3,4-dihydro-1(2h)-naphthalenone derivatives, methods of preparation and uses

Abstract

The application discloses a 3,4-dihydro-1(2H)-naphthalenone derivative, a preparation method and application thereof. The 3,4-dihydro-1(2H)-naphthalenone derivative of the application shows good anti-inflammatory activity and can be used for treating inflammatory diseases such as rheumatoid arthritis.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a 3,4-dihydro-1(2H)-naphthol derivative, its preparation method, and its uses. Background Technology

[0002] Inflammation is a non-specific response of the body to external stimuli and a protective response of the immune system, playing an important role in clearing harmful stimuli and activating immune cells. However, excessive inflammatory responses can lead to inflammatory diseases such as gouty arthritis, type 2 diabetes, and Alzheimer's disease. The NLRP3 inflammasome is an important regulatory center for the inflammatory response. Under the stimulation of various inducers such as adenosine triphosphate (ATP), lipopolysaccharide (LPS), and pathogens, the N-terminal pryin domain of NOD-like receptor protein 3 (NLRP3) interacts with the pyrin domain of apoptosis-associated speckled protein (ASC) to form a protein scaffold for ASC nucleation. Subsequently, pro-caspase-1 is recruited to form the protein complex NLRP3 inflammasome in the cytoplasm through CARD-CARD interactions. Studies have shown that activation of the nuclear factor-κB (NF-κB) signaling pathway releases the NF-κB complex into the nucleus, where it binds to DNA, promoting NLRP3 protein transcription and inflammasome assembly. Activated inflammasomes stimulate macrophages to migrate to the contact site and produce TNF-α, IL-6, IL-18 and IL-1β by mediating the conversion of pro-caspase-1 into active caspase-1, thus promoting the inflammatory response. Therefore, studies using macrophage activation can reflect the inflammatory process.

[0003] In recent years, ferns have received widespread attention for their medicinal value. Among them, fern metabolites such as erythritol Z, inosine 1, and inosine 2 possess potent smooth muscle relaxant effects. However, their structural instability, low bioavailability, and false positives limit their clinical application. Therefore, based on their pharmacodynamic structure and literature reports, the imidazole ring has been modified by ring expansion, with 3,4-dihydro-1(2H)-naphthone (DHN) being a typical example. This modification exhibits good anti-inflammatory activity by inhibiting the assembly and activation of the NLRP3 inflammasome. For example, methoxy-substituted 2-(4-methoxy-3-(trifluoromethyl)benzyl)-3,4-dihydronaphth-1(2H)-one (6m, Figure 1 It exhibits potent anti-inflammatory activity by inhibiting phosphorylation of IκB1 and NF-κB p65 in the NF-κB signaling pathway, and downregulating the secretion of inflammatory mediators IL-6, IL-1β, and ROS. The amino-substituted L1 shows anti-edema and anti-inflammatory effects, inhibits mast cell degranulation, and has a therapeutic effect on arachidonic acid-induced ear edema in mice. Summary of the Invention

[0004] Purpose of the invention: This invention provides 3,4-dihydro-1(2H)-naphthone derivatives, their preparation methods, and uses to solve the problems of the prior art.

[0005] Technical solution: This invention provides a 3,4-dihydro-1(2H)-naphthol derivative, the structural formula of which is as follows:

[0006]

[0007] Wherein, R1 is a halogen, methoxy, or hydroxyl group, and R2 is a phenyl or pyridyl group, wherein the phenyl or pyridyl group is reacted with one or more R groups. a Group substitution, the R a Selected from nitrogen-containing heterocycles.

[0008] Preferably, the R a It is selected from one or more of N-methylpiperazine, morpholino, and imidazole.

[0009] Preferably, the R2 is selected from one or more of the following groups:

[0010]

[0011] This invention further provides a method for preparing 3,4-dihydro-1(2H)-naphthol derivatives, comprising the following steps:

[0012] Add reactant 1R2-CHO and reactant 2 to the solvent In the presence of a catalyst

[0013] Under these conditions, the reaction yields the product, and the reaction formula is:

[0014]

[0015] In some specific embodiments, the synthetic route of the 3,4-dihydro-1(2H)-naphthone derivative of the present invention is as follows: Figure 6 As shown. The substituent groups of compounds 6a-u, 7a-e, and 8a-n are shown below:

[0016]

[0017]

[0018] Preferably, the product is purified by silica gel column chromatography, wherein the solvent for column chromatography is dichloromethane:methanol or dichloromethane:petroleum ether in a volume ratio of 5 to 15:1, to obtain the purified product.

[0019] Preferably, the solvent includes methanol.

[0020] Preferably, the catalyst is sodium hydroxide.

[0021] Preferably, the reaction temperature described above is room temperature. In this application, the room temperature can be 20℃±5℃.

[0022] Preferably, the molar ratio of reactant 1 to reactant 2 is (0.8–1.2):(0.8–1.2).

[0023] The present invention further provides a pharmaceutical composition comprising the 3,4-dihydro-1(2H)-naphthone derivative described above or the 3,4-dihydro-1(2H)-naphthone derivative prepared by the preparation method described above.

[0024] The pharmaceutical composition of the present invention comprises one or more of the above-mentioned 3,4-dihydro-1(2H)-naphthone derivatives. The pharmaceutical composition of the present invention may be composed of one or more of the above-mentioned 3,4-dihydro-1(2H)-naphthone derivatives combined with other compounds, or may be composed of one or more of the above-mentioned 3,4-dihydro-1(2H)-naphthone derivatives.

[0025] The present invention further provides the application of the 3,4-dihydro-1(2H)-naphthone derivative or the 3,4-dihydro-1(2H)-naphthone derivative prepared by the preparation method described above in the preparation of anti-inflammatory drugs.

[0026] The present invention further provides the use of the 3,4-dihydro-1(2H)-naphthone derivative or the 3,4-dihydro-1(2H)-naphthone derivative prepared by the preparation method described above in the preparation of a drug for treating rheumatoid arthritis.

[0027] Terminology Explanation

[0028] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. Unless otherwise stated, all patent documents, publicly disclosed materials, etc., referenced in this invention are included in the full text of the references. If the same term has multiple definitions in this invention, the definition in this section shall prevail.

[0029] It is important to understand that the general description preceding the text and the detailed description following the text are merely illustrative and explanatory, and are not restrictive on any of the claims. It should be noted that in the specification and appended claims, unless otherwise stated in the text, singular forms such as "a," "an," and "this" include plural forms. It should also be noted that unless otherwise stated, "or" means "and / or." Furthermore, terms such as "comprising" and "including" are not restrictive.

[0030] "Substitution" refers to the replacement of hydrogen atoms by substituents.

[0031] The terms "halogen" and "halogenated" used in this invention refer to fluorine, chlorine, bromine, and iodine, with fluorine and bromine being preferred.

[0032] The term "nitrogen-containing heterocycle" as used in this invention refers to N-methylpiperazine, morpholino, and imidazole.

[0033] The term "pyridinyl" as used in this invention includes pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, etc.

[0034] Unless otherwise stated, "%" in this invention refers to mass percentage.

[0035] Beneficial effects: The 3,4-dihydro-1(2H)-naphthol derivative of the present invention exhibits good anti-inflammatory activity and can be used to treat inflammatory diseases. Attached Figure Description

[0036] Figure 1 The figures show the anti-inflammatory activity test results of compound DHN 7a in this application. Figure (A) shows the structural diagram of compound DHN 7a; Figure (B) shows the inhibitory effect of compound DHN 7a on anti-inflammatory factor IL-6; Figure (C) shows the inhibitory effect of compound DHN 7a on anti-inflammatory factor TNF-α; Figure (D) shows the inhibitory effect of compound DHN 7a on anti-inflammatory factor IL-1β; and Figure (E) shows the inhibitory effect of compound DHN 7a on anti-inflammatory factor IL-18. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.

[0037] Figure 2 The images show the flow cytometry results of cells after incubation with the compound DHN 7a. The top left image shows the results of the blank control sample, the top right image shows the results after incubation with DHN 7a at a concentration of 1.5 μM, the bottom left image shows the results after incubation with DHN 7a at a concentration of 3.0 μM, and the bottom right image shows the results after incubation with DHN 7a at a concentration of 6.0 μM.

[0038] Figure 3 The results of the test on inflammatory mediators by compound DHN 7a of this application are shown in Figure 1. (A) shows the inhibitory effect of DHN 7a on ROS production in LPS-stimulated RAW246.7 cells, (B) shows the statistical analysis of the number of DCF-stained RAW264.7 cells by DHN 7a, and (C) shows the inhibitory effect of DHN 7a on NO production in LPS-stimulated RAW246.7 cells.

[0039] Figure 4 The effect of the compound of this application on the nuclear translocation of p65 protein of LPS in RAW246.7 cells by DHN 7a;

[0040] Figure 5 The results of this application's compound DHN 7a inhibiting the activation and expression of NF-κB and NLRP3 inflammasome-related proteins are shown in Figure (A). Figure (A) shows the effect of compound DHN 7a on the expression levels of inflammasome-related proteins. Figures (C) to (D) show the expression results of p-IκBα and IκBα, p-p65 and p65. Figures (B) and (E) show the expression results of NLRP3 inflammasome-related NLRP3 and ASC proteins induced by LPS in RAW264.7 cells. Compared with the LPS group: *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001;

[0041] Figure 6 This is a synthetic route diagram for the 3,4-dihydro-1(2H)-naphthone derivative of this application. Detailed Implementation

[0042] I. Experimental Materials and Testing Methods

[0043] 1.1 Chemical reagents were purchased from Leyan Technology Co., Ltd. (Beijing) and Bid Pharmaceutical Technology Co., Ltd. (Shanghai).

