 50.98, 50.09, 44.22, 28.96, 27.15, 9.22.

[0180] Compound 21 (DHN 21):

[0181] (E)-6-(4-Ethylpiperazin-1-yl)-2-(3-(trifluoromethyl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0182] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced with 3-trifluoromethylbenzaldehyde. The obtained product, Compound 21, was a yellow powder with a yield of 32.2%.

[0183] Mp: 185 - 188 °C;

[0184] 1HNMR(600MHz, DMSO-d6) δ 11.21 (s, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.82 (s, 1H), 7.80 (d, J = 7.9 Hz, 1H), 7.74 (d, J = 7.9 Hz, 1H), 7.72–7.66 (m, 2H), 7.03 (dd, J = 9.0, 2.5 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 4.12 (d, J = 13.9 Hz, 2H), 3.53 (d, J = 12.3 Hz, 2H), 3.38 (d, J = 12.1 Hz, 2H), 3.14 (q, J = 7.4, 6.6 Hz, 2H), 3.10–2.99 (m, 4H), 2.88 (t, J = 6.4 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0185] 13 C NMR(151MHz, DMSO-d6) δ 185.12, 153.24, 145.92, 138.17, 137.13, 133.96, 132.97, 130.09, 129.83 (q, J = 31.7 Hz), 129.28, 126.60 (q, J = 3.7 Hz), 125.31 (q, J = 3.9 Hz), 124.56 (q, J = 272.5 Hz), 124.39, 113.78, 112.74, 50.99, 50.11, 44.20, 28.94, 27.15, 9.24.

[0186] Compound 22 (DHN 22):

[0187] (E)-6-(4-Ethylpiperazin-1-yl)-2-(4-(trifluoromethyl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0188] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 4-trifluoromethylbenzaldehyde. The obtained product, Compound 22, was a yellow powder with a yield of 16.8%.

[0189] Mp: 172 - 174 °C;

[0190] 11H NMR (600 MHz, DMSO-d6) δ 11.21 (s, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.80 (d, J = 8.1 Hz, 2H), 7.71 (d, J = 8.1 Hz, 2H), 7.66 (s, 1H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.2 Hz, 2H), 3.38 (d, J = 11.6 Hz, 2H), 3.15 (q, J = 7.1 Hz, 2H), 3.07–3.00 (m, 4H), 2.88 (t, J = 6.5 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0191] 13 13C NMR (151 MHz, DMSO-d6) δ 185.12, 153.25, 145.97, 140.25, 138.65, 132.90, 130.84, 130.12, 128.78 (q, J = 31.8 Hz), 125.78 (q, J = 3.8 Hz), 124.64 (q, J = 272.1 Hz), 124.36, 113.79, 112.75, 51.00, 50.10, 44.19, 28.97, 27.23, 9.24.

[0192] Compound 23 (DHN 23):

[0193] (E)-2-(2,3-Difluorobenzylidene)-6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0194] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,3-difluorobenzaldehyde. The obtained product, Compound 23, was a yellow powder with a yield of 59.5%.

[0195] Mp: 162 - 166 °C;

[0196] 11H NMR (600 MHz, DMSO-d6) δ 11.20 (s, 1H), 7.89 (d, J = 8.8 Hz, 1H), 7.54 (s, 1H), 7.48 (q, J = 8.3 Hz, 1H), 7.37–7.26 (m, 2H), 7.04 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.9 Hz, 2H), 3.53 (d, J = 12.3 Hz, 2H), 3.38 (d, J = 13.0 Hz, 2H), 3.15 (q, J = 7.8 Hz, 2H), 3.05 (q, J = 11.6 Hz, 2H), 2.90 (s, 4H), 1.29 (t, J = 7.3 Hz, 3H).

[0197] 13 13C NMR (151 MHz, DMSO-d6) δ 184.72, 153.29, 150.41 (dd, J = 245.6, 12.5 Hz), 148.14 (dd, J = 248.3, 12.7 Hz), 146.18, 140.03, 130.17, 126.52 (d, J = 3.2 Hz), 126.11 (d, J = 10.8 Hz), 125.58 (d, J = 3.1 Hz), 125.30 (dd, J = 7.2, 4.5 Hz), 124.17, 117.97 (d, J = 16.9 Hz), 113.80, 112.75, 51.00, 50.10, 44.16, 28.96, 27.62, 9.24.

[0198] Compound 24 (DHN 24):

[0199] (E)-2-(2,4-Difluorobenzylidene)-6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0200] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,4-difluorobenzaldehyde. The obtained product, Compound 24, was a yellow powder with a yield of 11.4%.

