Use of an eight-membered selenium-containing benzazepine compound in the preparation of an antitumor drug

Abstract

The application belongs to the technical field of medicines, and particularly relates to application of an eight-membered selenium-containing benzazepine compound in preparation of an antitumor drug. The eight-membered selenium-containing benzazepine compound is prepared by an electrochemical method, and the reaction condition is mild and the operation is simple. The compound has certain antitumor and antibacterial activities, and particularly shows good inhibition on renal cancer cells 769-P and A498, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to the application of an eight-membered selenium-containing benzo[a]azine heterocyclic compound in the preparation of antitumor drugs. Background Technology

[0002] Selenium-containing organic compounds, due to their unique physicochemical properties, have broad research and application value in fields such as medicine, pesticides, and functional materials. Meanwhile, selenium, as one of the essential trace elements for the human body, participates in various redox-related physiological processes, playing an important role in antioxidation, anti-inflammation, anti-tumor activity, and immune regulation. Studies have shown that introducing selenium atoms into the backbone of organic molecules helps regulate the electronic structure and biocompatibility of molecules, enhancing their biological activity. Therefore, selenium-containing compounds have become one of the important directions in the research of novel drug lead structures.

[0003] Benzoazo heterocyclic compounds are an important class of pharmaceutical heterocyclic skeletons. The synergistic effect between the nitrogen atom and the benzene ring in their structure endows these compounds with excellent pharmacological activities, showing broad application prospects in antitumor, antibacterial and other fields. In the prior art, the structural modification of benzoazo heterocyclic compounds mainly focuses on five- or six-membered ring systems, usually through substituent modification or the introduction of other heteroatoms to improve their biological activity and drug-likeness.

[0004] However, the aforementioned structures still have certain limitations in terms of spatial configuration and molecular flexibility, making it difficult to meet the diverse requirements of complex biological targets for molecular three-dimensional structures. In contrast, eight-membered heterocycles, due to their larger ring strain adjustment space and higher conformational flexibility, show potential advantages in molecular recognition and target binding. However, due to factors such as complex synthetic routes and difficulty in controlling reaction conditions, there are few reports on eight-membered heterocyclic compounds in the current technology, especially compounds containing both selenium atoms and benzo[a]azine heterocyclic skeletons. The related structural types are relatively limited, and research on their biological activities and application prospects is still insufficient. Summary of the Invention

[0005] In view of the problems existing in the prior art, the present invention provides the application of an eight-membered selenium-containing benzo[a]azine heterocyclic compound in the preparation of antitumor drugs.

[0006] The technical solution of the present invention to solve the above-mentioned technical problems is as follows:

[0007] An eight-membered selenium-containing benzo[a]-azo heterocyclic compound, the structural formula of which is shown in Formula 3c:

[0008] .

[0009] This invention provides a method for preparing the compound of formula 3c.

[0010] .

[0011] The present invention also provides the application of the eight-membered selenium-containing benzo[a]azine heterocyclic compound in the preparation of antitumor drugs.

[0012] Furthermore, the tumor is kidney cancer.

[0013] Furthermore, the dosage form of the antitumor drug is capsule, tablet, granule, pill, powder, injection or spray.

[0014] The present invention also provides the use of the eight-membered selenium-containing benzo[a]azine heterocyclic compound in the preparation of antibacterial drugs.

[0015] Furthermore, the bacteria include one or a combination of Streptococcus pneumoniae, Escherichia coli, and Salmonella typhi.

[0016] Furthermore, the dosage form of the antibacterial drug is capsule, tablet, granule, pill, powder, injection, or spray.

[0017] The beneficial effects of this invention are: This invention uses an electrochemical method to construct the target compound under mild room temperature conditions. The reaction conditions are mild and the operation is simple. No additional strong oxidants or metal catalysts are required, which effectively reduces the environmental burden and improves the safety and controllability of the reaction.

[0018] Experimental results showed that the prepared compound 3c exhibited certain in vitro inhibitory activity against various renal cell carcinoma cells. Its inhibitory effect on 769-P and A498 cells was superior to that of the positive control drug cisplatin, demonstrating its potential as a candidate lead compound for further research and development in renal cell carcinoma. Furthermore, compound 3c also showed inhibitory effects against various bacterial strains, particularly against Streptococcus pneumoniae, which was superior to the positive control drug, indicating that this compound has potential applications in the antibacterial field and can be used to research and develop new antibacterial products. Attached Figure Description

[0019] Figure 1 Compound 3c of the present invention 1 H NMR spectrum;

[0020] Figure 2 Compound 3c of the present invention 13 C10 NMR spectrum. Detailed Implementation

[0021] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, all methods used in this invention are conventional in the art. Unless otherwise specified, the materials, reagents, methods, and instruments used are conventional in the art, and those skilled in the art can obtain them commercially or prepare the solutions using conventional methods in the art.