[0044] Reactant 1 in this application can be obtained by existing technology or synthesized in the laboratory. Taking 4-(4-methylpiperazin-1-yl)benzaldehyde as an example, the synthesis method is as follows:

[0045]

[0046] N-methylpiperazine (16.0256 g, 0.16 mol) and K₂CO₃ (27.64 g, 0.20 mol) were weighed and placed in a 100 mL three-necked flask. 30 mL of LDM was added, and the mixture was stirred at 25 °C for 6 hours. 4-fluorobenzaldehyde (2.482 g, 0.02 mol) was slowly added dropwise under nitrogen protection. The reaction was carried out at 100 °C and refluxed for 4-5 hours. The reaction system was filtered, and the filtrate was washed with distilled water and saturated brine. The solvent was removed by concentration under reduced pressure. The residue was eluented with dichloromethane:methanol (15:1, v / v) and column chromatography was used to obtain the pale yellow target product 4-(4-methylpiperazin-1-yl)benzaldehyde, with a yield of 79.2%. The reactant 1 used in the synthesis of compounds 6a-u, 7a-e, and 8a-n in this application can be obtained by the same synthetic method, except that the reactants are replaced with reactants corresponding to the corresponding groups of the compounds.

[0047] 1.2 Test Methods

[0048] (1) In dimethyl sulfoxide (DMSO-d6) solvent with tetramethylsilane (TMS) as an internal standard, using 1H NMR, Bruker Avance 400MHz or 600MHz spectrometer and 13 Spectra were recorded using C-NMR spectrometers at 100 MHz or 150 MHz using a Bruker Avance spectrometer. Chemical shifts (δ) are expressed in ppm, and coupling constants (J) in Hz. All melting factors were measured using a digital melting factor analyzer.

[0049] (2) Cytotoxicity assay: Mouse RAW264.7 cells were provided by Wuhan Pronosei Biotechnology Co., Ltd., and cultured in complete medium containing DMEM, 10% fetal bovine serum, and 1% streptomycin / penicillin antibiotics. Cells in the logarithmic growth phase were used for the following assays. Cytotoxicity was detected using CCK-8 assay according to the manufacturer's instructions. Cells were grown and treated in 96-well plates and incubated with CCK-8 reagent at 37°C for 1 hour. Absorbance was measured at 450 nm.

[0050] (3) Anti-inflammatory activity assay: RAW264.7 cells were randomly grouped. Cells were re-exposed for 22 h in the presence of 1.0 μg / mL LPS. After treatment, cells were collected and analyzed. TNF-α and IL-6 were measured according to the ELISA double-antibody sandwich kit instructions. Cells were re-exposed for 22 h in the presence of 1.0 μg / mL LPS, ATP was added to a final concentration of 5.0 mM, and incubated for 30 min, with the remaining steps being the same. IL-1β and IL-18 were measured according to the ELISA double-antibody sandwich kit instructions. After the assay, the absorbance at 450 nm was measured on an ELISA instrument. A standard curve was established for each measurement, with units of pg / mL.

[0051] (4) Western blotting experiment: RAW264.7 cells were placed in 2.0 × 10⁶ cells per well of a 6-well plate. 6Cells were exposed to the drug for 2 hours, followed by exposure to 1.0 μg / mL LPS for 24 hours. Lysis was performed using potent RIPA lysis buffer containing a phosphatase inhibitor (RIPA lysis buffer: phosphatase inhibitor = 99:1). After sonication on ice, the cells were centrifuged. Once protein concentration was determined, the protein solution was boiled. Protein samples were then subjected to electrophoresis on a 10% polyacrylamide gel. The proteins were transferred to a PVDF membrane and incubated with 5% skim milk for 2 hours. Subsequently, they were incubated overnight at 4°C with antibodies against IκB-α, p-IκB-α, p65, p-p65, NLRP3, ASC, and GAPDH (antibodies purchased from Cell Signaling Technology, USA). Incubation with goat anti-rabbit IgG / HRP antibody (Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 1.0 h was performed. Testing was performed using ECL developing solution (Shanghai Noren Biomedical Technology Co., Ltd.). Protein intensity was detected using ImageLab software and analyzed using ImageJ software.

[0052] (5) Apoptosis experiment: RAW264.7 cells were injected with 2×10⁻⁶ cells. 5 Cells were seeded at a density of cells / well in 12-well plates. They were then treated with 4kJ (0.275, 0.55, and 1.1 μM) for 24 hours. Cells were collected in centrifuge tubes and washed twice with pre-chilled PBS. After centrifugation with PBS, the cells were discarded. Cells were resuspended in 200 mL of binding buffer. Then, 5 mL of Annexin V-FITC and 5 mL of propidium iodide (BD Biosciences, San Jose, CA, USA) were added sequentially to the solution. Apoptosis was detected by flow cytometry (BD FACS Califur). Finally, the cells were analyzed by flow cytometry.

[0053] (6) RAW246.7 was converted to 2×10 5 Cells were seeded at a density in laser confocal culture dishes and incubated for 2 hours in a thermostatic incubator containing compound 7a (1.5, 3.0, and 6.0 μM), followed by incubation with the inducer LPS (1.0 μg / mL) for 2 hours. Cells were washed, fixed with 4% polybenzaldehyde at 37°C for 20 minutes, and then infiltrated for 30 minutes with 500 μL of 0.3% Triton-X100. Cells were then blocked with 5% BSA at room temperature for 2 hours and incubated overnight at 4°C with the primary antibody NF-κB / p65 (1:200). Cells were washed with PBS and then incubated with the secondary antibody (1:1000) for 30 minutes. Unbound secondary antibody was washed off, and the cells were then incubated in DAPI at room temperature in the dark for 30 minutes. Cells were then imaged using a Zeiss laser confocal microscope.

[0054] (7) Measurement of reactive oxygen species (ROS) and nitric oxide (NO) production: LPS (1.0 μg / mL) stimulation was used to detect ROS and NO production in RAW246.7 cells using a reactive oxygen species assay kit and Gress assay. Cells were seeded in 12-well plates, 1 × 10⁶ cells per well. 5 Cells were treated with 7a and LPS (1.0 μg / mL) for 24 hours. Cells were collected and resuspended by adding 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), incubated at 37°C, 5% CO2, and 95% humidity for 30 minutes, and analyzed by flow cytometry. Cells were treated in the same manner for 24 hours. The supernatant was collected, centrifuged, and equal volumes of Griess reagents A and B were mixed and added to the supernatant. OD was measured on a UV spectrophotometer at 540 nm after 15 minutes. A standard curve was established for the measurements, in μM.

[0055] II. Compound Synthesis

[0056] Example 1: Synthesis of compounds 6a-i, 6m-u, 7a-c, 7e, 8a-f and 8i-n

[0057] Compound 6a (DHN 6a):

[0058] (E)-7-bromo-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0059] 4-(4-methylpiperazin-1-yl)benzaldehyde (2.04 g, 0.01 mol) and 7-bromo-3,4-dihydronaphthyl-1(2H)-one (2.25 g, 0.01 mol) were dissolved in 3 mL of methanol and stirred in an ice-water bath. Then, 0.5 mL of 25% NaOH solution was added, and the reaction was carried out at room temperature with stirring for 40 min, monitored by TLC. After the reaction was complete, the suspension was filtered, and the residue was dissolved in dichloromethane, washed with distilled water and saturated brine, and the solvent was removed by pouring. The residue was then dissolved in dichloromethane and purified by silica gel column chromatography (dichloromethane:methanol = 20:1, v / v). Finally, the product was dissolved in 5 mL of dichloromethane and 5 mL of methanol and slowly evaporated at room temperature to obtain compound 6a as a yellow single crystal, with a yield of 78.6%.

[0060] MP: 87~89℃;

[0061] 1H NMR (600MHz, DMSO-d6) δ7.99(d,J=2.2Hz,1H),7.74(dd,J=8.1,2.3Hz,1H),7.67(d,J=2.1Hz,1H),7.45(d,J=8.6Hz,2H),7.36(d,J=8.2Hz ,1H),7.00(d,J=8.6Hz,2H),3.27(t,J=5.1Hz,4H),3.10(td,J=6.6,1.9Hz,2H),2.89(t,J=6.6Hz,2H),2.45(t,J=5.0Hz,4H),2.23(s,3H).

[0062] 13 C NMR(150MHz,DMSO-d6)δ185.72,151.73,142.73,137.93,135.99,135.49,132.45,1 31.38,131.25,129.90,125.10,120.38,114.70,54.82,47.38,46.19,27.64,26.88.

[0063] Compound 6b (DHN 6b):

[0064] (E)-7-bromo-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0065] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde. The resulting product was a yellow powder with a yield of 77.3%.

[0066] MP: 113~115℃;

[0067] 1 H NMR (600MHz, DMSO-d6) δ7.99(d,J=2.2Hz,1H),7.74(dd,J=8.1,2.2Hz,1H),7.69(s,1H),7.48(d,J=8.6Hz,2H),7.36(d,J=8 .2Hz,1H),7.01(d,J=8.5Hz,2H),3.74(t,J=4.8Hz,4H),3.23(t,J=4.9Hz,4H),3.11(t,J=6.8Hz,2H),2.90(t,J=6.6Hz,2H).

[0068] 13C NMR(150MHz,DMSO-d6)δ185.28,151.40,142.29,137.35,135.55,134.99,131.9 2,131.05,130.93,129.43,125.10,119.91,114.10,65.92,47.29,27.16,26.40.