[0201] Mp: 171 - 175 °C;

[0202] 11H NMR (600 MHz, DMSO-d6) δ 11.26 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.59 (q, J = 8.2 Hz, 1H), 7.52 (s, 1H), 7.41–7.34 (m, 1H), 7.20 (t, J = 8.2 Hz, 1H), 7.03 (d, J = 9.0 Hz, 1H), 6.90 (s, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.54 (d, J = 12.2 Hz, 2H), 3.38 (t, J = 13.1 Hz, 2H), 3.15 (q, J = 6.8 Hz, 2H), 3.05 (d, J = 11.5 Hz, 2H), 2.89 (s, 4H), 1.29 (t, J = 7.4 Hz, 4H).

[0203] 13 13C NMR (151 MHz, DMSO-d6) δ 184.35, 162.29 (dd, J = 250.8, 14.1 Hz), 160.27 (dd, J = 249.8, 12.3 Hz), 152.76, 145.58, 138.30, 132.07 (dd, J = 9.9, 4.1 Hz), 129.63, 125.48, 123.80, 119.82 (dd, J = 14.2, 3.7 Hz), 113.30, 112.28, 111.72 (dd, J = 21.2, 3.5 Hz), 104.35 (t, J = 26.1 Hz), 50.48, 49.58, 43.68, 28.48, 27.02, 8.72.

[0204] Compound 25 (DHN 25):

[0205] (E)-2-(2,5-Difluorobenzylidene)-6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0206] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,5-difluorobenzaldehyde. The obtained product, Compound 25, was a yellow powder with a yield of 53.5%.

[0207] Mp: 171 - 174 °C;

[0208] 11H NMR (600 MHz, DMSO-d6) δ 11.18 (s, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.50 (s, 1H), 7.37 (qd, J = 9.1, 3.9 Hz, 2H), 7.30 (td, J = 8.8, 8.3, 3.7 Hz, 1H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.9 Hz, 2H), 3.53 (d, J = 12.3 Hz, 2H), 3.38 (d, J = 13.3 Hz, 2H), 3.14 (q, J = 7.3 Hz, 2H), 3.04 (q, J = 12.4 Hz, 2H), 2.90 (q, J = 5.4, 5.0 Hz, 4H), 1.28 (t, J = 7.3 Hz, 3H).

[0209] 13 13C NMR (151 MHz, DMSO-d6) δ 184.70, 159.11 (d, J = 242.0 Hz), 156.71 (d, J = 242.0 Hz), 153.29, 146.19, 139.84, 130.16, 125.81, 125.24 (dd, J = 16.8, 8.7 Hz), 124.18, 117.69 (dd, J = 25.1, 9.2 Hz), 117.41 (d, J = 9.06 Hz), 117.41 (dd, J = 24.6, 3.8 Hz), 113.79, 112.74, 51.00, 50.11, 44.17, 28.88, 27.46, 9.24.

[0210] Compound 26 (DHN 26):

[0211] (E)-2-(2,6-Difluorobenzylidene)-6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0212] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,6-difluorobenzaldehyde. The obtained product, Compound 26, was a yellow powder with a yield of 17.0%.

[0213] Mp: 154 - 157 °C;

[0214] 11H NMR (600 MHz, DMSO-d6) δ 11.23 (s, 1H), 7.89 (d, J = 8.9 Hz, 1H), 7.52 (p, J = 8.2, 7.4 Hz, 1H), 7.29 (s, 1H), 7.21 (t, J = 8.0 Hz, 2H), 7.04 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.4 Hz, 2H), 3.38 (t, J = 11.7 Hz, 2H), 3.18–3.11 (m, 2H), 3.04 (q, J = 9.1 Hz, 2H), 2.88 (t, J = 6.4 Hz, 2H), 2.66 (t, J = 6.8 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0215] 13 13C NMR (151 MHz, DMSO-d6) δ 184.41, 160.03 (dd, J = 248.0, 7.5 Hz), 153.31, 146.43, 141.76, 131.53 (t, J = 10.4 Hz), 130.20, 124.03, 120.80, 113.82, 113.15 (t, J = 19.8 Hz), 112.80, 112.39 (dd, J = 21.0, 4.5 Hz), 50.98, 50.09, 44.13, 29.03, 28.48, 9.22.

[0216] Compound 27 (DHN 27):

[0217] (E)-2-(3,5-Difluorobenzylidene)-6-(4-ethylpiperazin-1-yl)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0218] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced with 3,5-difluorobenzaldehyde. The obtained product, Compound 27, was a yellow powder with a yield of 61.0%.

[0219] Mp: 168 - 173 °C;

[0220] 11H NMR (600 MHz, DMSO-d6) δ 11.26 (s, 1H), 7.86 (d, J = 8.9 Hz, 1H), 7.56 (s, 1H), 7.27 (dd, J = 9.3, 2.3 Hz, 1H), 7.24 (d, J = 6.7 Hz, 2H), 7.02 (dd, J = 8.9, 2.5 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.3 Hz, 2H), 3.38 (d, J = 11.7 Hz, 2H), 3.14 (q, J = 7.2, 6.2 Hz, 2H), 3.08–2.98 (m, 4H), 2.88 (t, J = 6.5 Hz, 2H), 1.28 (t, J = 7.3 Hz, 3H).

