A 3-selenobenzofuran derivative and its preparation method and application

Abstract

The invention discloses a 3-selenobenzofuran derivative and a preparation method and application thereof. The 3-selenobenzofuran derivative has a structure of formula I, and the preparation method thereof is obtained by amidation reaction of benzoselenol and an indolone derivative. The 3-selenobenzofuran derivative of the invention has better anti-tumor efficacy, and an in vitro cell activity inhibition test shows that the 3-selenobenzofuran derivative of the invention has excellent anti-tumor activity and low toxic and side effects, can be prepared into an anti-tumor drug for clinical anti-tumor treatment, and is of great significance to the research of anti-tumor active drugs.

Description

Technical Field

[0001] The invention belongs to the field of drug synthesis, and specifically relates to a 3-selenobenzofuran derivative and a preparation method and application thereof. Background Art

[0002] Selenium (element 34) is one of the 14 essential trace elements for the human body. It plays an important role in human health and enjoys the reputation of "longevity element" and "king of anti-cancer". Organic selenium is a trace element and an essential nutrient for the human body. Organic selenium can be converted into selenoprotein in the human body, which can enhance the body's immunity and promote metabolism in the body, which is beneficial to health. If the body lacks organic selenium, it may lead to a decrease in the body's immunity and may also weaken the body's disease resistance. Taking an appropriate amount of organic selenium can supplement the selenium element needed by the body, promote metabolism in the body, improve the body's immunity, and have certain benefits to the body.

[0003] When the human body is in a state of selenium deficiency for a long time, it will lead to the occurrence of more than 40 diseases, including cancer, cardiovascular disease, liver disease, diabetes, thyroid disease, gastrointestinal disease, Keshan disease, respiratory disease, etc. Aryl selenides and heteroaryl selenides have been found to be useful for treating a variety of human diseases, and therefore have been used in the human body as potential therapeutic means. Among heteroaryl selenides, N-heteroaryl selenides (such as selenoindole and selenoimidazo[1,2-a]pyridine) are considered to be the most likely to prevent and treat human diseases because N-heterocycles have important biological significance.

[0004] Selenium-containing compounds, such as selenols, selenoxides, selenoesters, selenoamides, and selenoic anhydrides, generally exhibit stronger biological activity than their sulfur-containing counterparts. The subtle chemical differences between Se and S elements increase the activity of Se-containing compounds. For example, the size of Se is larger than that of S, resulting in looser binding of the outer valence electrons. At the same time, the larger hybrid orbital and longer bond length of the Se=O double bond lead to weaker π overlap and are more easily reduced. In this way, selenium-containing compounds have stronger antioxidant and free radical scavenging abilities. The chemical properties of Se affect its pharmacological activity, so understanding these relationships is of great significance for the rational design and development of selenium-containing drugs.

[0005] Selenium-containing heterocycles represent a large class of Se compounds that have been endowed with various promising biological properties, such as anticancer, anti-inflammatory, and antiviral activities. In addition, the cyclic structure of selenium heterocycles (in which Se is sp2 hybridized) prevents them from releasing elemental Se and therefore generally exhibits better metabolic stability. Selenazoles, selenadiazoles, thioselenazoles, and their related fusion analogs are important pharmacophores. Some of these compounds have become dominant structures and have been used to produce various promising pharmacological effects. In recent years, the research on selenium-containing drugs has become a hot topic. In addition to prostate cancer, selenium-containing drugs also have good effects on breast cancer and lung cancer. Summary of the invention

[0006] The present invention provides a 3-selenobenzofuran derivative and a preparation method and application thereof. One of the purposes of the present invention is to provide a class of 3-selenobenzofuran derivatives with anti-tumor activity; the second purpose of the present invention is to provide a method for preparing and purifying the above-mentioned 3-selenobenzofuran derivatives; the third purpose of the present invention is to apply the above-mentioned 3-selenobenzofuran derivatives to prepare clinical anti-tumor drugs; the above-mentioned purposes of the present invention are achieved by the following technical solutions:

[0007] A 3-selenobenzofuran derivative with anti-tumor activity has a structural formula as shown in Formula I:

[0008]

[0009] Wherein the R group is: Wherein R is: Wherein R is: one of H, CH3, OCH3, F, Cl, Br.

[0010] In the above steps, the catalyst is one of p-toluenesulfonic acid, methanesulfonic acid, boron trifluoride etherate, trifluoroacetic acid, trifluoromethanesulfonic acid, and trifluoromethanesulfonic anhydride, preferably p-toluenesulfonic acid; the reaction solvent is one of 1,2-dichloroethane, 1,4-dioxane, and o-dichlorobenzene, preferably 1,2-dichloroethane. The mass of the reaction solvent is 5 times that of indolone; the molar ratio of indolone to phenylselenol is 1:1.2-1.5; the amount of catalyst added is that the amount of water added is 10-15% of the molar amount of indolone.