[0069] Compound 6c (DHN 6c):

[0070] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-7-bromo-3,4-dihydronaphthalen-1(2H)-one

[0071] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl). The product obtained was a yellow powder with a yield of 73.5%.

[0072] MP: 119~121℃;

[0073] 1 H NMR (600MHz, DMSO-d6) δ8.37(s,1H),8.03(d,J=2.2Hz,1H),7.84(t,J=1.5Hz,1H),7.78(d,J=8.4Hz,4H) ,7.70(d,J=8.6Hz,2H),7.39(d,J=8.2Hz,1H),7.15(s,1H),3.12(t,J=5.6Hz,2H),2.94(t,J=6.5Hz,2H).

[0074] 13 C NMR(150MHz,DMSO-d6)δ186.00,143.11,137.41,136.47,136.06,135.94,135.47,135 .05,133.95,132.14,131.60,130.53,130.05,120.59,120.50,118.34,27.67,26.81.

[0075] Compound 6d (DHN 6d):

[0076] (E)-7-fluoro-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-on e

[0077] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 83.5%.

[0078] MP: 77~78℃;

[0079] 1 H NMR (600MHz, Chloroform-d) δ7.94(d,J=1.9Hz,1H),7.87(dd,J=9.2,2.8Hz,1H),7.52(d,J=8.8Hz,2H),7.35–7.29(m,1H),7.27(td,J=8.3,2. 8Hz,1H),7.03(d,J=8.9Hz,2H),3.45(t,J=5.1Hz,4H),3.25(td,J=6.6,1.9Hz,2H),3.01(t,J=6.5Hz,2H),2.73(t,J=5.0Hz,4H),2.50(s,3H).

[0080] 13 C NMR(150MHz,Chloroform-d)δ186.88,161.89(d,J=245.3Hz),151.18,138.68(d,J=3.1Hz),137.96,135.45(d,J=5.7Hz),13 1.72, 129.78 (d, J = 7.5Hz), 126.35, 120.10 (d, J = 21.8Hz), 114.84, 114.12 (d, J = 21.9Hz), 54.78, 47.79, 45.97, 28.04, 27.27.

[0081] Compound 6e (DHN 6e):

[0082] (E)-7-fluoro-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0083] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 78.4%.

[0084] MP: 87~89℃;

[0085] 1 H NMR(600MHz,Chloroform-d)δ7.84(s,1H),7.78(dd,J=9.2,2.8Hz,1H),7.44(d,J=8.8Hz,2H),7.25–7.20(m,1H),7.18(td,J=8.2, 2.8Hz, 1H), 6.99 (d, J = 7.9Hz, 2H), 3.91 (t, J = 4.8Hz, 4H), 3.28 (t, J = 4.9Hz, 4H), 3.14 (td, J = 6.9, 1.8Hz, 2H), 2.91 (t, J = 6.6Hz, 2H).

[0086] 13 C NMR(150MHz,Chloroform-d)δ186.97,162.02(d,J=246.2Hz),138.81(d,J=2.8Hz),137.73,135.49(d,J=6.5Hz),132 .39,131.97,129.95(d,J=6.7Hz),120.33(d,J=21.9Hz),115.13,114.27(d,J=22.6Hz),66.65,48.83,28.16,27.37.

[0087] Compound 6f (DHN 6f):

[0088] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-7-fluoro-3,4-dihydronaphthalen-1(2H)-one

[0089] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl), and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 80.2%.

[0090] MP: 99~101℃;

[0091] 1H NMR(600MHz,DMSO-d6)δ8.36(s,1H),7.83(s,1H),7.78(s,1H),7.77(s,2H),7.70(d,J=8.4Hz,2H), 7.65(d,J=9.7Hz,1H),7.49–7.44(m,2H),7.14(s,1H),3.12(t,J=6.5Hz,2H),2.95(t,J=6.4Hz,2H).

[0092] 13 C NMR (150MHz, DMSO-d6) δ186.30, 161.62 (d, J = 243.4Hz), 140.13 (d, J = 3.1Hz), 137.39, 136.06, 135.84, 135.51, 134.93 (d, J = 6.5 Hz), 133.97, 132.11, 131.48 (d, J = 7.5Hz), 130.60, 121.23 (d, J = 21.9Hz), 120.58, 118.32, 113.39 (d, J = 21.9Hz), 27.50, 27.07.

[0093] Compound 6g (DHN 6g):

[0094] (E)-7-methoxy-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0095] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 42%.

[0096] MP: 76~78℃;

[0097] 1H NMR(600MHz,Chloroform-d)δ7.82(s,1H),7.61(d,J=2.8Hz,1H),7.42(d,J=8.8Hz,2H),7.15(d,J=8.3Hz,1H),7.05(dd,J=8.3,2.9Hz,1H),6. 93(d,J=8.9Hz,2H),3.87(s,3H),3.33(t,J=5.1Hz,4H),3.14(td,J=6.6,1.8Hz,2H),2.88(t,J=6.2Hz,2H),2.61(t,J=5.0Hz,4H),2.39(s,3H).

[0098] 13 C NMR(150MHz,Chloroform-d)δ187.26,158.13,150.57,136.72,135.21,134.11,132.05,13 1.25,128.77,126.08,120.63,114.33,109.81,55.06,54.32,47.40,45.52,27.44,26.99.

[0099] Compound 6h (DHN 6h):

[0100] (E)-7-methoxy-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0101] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 50.3%.

[0102] Mp: 111~112℃;

[0103] 1H NMR(600MHz,Chloroform-d)δ7.83(s,1H),7.61(d,J=2.7Hz,1H),7.44(d,J=8.8Hz,2H),7.15(d,J=8.4Hz,1H),7.06(dd,J=8.3,2.8H z,1H),6.93(d,J=8.3Hz,2H),3.88(t,J=4.9Hz,4H),3.87(s,3H),3.25(t,J=5.0Hz,4H),3.14(t,J=6.5Hz,2H),2.88(t,J=6.6Hz,2H).

[0104] 13 C NMR(150MHz,Chloroform-d)δ187.78,158.66,151.14,137.08,135.73,134.61,132.85 ,131.74,129.31,127.10,121.20,114.64,110.35,66.72,55.58,48.40,27.96,27.51.

[0105] Compound 6i (DHN 6i):

[0106] (E)-7-methoxy-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0107] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl), and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 78.6%.

[0108] MP: 90~92℃;

[0109] 1H NMR(600MHz,Chloroform-d)δ7.96(s,1H),7.85(s,1H),7.63(d,J=2.9Hz,1H),7.56(d,J=8.5Hz,2H),7.46(d,J=8.5Hz,2H),7.33(s, 1H),7.25(s,1H),7.18(d,J=8.3Hz,1H),7.09(dd,J=8.3,2.9Hz,1H),3.88(s,3H),3.12(td,J=6.5,1.8Hz,2H),2.92(t,J=6.3Hz,2H).

[0110] 13 C NMR(150MHz,Chloroform-d)δ187.55,158.79,136.95,136.39,135.84,135.42,135.30,134 .98,134.13,131.47,130.43,129.54,121.77,121.26,118.10,110.38,55.60,27.98,27.45.

[0111] Example 2: Synthesis of compounds 6j-l, 7d and 8g-h

[0112] Compound 6j (DHN 6j):

[0113] (E)-7-hydroxy-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0114] 4-(4-methylpiperazin-1-yl)benzaldehyde (2.04 g, 0.01 mol) and 7-hydroxy-1-tetrahydronaphthone (1.62 g, 0.01 mol) were placed in a 25 mL single-necked round-bottom flask, and 2 mL of MeOH was added. The mixture was stirred in an ice-water bath, and 0.5 mL of 25% sodium hydroxide solution was added dropwise. The reaction progress was monitored by thin-layer chromatography. The reaction time was approximately 25 min. The reaction was then terminated, and the mixture was passed successively through a mixed solution of n-hexane and acetone, and then through a mixed solution of dichloromethane and petroleum ether. The mixture was sonicated for 20 min, and the system was filtered. The filter residue was purified by column chromatography (DCM:MeOH = 15:1). The solvent was removed under vacuum to obtain a yellow powder with a yield of 76.4%.

[0115] MP: 160~161℃;

[0116] 1H NMR (600MHz, DMSO-d6) δ9.58(s,1H),7.61(s,1H),7.43(d,J=8.6Hz,2H),7.31(d,J=2.7Hz,1H),7.17(d,J=8.2Hz,1H),6.99(d,J=8.8Hz,2H) ,6.96(dd,J=8.2,2.7Hz,1H),3.25(t,J=5.1Hz,4H),3.05(td,J=6.5,1.8Hz,2H),2.80(t,J=6.5Hz,2H),2.45(t,J=5.0Hz,4H),2.22(s,3H).

[0117] 13 C NMR(150MHz,DMSO-d6)δ185.86,155.55,150.44,135.70,133.46,133.11,131.27,1 31.07,128.89,124.36,120.30,113.69,111.97,53.77,46.42,45.13,26.48,26.41.

[0118] Compound 6k (DHN 6k):

[0119] (E)-7-hydroxy-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0120] The synthesis method was the same as in Example 2, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinebenzaldehyde. The resulting product was a yellow powder with a yield of 78.9%.

[0121] Mp: 142~144℃;

[0122] 1 H NMR (600MHz, DMSO-d6) δ9.59(s,1H),7.63(s,1H),7.45(d,J=8.3Hz,2H),7.32(s,1H),7.17(d,J=8.2Hz,1H),7.00(d,J=8. 3Hz, 2H), 6.97 (d, J = 8.3Hz, 1H), 3.74 (t, J = 4.9Hz, 4H), 3.21 (t, J = 4.9Hz, 4H), 3.06 (t, J = 6.6Hz, 2H), 2.80 (t, J = 6.3Hz, 2H).