[0221] 13 13C NMR (151 MHz, DMSO-d6) δ 184.95, 162.78 (dd, J = 246.2, 13.6 Hz), 153.25, 146.00, 139.63 (t, J = 9.9 Hz), 138.70, 132.25, 130.12, 124.30, 113.77, 113.18 (dd, J = 20.1, 5.2 Hz), 112.71, 104.23 (t, J = 25.9 Hz), 50.99, 50.09, 44.17, 28.82, 27.18, 9.23.

[0222] Compound 28 (DHN 28):

[0223] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2,3,4-trifluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0224] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,3,4-trifluorobenzaldehyde. The obtained product, compound 28, was a purple powder with a yield of 32.6%.

[0225] Mp: 195 - 198 °C;

[0226] 11H NMR (600 MHz, DMSO-d6) δ 11.06 (s, 1H), 7.88 (dd, J = 8.9, 2.9 Hz, 1H), 7.48 (s, 1H), 7.40 (q, J = 9.2 Hz, 2H), 7.04 (d, J = 9.0 Hz, 1H), 6.90 (s, 1H), 4.14 (d, J = 13.8 Hz, 2H), 3.54 (d, J = 12.2 Hz, 2H), 3.38 (s, 2H), 3.16 (t, J = 6.9 Hz, 2H), 3.05 (q, J = 11.7 Hz, 2H), 2.89 (d, J = 3.0 Hz, 4H), 1.29 (t, J = 7.9 Hz, 3H).

[0227] 13 13C NMR (151 MHz, DMSO-d6) δ 184.61, 153.31, 150.57 (ddd, J = 240.1, 38.4, 12.6 Hz), 148.19 (ddd, J = 240.1, 38.4, 12.6 Hz), 146.14, 140.11, 139.75 (dt, J = 248.9, 15.8 Hz), 130.16, 125.73 (d, J = 5.4 Hz), 124.89, 124.11, 121.91 (dd, J = 10.57, 3.5 Hz), 113.80, 113.21 (dd, J = 17.1, 3.5 Hz), 112.73, 50.97, 50.07, 44.13, 28.91, 27.56, 9.21.

[0228] Compound 29 (DHN 29):

[0229] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2,3,5-trifluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0230] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,3,5-difluorobenzaldehyde. The obtained product, Compound 29, was a yellow powder with a yield of 39.0%.

[0231] Mp: 160 - 163 °C;

[0232] 1HNMR(600MHz, DMSO-d6) δ 11.11(s, 1H), 7.89(d, J = 8.9Hz, 1H), 7.60(dddd, J = 11.2, 9.0, 6.2, 3.1Hz, 1H), 7.48(s, 1H), 7.28(dq, J = 7.7, 2.4Hz, 1H), 7.04(dd, J = 8.9, 2.6Hz, 1H), 6.91(d, J = 2.5Hz, 1H), 4.14(d, J = 13.8Hz, 2H), 3.54(d, J = 12.2Hz, 2H), 3.38(d, J = 11.9Hz, 2H), 3.19–3.12(m, 2H), 3.05(q, J = 9.1Hz, 2H), 2.90(s, 4H), 1.29(t, J = 7.3Hz, 3H).

[0233] 13 C NMR(151MHz, DMSO-d6) δ 184.45, 157.38(dd, J = 242.2, 11.3Hz), 153.35, 150.24(dt, J = 248.0, 14.3Hz), 146.26, 144.61(ddd, J = 244.7, 16.1, 3.8Hz), 141.00, 130.21, 127.50–125.96(m), 124.70, 124.01, 113.79, 112.90(d, J = 3.1Hz), 112.73(d, J = 5.8Hz),, 106.38(dd, J = 28.5, 21.5Hz), 50.97, 50.06, 44.10, 28.83, 27.56, 9.20.

[0234] Compound 30 (DHN 30):

[0235] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2,4,6-trifluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0236] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced with 2,4,6-difluorobenzaldehyde. The obtained product, Compound 30, was a purple powder with a yield of 41.3%.

[0237] Mp: 181 - 184 °C;

[0238] 1HNMR(600MHz, DMSO-d6) δ 11.55 (s, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.32 (t, J = 8.8 Hz, 2H), 7.21 (s, 1H), 7.03 (d, J = 8.9 Hz, 1H), 6.89 (s, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.52 (d, J = 12.2 Hz, 2H), 3.41 (t, J = 13.1 Hz, 2H), 3.14 (p, J = 6.9 Hz, 2H), 3.05 (dd, J = 24.4, 12.0 Hz, 2H), 2.87 (t, J = 6.4 Hz, 2H), 2.65 (t, J = 6.6 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0239] 13 C NMR(151MHz, DMSO-d6) δ 184.31, 162.27 (dt, J = 248.7, 15.9 Hz), 160.11 (ddd, J = 248.9, 15.4, 10.7 Hz), 153.33, 146.41, 141.95, 130.19, 123.95, 119.95, 113.81, 112.77, 110.03 (dd, J = 20.3, 4.6 Hz), 102.32–100.51 (m), 50.93, 50.03, 44.07, 28.99, 28.42, 9.17.

