阅读说明：本技术 多取代异苯并呋喃化合物及其用途 (Polysubstituted isobenzofuran compounds and uses thereof ) 是由 徐华栋 陶鹏 沈美华 王燕 于 2021-07-07 设计创作，主要内容包括：本发明属于药物化学技术领域,具体涉及一种多取代异苯并呋喃化合物及其用途。以2-芳基甲酰基芳醛为原料,通过与三甲基硅烷甲酰胺试剂反应,现场获得醛基甲酰胺化中间产物,再在酸性条件下分子内关环缩合获得多取代异苯并呋喃化合物。用MTT法测定了多取代异苯并呋喃部分化合物对人肾癌细胞增殖的抑制作用,结果显示了多取代异苯并呋喃化合物的潜在抗肿瘤活性。(The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a polysubstituted isobenzofuran compound and application thereof. Taking 2-aryl formyl aromatic aldehyde as a raw material, reacting with a trimethylsilane formamide reagent to obtain an aldehyde formyl amidation intermediate product on site, and performing intramolecular cyclization condensation under an acidic condition to obtain a polysubstituted isobenzofuran compound. The MTT method is used for measuring the inhibition effect of the multi-substituted isobenzofuran partial compound on the proliferation of human renal cancer cells, and the result shows the potential anti-tumor activity of the multi-substituted isobenzofuran compound.)

1. A polysubstituted isobenzofuran compound characterized in that: the general structural formula of the polysubstituted isobenzofuran compound is as follows:

wherein R is₁Selected from hydrogen, methyl, ethyl, fluorine, chlorine; r₂Selected from hydrogen, methyl, ethyl, fluorine, chlorine; r₃Selected from hydrogen, fluorine, chlorine; r₄Selected from methyl, ethyl, propyl, isopropyl, tert-butyl; r₅Selected from methyl, ethyl, propyl, isopropyl, tert-butyl.

2. A method for preparing a polysubstituted isobenzofuran compound, which is characterized by comprising the following steps: the preparation method comprises the following steps: under the protection of nitrogen, sequentially adding 2-aryl formyl aromatic aldehyde, anhydrous tetrahydrofuran and trimethylsilylcarboxamide reagents into a closed tube reactor, sealing, placing in an oil bath, heating to reflux reaction under stirring, cooling to room temperature after 12 hours, adding a hydrochloric acid aqueous solution with the concentration of 4M, stirring to react for 2 hours at room temperature, adding a saturated sodium bicarbonate solution to neutralize, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, separating and purifying the obtained residue by silica gel column chromatography by taking ethyl acetate and petroleum ether as eluent to obtain the polysubstituted isobenzofuran product.

3. The process for producing a polysubstituted isobenzofuran compound according to claim 2, wherein said 2-arylformylaromatic aldehyde has the formula:

4. the process for producing a polysubstituted isobenzofuran compound according to claim 2, wherein said trimethylsilane carboxamide has the formula:

5. the method for producing a polysubstituted isobenzofuran compound according to claim 2, wherein the molar ratio of the 2-arylformylarylaldehyde to the trimethylsilane carboxamide to the 4M aqueous hydrochloric acid solution is 1:1.1 to 1.5:2.0 to 5.0.

6. Use of a polysubstituted isobenzofuran compound according to claim 1, wherein: the polysubstituted isobenzofuran compound is applied to antitumor drugs.

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a polysubstituted isobenzofuran compound and application thereof.

Background

The isobenzofuran compounds are important heterocyclic compounds, serve as high-activity multifunctional synthetic intermediates, and are widely applied to the fields of chemical industry and materials. Meanwhile, many isobenzofuran derivatives have good biological activity and are developed into medicines for sale. For example, the phenolphthalein tablet used for treating constipation contains isobenzofuranone structure, and for example, citalopram hydrobromide with unique antidepressant effect contains dihydroisobenzofuran structure. However, up to now, the isobenzofuran skeleton itself has not been found to be present in active compounds, and various substituted isobenzofurans have been reported more as a base material or a pharmaceutical intermediate.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: based on the importance of the structure of the polysubstituted isobenzofuran and the blank of the biological activity, the invention provides a polysubstituted isobenzofuran compound which can effectively inhibit the growth of renal cancer cells.

The technical scheme adopted by the invention for solving the technical problems is as follows: provided is a polysubstituted isobenzofuran compound represented by the chemical structural formula (I):

in the chemical formula (I), R₁Selected from hydrogen, methyl, ethyl, fluorine, chlorine; r₂Selected from hydrogen, methyl, ethyl, fluorine, chlorine; r₃Selected from hydrogen, fluorine, chlorine; r₄Selected from methyl, ethyl, propyl, isopropyl, tert-butyl; r₅Selected from methyl, ethyl, propyl, isopropyl, tert-butyl.

The specific synthetic route of the invention is shown in equation (1):

wherein R is₁、R₂、R₃、R₄、R₅The definition of (1) is as before.

