Application of a nitrogen-containing heterocyclic compound in the preparation of antibacterial drugs

Abstract

The present invention belongs to the field of pharmacy, and is specifically an application of a nitrogen-containing heterocyclic compound in the preparation of an antibacterial drug. The compound of the present invention is prepared by electrochemical driving under metal-free and oxidant-free conditions, is a yellow solid, and a high-resolution mass spectrum gives a quasi-molecular ion peak [M+Na]+: 632.1101. The application involved is about the application of (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring in the preparation of an antibacterial drug.

Description

[0001] This application is a divisional application of application number 202410317783.X, application date March 20, 2024, and the invention name at the time of application is “(E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azepine ring”. Technical Field

[0002] The invention belongs to the field of pharmacy, and specifically relates to a (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring compound and application thereof. Background Art

[0003] Nitrogen-containing heterocyclic compounds are an important class of heterocyclic compounds, which are inseparable from people's lives and occupy an important position in the fields of food, medicine, agricultural chemistry, etc., and the eight-membered ring skeleton is widely present in a variety of natural products and biologically active molecules. This special structure has a high application value in drug preparation. The present invention studies the activity of the product on the basis of the synthesis method disclosed in CN202211259321.4, and finds that (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring compound has certain anti-tumor and antibacterial activities. Summary of the invention

[0004] The object of the present invention is to provide a (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring compound and its application.

[0005] The technical solution to solve the above technical problems is as follows:

[0006] The preparation method of (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine (3m) prepared by electrochemical driving under metal-free and oxidant-free conditions is as follows:

[0007] At room temperature, N-(but-3-en-1-yl)-N-(4-chloro-2-(1-(2-fluorophenyl)vinyl)phenyl)-4-methylbenzenesulfonamide 1m (0.3mmol), TsSePh 2a (1.1equiv, 0.33mmol), Ru(bpy)3Cl2·6H2O (0.006mmol) and DCM (5.0mL) were added to the reaction bottle in sequence, and a fan was provided outside the reaction bottle. The photolysis was carried out under an air atmosphere at 25°C using a 3W blue LED until the starting material was completely consumed (TLC monitoring, about 12h). After the reaction was completed, the mixture was quenched with NaHCO3 (sat.aq. 15mL) and extracted with CH2Cl2 (3×5mL). The organic solvent was then concentrated in vacuo. The residue was purified by flash column chromatography using ethyl acetate and petroleum ether as eluents (volume ratio of petroleum ether to ethyl acetate was 5:1) to obtain (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azepine (3m).

[0008]

[0009] The use of (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring compound in the preparation of anti-tumor drugs, the structural formula of which is shown in Formula 3m:

[0010]

[0011] The use of (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine ring compound in the preparation of antibacterial drugs, the structural formula of which is shown in Formula 3m:

[0012]

[0013] Beneficial effects of the present invention: The (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine prepared by electrochemical driving under metal-free and oxidant-free conditions has anti-tumor and antibacterial activities, providing a lead compound for the research and development of new anti-tumor drugs and antibacterial drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 For compound 3m 1 H NMR spectrum;

[0015] Figure 2 For compound 3m 13 C NMR spectrum;

[0016] Figure 3 For compound 3m 19 F NMR spectrum;

[0017] Figure 4 This is the high-resolution mass spectrum of compound 3m. DETAILED DESCRIPTION

[0018] The present invention is further described in detail below in conjunction with embodiments so that those skilled in the art can implement the invention with reference to the description.