[0123] 13C NMR(150MHz,DMSO-d6)δ186.98,156.65,151.66,136.69,134.53,134.22,132.6 1,132.11,129.99,125.94,121.43,114.63,113.06,66.41,47.88,27.56,27.49.

[0124] Compound 6l (DHN 6l):

[0125] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-7-hydroxy-3,4-dihydronaphthalen-1(2H)-one

[0126] The synthesis method was the same as in Example 2, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl). The product obtained was a yellow powder with a yield of 77.6%.

[0127] Mp: 194~196℃;

[0128] 1 H NMR (600MHz, DMSO-d6) δ9.66(s,1H),8.35(s,1H),7.83(s,1H),7.76(d,J=8.7Hz,2H),7.71(d,J=2.0Hz,1H),7.67(d,J=8.7Hz,2H),7.3 5(d,J=2.7Hz,1H),7.20(d,J=8.2Hz,1H),7.14(s,1H),7.00(dd,J=8.2,2.7Hz,1H),3.07(td,J=6.5,1.8Hz,2H),2.84(t,J=6.4Hz,2H).

[0129] 13 C NMR(150MHz,DMSO-d6)δ186.66,156.27,136.75,135.94,135.60,134.44,134.12,133 .78,133.68,131.50,130.10,129.76,121.48,120.12,117.88,112.60,27.05,27.02.

[0130] Compound 6m (DHN 6m):

[0131] (E)-6-bromo-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0132] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 74.8%.

[0133] Mp: 81~83℃;

[0134] 1 H NMR (600MHz, DMSO-d6) δ7.83(d,J=8.3Hz,1H),7.65(d,J=11.8Hz,2H),7.59(dd,J=8.3,2.0Hz,1H),7.45(d,J=8.6Hz,2H),7. 00(d,J=8.8Hz,2H),3.27(t,J=5.1Hz,4H),3.10(t,J=5.7Hz,2H),2.93(t,J=6.6Hz,2H),2.46(t,J=5.2Hz,4H),2.24(s,3H).

[0135] 13 C NMR(150MHz,DMSO-d6)δ186.21,151.66,145.80,137.53,132.81,132.39,131.55,1 31.46,130.51,129.86,127.62,125.19,114.74,54.80,47.37,46.14,27.84,26.97.

[0136] Compound 6n (DHN 6n):

[0137] (E)-6-bromo-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0138] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 68.6%.

[0139] MP: 99~101℃;

[0140] 1 H NMR(600MHz,Chloroform-d)δ7.77(s,1H),7.70(dd,J=9.2,2.8Hz,1H),7.36(d,J=8.8Hz,2H),7.15(dd,J=8.4,5.2Hz,1H),7.09(td,J= 8.3, 2.8Hz, 1H), 6.86 (d, J = 8.5Hz, 2H), 3.80 (t, J = 4.9Hz, 4H), 3.18 (t, J = 4.8Hz, 4H), 3.07 (td, J = 6.6, 1.8Hz, 2H), 2.83 (t, J = 6.5Hz, 2H).

[0141] 13 C NMR(150MHz,Chloroform-d)δ187.33,145.01,137.93,132.95,132.19,132.18,132 .17,131.32,131.31,130.74,130.24,128.41,115.10,66.97,48.80,28.85,27.42.

[0142] Compound 6o (DHN 6o):

[0143] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-6-bromo-3,4-dihydronaphthalen-1(2H)-one

[0144] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl), and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 72.8%.

[0145] MP: 85~86℃;

[0146] 1H NMR (600MHz, DMSO-d6) δ8.36(s,1H),7.88(d,J=8.3Hz,1H),7.83(t,J=1.4Hz,1H),7.78(s,1H),7.76(d,J=5.8Hz,2H),7. 70(s,1H),7.68(d,J=2.8Hz,2H),7.62(dd,J=8.3,2.0Hz,1H),7.14(s,1H),3.12(t,J=5.6Hz,2H),2.97(t,J=6.5Hz,2H).

[0147] 13 C NMR(150MHz,DMSO-d6)δ186.45,146.11,137.38,136.06,135.70,135.59,133.98,132 .39,132.10,131.69,130.68,130.61,130.02,128.16,120.57,118.31,27.86,26.89.

[0148] Compound 6p (DHN 6p):

[0149] (E)-6-fluoro-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0150] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 69.3%.

[0151] MP: 79~81℃;

[0152] 1 H NMR(600MHz,Chloroform-d)δ8.14(dd,J=8.7,6.0Hz,1H),7.83(s,1H),7.41(d,J=8.8Hz,2H),7.02(td,J=8.6,2.6Hz,1H),6.93(d,J=8.8H z,2H),6.91(d,J=2.6Hz,1H),3.33(t,J=5.1Hz,4H),3.16(td,J=6.6,1.8Hz,2H),2.92(t,J=6.5Hz,2H),2.60(t,J=5.0Hz,4H),2.38(s,3H).

[0153] 13 C NMR (150MHz, DMSO-d6) δ186.67,165.63(d,J=254.1Hz),151.32,146.11(d,J=8.7Hz),137.66,132.03,131.91,131.32(d,J=9 .1Hz),130.54(d,J=3.1Hz),126.46,114.93,114.61(d,J=21.3Hz),114.57(d,J=21.8Hz),54.97,48.02,46.19,29.00,27.30.

[0154] Compound 6q (DHN 6q):

[0155] (E)-6-fluoro-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0156] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 78.2%.

[0157] MP: 85~87℃;

[0158] 1 H NMR (600MHz, DMSO-d6) δ7.69(s,1H),7.61(dd,J=9.3,2.5Hz,1H),7.48(d,J=8.9Hz,2H),7.46–7.40(m,2H),7. 01(d,J=8.9Hz,2H),3.74(t,J=4.9Hz,4H),3.23(t,J=4.8Hz,4H),3.11(t,J=5.7Hz,2H),2.91(t,J=6.5Hz,2H).

[0159] 13C NMR (150MHz, DMSO-d6) δ186.07,161.61(d,J=243.9Hz),151.85,139.76,137.71,135.36(d,J=6.4Hz),132.35,131. 61,131.23(d,J=7.5Hz),125.63,120.73(d,J=21.9Hz),114.59,113.27(d,J=21.8Hz),66.40,47.78,27.47,27.14.

[0160] Compound 6r (DHN 6r):

[0161] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-6-fluoro-3,4-dihydronaphthalen-1(2H)-one

[0162] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl), and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 71.3%.

[0163] MP: 119~121℃;

[0164] 1 H NMR (600MHz, Chloroform-d) δ8.17(dd,J=8.7,5.9Hz,1H),7.99(s,1H),7.86(d,J=1.8Hz,1H),7.56(d,J=8.5Hz,2H),7.47(d,J=8. 5Hz,2H),7.34(s,1H),7.06(td,J=8.5,2.6Hz,1H),6.95(dd,J=8.9,2.5Hz,1H),3.14(td,J=6.5,1.8Hz,2H),2.97(t,J=6.5Hz,2H).

[0165] 13C NMR (150MHz, Chloroform-d) δ 186.26, 165.81 (d, J = 256.1Hz), 146.12 (d, J = 8.7Hz), 137.00, 135.83, 135.38, 135.27, 135.17, 131. 49(d,J=9.7Hz),131.48,130.28,129.93(d,J=3.1Hz),121.32,118.12,114.82(d,J=21.9Hz),114.75(d,J=21.7Hz),28.86,27.10.

[0166] Compound 6s (DHN 6s):

[0167] (E)-6-methoxy-2-(4-(4-methylpiperazin-1-yl)benzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0168] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 57.3%.

[0169] MP: 73~74℃;

[0170] 1 H NMR (600MHz, DMSO-d6) δ7.86(d,J=8.6Hz,1H),7.57(s,1H),7.40(d,J=8.6Hz,2H),6.96(d,J=8.7Hz,2H),6.89(dd,J=8.7,2.5Hz,1H),6. 85(d,J=2.5Hz,1H),3.80(s,3H),3.70(t,J=5.1Hz,4H),3.17(t,J=5.0Hz,4H),3.03(t,J=6.5Hz,2H),2.86(t,J=6.6Hz,2H),2.46(s,3H).

[0171] 13 C NMR (150MHz, DMSO-d6) δ186.08,158.18,150.65,138.68,135.56,133.96,133.68,132. 55,129.74,120.55,120.52,110.19,106.32,65.90,55.29,44.72,40.06,27.01,26.92.

[0172] Compound 6t (DHN 6t)

[0173] (E)-6-methoxy-2-(4-morpholinobenzylidene)-3,4-dihydronaphthalen-1(2H)-one

[0174] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 4-morpholinobenzaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 65.8%.

[0175] MP: 87~89℃;

[0176] 1 H NMR (600MHz, DMSO-d6) δ7.90(d,J=8.6Hz,1H),7.61(s,1H),7.44(d,J=8.6Hz,2H),7.00(d,J=8.7Hz,2H),6.93(dd,J=8.7,2.5Hz,1 H), 6.89 (d, J = 2.5Hz, 1H), 3.84 (s, 3H), 3.74 (t, J = 4.9Hz, 4H), 3.21 (t, J = 4.8Hz, 4H), 3.07 (t, J = 6.5Hz, 2H), 2.90 (t, J = 6.6Hz, 2H).

[0177] 13 C NMR (150MHz, DMSO-d6) δ185.78,163.52,151.55,146.13,136.05,132.70,132.00,130. 23,127.09,126.08,114.70,114.02,112.71,66.42,55.99,47.95,40.52,28.70,27.28.