In equation (1), acid chloride compound 1 was purchased commercially directly or prepared from the corresponding carboxylic acid purchased directly, compound 3 was purchased commercially directly, and trimethylsilylcarboxamide reagent 6 was purchased commercially directly or prepared with reference to known literature (Cunico R.F., Chen J., Synthetic Communications,2003,33(11), 1963-. The preparation of compound 5 is described in the known literature: jacq J., Einhorn C., Einhorn J., Organic Leters,2008,10(17), 3757-.

The synthesis method of the chemical structural formula (I) is as follows: under the protection of nitrogen, 2-aryl formyl aromatic aldehyde (compound 5), anhydrous tetrahydrofuran and trimethyl silane formamide reagent 6 are added into a closed tube reactor in sequence, sealed and then placed in an oil bath, and heated to reflux reaction under stirring. After 12 hours, the reaction mixture was cooled to room temperature, and a 4M aqueous hydrochloric acid solution was added thereto, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction solution with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue with ethyl acetate and petroleum ether as eluent by silica gel column chromatography to obtain the polysubstituted isobenzofuran product with the chemical structural formula (I).

Wherein the molar ratio of the 2-arylformyl aromatic aldehyde to the trimethylsilane formamide to the 4M aqueous hydrochloric acid solution is 1: 1.1-1.5: 2.0 to 5.0.

Selecting part of compounds in the polysubstituted isobenzofuran products, and determining the inhibition effect of the compounds on the proliferation of cancer cells by using an MTT method. The invention adopts MTT method to measure the inhibition effect of the target compound on the proliferation of human cancer cells. Selecting OSRC-2 type renal cancer cells as test cell strains, carrying out in-vitro antitumor activity evaluation on the synthesized target compound by using an MTT method, and taking a blank as a control. Taking tumor cells in logarithmic growth phase, centrifuging, and diluting to 5 × 10 with RPMI1640 culture solution⁴one/mL, seeded in 96-well plates. After overnight incubation at 37 ℃ samples of different concentrations were added and incubated for a further 72h, 10 uL/well of MTT solution (5mg/mL) was added and after incubation at 37 ℃ for 4h 100uL DMSO was added per well. After 10min, shaking, the well plate was placed in an automated microplate spectrophotometer, absorbance values were measured at 570nm and 630nm, and the median effective Inhibitory Concentration (IC) was calculated by the Bliss method₅₀). Each set of samples was tested in 3 replicates.

The MTT method is adopted to test that: when R is₁、R₂、R₃Are all chlorine, R₄、R₅When the compounds are methyl, the compound has obvious inhibition effect on cancer cell proliferation; when R is₁、R₂、R₃Are each hydrogen, R₄、R₅Are all made ofMethyl, or R₁、R₂Is hydrogen, R₃Is chlorine, R₄、R₅Are both methyl or R₁Is chlorine, R₂Is hydrogen, R₃Is fluorine, R₄、R₅When both are methyl, the compound has anticancer activity on cancer cells; the compounds with other structures have potential anticancer activity and can be applied to antitumor drugs.

The invention has the beneficial effects that: the invention provides a polysubstituted isobenzofuran compound with a novel structure; MTT method activity test shows that the compound has potential biological activity and can be used for preparing anticancer drugs.

Detailed Description

The invention will now be further illustrated by reference to specific examples, which are intended to be illustrative of the invention and are not intended to be a further limitation of the invention.

The reagent used in the invention is prepared by reference to relevant documents, and the solvent is purified and refined.

Example 1

Taking 10mL of sealed tube, weighing 5a 105mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and changing nitrogen, and reacting for 12 hours at 70 ℃ in an oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue with ethyl acetate and petroleum ether as eluent by silica gel column chromatography to obtain light yellow solid A with the yield of 68%.¹H NMR(300MHz,CDCl₃)δ8.16–8.13(m,1H),7.92–7.82(m,3H),7.54–7.45(m,2H),7.39–7.33(m,1H),7.19–7.06(m,2H),3.36(br,6H).¹³C NMR(75MHz,CDCl₃) Delta 160.70,145.89,137.04,130.86,130.32,129.06,128.15,127.26,125.83,125.48,121.63,121.10,119.54,29.65 HRMS (ESI) m/z theory C₁₇H₁₆NO₂ ⁺[M+H]⁺266.1176, found 266.1162.

Example 2

Taking 10mL of sealed tube, weighing 5b 112mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and replacing nitrogen, and reacting for 12 hours at 70 ℃ in an oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue with ethyl acetate and petroleum ether as eluent by silica gel column chromatography to obtain light yellow solid B with the yield of 60%.¹H NMR(300MHz,CDCl₃)δ8.13(dt,J＝8.9,0.9Hz,1H),7.61–7.50(m,2H),7.39–7.28(m,3H),7.16(ddd,J＝8.9,6.4,0.8Hz,1H),7.03(ddd,J＝8.8,6.4,0.9Hz,1H),3.30(br,6H),2.45(s,3H).¹³C NMR(75MHz,CDCl₃) δ 160.58,147.35,136.64,131.16,129.77,129.59,129.27,128.84,127.18,125.84,125.19,125.08,122.22,121.09,119.52,29.48,20.38 HRMS (ESI) m/z theoretical value C₁₈H₁₈NO₂ ⁺[M+H]⁺280.1332, found 280.1337.