[0011] The 3-selenobenzofuran derivatives prepared by the present invention have excellent anti-tumor properties and can be well applied to the preparation of anti-tumor disease drugs, among which the therapeutic effects on tumor diseases such as breast cancer, ovarian cancer, cervical cancer, prostate cancer, colon cancer, bronchial cancer, lung cancer, etc. are particularly significant.

[0012] Furthermore, the 3-selenobenzofuran derivatives obtained by the present invention or their pharmaceutically acceptable salts, solvates and hydrates can be used in combination with pharmaceutically acceptable carriers or diluents to form pharmaceutical preparations. Pharmaceutically acceptable carriers include inert solid fillers or diluents and sterile aqueous or organic solutions.

[0013] Beneficial effects of the present invention:

[0014] (1) The preparation method of the 3-selenobenzofuran derivative provided by the present invention is simple, has a high reaction yield, and the obtained product has high purity and is easy to purify, thus having industrial feasibility.

[0015] (2) Experiments have shown that the 3-selenobenzofuran derivatives or pharmaceutically acceptable salts thereof provided by the present invention can effectively inhibit cancer cells, especially lung cancer cells, and have a significant effect in reducing the level of intracellular anti-apoptotic mediator Bcl-2 and enhancing the apoptosis of cancer cells. DETAILED DESCRIPTION

[0016] The present invention is further described below by way of examples. It should be correctly understood that the examples of the present invention are only used to illustrate the present invention, rather than to limit the present invention, so simple improvements to the present invention based on the method of the present invention are within the scope of protection claimed by the present invention.

[0017] In this example, the NMR data used tetramethylsilane as an internal standard, and the 1H NMR and 13C NMR spectra were measured in DMSO-d6 on a Bruker AV 600. The HRMS data were measured on a Varian QFT-ESI.

[0018] I. Preparation of compounds

[0019] The general structural formula I is:

[0020] Preparation of 3-selenobenzofuran derivatives, wherein R is: wherein R is: H, CH3, OCH3, F, Cl, Br.

[0021] The preparation method is:

[0022] 3-selenobenzofuran derivative I is obtained by reacting benzoselenol and indole derivative in a solvent under the action of a catalyst. After the reaction, an NH4Cl aqueous solution is added, and dichloromethane is added to obtain the crude product. The organic phase is dried and the solvent is concentrated in vacuo to obtain a crude product. The crude product is purified by a silica gel column to obtain a 3-selenobenzofuran derivative I, wherein the structure of the indole derivative used as the raw material is:

[0023] Wherein R is: H, CH3, OCH3, F, Cl, Br.

[0024] Example 1

[0025] When R is H, the structural formula of the raw material indole derivative used is:

[0026]

[0027] The obtained compound 3-selenobenzofuran derivative has the structural formula

[0028]

[0029] To a solution of 1,2-dichloroethane (2 mL) containing 3-hydroxy-3-((2-hydroxyphenyl)ethynyl)-1-methylindole-2-one (0.2 mmol) and phenylselenol (0.3 mmol) was added p-toluenesulfonic acid (0.24 mmol). The mixture was stirred at 60° C. for 6 hours. The resulting solution was then quenched with a saturated NH4Cl aqueous solution. The mixture was then extracted with DCM, the organic layer was separated and washed with brine respectively. The organic layer was then dried over Na2SO4, filtered and concentrated in vacuo to give a crude product with a product yield of 97.2% and a purity of 96.5%. The product 1-methyl-3-(3-(phenylselenyl)benzofuran-2-yl)indole-2-one was purified by column chromatography (petroleum ether / EtOAc=4:1) with a product yield of 92.8% and a product purity of 99.4%. The obtained compound 1-methyl-3-(3-(phenylseleno)benzofuran-2-yl)indol-2-one has the following characterization data: melting point 178-79°C. 1H NMR (CDCl3, 400MHz): δ7.47-7.42 (m, 1H), 7.37 (d, J = 8.2 Hz, 1H), 7.31 (dd, J = 8.0, 3.2 Hz, 1H), 7.29-7.19 (m, 6H), 7.14 (d, J = 7.2 Hz, 1H), 7.01-6.95 (m, 2H), 6.89 (d, J = 7.6 Hz, 1H), 5.33 (s, 1H), 3.29 (s, 3H); 13 C NMR(CDCl3,100MHz)δ172.7,156.3,154.7,144.4,135.9,129.0(twopeaks),128.9,128.8(two peaks),127.2,126.0,125.8,125.2,124.4,123.5,122.8,120.2,111.6,109.6,108.5,45.0,26.7; HRMS-ESI(m / z):calcd for C 23 H 17 NO2Se[M+H] +:420.0503; found420.0509