[0178] Compound 6u (DHN 6u):

[0179] (E)-2-(4-(1H-imidazol-1-yl)benzylidene)-6-methoxy-3,4-dihydronaphthalen-1(2H)-one

[0180] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 4-(1H-imidazol-1-yl), and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 71.5%.

[0181] MP: 104~106℃;

[0182] 1 H NMR (600MHz, DMSO-d6) δ8.35(s,1H),7.94(d,J=8.6Hz,1H),7.82(s,1H),7.76(d,J=8.8Hz,2H),7.69(s,1H),7.66(d,J=8.3Hz, 2H),7.14(s,1H),6.96(dd,J=8.7,2.5Hz,1H),6.92(d,J=2.5Hz,1H),3.85(s,3H),3.09(t,J=6.3Hz,2H),2.93(t,J=6.5Hz,2H).

[0183] 13 C NMR(150MHz,DMSO-d6)δ185.80,163.83,146.49,137.15,136.44,136.06,134.33,13 1.90,130.57,130.43,126.76,120.58,118.34,114.26,112.83,56.05,28.71,27.22.

[0184] Compound 7a (DHN 7a):

[0185] (E)-7-bromo-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphthal en-1(2H)-one

[0186] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl) was replaced with 5-(4-methylpiperazin-1-yl)pyridinecarboxaldehyde. The product obtained was a yellow powder with a yield of 70.2%.

[0187] Mp: 78~79℃;

[0188] 1H NMR(600MHz,Chloroform-d)δ8.40(d,J=3.0Hz,1H),8.23(d,J=2.1Hz,1H),7.68(s,1H),7.57(dd,J=8.1,2.2Hz,1H),7.35(d,J=8.7Hz,1 H),7.14(dd,J=8.6,3.2Hz,2H),3.62(t,J=6.5Hz,2H),3.36(t,J=5.1Hz,4H),2.92(t,J=6.5Hz,2H),2.61(t,J=5.1Hz,4H),2.38(s,3H).

[0189] 13 C NMR(150MHz,Chloroform-d)δ187.16,145.57,145.35,142.64,137.41,135.69,135.19,13 4.95,134.45,130.94,130.03,128.43,120.92,120.78,54.56,47.38,46.06,28.25,26.26.

[0190] Compound 7b (DHN 7b):

[0191] (E)-7-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphthal en-1(2H)-one

[0192] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 5-(4-methylpiperazin-1-yl)pyridinecarboxaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 64.3%.

[0193] MP: 65~66℃;

[0194] 1H NMR(600MHz,Chloroform-d)δ8.40(d,J=3.0Hz,1H),7.78(dd,J=9.2,2.8Hz,1H),7.68(s,1H),7.35(d,J=8.6Hz,1H),7.24(dd,J=8.5,5.3Hz,1 H),7.17(ddd,J=19.1,8.5,2.9Hz,2H),3.62(t,J=6.4Hz,2H),3.36(t,J=5.1Hz,4H),2.94(t,J=6.5Hz,2H),2.62(t,J=5.0Hz,4H),2.39(s,3H).

[0195] 13 C NMR (150MHz, DMSO-d6) δ 186.29, 161.11 (d, J = 243.7Hz), 145.65, 143.63, 140.08 (d, J = 2.2Hz), 136.95, 134.78 (d, J = 6.4Hz), 134.2 7,133.82,130.91(d,J=7.4Hz),128.72,120.42,120.41(d,J=21.8Hz),112.91(d,J=21.8Hz),54.17,46.50,45.77,27.10,26.09.

[0196] Compound 7c (DHN 7c):

[0197] (E)-7-methoxy-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0198] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 5-(4-methylpiperazin-1-yl)pyridinecarboxaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 70.1%.

[0199] Mp: 78~79℃;

[0200] 1H NMR (600MHz, DMSO-d6) δ8.45(d,J=3.0Hz,1H),7.54(d,J=1.8Hz,1H),7.51(d,J =8.7Hz,1H),7.44(d,J=2.9Hz,1H),7.33(dd,J=8.8,3.1Hz,1H),7.30(d,J=8.4H z,1H),7.16(dd,J=8.3,2.8Hz,1H),3.81(s,3H),3.54(td,J=6.6,1.8Hz,2H),3 .33(t,J=5.4Hz,4H),2.87(t,J=6.5Hz,2H),2.46(t,J=5.1Hz,4H),2.23(s,3H).

[0201] 13 C NMR (150MHz, DMSO-d6) δ187.41,158.59,145.98,144.31,137.36,136.76,135.02,134.40,134.05,130.26,12 8.86,121.05,120.93,110.66,55.75,54.61,46.97,46.19,27.51,26.84.124.26,56.45(d,J=6.3Hz),45.56.

[0202] Compound 7d (DHN 7d):

[0203] (E)-7-hydroxy-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0204] The synthesis method was the same as in Example 2, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 5-(4-methylpiperazin-1-yl)pyridinecarboxaldehyde. The resulting product was a yellow powder with a yield of 62.1%.

[0205] MP: 143~144℃;

[0206] 1H NMR (600MHz, DMSO-d6) δ9.59(s,1H),8.44(d,J=3.0Hz,1H),7.50(d,J=1.9Hz,1H),7.48(d,J=8.7Hz,1H),7.33(dd,J=8.5,3.0Hz,2H),7.18(d,J=8. 2Hz,1H),6.97(dd,J=8.2,2.7Hz,1H),3.51(td,J=6.5,1.8Hz,2H),3.31(d ,J=4.7Hz,4H),2.82(t,J=6.5Hz,2H),2.45(t,J=5.1Hz,4H),2.22(s,3H).

[0207] 13 C NMR(150MHz,DMSO-d6)δ186.50,155.52,144.88,143.32,136.28,134.17,133.91,133.3 1,132.74,129.00,127.67,120.47,119.89,112.00,53.55,45.94,45.14,26.46,25.87.

[0208] Compound 7e (DHN 7e):

[0209] (E)-6-bromo-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphthal en-1(2H)-one

[0210] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 5-(4-methylpiperazin-1-yl)pyridinecarboxaldehyde, and 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one. The product obtained was a yellow powder with a yield of 65.5%.

[0211] MP: 103~105℃;

[0212] 1H NMR (600MHz, Chloroform-d) δ8.40(d,J=3.0Hz,1H),7.98(d,J=8.3Hz,1H),7.67(t,J=1.4Hz,1H),7.48(dd,J=8.4,1.9Hz,1H),7.44(d,J=1.9Hz,1H),7.34( d,J=8.7Hz,1H),7.14(dd,J=8.6,3.0Hz,1H),3.63(td,J=6.6,1.8Hz,2H),3.3 5(t,J=4.9Hz,4H),2.94(t,J=6.5Hz,2H),2.61(t,J=5.0Hz,4H),2.38(s,3H).

[0213] 13 C NMR(150MHz,Chloroform-d)δ187.63,145.66,145.59,145.39,137.42,135.14,134.22,13 2.50,131.10,130.25,129.96,128.38,128.14,120.94,54.59,47.43,46.10,28.53,26.34.

[0214] Compound 8a (DHN 8a):

[0215] (E)-7-bromo-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphthal en-1(2H)-one

[0216] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The resulting product was a yellow powder with a yield of 70.8%.

[0217] MP: 75~77℃;

[0218] 1H NMR (600MHz, DMSO-d6) δ8.36(d,J=2.4Hz,1H),7.99(d,J=2.2Hz,1H),7.78(dd,J=9.1,2.5Hz,1H),7.74(dd,J=8.1,2.2Hz,1H),7.64(t,J=1.8Hz,2H),7. 37(d,J=8.2Hz,1H),6.91(d,J=9.0Hz,1H),3.61(t,J=4.9Hz,4H),3.08(td,J =6.6,1.8Hz,2H),2.90(t,J=6.5Hz,2H),2.41(t,J=5.0Hz,4H),2.23(s,3H).

[0219] 13 C NMR(150MHz,DMSO-d6)δ185.05,158.22,151.13,142.33,138.73,135.61,134.93,134.7 0,131.39,130.95,129.46,119.95,119.91,106.35,54.27,45.69,44.16,27.12,26.47.

[0220] Compound 8b (DHN 8b):

[0221] (E)-7-bromo-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0222] The synthesis method was the same as in Example 1, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The resulting product was a yellow powder with a yield of 62.8%.

[0223] MP: 97~98℃;

[0224] 1H NMR (600MHz, DMSO-d6) δ8.38(d,J=2.5Hz,1H),7.99(d,J=2.2Hz,1H),7.81(dd,J=9.0,2.5Hz,1H),7.75(dd,J=8.1,2.3Hz,1H),7.66(d,J=1.9Hz,1 H),7.37(d,J=8.2Hz,1H),6.91(d,J=8.9Hz,1H),3.70(t,J=4.8Hz,4H),3.57(t,J=4.9Hz,4H),3.09(td,J=6.6,1.9Hz,2H),2.91(t,J=6.5Hz,2H).

[0225] 13 C NMR(150MHz,DMSO-d6)δ185.05,158.38,150.94,142.33,138.76,135.63,134.88,13 4.56,131.67,130.95,129.46,120.34,119.93,106.33,65.89,44.69,27.09,26.44.

[0226] Compound 8c (DHN 8c):

[0227] (E)-7-fluoro-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0228] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The product obtained was a yellow powder with a yield of 71.0%.