[0019] Example 1

[0020] The method for preparing (E)-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[b]azocine (3m) by electrochemical drive under metal-free and oxidant-free conditions is as follows:

[0021] N-(But-3-en-1-yl)-N-(4-chloro-2-(1-(2-fluorophenyl)vinyl)phenyl)-4-methylbenzenesulfonamide (1m) (0.3mmol), TsSePh (2a) (1.1equiV, 0.33mmol), Ru(bpy)3Cl2·6H2O (0.006mmol) and DCM (5.0mL) were added to the reaction bottle in sequence at room temperature, and a fan was provided outside the reaction bottle. The photolysis was carried out under air atmosphere at 25°C using a 3W blue LED until the starting material was completely consumed (TLC monitoring, about 12h). After the reaction was completed, the mixture was quenched with NaHCO3 (sat.aq. 15mL) and extracted with CH2Cl2 (3×5mL). The organic solvent was then concentrated in vacuo. The residue was purified by flash column chromatography using ethyl acetate and petroleum ether as eluents (the volume ratio of petroleum ether to ethyl acetate was 5:1) to give the product 3m with a yield of 66.9%.

[0022]

[0023] According to compound 3m 1 H NMR, 13 C NMR, 19 The structure of the compound was determined by F NMR spectroscopy ( Figure 1 , Figure 2 , Figure 3 ). Specifically: Compound 3m was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:2, Rf = 0.35), yellow solid (122.3 mg, 66.9%): mp: 103-104 °C. 1H NMR (500MHz, CDCl3) δ7.64 (t, J=6.9Hz, 4H), 7.26-7.20 (m, 7H), 7.08-6.99 (m, 3H), 6.84 (d, J=8.5Hz, 1H), 5.99 (d, J=8.6Hz, 1H), 4.21 (d, J=13.5Hz, 1H), 3.29-3.18 (m, 2H), 2.91 (t, J=12.6Hz, 1H), 2.40 (d, J=8.6Hz, 6H), 2.3 1(t, J=15.0Hz, 1H), 1.93 (d, J=13.4Hz, 1H), 1.79 (td, J=14.0, 4.3Hz, 1H); 13 C NMR(126MHz, CDCl3)δ 161.29, 159.31, 144.91, 143.71, 142.92, 138.61, 137.14, 136.14, 135.37 (d, J=6.5Hz), 134.28, 132.59, 130.85, 130.32, 129.89, 12 9.69, 129.41, 129.12 (d, J=8.5Hz), 127.91, 127.66, 124.09 (d, J=3.4Hz), 115.85, 115.67, 61.81, 50.47, 34.25, 32.26, 21.62, 21.57; 19 FNMR (471MHz, CDCl3) δ-115.22. High-resolution mass spectrometry determined that its molecular formula is C 32 H 29 CIFNO4S2, mass-to-nuclear ratio m / z: 632.1103[M+Na]+( Figure 4 ).

[0024] Example 2

[0025] Anti-tumor experiment:

[0026] Human lung cancer cells A549 and human breast cancer cells MCF-7 were cultured in RPMI-1640 medium containing 10% fetal bovine serum; human liver cancer cells HepG-2, human colon cancer cells HCT-116 and human gastric cancer cells MGC-803 were cultured in DMEM medium containing 10% fetal bovine serum, digested and subcultured using trypsin, and cells in the logarithmic growth phase were selected for experiments.

[0027] The drug was dissolved in DMSO and prepared into a 1 mg / mL stock solution, which was stored at -80°C for future use. Before the experiment, the prepared culture medium was used to dilute the solution to the concentration required for the experiment (the final DMSO concentration was <0.01%).

[0028] The MTT colorimetric method was used to detect the inhibitory effect of compound 3m on the proliferation of various tumor cells. The test cells in the logarithmic growth phase were taken and (5-8)×10 4 The cells were inoculated at a density of 100 μL / mL in a 96-well culture plate, with 100 μL in each well. After incubation for 24 hours, 10 μL of compound 3m with final concentrations of 5, 10, 20, 40, and 80 μmol / L was added to each well. The positive control was cisplatin, and a blank control was set up. Each group had 6 replicate wells. After culturing in a 37°C constant temperature CO2 incubator for 48 hours, 10 μL of MTT solution (5 mg / mL) was added to each well and continued to be cultured in the incubator for 4 hours. The supernatant was discarded, 150 μL of DMSO was added to each well, and the plate was shaken for 10 minutes. The absorbance (OD) was detected at a wavelength of 490 nm using an enzyme reader, and its half-inhibitory concentration (IC 50 ), and the experiment was repeated 3 times.

