Use of 3-azido-N-(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide in the preparation of anti-tumor drugs

Abstract

The present invention belongs to the field of pharmacy, and specifically relates to 3-azido-N-(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide compounds and their use in the preparation of anti-tumor drugs. The compounds of the present invention are obtained by a metal-free catalytic selenazidation method of olefins, and are red liquids with the molecular formula C 17 H 14 ClN5OSe.

Description

[0001] This application is a divisional application of the application with the application number 202110512784.6, the application date of May 11, 2021, and the invention name of "3-azido- N -(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide" at the time of application. Technical Field

[0002] The present invention relates to the field of pharmacy, and in particular to the use of 3-azido- N -(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide in the preparation of anti-tumor drugs. Background Art

[0003] Organic azide compounds have a wide range of applications in the fields of fine chemicals and the pharmaceutical industry. The azide group is not only a very good functional group for organic synthesis transformation, but also a functional group in drugs. The carbon-N 3 bond is an important synthetic method, which provides a method for introducing nitrogen atoms into various organic molecules. Therefore, how to introduce N 3 into organic molecules has always been a research hotspot for organic chemists. Therefore, in synthetic organic chemistry, there is a continuous need to develop sustainable and practical methods to form carbon-N 3 bonds. On the other hand, nitrogen-containing heterocycles are the key backbone structures of many drug molecules. In particular, the structurally diverse quinoline-2,4-diones are widely present in many natural products, drugs, and agrochemicals. Although some methods for constructing quinoline-2,4-diones have been reported, the metal-free catalytic radical tandem carbocyclization reaction is undoubtedly one of the best choices for obtaining atom and step economy and thus obtaining these complex molecules.

[0004] Organic selenium compounds are important intermediates in synthetic chemistry and exist as the core structural backbone in a considerable number of bioactive drug molecules. Organic selenium compounds have a wide range of applications in the fields of organic synthesis, drugs, organic materials, etc. Therefore, the development of green, novel, efficient, and practical synthesis methods for organic selenium compounds has attracted extensive attention from chemists. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide the use of the compound 3-azido- N -(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide in the preparation of anti-tumor drugs.

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

[0007] One of the purposes of the present invention is to provide the compound 3-azido- N- (3-Chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide, whose chemical structural formula 2 is as follows:

[0008] 。

[0009] The preparation method of the above compound is as follows: Using N -(3-Chloro-2-cyanophenyl)methacrylamide (1) as the substrate, the reaction of (PhSe) 2 and PhI(OAc) 2 pre-activated TMSN 3 effectively obtains the azidation-selenation product of the olefin through the azidation-selenation process in DMSO. After the reaction is completed by shaking for 12 hours, the mixture is quenched with H 2 O and extracted with CH 2 Cl 2 Then the organic solvent is concentrated in vacuo, and the residue is purified by flash column chromatography, and 3-azido-N-(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide (2) is obtained using petroleum ether and ethyl acetate as eluents.

[0010] Furthermore, the equation of the reaction is:

[0011] 。

[0012] Furthermore, the application of the compound 3-azido- N -(3-Chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide (2) in the preparation of anti-tumor drugs.

[0013] Furthermore, the application of the compound 3-azido- N -(3-Chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide (2) in the preparation of tyrosinase inhibitor drugs.

[0014] The beneficial effect of the present invention is: A method for azidation-selenation of olefins without metal catalysis, and the application of the obtained new compound in the preparation of anti-tumor and tyrosinase inhibitor drugs.

[0015] 3-Azido- N -(3-Chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide (2) structure analysis:

[0016] As Figures 1-3 shown, the 1 H NMR of compound 2 of the present invention, 13The structure of the compound was determined by 1 1H NMR, 3 13C NMR and HR-ESI-MS spectra. Specifically, compound 2 was a red liquid with Rf (petroleum ether: ethyl acetate) = 0.42. δ 1H NMR (600 MHz, CDCl J 3): δ 8.79 (s, 1H), 8.27 (dd, J J = 8.5, 0.5 Hz, 1H), 7.60 - 7.58 (m, 2H), 7.50 (t, J J = 8.3 Hz, 1H), 7.42 - 7.39 (m, 1H), 7.32 - 7.30 (m, 2H), 7.23 (dd, J J = 8.1, 0.8 Hz, 1H), 3.92 (d, J J = 12.6 Hz, 1H), 3.69 (d, 13 J = 12.6 Hz, 1H), 1.69 (s, 3H). 3 13C NMR (150 MHz, CDCl δ 3): δ 170.76, 142.11, 137.59, 136.73, 134.51, 130.07, 129.48, 125.21, 124.80, 118.60, 113.76, 103.24, 58.04, 51.18, 22.45. HRMS (ESI) calcd for C 17 13 14 H 5 6 + ClN OSe [M + Na]+: 441.9944, found: 441.9944.

[0017] Figure 1 BRIEF DESCRIPTION OF THE DRAWINGS 1 1H NMR spectrum of compound 2;

[0018] Figure 2 13C NMR spectrum of compound 2; 13 HR-ESI-MS spectrum of compound 2.

[0019] Figure 3 DETAILED DESCRIPTION OF THE INVENTION

[0020] The present invention will be further described below by way of examples, so that those skilled in the art can understand the present invention more comprehensively, but the present invention is not limited in any way.