[0240] Compound 31 (DHN 31):

[0241] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2,4,5-trifluorobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0242] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2,4,5-difluorobenzaldehyde. The obtained product, Compound 31, was a yellow powder with a yield of 55.3%.

[0243] Mp: 174 - 177 °C;

[0244] 11H NMR (600 MHz, DMSO-d6) δ 11.38 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.68 (td, J = 10.6, 5.0 Hz, 2H), 7.46 (s, 1H), 7.03 (d, J = 9.0 Hz, 1H), 6.91 (s, J = 2.7 Hz, 1H), 4.14 (d, J = 13.8 Hz, 2H), 3.54 (d, J = 12.2 Hz, 2H), 3.40 (t, J = 14.1 Hz, 2H), 3.15 (q, J = 7.2, 6.4 Hz, 2H), 3.04 (q, J = 11.3 Hz, 2H), 2.90 (s, 4H), 1.30 (t, J = 7.3 Hz, 3H).

[0245] 13 13C NMR (151 MHz, DMSO-d6) δ 184.13, 155.46 (dd, J = 246.6, 9.8 Hz), 152.82, 149.17 (dt, J = 251.1, 13.6 Hz), 145.90 (ddd, J = 242.5, 12.8, 3.02 Hz), 145.68, 139.32, 129.68, 124.52, 123.65, 120.22 (d, J = 16.8 Hz), 118.49 (dd, J = 19.6, 4.2 Hz), 113.30, 112.25, 106.38 (dd, J = 28.9, 21.5 Hz), 50.50, 49.59, 43.66, 28.36, 26.94, 8.73.

[0246] Compound 32 (DHN 32):

[0247] (E)-6-(4-Ethylpiperazin-1-yl)-2-(2-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0248] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 2-nitrobenzaldehyde. The obtained product, Compound 32, was a yellow powder with a yield of 28.2%.

[0249] Mp: 169 - 172 °C;

[0250] 11H NMR (600 MHz, DMSO-d6) δ 11.20 (s, 1H), 8.17 (d, J = 8.2 Hz, 1H), 7.89 (d, J = 8.9 Hz, 1H), 7.82 (t, J = 7.6 Hz, 1H), 7.75 (s, 1H), 7.66 (t, J = 7.8 Hz, 1H), 7.58 (d, J = 7.7 Hz, 1H), 7.04 (dd, J = 9.0, 2.5 Hz, 1H), 6.89 (d, J = 2.7 Hz, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.5 Hz, 3H), 3.37 (t, J = 13.1 Hz, 2H), 3.15 (p, J = 7.2 Hz, 2H), 3.09–3.00 (m, 2H), 2.87 (t, J = 6.4 Hz, 2H), 2.78 (t, J = 6.5 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0251] 13 13C NMR (151 MHz, DMSO-d6) δ 185.01, 153.27, 148.52, 146.18, 137.92, 134.25, 131.94, 131.52, 131.01, 130.12, 130.02, 125.27, 124.27, 113.81, 112.82, 50.99, 50.09, 44.16, 29.16, 27.16, 9.23.

[0252] Compound 33 (DHN 33):

[0253] (E)-6-(4-Ethylpiperazin-1-yl)-2-(3-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0254] The synthesis method was the same as that of Example 2, except that 2-fluorobenzaldehyde was replaced by 3-nitrobenzaldehyde. The obtained product, Compound 33, was a yellow powder with a yield of 66.8%.

[0255] Mp: 163 - 167 °C;

[0256] 11H NMR (600 MHz, DMSO-d6) δ 11.18 (s, 1H), 8.29 (s, 1H), 8.22 (dd, J = 8.2, 2.3 Hz, 1H), 7.95 (d, J = 7.8 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.75 (t, J = 8.0 Hz, 1H), 7.69 (s, 1H), 7.03 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (d, J = 2.6 Hz, 1H), 4.13 (d, J = 13.9 Hz, 2H), 3.53 (d, J = 12.2 Hz, 2H), 3.38 (d, J = 14.4 Hz, 2H), 3.15 (p, J = 7.5, 6.9 Hz, 2H), 3.04 (t, J = 4.9 Hz, 4H), 2.89 (t, J = 6.5 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0257] 13 13C NMR (151 MHz, DMSO-d6) δ 185.00, 153.26, 148.40, 145.94, 138.80, 137.71, 136.57, 132.24, 130.54, 130.13, 124.43, 124.31, 123.40, 113.80, 112.74, 51.00, 50.11, 44.19, 28.91, 27.15, 9.24.