[0030] Example 2

[0031] Boron trifluoride etherate (0.24 mmol, 1.2 equivalents) was added to a solution of 3-hydroxy-3-((2-hydroxyphenyl)ethynyl)-1-methylindol-2-one (0.2 mmol) and phenylselenol (0.3 mmol) in 1,4-dioxane (2 mL). The mixture was stirred at 50 ° C for 6 hours. The resulting solution was then quenched with a saturated NH4Cl aqueous solution. The mixture was then extracted with DCM, the organic layer was separated and washed with brine respectively. The organic layer was then dried over Na2SO4, filtered and concentrated in vacuo to give a crude product with a product yield of 94.1% and a purity of 95.8%. The product 1-methyl-3-(3-(phenylselenyl)benzofuran-2-yl)indol-2-one was purified by column chromatography (petroleum ether / EtOAc=4:1) with a product yield of 91.2% and a product purity of 99.6%.

[0032] Example 3

[0033] To a solution of 3-hydroxy-3-((2-hydroxyphenyl)ethynyl)-1-methylindole-2-one (0.2mmol) and phenylselenol (0.3mmol) in 1,2-dichloroethane (2mL) was added copper trifluoromethanesulfonate (0.24mmol). The mixture was stirred at 60°C for 6 hours. The resulting solution was then quenched with a saturated NH4Cl aqueous solution. The mixture was then extracted with DCM, the organic layer was separated and washed with brine respectively. The organic layer was then dried over Na2SO4, filtered and concentrated in vacuo to give a crude product with a product yield of 96.8% and a purity of 94.6%. The product 1-methyl-3-(3-(phenylselenyl)benzofuran-2-yl)indole-2-one was purified by column chromatography (petroleum ether / EtOAc=4:1) with a product yield of 90.7% and a product purity of 99.3%.

[0034] Taking the above embodiments as an example, indole ketone derivatives wherein R is CH3, OCH3, F, Cl, or Br are reacted to obtain corresponding 3-selenobenzofuran derivatives wherein R is CH3, OCH3, F, Cl, or Br. The specific preparation method is not described in detail.

[0035] 2. Cell Experiment Data

[0036] The RPMI-1640 culture medium, trypsin and PBS used in the cell experiment of the present invention were purchased from Gibco (USA), and CCK-8 was purchased from Abmole (USA). PC-3 (human prostate cancer cells), NCIH460 (human large cell lung cancer cells), 16HBE (human airway epithelial cancer cells), A549 (human non-small cell lung cancer cells), and SKMES1 (human squamous cell lung cancer cells) were from ATCC China.

[0037] The components of the culture medium used were

[0038] PC-3 (prostate cancer cells) 1640 culture medium + 10% fetal bovine serum + 1% double antibody (penicillin and streptomycin)

[0039] NCIH460 (human large cell lung cancer cells) 1640 culture medium + 10% fetal bovine serum + 1% double antibody (penicillin and streptomycin)

[0040] A549 (human non-small cell lung cancer cells) DMEM medium + 10% fetal bovine serum + 1% double antibody (penicillin and streptomycin)

[0041] The ELISA reader is the Thermo Fisher Varioskan LUX multifunctional ELISA reader

[0042] (1) Cell recovery

[0043] Small dish: add 3 mL of culture medium to the melted cell solution; after the cells adhere to the wall, change the medium in the middle, aspirate the liquid in the dish, and add 4 mL of culture medium.

[0044] (2) Cell passaging

[0045] Large dish: Take out the cells from the small dish, aspirate and discard the 4mL culture medium in the dish, add 1mL of PBS buffer, then aspirate and discard, add 1mL of trypsin to digest for 1-2min (during this waiting process, you can take a new large dish and add 7mL of culture medium for standby use), aspirate and discard, add 1mL of culture medium, wash the adherent cells, blow and suspend them, transfer them to a new dish that has been added with 7mL of culture medium, shake it evenly, observe it under a microscope, and put it in the incubator.

[0046] Take out the cells in the large dish, aspirate and discard the 8mL culture medium in the dish, add 3mL of PBS buffer, then aspirate and discard, add 2mL of trypsin to digest for 1-2min (during this waiting process, you can take 3 new dishes and add 7mL of culture medium each for standby), aspirate and discard, add 3mL of culture medium, wash the adherent cells, blow to suspend them, transfer to the new dish that has been added with 7mL of culture medium, transfer 1mL each time, shake it slightly to evenly distribute, observe under a microscope, and put it into the incubator.