[0029] The calculation formula of the inhibition rate of compound 3m on tumor cell proliferation is:

[0030] Inhibition rate (%) = (1-OD value of experimental group / OD value of blank control group) * 100%

[0031] Experimental Results

[0032] IC values ​​of compound 3m and positive control drug cisplatin on five tumor cell lines 50 The values ​​are shown in Table 1. From the results in the table, it can be seen that the compound 3m of the present invention has an inhibitory effect on the proliferation of five different tumor cells through in vitro anti-tumor activity tests, among which the inhibitory effect on the proliferation of human liver cancer cells HepG-2 is slightly higher than that of the positive drug, and has the potential to be developed into an anti-tumor drug.

[0033] Table 1 Inhibitory effect of compound 3m on tumor cell proliferation (IC 50 μmol / L)

[0034]

[0035] Example 3

[0036] Antibacterial activity test:

[0037] The tested bacterial strains were: Bacillus subtilis (CMCC(B)63501), Staphylococcus aureus (CMCC(B)26003), Salmonella paratyphi B (CMCC(B)50094), and Escherichia coli (CMCC(B)44102).

[0038] Experimental methods

[0039] The micro-dilution method was used to evaluate the antibacterial activity of the samples to be tested against each test bacteria. The bacteria were activated with LB medium, cultured on a shaker at 37°C and 170 rpm / min until the logarithmic growth phase, and the bacteria were diluted with LB medium to 5×10 5 CFU / mL. 100 μL was inoculated into each well of a sterile 96-well plate. Compound 3m was prepared into a 50 mg / mL stock solution with DMSO, and the stock solution was diluted to 512 μg / mL with LB medium, and added to the first column of wells of the 96-well plate, 100 μL per well, and 100 μL of liquid culture medium was added to each well after the first column, and diluted to an antibiotic concentration gradient to 100 μL per well with a lance, and set aside. 100 μL of bacterial solution was added to the 96-well plate in sequence, so that the final concentration of the sample was 512, 256, 128, 64, 32, and 16 μg / mL, respectively. Streptomycin sulfate was used as a positive control, and LB medium containing 0.5% DMSO was used as a negative control. The 96-well plate was incubated at 37°C in the dark for 16-18 hours, and its growth was observed. Each group of experiments was repeated 3 times. The minimum concentration corresponding to the well with sterile growth is the minimum inhibitory concentration (MIC) of the sample.

[0040] 10 μL of the growth-inhibited bacteria was applied to nutrient agar medium from each well and cultured in a 37°C constant temperature incubator for 24 hours. Each experiment was repeated 3 times, and the minimum bactericidal concentration (MBC) was taken as the lowest concentration at which the bacteria were completely killed.

[0041] Experimental Results

[0042] The MIC and MBC values ​​of compound 3m and the positive control drug streptomycin sulfate against four standard bacterial strains are shown in Table 2. From the results in the table, it can be seen that compound 3m has an inhibitory effect on the three selected standard strains, among which the antibacterial activity of compound 3m against Bacillus subtilis is better than that of the positive drug streptomycin sulfate, with a MIC of 128 μg / mL and an MBC of 256 μg / mL.

[0043] Table 2 MIC and MBC values ​​of compound 3m against four tested bacterial strains (μg / mL)

[0044]

[0045] In the table, "-" means that the corresponding MIC and MBC values ​​were not detected within the experimental concentration range.

[0046] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to specific details.

Claims

1.( E )-8-chloro-6-(2-fluorophenyl)-1-p-methyl-4-(p-methyl)-1,2,3,4-tetrahydrobenzo[ b ] The use of an azooctane compound in the preparation of an antibacterial drug, characterized in that: The bacteria are selected from one or a combination of Bacillus subtilis, Staphylococcus aureus, and Escherichia coli; the structure of the compound is shown in Formula 3m: 。

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