[0229] MP: 70~71℃;

[0230] 1H NMR (600MHz, DMSO-d6) δ8.36(d,J=2.5Hz,1H),7.78(dd,J=9.0,2.5Hz,1H),7.65(s,1H),7.60(dd,J=9.3,2.7Hz,1H),7.48–7.31(m, 2H), 6.91 (d, J = 9.0Hz, 1H), 3.60 (t, J = 5.0Hz, 4H), 3.08 (t, J = 6.3Hz, 2H), 2.92 (t, J = 6.5Hz, 2H), 2.39 (t, J = 5.0Hz, 4H), 2.22 (s, 3H).

[0231] 13 C NMR (150MHz, DMSO-d6) δ185.32, 161.12 (d, J = 243.5Hz), 158.21, 151.04, 139.29 (d, J = 3.2Hz), 138.68, 134.79 (d, J = 6.1Hz), 134.5 6,131.43,130.76(d,J=7.5Hz),120.29(d,J=22.1Hz),119.90,112.81(d,J=21.8Hz),106.31,54.31,45.76,44.20,26.93,26.71.

[0232] Compound 8d (DHN 8d):

[0233] (E)-7-fluoro-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0234] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-fluoro-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The product obtained was a yellow powder with a yield of 73.1%.

[0235] MP: 106~108℃;

[0236] 1H NMR(600MHz,Chloroform-d)δ8.37(d,J=2.4Hz,1H),7.82–7.70(m,2H),7.65(dd,J=8.9,2.4Hz,1H),7.23(dd,J=8.4,5.2Hz,1H),7.18(td, J=8.2,2.8Hz,1H),6.67(d,J=9.0Hz,1H),3.83(t,J=4.9Hz,4H),3.62(t,J=4.9Hz,4H),3.13(td,J=6.6,1.8Hz,2H),2.93(t,J=6.6Hz,2H).

[0237] 13 C NMR (150MHz, Chloroform-d) δ 186.96, 161.99 (d, J = 245.4Hz), 151.39, 138.79 (d, J = 3.2Hz), 137.92, 135.52 (d, J = 6.5Hz), 132.14 ,132.05,131.96,129.92(d,J=6.9Hz),126.87,120.25(d,J=22.6Hz),114.69,114.22(d,J=21.9Hz),66.80,48.39,28.13,27.36.

[0238] Compound 8e (DHN 8e):

[0239] (E)-7-methoxy-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0240] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The product obtained was a yellow powder with a yield of 69.3%.

[0241] MP: 79~81℃;

[0242] 1H NMR(600MHz,Chloroform-d)δ8.36(d,J=2.4Hz,1H),7.75(d,J=1.7Hz,1H),7.62(dd ,J=8.9,2.5Hz,1H),7.60(d,J=2.8Hz,1H),7.26(s,1H),7.15(d,J=8.3Hz,1H),7.06( dd,J=8.3,2.8Hz,1H),6.67(d,J=8.9Hz,1H),3.87(s,3H),3.68(t,J=4.9Hz,4H),3. 11(td,J=6.5,1.8Hz,2H),2.89(t,J=6.5Hz,2H),2.56(t,J=5.1Hz,4H),2.38(s,3H).

[0243] 13 C NMR(150MHz,Chloroform-d)δ187.54,158.81,158.52,150.72,139.06,135.76,134.62,134.44,1 33.13,129.44,121.38,121.27,110.49,106.25,55.71,54.86,46.22,44.83,29.84,28.01,27.75.

[0244] Compound 8f (DHN 8f):

[0245] (E)-7-methoxy-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0246] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The product obtained was a yellow powder with a yield of 59.3%.

[0247] MP: 69~70℃;

[0248] 1H NMR (600MHz, DMSO-d6) δ8.36(d,J=2.4Hz,1H),7.79(dd,J=9.0,2.5Hz,1H),7.62(s,1H),7.41(d,J=2.8Hz,1H),7.30(d,J=8.4Hz,1H),7.15(dd,J=8 .3,2.9Hz,1H),6.90(d,J=8.9Hz,1H),3.80(s,3H),3.70(t,J=4.9Hz,4H), 3.55(t,J=4.9Hz,4H), 3.05(td,J=6.5,1.8Hz,2H), 2.86(t,J=6.5Hz,2H).

[0249] 13 C NMR(150MHz,DMSO-d6)δ186.11,158.34,158.19,150.66,138.69,135.58,133.97,133.7 0,132.56,129.75,120.57,120.53,110.21,106.33,65.91,55.30,44.74,27.03,26.93.

[0250] Compound 8g (DHN 8g):

[0251] (E)-7-hydroxy-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0252] The synthesis method was the same as in Example 2, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The resulting product was a yellow powder with a yield of 55.3%.

[0253] MP: 147~148℃;

[0254] 1H NMR (600MHz, DMSO-d6) δ9.58(s,1H),8.32(d,J=2.5Hz,1H),7.74(dd,J=9.0,2.5Hz,1H),7.58(s,1H),7.31(d,J=2.7Hz,1H),7.16(d,J=8.3Hz,1H),6.9 6(dd,J=8.2,2.8Hz,1H),6.89(d,J=9.0Hz,1H),3.58(t,J=5.1Hz,4H),3.02 (t,J=6.6Hz,2H),2.80(t,J=6.4Hz,2H),2.39(t,J=5.1Hz,4H),2.22(s,3H).

[0255] 13 C NMR(150MHz,DMSO-d6)δ186.22,158.13,156.17,150.71,138.58,133.97,133.76,133.5 1,132.47,129.53,120.99,120.14,112.58,106.31,54.31,45.76,44.23,27.13,26.94.

[0256] Compound 8h (DHN 8h):

[0257] (E)-7-hydroxy-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0258] The synthesis method was the same as in Example 2, except that 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The resulting product was a yellow powder with a yield of 53.7%.

[0259] Mp: 102~103℃;

[0260] 1H NMR (600MHz, DMSO-d6) δ9.60(s,1H),8.35(d,J=2.5Hz,1H),7.78(dd,J=9.0,2.5Hz,1H),7.60(d,J=2.0Hz,1H),7.32(d,J=2.7Hz,1H),7.18(d,J=8.2H z,1H),6.97(dd,J=8.2,2.8Hz,1H),6.90(d,J=9.0Hz,1H),3.70(t,J=4.9H z, 4H), 3.55 (t, J = 4.9Hz, 4H), 3.04 (t, J = 5.9Hz, 2H), 2.82 (t, J = 6.5Hz, 2H).

[0261] 13 C NMR(150MHz,DMSO-d6)δ186.73,158.78,156.67,151.04,139.11,134.42,134.27,13 3.87,133.26,130.02,121.51,121.10,113.07,106.78,66.37,45.21,27.60,27.42.

[0262] Compound 8i (DHN 8i):

[0263] (E)-6-bromo-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0264] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The product obtained was a yellow powder with a yield of 75.7%.

[0265] MP: 96~98℃;

[0266] 1H NMR(600MHz,Chloroform-d)δ8.34(d,J=2.3Hz,1H),7.96(d,J=8.4Hz,1H),7.75(s,1H),7.61(dd,J=8.9,2.5Hz,1H),7.48(dd,J=8.4,2.0Hz,1H),7.42 (d,J=1.9Hz,1H),6.66(d,J=8.9Hz,1H),3.68(t,J=5.1Hz,4H),3.12(td,J= 6.5,1.8Hz,2H),2.92(t,J=6.6Hz,2H),2.55(t,J=5.1Hz,4H),2.37(s,3H).

[0267] 13 C NMR(150MHz,Chloroform-d)δ186.98,158.79,151.12,144.90,139.28,135.29,132.87,13 2.49,131.31,130.77,130.24,128.46,121.18,106.46,55.05,46.42,45.00,28.75,27.52.

[0268] Compound 8j (DHN 8j):

[0269] (E)-6-bromo-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0270] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-bromo-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The product obtained was a yellow powder with a yield of 58.2%.

[0271] MP: 117~118℃;

[0272] 1H NMR(600MHz,Chloroform-d)δ8.36(d,J=2.4Hz,1H),7.96(d,J=8.3Hz,1H),7.75(s,1H),7.64(dd,J=8.9,2.4Hz,1H),7.49(dd,J=8.4,1.9H z,1H),7.43(s,1H),6.67(d,J=8.9Hz,1H),3.83(t,J=4.9Hz,4H),3.62(t,J=4.8Hz,4H),3.12(td,J=6.5,1.8Hz,2H),2.93(t,J=6.5Hz,2H).

[0273] 13 C NMR(150MHz,DMSO-d6)δ186.73,158.68,150.71,144.68,139.18,134.82,132.5 9,131.13,130.60,130.05,128.33,121.42,106.21,66.78,45.37,28.53,27.32.

[0274] Compound 8k (DHN 8k):

[0275] (E)-6-fluoro-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphthal en-1(2H)-one

[0276] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The product obtained was a yellow powder with a yield of 69.5%.

[0277] MP: 76~77℃;

[0278] 1H NMR (600MHz, DMSO-d6) δ8.34(d,J=2.4Hz,1H),7.99(dd,J=8.6,6.1Hz,1H),7.76(dd,J=9.0,2.5Hz,1H),7.62(s,1H),7.28–7.17(m,2H ), 6.90 (d, J = 9.0Hz, 1H), 3.60 (t, J = 4.9Hz, 4H), 3.08 (td, J = 6.6, 1.9Hz, 2H), 2.95 (t, J = 6.5Hz, 2H), 2.40 (t, J = 5.0Hz, 4H), 2.22 (s, 3H).

[0279] 13 C NMR(150MHz,DMSO-d6)δ185.47,165.26(d,J=252.4Hz),158.65,151.34,147.01(d,J=9.8Hz),139.12,134.47,132.27,131 .07(d,J=9.8Hz),130.51,120.47,115.22(d,J=21.7Hz),114.82(d,J=21.9Hz),106.80,54.75,46.19,44.66,28.17,27.10.