[0047] (3) 96-well plate

[0048] Take the cells that have been passaged and count them using a cell counter (usually 25×10 5 Take an appropriate amount into the sample dilution tank and add culture medium to dilute to 2-5×10 5 Use a pipette to draw 100 μl into each well of a 96-well plate. The second row is not added as a blank. The surrounding wells are added to prevent edge effects. No drugs are added later. Each 96-well plate needs 10 mL of liquid.

[0049] (4) Dosing

[0050] 5 mg of the sample prepared in the embodiment of the present invention and the comparative sample used were taken in sequence, added to 5 mL centrifuge tubes respectively, put into sealed bags, and sterilized in a UV box in a cell room (20 min). According to the sampling amount and molecular weight, the culture medium (first dissolved with 30-50 μl DMSO, then added with culture medium) was configured into a 2 mmol / L mother solution for standby use; 100 μl of the mother solution was taken and placed in a 5 mL centrifuge tube, 1.9 mL of culture medium was added to form a 100 μmol / L sample solution, 500 μl was taken and placed in a 2 mL tube, 500 μl of culture medium was added to form a 50 μmol / L sample solution, and the sample solution was diluted to 25, 12.5, 6.25, 3.125, and 1.5625 μmol / L in sequence, and the negative control was 2 μl DMSO + 2mL culture medium; after preparing the sample solution, take out the 96-well plate, aspirate and discard the liquid in rows 4 to 11, and add 100μl of the above sample solution and negative solution in turn, adding 3 to 6 wells for each concentration solution.

[0051]

[0052] (5) CCK-8 test

[0053] Take CCK-8 solution and culture medium in a ratio of 1:9, prepare a test solution, and set aside (15 mL of test solution for every 2 plates); aspirate and discard the liquid in rows 4 to 11, add 100 μl of CCK-8 test solution to rows 2 and 4 to 11 (the third row and the surrounding circle are left untouched) with a pipette, and measure the absorbance with an ELISA reader after 2 hours, and calculate the compound IC of the samples prepared in the example respectively. 50 The specific results are shown in Table 1.

[0054] Table 1 IC of compounds against cancer cells 50 value

[0055]

[0056] According to the results obtained in Table 1, the compounds obtained by the present invention have more advantages in treating lung cancer and prostate cancer than existing drugs.

[0057] Although the above describes the specific implementation methods of the present invention, it is not intended to limit the scope of protection of the present invention. Those skilled in the art should understand that various modifications or variations that can be made by those skilled in the art on the basis of the technical solution of the present invention without creative work are still within the scope of protection of the present invention.

Claims

1. A 3-selenobenzofuran derivative, wherein the 3-selenobenzofuran derivative has a general formula I structure, characterized in that: The general formula I structural formula is: Wherein R is: H, CH3, OCH3, F, Cl, Br.

2. A method for preparing the 3-selenobenzofuran derivative according to claim 1, characterized in that: The 3-selenobenzofuran derivative I is obtained by reacting phenylselenol and an indolone derivative in a solvent under the action of a catalyst, adding an NH4Cl aqueous solution after the reaction, adding dichloromethane for extraction, drying the organic phase, and concentrating the solvent in vacuo to obtain a crude product, and the crude product is purified by a silica gel column to obtain a 3-selenobenzofuran derivative I, wherein the indolone derivative has the following structure; Wherein R is: H, CH3, OCH3, F, Cl, Br, the catalyst is one of p-toluenesulfonic acid, methanesulfonic acid, boron trifluoride ether, trifluoroacetic acid, trifluoromethanesulfonic acid, trifluoromethanesulfonic anhydride; the reaction solvent is one of 1,2-dichloroethane, 1,4-dioxane, o-dichlorobenzene.

3. The method for preparing a 3-selenobenzofuran derivative according to claim 2, characterized in that: The mass of the reaction solvent is 5 times that of the indolinone derivative; the molar ratio of the indolinone derivative to the phenylselenol is 1:1.2-1.5; and the amount of the catalyst added is 10-15% of the molar amount of the indolinone derivative.

4. Use of the 3-selenobenzofuran derivative or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a drug for treating tumor diseases, wherein the tumor diseases are prostate cancer and lung cancer. 5 . A pharmaceutical composition comprising the 3-selenobenzofuran derivative or a pharmaceutically acceptable salt thereof according to claim 1 , and a pharmaceutically acceptable carrier.

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