[0258] Compound 34 (DHN 34):

[0259] (E)-6-(4-Ethylpiperazin-1-yl)-2-(4-nitrobenzylidene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0260] The synthesis method was the same as that of Example 22, except that 2-fluorobenzaldehyde was replaced by 3-nitrobenzaldehyde. The obtained product, Compound 34, was a yellow powder with a yield of 83.5%.

[0261] Mp: 183 - 188 °C;

[0262] 1HNMR(600MHz, DMSO-d6) δ 11.16 (s, 1H), 8.28 (d, J = 8.2 Hz, 2H), 7.88 (d, J = 8.8 Hz, 1H), 7.77 (d, J = 8.3 Hz, 2H), 7.67 (s, 1H), 7.04 (d, J = 8.9 Hz, 1H), 6.90 (s, 1H), 4.13 (d, J = 13.8 Hz, 2H), 3.53 (d, J = 12.2 Hz, 2H), 3.38 (d, J = 16.2 Hz, 3H), 3.15 (s, 2H), 3.04 (s, 4H), 2.90 (s, 1H), 1.29 (t, J = 7.4 Hz, 3H).

[0263] 13 C NMR(151MHz, DMSO-d6) δ 184.93, 153.29, 147.19, 146.01, 142.96, 139.67, 132.28, 131.34, 130.17, 124.28, 124.05, 113.82, 112.72, 51.03, 50.13, 44.19, 28.89, 27.33, 9.26.

[0264] Compound 35 (DHN 35):

[0265] (E)-6-(4-Ethylpiperazin-1-yl)-2-((5-fluoropyridin-2-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one hydrochloride

[0266] The synthesis method is the same as that of Example 2, except that 2-fluorobenzaldehyde is replaced by 5-fluoropyridine-2-carbaldehyde. The obtained product, Compound 35, is a yellow powder with a yield of 43.7%.

[0267] Mp: 183 - 188 °C;

[0268] 11H NMR (600 MHz, DMSO-d6) δ 11.41 (s, 1H), 8.69 (d, J = 2.9 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.81 (td, J = 8.7, 2.9 Hz, 1H), 7.75 (dd, J = 8.7, 4.7 Hz, 1H), 7.54 (s, 1H), 7.02 (dd, J = 8.9, 2.5 Hz, 1H), 6.90 (s, 1H), 4.12 (d, J = 13.8 Hz, 2H), 3.52 (d, J = 12.1 Hz, 2H), 3.45 (t, J = 6.5 Hz, 2H), 3.42–3.36 (m, 2H), 3.14 (dt, J = 12.3, 6.3 Hz, 2H), 3.04 (q, J = 9.2 Hz, 2H), 2.90 (t, J = 6.5 Hz, 2H), 1.29 (t, J = 7.3 Hz, 3H).

[0269] 13 13C NMR (151 MHz, DMSO-d6) δ 185.63, 158.44 (d, J = 256.7 Hz), 153.16, 151.91 (d, J = 4.0 Hz), 146.37, 139.62, 138.08 (d, J = 24.1 Hz), 130.92, 130.14, 124.38 (d, J = 6.4 Hz), 124.24, 113.75, 112.74, 50.97, 50.06, 44.19, 28.90, 26.85, 9.21.

[0270] III. Result Evaluation

[0271] 3.1 Anti-hepatocellular carcinoma activity

[0272] The anti-hepatocellular carcinoma evaluation was performed using the CCK-8 method. All experiments were repeated in parallel 3 times, and the survival mean was taken. HepG2 cells were treated with Compounds 1 - 35 (DHNs1 - 35). After treating HepG2 cells with different concentrations of DHNs, the cytotoxicity IC50 of each compound was calculated after fitting the curve. The IC50 values of the DHNs synthesized in the examples of this application are shown in Table 1.

[0273] The experimental results show that the N-methylpiperazine-substituted compounds 1-35 synthesized in the examples of this application have anti-hepatocellular carcinoma cell activity. However, as can be seen from the results in Table 1, the overall anti-hepatocellular carcinoma activity of the compounds substituted with N-methylpiperazine is lower than that of the compounds substituted with N-ethylpiperazine. The compounds with fluorine atom substitution at the ortho and meta positions have better overall anti-hepatocellular carcinoma activity, probably because the ortho and meta substitutions can better bind to proteins and exert their pharmacological effects. Among them, compounds 20, 21, 26, and 27 are comparable to the positive drug DOX, and the anti-hepatocellular carcinoma activity of compound 31 is even better than that of DOX. It can be seen that DHN derivatives have high anti-hepatocellular carcinoma activity.

[0274] Table 1 IC50 of Compounds 1-35 against HepG2 Cells

[0275]

[0276]

[0277] DOX is doxorubicin hydrochloride, and its CAS number is 25316-40-9.