[0022] Example 1

[0023] At room temperature, add the following ingredients sequentially to the reaction flask: N -(but-3-en-1-yl)-4-methyl-N-(2-(1-phenylvinyl)phenyl)benzenesulfonamide 1a (0.2 mmol), 1,2-bis(3-methoxyphenyl)diselenoether 2c (1.2 equiv, 0.24 mmol), n Bu4NPF6 (0.4 mmol) and DCM (5 mL) were used in a reaction flask equipped with two Pt electrodes (1 cm × 1 cm). Electrolysis was carried out at room temperature (25 °C) using a constant current of 2 mA until the starting material was completely consumed (TLC monitoring, approximately 12 h). After the reaction was complete, the mixture was quenched with NaHCO3 (sat. aq. 15 mL) and extracted with CH2Cl2 (3 × 5 mL), and the organic solvent was concentrated under vacuum. The residue was purified by rapid column chromatography, and product 3c was obtained by washing with a mixture of ethyl acetate and petroleum ether.

[0024] .

[0025] Appendix Figure 1 and attached Figure 2 Compound 3c of the present invention 1 H NMR and 13 C10 NMR spectrum. Rf (petroleum ether: ethyl acetate = 10:1): 0.11; yellow liquid (72 mg, yield 61%). 1 H NMR (500 MHz, CDCl3) δ 7.64 (d, J = 8.1Hz, 2H), 7.35 (d, J = 7.1 Hz, 2H), 7.29-7.18 (m, 7H), 7.11 (dd, J = 6.9, 2.2 Hz, 1H), 7.07 (t, J = 7.9 Hz, 1H), 6.98-6.90 (m, 3H), 6.71 (dd, J = 8.1, 1.7 Hz, 1H), 5.96 (d,J = 8.7 Hz, 1H), 4.24 (dd, J = 14.6, 3.5 Hz, 1H), 3.72 (s, 3H), 3.09 (dd, J = 11.4, 8.5 Hz, 1H), 3.01-2.87 (m, 2H), 2.40 (s, 3H), 2.08-1.98 (m,1H), 1.85-1.75 (m, 1H), 1.65 (d, J = 13.5 Hz, 1H);

[0026] 13 C NMR (125 MHz, CDCl3) δ 159.67, 143.17, 142.11, 141.97, 140.30, 138.50, 137.73, 133.01, 131.05, 130.95, 129.75, 129.44, 128.87, 128.59, 128.54, 128.23, 127.97, 127.74, 127.24, 125.11, 118.29, 112.67, 55.25, 50.98, 38.68, 35.68, 33.69, 21.54.

[0027] Experimental Example

[0028] 1. Antitumor activity experiment

[0029] Cell preparation: Cells were removed from liquid nitrogen, revived, and cultured. Human renal cell adenocarcinoma cells (786-O) and (769-P) were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin antibiotic solution; human clear cell renal cell carcinoma skin metastases (CAKI-1) were cultured in McCoy's 5A medium containing 10% fetal bovine serum and 1% penicillin-streptomycin antibiotic solution; and human renal cell carcinoma cells (A498) were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin antibiotic solution. All cells were grown at 37°C and 5% CO2. When the cells reached 80% of the bottom area of ​​the culture dish, they were digested with trypsin and passaged. Cells in good logarithmic growth phase were selected for subsequent experiments.

[0030] MTT solution preparation: Weigh MTT powder using an analytical balance and prepare a 5 mg / mL MTT working solution with PBS buffer. Prepare fresh before use.

[0031] Compound 3c was dissolved in DMSO (final DMSO concentration <0.1%). It was then serially diluted to the required working concentration using culture medium before the experiment. Tumor cells in the logarithmic growth phase were used at a concentration of 5 × 10⁻⁶ cells / mL. 3 Cells were seeded at a density of 1 / mL in 96-well plates and incubated at 37°C in a 5% CO2 incubator for 24 hours. After cell attachment, the original culture medium was discarded, and 100 μL of culture medium containing compound 3c was added to each well as the experimental group. The concentration gradients of the experimental groups were 0.78, 1.56, 3.125, 6.25, 12.5, 25, 50, and 100 μM. Cisplatin was used as a positive control, and each well was divided into 6 replicates. After culturing for another 48 hours, 10 μL of MTT solution (5 mg / mL) was added to each well and incubated for 4 hours. The supernatant was discarded, and then 150 μL of DMSO was added to each well. The plates were shaken at room temperature for 10 minutes, and the absorbance of each well was measured at 570 nm using a microplate reader.