[0021] ​Example 1

[0022] At room temperature, add to the reaction tube N -(3-chloro-2-cyanophenyl)methacrylamide (1) (0.3 mmol, 66 mg) and azidotrimethylsilane TMSN pre-activated by diphenyldiselenide (PhSe) 2 (0.3 mmol, 93.6 mg) and diethyl iodobenzene PhI(OAc) 2 (0.45 mmol, 145 mg) (93%, 3.0 equiv, 0.9 mmol, 127 μL) The reaction was carried out in DMSO (2 mL). Then the reaction tube was stirred at room temperature for 12 hours until the raw materials were completely consumed as monitored by TLC analysis. After the reaction was completed, the mixture was quenched with H 3 O (15 mL) and extracted with CH 2 Cl 2 Cl 2 (3 × 5 mL). Then, the organic solvent was concentrated in vacuo. The residue was purified by flash column chromatography using petroleum ether and ethyl acetate as eluents, Rf = 0.42, to obtain 3-azido-N-(3-chloro-2-cyanophenyl)-2-methyl-2-(phenylseleno)propanamide (2) with a yield of: 55%, 69 mg. The equation for the reaction is:

[0023]

[0024] Example 2

[0025] 1. Tumor cell culture: Human hepatocellular carcinoma cells (HepG-2), human lung cancer cells (A549), human breast cancer cells (MCF-7), human gastric cancer cells (SGC-7901), and human ovarian cancer cells (SKOV-3) were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin double antibody, digested and passaged with trypsin, and cells in the logarithmic growth phase were selected for experiments.

[0026] 2. Drug preparation: The drug was dissolved in DMSO to prepare a 1 mg / mL stock solution and stored at -80 o °C for later use. Before the experiment, it was diluted to the required concentration with the prepared medium (final DMSO concentration < 0.01%).

[0027] 3. Detection of cell proliferation inhibition

[0028] The MTT colorimetric method was selected to detect the inhibitory effect of compound 2 on the proliferation of various tumor cells. Tumor cells in the logarithmic growth phase were inoculated into 96-well plates at 100 μL, and the cell amount was about 1×10 5cells / mL. After incubation for 24 h, 10 μL of Compound 2 with final concentrations of 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL, 10 μg / mL, and 20 μg / mL was added to each well, and the negative control group was added with an equal volume of RPMI 1640 medium. Three parallel wells were set in each group. The 96-well plate was placed in a 37 o °C constant temperature CO 2 incubator and cultured for 48 h. 20 μL of 0.5% MTT solution was added to each well and incubated in the incubator for another 4 - 6 h. The supernatant was discarded, 100 μL of DMSO was added to each well, and after sufficient oscillation, the absorbance value (OD value) was measured at 570 nm, and the half-maximal inhibitory concentration (IC 50 ) was calculated. The experiment was repeated 3 times.

[0029] The formula for calculating the proliferation inhibition rate of Compound 2 on tumor cells is: Proliferation inhibition rate (%) = (1 - OD value of experimental group / OD value of blank control group) * 100%.

[0030] 4. Experimental results

[0031] The experimental results are shown in Table 1. After 72 h of treatment with different concentrations of Compound 2 on human hepatocellular carcinoma HepG-2 cells, human lung cancer A549 cells, human breast cancer MCF-7 cells, human gastric cancer SGC-7901 cells, and human ovarian cancer SKOV-3 cells, the proliferation of the above tumor cells was inhibited to varying degrees. As the drug concentration increased, the proliferation inhibition rate of each tumor cell gradually increased, showing concentration dependence.

[0032] Table 1 Inhibitory effect of Compound 2 on the proliferation of different tumor cells

[0033]

[0034] Example 3: Determination of tyrosinase inhibitory activity

[0035] 1. Preparation of sample solution and reference solution: Compound 2 and positive reference substances citric acid and arbutin were prepared into a stock solution of 2 mg / mL with a phosphate buffer-dimethyl sulfoxide (PBS-DMSO) mixture, and then diluted successively into solutions of 1, 0.5, 0.25, and 0.125 mg / mL.

[0036] 2. Take 80 μL of sample solutions with different concentrations, as well as citric acid and arbutin solutions, and successively add 280 μL of PBS (pH 6.8) and 80 μL of L-tyrosine solution at 0.1 mg / mL. Mix evenly and react at a constant temperature in a water bath at 25°C for 10 min. Then add 40 μL of tyrosinase (100 U / mg), mix evenly, react at a constant temperature in a water bath at 25°C for 10 min, take out, and use an ultraviolet spectrophotometer to measure the absorbance at 475 nm. Taking each blank control as a reference, calculate the inhibition rate of the sample solution, citric acid, and arbutin solutions on tyrosinase, and calculate the half-inhibitory concentration (IC 50 ) according to the dose-effect curve.

[0037] Inhibition rate (%) = (A 0 - A 1 ) / A 0 ×100%

[0038] Where: A 0 is the absorbance of the standard control, that is, the absorbance of the standard control solution composed of PBS buffer, L-tyrosine solution, and tyrosinase; A 1 is the absorbance of the sample solution or citric acid and arbutin.

[0039] 3. The measurement results are shown in Table 2. The inhibitory effects of Compound 2, citric acid, and arbutin on tyrosinase increase with the increase in concentration. The IC 50 value of Compound 2 is 0.865 mg / mL, the IC 50 value of citric acid is 0.813 mg / mL, and the IC 50 value of arbutin is 0.485 mg / mL. The inhibitory effect of Compound 2 on tyrosinase is equivalent to that of the positive control citric acid and slightly worse than that of the positive control arbutin.

[0040] Table 2 Inhibitory effects of Compound 2, citric acid, and arbutin on tyrosinase

[0041]

[0042] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims

1. 3 - azido - N - (3 - chloro - 2 - cyanophenyl) - 2 - methyl - 2 - (phenylseleno) propanamide compound in the preparation of a drug for treating one or more of human breast cancer cells, human gastric cancer cells, and human ovarian cancer cells, and the structural formula of the compound is as shown in 2: .

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