[0278] Research shows that when a single substituent is replaced, the compounds with fluorine atom substitution on the benzene ring have better anti-hepatocellular carcinoma effects than the compounds with nitro substitution. However, this effect was not found in the compounds synthesized in this application. In addition, as can be seen from the results in Table 1, replacing the benzene ring of the compounds in this application with a pyridine ring has higher anti-hepatocellular carcinoma efficacy. Then, we added a trifluoromethyl group as a substituent. Compared with compound 1 (11.33 μM) and compound 3 (62.06 μM), the IC50 value of compound 4 is 6.37 μM, confirming its higher anti-hepatocellular carcinoma efficacy. However, when we used two trifluoromethyl groups as substituents, the IC50 value of compound 5 is 5.75 μM, which is comparable to that of compound 2 (5.49 μM) and shows no obvious increase in the anti-hepatocellular carcinoma effect.

[0279] When more than one fluorine atom is used for substitution, such as in the series of compounds 6 - 9, the anti-cancer efficiency decreases instead. The IC50 values are 13.06 μM, 16.32 μM, 24.85 μM, and 13.42 μM in sequence. However, it is worth noting that compound 10 substituted at the 3- and 5-positions of the benzene ring achieved remarkable anti-hepatocellular carcinoma efficiency, with an IC50 value of 2.63 μM. When the number of substituents increases and three fluorine groups are substituted, the anti-cancer efficiency of the series of compounds 11 - 13 did not increase significantly, and the IC50 values are 10.20 μM, 12.39 μM, and 12.26 μM in sequence. When a nitro group substitutes a hydrogen atom on the benzene ring, the therapeutic effect is the worst for para-substitution, and the IC50 value of compound 16 is 156.19 μM. When substitution occurs at the ortho- and meta-positions, the effects are better. The IC50 values of compounds 14 and 15 are 11.40 μM and 8.09 μM in sequence, but the anti-hepatocellular carcinoma potency is still lower than that when fluorine substitution occurs at the meta-position.

[0280] We also synthesized a series of compounds 18 - 35 with N - ethylpiperazine as the side arm of 3,4 - dihydronaphthalen - 1(2H) - one. To our delight, the anti - liver cancer potency of these compounds was significantly improved. The IC50 values of compounds 18 - 20 with a single fluorine group as the substituent on benzene ring B were 2.15 μM, 2.46 μM, and 3.38 μM respectively, which were generally lower than the IC50 values of compounds 1 - 3. Interestingly, although the anti - cancer potency of ortho - and para - nitro - substituted compounds was worse than that of single - fluorine - substituted compounds, with the IC50 values of compounds 32 and 34 being 5.70 μM and 19.12 μM respectively, the IC50 value of compound 33 reached 1.92 μM, lower than that of the positive drug DOX (2.04 μM). In addition, when the pyridine ring substituted benzene ring B, the anti - cancer efficiency of compound 35 did not increase, and the IC50 value was 19.74 μM. When N - ethylpiperazine was used as the side arm of 3,4 - dihydronaphthalen - 1(2H) - one, the anti - cancer potency of compounds 23 - 31 with multiple fluorine groups substituting benzene ring B was generally inferior to that of single - fluorine - substituted compounds, but some compounds still showed anti - cancer effects comparable to those of the positive drug DOX. For example, the IC50 value of compound 23 reached 2.47 μM. Surprisingly, the anti - cancer potencies of compounds 24 and 28 against liver cancer were higher than those of the effective drug DOX, reaching 2.03 μM and 0.92 μM respectively. On this basis, we evaluated the DHN series of compounds 1 - 35. Compared with the substitution of benzene ring B, the substitution of benzene ring A had a greater impact on the anti - hepatocellular carcinoma efficacy. When N - ethylpiperazine was used as the side arm of DHN compounds, the anti - liver cancer effect was greatly improved. However, different substituents and their positions also affected the anti - cancer effects of DHN compounds. For example, the anti - liver cancer efficiency of benzene ring B substituted with a single fluorine group was generally higher than that of benzene ring B substituted with multiple fluorine groups. However, when benzene ring B was substituted with three fluorine groups, the anti - liver cancer potency of compounds with fluorine substituents at the 2 - and 3 - positions was better. Therefore, we selected the compound DHN28 with a longer carbon chain in piperazine compounds and fluorine substitutions at the 2nd, 3rd, and 4th positions of benzene ring B, which had the best anti - cancer effect, as the lead compound for subsequent research.

[0281] 3.2 Effects of Compound 28 on Apoptosis and Cell Cycle Arrest of HepG2 Cells

[0282] We used the Annexin V-FITC / PI apoptosis kit to detect the apoptosis pattern of compound 28-induced HepG2 cells. Apoptotic cells can be distinguished by staining the cells with propidium iodide (PI) (which stains DNA) and an Annexin dye (which can stain phosphatidylserine (PS), a major component in the cell membrane). During apoptosis, PS located inside the cell membrane translocates to the outside of the cell membrane, exposing it to staining. At the same time, the cell is more easily embedded with PI. Therefore, PI / Annexin double staining is considered an established method for apoptosis detection. Since compound 28 is the most promising anti-hepatocellular carcinoma agent, the ability of compound 28 to induce apoptosis and cell cycle arrest was investigated by flow cytometry.