[0032] Data processing: All data were analyzed for standard deviation using GraphPad Prism 9.0, and the results are expressed as mean ± standard deviation. ±SD).

[0033] Table 1. Inhibitory effect of compound 3c on tumor cell proliferation (IC50) 50 (μM)

[0034] 786-O 769-P CAKI-1 A498 3c 30.42±2.01 10.61±0.79 18.14±1.28 6.15±0.41 Cisplatin 6.08±0.18 19.20±0.48 3.32±0.099 15.13±0.38

[0035] Experimental results

[0036] The results are shown in Table 1. Compound 3c exhibited certain in vitro inhibitory activity against all four renal cell carcinoma lines, but there were significant differences in sensitivity among the different cell lines. Among them, A498 cells were the most sensitive to compound 3c, with an IC50 value of [missing value]. 50 The value was 6.15 ± 0.41 μM. Compared with the positive control cisplatin, compound 3c showed stronger inhibitory activity in 769-P and A498 cells.

[0037] The above results provide experimental basis for further structural optimization and mechanism research of compound 3c as a candidate lead compound for renal cell carcinoma.

[0038] 2. Antibacterial activity test

[0039] The tested bacterial strains were: Streptococcus pneumoniae (CICC 24947), Staphylococcus aureus (CMCC(B)26003), Escherichia coli (CMCC(B)44102) and Salmonella typhi (CMCC(B)50071).

[0040] Compound 3c was prepared into a stock solution with a concentration of 5 mg / mL using DMSO (DMSO concentration ≤ 0.5%). The bacterial strains used in the activity test were activated in a 37℃ incubator for 2-3 h. A small amount of the activated strain was then picked, dissolved in sterile water, and shaken well to prepare a concentration of 5 × 10⁻⁶ mg / mL. 5 CFU / mL bacterial suspension. In a 96-well plate, add 188 μL of the corresponding culture medium and 2 μL of the stock solution of the test compound to the first well. Perform a 2-fold serial dilution in the 96-well plate, and finally add 10 μL of the test bacterial suspension to each well, so that the final concentrations of the test compound added to the 96-well plate are 128, 64, 32, 16, 8, 4, 2, and 1 μg / mL, respectively. Chloramphenicol was used as a positive control, 0.5% DMSO as a negative control, and empty culture medium as a blank control. Each experiment was repeated three times. After incubation at 37℃ for 3 days, observe the turbidity of the bacterial suspension in the 96-well plate. The minimum concentration at which the culture medium becomes clear is the minimum inhibitory concentration (MIC) of the compound.

[0041] Table 2. MIC values ​​(μg / mL) of compound 3c against four tested bacterial strains.

[0042] Streptococcus pneumoniae Staphylococcus aureus Escherichia coli Salmonella typhi 3c 4 -- 8 32 Chloramphenicol 8 16 8 4

[0043] Note: "--" indicates that no inhibitory activity was detected within the tested concentration range.

[0044] Experimental results

[0045] The results of the antibacterial activity experiments of compound 3c against four tested bacterial strains are shown in Table 2. The results indicate that it exhibits certain inhibitory effects against the other three tested bacteria, excluding Staphylococcus aureus. Among them, the inhibitory effect against Streptococcus pneumoniae is more significant, with a MIC value of 4 μg / mL, and its antibacterial effect is superior to that of the positive control drug chloramphenicol. The antibacterial activity of compound 3c against Escherichia coli is comparable to that of the positive control drug chloramphenicol.

[0046] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An eight-membered selenium-containing benzo[a]azine heterocyclic compound, characterized in that, The structural formula of the compound is shown in Formula 3c: 。 2. The use of the eight-membered selenium-containing benzo[a]azine heterocyclic compound of claim 1 in the preparation of antitumor drugs.

3. The application of the eight-membered selenium-containing benzo[a]azine heterocyclic compound according to claim 2 in the preparation of antitumor drugs, characterized in that, The tumor is kidney cancer.

4. The application of the eight-membered selenium-containing benzo[a]azine heterocyclic compound according to claim 2 in the preparation of antitumor drugs, characterized in that, The dosage forms of the antitumor drugs are capsules, tablets, granules, pills, powders, injections, or sprays.

Images