[0280] Compound 8l (DHN 8l):

[0281] (E)-6-fluoro-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0282] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 6-fluoro-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The product obtained was a yellow powder with a yield of 72.5%.

[0283] Mp: 91~92℃;

[0284] 1H NMR (600MHz, DMSO-d6) δ8.37(d,J=2.5Hz,1H),8.00(dd,J=8.7,6.0Hz,1H),7.79(dd,J=9.0,2.5Hz,1H),7.63(s,1H),7.30–7.16 (m,2H),6.91(d,J=8.9Hz,1H),3.70(t,J=4.9Hz,4H),3.56(t,J=5.0Hz,4H),3.09(td,J=6.6,1.9Hz,2H),2.95(t,J=6.6Hz,2H).

[0285] 13 C NMR(150MHz,DMSO-d6)δ185.50,165.28(d,J=252.0Hz),158.83,151.19,147.05(d,J=9.0Hz),139.18,134.35,132.57, 131.09(d,J=9.9Hz),130.50,120.93,115.23(d,J=21.7Hz),114.84(d,J=21.9Hz),106.80,66.37,45.19,28.17,27.10.

[0286] Compound 8m (DHN 8m):

[0287] (E)-6-methoxy-2-((6-(4-methylpiperazin-1-yl)pyridin-3-yl)methylene)-3,4-dihydronaphth alen-1(2H)-one

[0288] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-(4-methylpiperazin-1-yl)nicotinaldehyde. The product obtained was a yellow powder with a yield of 69.7%.

[0289] MP: 85~87℃;

[0290] 1H NMR (600MHz, DMSO-d6) δ8.32(s,1H),7.90(d,J=8.6Hz,1H),7.74(d,J=9.0Hz,1H),7.57(s,1H),6.93(dd,J=8.6,2.5Hz,1H),6.89(d ,J=8.9Hz,2H),3.84(s,3H),3.58(t,J=4.8Hz,4H),3.05(t,J=6.3Hz,2H),2.91(t,J=6.5Hz,2H),2.39(t,J=4.9Hz,4H),2.22(s,3H).

[0291] 13 C NMR(150MHz,DMSO-d6)δ185.50,163.55,158.58,151.02,146.15,139.03,133.34,133.05,1 30.25,127.02,120.69,114.03,112.73,106.78,55.99,54.77,46.22,44.71,28.63,27.33.

[0292] Compound 8n (DHN 8n):

[0293] (E)-6-methoxy-2-((6-morpholinopyridin-3-yl)methylene)-3,4-dihydronaphthalen-1(2H)-one

[0294] The synthesis method was the same as in Example 1, except that 7-bromo-3,4-dihydronaphthyl-1(2H)-one was replaced with 7-methoxy-3,4-dihydronaphthyl-1(2H)-one, and 4-(4-methylpiperazin-1-yl)benzaldehyde was replaced with 6-morpholinoaldehyde. The product obtained was a yellow powder with a yield of 62.7%.

[0295] MP: 79~80℃;

[0296] 1H NMR (600MHz, DMSO-d6) δ8.34(d,J=2.4Hz,1H),7.90(d,J=8.6Hz,1H),7.77(dd,J=9.0,2.5Hz,1H),7.58(s,1H),6.94(dd,J=8.6,2.6Hz ,1H),6.92–6.88(m,2H),3.84(s,3H),3.70(t,J=4.8Hz,4H),3.55(t,J=4.9Hz,4H),3.06(td,J=6.6,1.9Hz,2H),2.91(t,J=6.5Hz,2H).

[0297] 13 C NMR(150MHz,DMSO-d6)δ185.50,163.57,158.75,150.87,146.18,139.08,133.35,133.22,130.27, 126.99,121.18,114.05,112.75,106.79,66.38,56.00,45.23,28.63,27.33.HRMS(ESI):calcd.for C 21 H 22 N₂O₃, [M+H] + ,351.1708;found:351.1709.

[0298] III. Results Evaluation

[0299] 3.1 Cytotoxic and anti-inflammatory activities

[0300] In vitro cytotoxicity was assessed using the CCK-8 assay. All experiments were performed in triplicate, and the mean survival rate was used. The survival rates of RAW264.7 cells treated with compounds 6a-u, 7a-e, and 8a-n (DHNs, 2.5 μM) are shown in Table 1. After treatment with 2.5 μM DHNs, the survival rate of RAW264.7 cells was greater than 80%, demonstrating that the synthesized DHNs had no significant toxic effect on RAW264.7 cells.

[0301] TNF-α and IL-6 are key pro-inflammatory factors inducing inflammatory responses, apoptosis, and immune cell activation. LPS stimulation transforms macrophages into the M1 phenotype, leading to the secretion of inflammatory factors and triggering an inflammatory response. The level of inflammatory factors can serve as an indicator of the severity of the inflammatory response. This study used ELISA to detect LPS-pretreated experimental cell lines. The NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) was used as a positive control. The average inhibition rates are shown in Table 2. At a concentration of 2.5 μM, all DHNs effectively inhibited LPS-induced production of TNF-α and IL-6 in RAW264.7 cells. Compounds 6a, 7a, 8a, and 8g showed inhibition rates of 49.8%, 81.4%, 51.0%, and 55.9% against TNF-α, respectively, significantly higher than the positive control PDTC. The order of anti-inflammatory activity was 7a > 8g > 8a > 6a. Furthermore, most synthetic DHN derivatives also showed significant inhibitory effects on IL-6. PDTC (10 μM) inhibited IL-6 secretion by 48.0%. Encouragingly, 11 compounds showed stronger inhibitory effects than PDTC, with compound 7a exhibiting the strongest inhibition at 69.2%.

[0302] Table 1. Results of the detection of the survival rate of RAW264.7 cells by compounds 6a-u, 7a-e, and 8a-n.

[0303]

[0304] Table 2. Results of anti-inflammatory activity tests for compounds 6a-u, 7a-e, and 8a-n.

[0305]

[0306]

[0307]

[0308] PDTC stands for: Ammonium pyrrolidine dithiocarbamate, CAS number: 5108-96-3

[0309] First, using the 3,4-dihydronaphthyl-1(2H)-one group as a constant main active center, different substituents were introduced to explore their structure-activity relationships. We found that the 7-OH-substituted DHN 6j-l exhibited moderate anti-inflammatory activity, with inhibition rates of 56.3%, 48.9%, and 50.2% against the cytokine IL-6, respectively. To obtain compounds with better anti-inflammatory activity, the inventors introduced a methoxy group at site 7 to obtain 6g-i. Unfortunately, compared with 6j-l, their anti-inflammatory activity was not significantly improved, with inhibition rates against IL-6 only 50.1%, 47.2%, and 55.3%, respectively.

[0310] Secondly, considering the electron-donating properties of methoxy and hydroxyl groups, we attempted to introduce electron-withdrawing substituents to obtain bromine- and fluorine-substituted DHN 6a-f. Notably, the inhibitory activity of 6a substituted with 7-Br and N-methylpiperazine was significantly enhanced. The inhibition rates of TNF-α and IL-6 were as high as 49.8% and 62.6%, respectively, superior to 6d, 6g, and 6j. However, the inhibition rate of 6b was significantly reduced when the N-methyl group was substituted with an isotopic oxygen. Interestingly, the inhibition rates of 6c substituted with imidazole for TNF-α and IL-6 were 39.8% and 55.3%, respectively.

[0311] Third, DHN 8a-h were synthesized by substituting the benzene ring with a 3-pyridine substituent. After screening for anti-inflammatory activity, we found that the inhibitory effects of 8a and 8g substituted with 7-Br, 7-OH, and N-methylpiperazine were significantly improved, with IL-6 inhibition rates of 63.4% and 66.6%, respectively. The anti-inflammatory effects of morphine-substituted 8b, 8d, 8f, and 8h were almost indistinguishable from those of 6b, 6e, 6h, and 6k. After replacing the 3-pyridine group with a 2-pyridine group (7a-d), their anti-inflammatory activities were unexpectedly significantly enhanced, especially the 7-Br and N-methylpiperazine-substituted compounds 7a, which showed the strongest anti-inflammatory activity compared to the phenyl-substituted compound 6a and the 3-pyridine-substituted compound 8a. The inhibition rate of compound 7a against the cytokine TNF-α increased from 49.8% and 51.0% to 81.4%, respectively.

[0312] To investigate the effect of substitution position on the anti-inflammatory activity of DHN derivatives, we synthesized compounds 6m-u, 7e, and 8i-n, which were substituted with electron-donating substituents (-OCH3) or electron-withdrawing substituents (-Br, -F). The results showed that the 6-Br and N-methylpiperazine substituted compounds 6m, 7e, 8i, and 6-OCH3, as well as the N-methylpiperazole substituted compound 8m, exhibited high inhibition rates of IL-6 release, exceeding 53%. However, compared to compound 7a, these structural changes reduced the anti-inflammatory activity.

[0313] As shown in Table 2, the 3,4-dihydronaphthyl-1(2H)-one prepared in this invention exhibits significant anti-inflammatory activity as the parent nucleus, and the presence of the naphthalene ring increases the structural stability of the molecule. 7-Br substitution showed a more significant inhibitory effect than other substituents. The anti-inflammatory activity of nitrogen-containing heterocyclic compounds was N-methylpiperazine > imidazole > morpholine. Notably, the anti-inflammatory activity was significantly enhanced when the benzene ring was substituted with 2-pyridyl or 3-pyridyl groups. SAR analysis showed that compound 7a, with 7-Br and (5-(4-methylpiperazin-1-yl)pyridin-2-yl)methylene substitution, exhibited the strongest inhibitory effect on cytokines IL-6 and TNF-α, significantly stronger than the positive control PDTC, and lower cytotoxicity. Comprehensive analysis indicates that DHN 7a can serve as a lead compound for further research.