[0283] As shown in Figure 1 Figure A in Figure 1 , compound 28 can induce apoptosis of HepG2 cells from early to late stage in a dose-dependent manner. As shown in Figure A and Figure B in

[0284] As shown in Figure 2 , the cell cycle distribution results of HepG2 cells treated with compound 28 (0 - 1.84 μM) for 24 h. Compared with the control group, the proportion of cells in the G2 / M phase of compound 28-treated cells increased significantly, inhibiting the mitotic phase of the cells. As the concentration of compound 28 increased from 0.46 μM to 0.92 μM, the proportion of HepG2 cells in the G2 / M phase was observed to change from 22.43% to 38.04%. At a concentration of 1.84 μM, the proportion of the G2 / M phase was 76.22%. This demonstrated that the cell cycle arrest effect of compound 28 is concentration-dependent, making it a potential anti-hepatocellular carcinoma cell proliferation drug.

[0285] 3.3 Effects of compound 28 on the expression of apoptosis-related genes

[0286] At the gene level, the expression of apoptosis-related genes plays a crucial role in anti-hepatocellular carcinoma. In cells, the apoptosis program can be initiated through the participation of BAX and BCL-2 (pro-apoptotic and anti-apoptotic protein families), thereby playing a role in anti-hepatocellular carcinoma cell proliferation. C-caspase-3 (Cleaved-caspase-3) is a member of the caspase family and is a key execution enzyme during apoptosis. As shown in Figure 3As shown, compared with the control group, the relative expression levels of BAX and C-caspase-3 in HepG2 cells treated with compound 28 were significantly increased, while the relative expression level of BCL-2 was significantly decreased. With the increase in the concentration of compound 28, the expression of apoptosis-related genes showed an obvious dose-response relationship, indicating that compound 28 has a concentration-dependent effect on the expression of apoptosis genes in HepG2 cells.

[0287] 3.4 Compound 28 inhibits NF-κB nuclear translocation in HepG2 cells

[0288] Tumor necrosis factor-α (TNF-α) can promote malignant behaviors such as the proliferation, survival, and migration of hepatocellular carcinoma cells by activating multiple intracellular signaling pathways. Among them, the NF-κB signaling pathway is one of the key downstream pathways for TNF-α to exert its effects, and P65 is an important subunit of the NF-κB complex. The activation of the NF-κB signaling pathway causes the nuclear translocation of the P65 subunit into the nucleus, binds to specific DNA sequences, and initiates the transcription of a series of genes related to tumor progression, such as anti-apoptosis genes, cell cycle regulatory genes, and inflammation-related genes. To study the inhibitory effect of compound 28 on NF-κB nuclear translocation in HepG2 cells, immunofluorescence was used for detection. As Figure 4 shown, the control group was DMSO. It can be seen that the p65 protein was almost entirely concentrated in the cytoplasm. After induction with 10 ng / mL TNF-α, some P65 protein could appear in the nucleus. In HepG2 cells treated with 1.84 μM compound 28 and TNF-α (10 ng / mL), nuclear translocation was inhibited, and the P65 protein was expressed in the cytoplasm. In summary, compound 28 can effectively inhibit the nuclear translocation of the P65 protein, thereby blocking the activation of the NF-κB signaling pathway and playing an anti-hepatocellular carcinoma role.

[0289] 3.5 DHN 28 inhibits the migration of HepG2 cells

[0290] The distant metastasis of cancer cells is the ultimate cause of death for most cancer patients. Therefore, we used the scratch assay to explore the effect of compound 28 on the migration ability of HepG2 cells. As Figure 5As shown, after 24 hours of culture, compared with the control group, the wound healing degree of HepG2 cells treated with compound 28 was significantly inhibited. There were also significant differences among the treatment groups with different concentrations of compound 28. Although the cells treated with low concentrations of compound 28 were inhibited, there was still partial migration. Interestingly, when the concentration of compound 28 reached 1.84 μM, the scratch width hardly changed, indicating that the cell migration ability was completely inhibited, reflecting that the effect of compound 28 on the migration ability of HepG2 cells was dose-dependent. We used the Transwell assay to further verify the effect of compound 28 on the migration ability of HepG2 cells, and the results were as Figure 6 shown. Among them, Panel A shows the results of crystal violet staining. From the crystal violet staining results in Panel A, it can be seen that the number of cells that penetrated the membrane in the experimental group was significantly reduced compared with the control group, and the number of cells that penetrated the membrane decreased with the increase in the concentration of compound 28. As Figure 6 shown in Panel B of

[0291] 3.6 DHN 28 inhibits the activation of NF-κB and MAPK

[0292] We used Western blot analysis to further explore the inhibitory mechanism of compound 28 on the NF-κB, p38, and ERK signaling pathways, and the results were as Figure 7 shown. When cells are stimulated by the external environment, the IκB kinase complex (IKK) is activated. The activated IKK phosphorylates the IκB protein, promoting its ubiquitination and proteasome-mediated degradation, thereby leading to the nuclear localization of p65 and initiating the expression of a series of genes related to tumor progression: inducing the expression of anti-apoptotic genes, cyclins, and proto-oncogenes, etc. Among them, the abnormal increase in the phosphorylation level of IκB-α leads to its rapid degradation, resulting in the continuous activation of NF-κB. TNF-α (10 ng / mL) can effectively activate the NF-κB signaling pathway in HepG2 cells. After treating the cells with different concentrations of compound 28 (0.46 μM, 0.92 μM, 1.84 μM), the phosphorylation levels of P65 protein and IκB-α protein were significantly reduced, and the concentration of compound 28 was inversely proportional to the protein phosphorylation level.

[0293] In liver cancer cells, there is extensive crosstalk between the NF-κB pathway and the MAPK (mitogen-activated protein kinase) pathway. For example, p38 in the MAPK pathway can phosphorylate and activate NF-κB, thereby enhancing the transcriptional activity of NF-κB. The MAPK pathway plays a key role in the development and progression of cancer, including extracellular regulated kinase (ERK) and p38 kinase. ERK is activated by growth factors and plays an important role in cell proliferation and survival, while the activation of p38 signaling is generally considered to promote apoptosis. After pretreatment of HepG2 cells with compound 28 at 0.46 μM, 0.92 μM, and 1.84 μM for two hours, TNF-α (10 ng / mL) was used for treatment. The relative expression levels of P-p38 and P-ERK were significantly decreased, showing an obvious dose-response relationship. Based on these results, we believe that compound 28 exerts an anti-liver cancer effect by inhibiting the phosphorylation levels of p65, IκB-α, p38, and ERK proteins, thereby inhibiting the NF-κB and MAPK signaling pathways.

[0294] As can be seen from the above results, the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives of the present application have potential anti-liver cancer effects and are expected to become potential multifunctional drugs for the clinical treatment of primary liver cancer.

[0295] The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivatives, preparation methods, and uses provided in the embodiments of the present application have been introduced in detail above. Specific examples have been used in this article to elaborate on the principles and implementation manners of the present application. The descriptions of the above embodiments are only used to help understand the method and its core idea of the present application; at the same time, for those skilled in the art, according to the idea of the present application, there will be changes in the specific implementation manners and application scopes. In summary, the content of this specification should not be construed as a limitation to the present application.

Claims

1. A piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, the structural formula of which is shown in formula (I): Among them, R1 is N-methylpiperazine or N-ethylpiperazine, R2 is phenyl or pyridyl, and the phenyl or pyridyl is substituted by one or more R a groups, and the R a is selected from one or more of a single or multiple fluoro groups, nitro groups, methyl groups, and trifluoromethyl groups.

2. The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 1, wherein, The structural formula is as follows: Among them, R1 is N-methylpiperazine or N-ethylpiperazine, and the R a is selected from one or more of a single or multiple fluorine groups, nitro groups, methyl groups, and trifluoromethyl groups.

3. The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 1, wherein, Ra is selected from the following groups: 2-F, 3-F, 4-F, 2-CF3, 3-CF3, 4-CF3, 3,5-CF3, 2,3-F, 2,4-F, 2,5-F, 2,6-F, 3,4-F, 3,5-F, 2,3,4-F, 2,3,5-F, 2,4,6-F, 2,4,5-F, 2-NO2, 3-NO2, 4-NO2.

4. The piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 1, characterized in that, The structural formula is as follows:

5. A method for preparing a piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative, characterized in that, Comprising the following steps: Add the first reactant to the solvent and the second reactant R2-CHO, and react to obtain a product in the presence of a catalyst; Among them, R1 is N-methylpiperazine or N-ethylpiperazine, R2 is phenyl or pyridyl, and the phenyl or pyridyl is substituted by one or more R a groups, and the R a is selected from one or more of a single or multiple fluoro groups, nitro groups, methyl groups, and trifluoromethyl groups.

6. The preparation method of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 5, characterized in that, The product is purified by acetone washing to obtain the purified product.

7. The preparation method of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 5, characterized in that, The solvent includes methanol.

8. The preparation method of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to claim 5, characterized in that, The catalyst is hydrochloric acid; and / or The reaction temperature is -5°C to 5°C.

9. A pharmaceutical composition, characterized in that, Comprising the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to any one of claims 1 to 4 or the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative prepared by the preparation method according to any one of claims 5 to 8.

10. Use of the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative according to any one of claims 1 to 4 or the piperazine-substituted 3,4-dihydro-1(2H)-naphthalenone derivative prepared by the preparation method according to any one of claims 5 to 8 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for treating tumors.

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