[0314] The anti-IL-6 and anti-TNF-α effects of compound 7a, such as Figure 1 As shown in Figures (B) and (C), IL-6 expression was inhibited by 48.0% after exposure to PDTC (10 μM). The lead compound L10 (2.5 μM), with good anti-inflammatory activity, was used as a control. Post-exposure inhibition rate decreased to 44.2%, lower than PDTC. Post-exposure inhibition rate of compound DHN 7a increased to 69.2%, superior to L10 and PDTC. LPS- and ATP-stimulation increased the secretion of inflammatory cytokines IL-18 and IL-1β. Figure 1 As shown in Figures (D) and (E), compound 7a, in contrast, reversed the release of macrophage inflammatory cytokines TNF-α, IL-6, IL-18, and IL-1β under stimulation with low toxic concentrations of LPS in a dose-dependent manner. This indicates that compound 7a can inhibit the production of multiple inflammatory cytokines, thereby exerting an anti-inflammatory effect.

[0315] The results of this experiment show that compound 7a can effectively reduce the concentrations of TNF-α and IL-6, and has a good anti-inflammatory effect.

[0316] 3.2 Effect of compound 7a on apoptosis in RAW246.7 cells

[0317] The Annexin V-FITC / PI apoptosis kit was used to detect the apoptosis pattern induced by compound 7a in RAW246.7 cells. Annexin V is a FITC-labeled protein that binds to phosphatidylserine (PS) on the surface of apoptotic cell membranes, thereby detecting apoptotic cells. Furthermore, in late-stage apoptotic or necrotic cells, propidium iodide (PI) can cross the cell membrane and bind to DNA in the cell nucleus due to loss of cell membrane integrity. This kit was used to detect late-stage apoptotic or necrotic cells. The interaction between these two substances leads to differentiation at different apoptotic stages. Detection was performed using flow cytometry (FITC) channels. Figure 2As shown, almost no cell apoptosis was observed in the blank group. After treatment with different doses of compound 7a (1.5, 3.0, and 6.0 μM), the cell apoptosis rates in the three dose groups were 3.8%, 5.6%, and 6.8%, respectively. The apoptosis rate of RAW264.7 cells was dose-dependent with increasing dose, which also indicates that compound 7a has low toxicity to RAW246.7 cells.

[0318] 3.3 Compound 7a inhibits the production of NO and ROS.

[0319] RAW246.7 cells stimulated by the inducer LPS underwent a series of cellular physiological and pathological changes, including mitochondrial dysfunction, increased oxidative stress, and lysosomal destruction, leading to excessive intracellular ROS and nitric oxide (NO) production. This, in turn, induced the activation of NLRP3 inflammatory vesicles and exacerbated the inflammatory response. We used a 2,7-dichlorodiacetate (DCFH-DA) fluorescent probe to monitor ROS production. During oxidative stress, DCFH-DA was oxidized to dichlorofluorescein (DCF) by intracellular reactive oxygen species, detected by flow cytometry (FACS). ROS production was quantified by counting the number of cells containing DCF. Figure 3 As shown in Figure (A). Furthermore, to assess the amount of NO produced, we used the Griess reagent method. By converting NO to nitrite and reacting it with N,N-dimethylphenylsulfonamide in the Griess reagent to produce a deep purple organic dye, the intensity of which is proportional to the NO concentration, we assessed the amount of intracellular NO produced by measuring the absorbance of the product. (See Figure A). Figure 3 Chinese (B) map and Figure 3 As shown in Figure (C), after LPS stimulation, the levels of ROS and NO in the model group increased, but compound 7a could inhibit the activation of NLRP3 inflammatory vesicles by dose-dependently scavenging intracellular ROS and nitric oxide production.

[0320] 3.4 Compound 7a inhibits LPS-induced NF-κB nuclear translocation in RAW246.7 cells.

[0321] As previously mentioned, activation of the NF-κB signaling pathway releases p65 from the cytoplasm, which translocates to the nucleus and binds to DNA, thereby mediating NLRP3 transcription and the production of inflammatory mediator precursors, accelerating the inflammatory response. To investigate the inhibitory effect of compound 7a on NF-κB nuclear translocation, we used immunofluorescence to detect p65 nuclear translocation in RAW246.7 cells. Dimethyl sulfoxide was used as a control. Figure 4As shown, most p65 subunits remained in the cytoplasm in the control group. After stimulation of RAW246.7 cells with LPS (1.0 μg / mL), most of the p65 subunits in the experimental cells translocated to the nucleus. However, pretreatment with compound 7a (6.0 μM) concurrently with stimulation resulted in the blocking of p65 nuclear translocation in RAW246.7 cells, with most p65 subunits remaining in the cytoplasm. These results indicate that compound 7a can block the nuclear translocation of p65 subunits, suggesting that compound 7a can inhibit NLRP3 transcription and the production of inflammatory mediator precursors, thus possessing potential anti-inflammatory activity.

[0322] 3.5DHN 7a inhibits the activation of NF-κB and NLRP3.

[0323] The NLRP3 inflammasome plays a crucial role in the body's immune and inflammatory responses. Its activation primarily occurs through two pathways. First, under inflammatory stimulation, IκBα phosphorylation leads to ubiquitination and degradation by proteases. The proteases release the NF-κB / Rel complex, which translocates to the nucleus and binds to DNA, mediating NLRP3 inflammasome activation. Second, under the stimulation of various inducers, NOD-like receptor protein 3 (NLRP3), apoptosis-associated speckled protein (ASC), and procysteine ​​protease-1 (caspase-1) assemble into the NLRP3 inflammasome. This assembly process activates the active factor of pro-caspase-1, thereby converting pro-caspase-1 into active caspase-1 via protease cleavage. These processes collectively lead to the activation of the NLRP3 inflammasome, triggering an inflammatory response. Therefore, we verified whether compound 7a exerts its anti-inflammatory effect by inhibiting NF-κB activation and NLRP3 inflammasome activation, such as by suppressing the phosphorylation levels of IκBα and p65 and the expression of NLRP3 inflammasome-related proteins including NLRP3 and ASC. Figure 5 As shown in Figure (A). Figure 5Figures (B) to (E) show that RAW246.7 cells were treated with different concentrations of compound 7a (1.5 μM, 3.0 μM, and 6.0 μM), followed by stimulation with LPS (1.0 μg / mL) after 2 hours. Cells were collected for Western blot analysis after 22 hours. The control group showed low phosphorylation expression. LPS stimulation significantly upregulated the phosphorylation expression of IκBα and p65 proteins in RAW246.7 cells, indicating that the dose-dependent inhibition of IκBα and p65 phosphorylation by compound 7a (1.5, 3.0, and 6.0 μM) inhibited the activation of the NF-κB signaling pathway. NLRP3 and ASC are well-known components of the NLRP3 inflammasome. Therefore, investigating the relationship between NLRP3 and ASC is crucial to determining whether compound 7a exerts its anti-inflammatory effect by inhibiting the NLRP3 inflammasome. Within 30 minutes of LPS stimulation, the protein expression levels of NLRP3 and ASC in RAW246.7 cells were significantly increased. Pretreatment with different concentrations of compound 7a (1.5, 3.0, and 6.0 μM) significantly reduced the expression levels of NLRP3 and ASC. These results indicate that compound 7a inhibits the assembly of the NLRP3 inflammasome by suppressing the expression of NLRP3 and ASC. In summary, compound 7a exerts its anti-inflammatory activity by inhibiting the NF-κB signaling pathway through downregulation of p65 and IκBα phosphorylation levels, and by inhibiting the assembly of the inflammasome, namely the expression of NLRP3 and ASC, thereby inhibiting the activation of the NLRP3 inflammasome.

[0324] The results above show that the 3,4-dihydro-1(2H)-naphthol derivative of the present invention has potential anti-inflammatory effects and is expected to become a potential multifunctional drug for the clinical treatment of inflammatory diseases.

Claims

1. A 3,4-dihydro-1(2H)-naphthone derivative, the structural formula of which is as follows: 。 2. A method for preparing the 3,4-dihydro-1(2H)-naphthone derivative as described in claim 1, characterized in that, Includes the following steps Add to solvent and Under the condition of a catalyst, the reaction yields the product, and the reaction formula is: Where R2 is R1 is a bromine group; When R2 is R1 is a bromine group or a hydroxyl group.

3. The preparation method according to claim 2, characterized in that, The product was purified by silica gel column chromatography, wherein the solvent for column chromatography was dichloromethane and methanol in a volume ratio of (5~15):1 or dichloromethane and petroleum ether in a volume ratio of (1~10):1, to obtain the purified product.

4. The preparation method according to claim 2, characterized in that, The solvent is methanol; the catalyst is sodium hydroxide.

5. A pharmaceutical preparation, characterized in that, Includes the 3,4-dihydro-1(2H)-naphthone derivative as described in claim 1 or the 3,4-dihydro-1(2H)-naphthone derivative prepared by the preparation method as described in any one of claims 2 to 4.

6. The use of the 3,4-dihydro-1(2H)-naphthone derivative as described in claim 1 or the 3,4-dihydro-1(2H)-naphthone derivative prepared by any one of the preparation methods described in claims 2 to 4 in the preparation of anti-inflammatory drugs.

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