Example 3

Taking 10mL of sealed tube, weighing 5c 122mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and replacing nitrogen, and reacting for 12 hours at 70 ℃ in an oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue by silica gel column chromatography with ethyl acetate and petroleum ether as eluent to obtain light yellow solid C with the yield of 64%.¹H NMR(400MHz,CDCl₃)δ8.16(d,J＝8.9Hz,1H),7.64–7.60(m,1H),7.54–7.51(m,2H),7.39–7.33(m,2H),7.14(dd,J＝8.7,6.4Hz,1H),7.04(dd,J＝8.5,6.7Hz,1H),3.32(br,6H).¹³C NMR(100MHz,CDCl₃) δ 160.53,143.73,138.24,132.63,131.36,130.83,129.94,129.48,129.11,127.15,126.88,125.57,122.96,121.39,119.43,29.59 HRMS (ESI) m/z theory C₁₇H₁₅ClNO₂ ⁺[M+H]⁺300.0786, found 300.0798.

Example 4

Taking 10mL of sealed tube, weighing 5d 157mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and replacing nitrogen, and reacting for 12 hours at 70 ℃ in an oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue with ethyl acetate and petroleum ether as eluent by silica gel column chromatography to obtain light yellow solid D with the yield of 55%.¹H NMR(400MHz,CDCl₃)δ8.18(s,1H),7.46–7.44(m,2H),7.37–7.34(m,1H),7.20(dd,J＝8.0,0.5Hz,1H),7.04(dd,J＝8.2,1.7Hz,1H),3.28(br,6H).¹³C NMR(100MHz,CDCl₃) δ 160.24,141.21,138.40,136.92,133.41,131.66,129.11,128.59,127.92,127.53,122.42,120.84,120.09,29.72 HRMS (ESI) m/z theory C₁₇H₁₃Cl₃NO₂ ⁺[M+H]⁺368.0006, found 368.0007.

Example 5

Taking 10mL of sealed tube, weighing 5e 114mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and changing nitrogen, and reacting for 12 hours at 70 ℃ in oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue by silica gel column chromatography with ethyl acetate and petroleum ether as eluent to obtain light yellow solid E with the yield of 70%.¹H NMR(300MHz,CDCl₃)δ7.84–7.81(m,1H),7.79–7.76(m,1H),7.65–7.61(m,1H),7.50–7.43(m,2H),7.37–7.32(m,1H),7.25–7.21(m,1H),3.32(br,6H).¹³C NMR(75MHz,CDCl₃) δ 163.82(d, J ═ 252.8Hz),159.79,146.63(d, J ═ 1.5Hz),138.38(d, J ═ 27.8Hz),131.64(d, J ═ 8.2Hz),130.45(d, J ═ 25.5Hz),130.42,129.92,128.62,128.47,121.63,122.25(d, J ═ 10.5Hz),103.58(d, J ═ 24.8Hz),29.62 hrms (esi) m/z theory C₁₇H₁₅FNO₂ ⁺[M+H]⁺284.1081, found 284.1083.

Example 6

Taking 10mL of sealed tube, weighing 5f 131mg, adding 1mL of tetrahydrofuran and 290mg of N, N,1,1, 1-pentamethylsilane formamide reagent 6a, vacuumizing and replacing nitrogen, and reacting for 12 hours at 70 ℃ in an oil bath. The reaction was stopped, cooled, and then 0.5mL of 4M aqueous hydrochloric acid was added, followed by stirring at room temperature for 2 hours. Adding saturated sodium bicarbonate solution for neutralization, extracting the obtained reaction liquid with ethyl acetate, drying the organic phase, concentrating, and separating and purifying the obtained residue by silica gel column chromatography with ethyl acetate and petroleum ether as eluent to obtain light yellow solid F with the yield of 75%.¹H NMR(400MHz,CDCl₃)δ7.73–7.69(m,1H),7.57–7.49(m,3H),7.39–7.36(m,2H),6.86–6.82(m,1H),3.30(br,6H).¹³C NMR(100MHz,CDCl₃) δ 161.52(d, J ═ 235.0Hz),160.21,141.52,138.19,136.79,132.77,131.38,130.93,130.37,129.73,127.02,122.31(d, J ═ 10.5Hz),120.82,118.56(d, J ═ 30.1Hz),117.68,103.38(d, J ═ 25.2Hz),29.62 hrms (esi) m/z theoretical value C₁₇H₁₄ClFNO₂ ⁺[M+H]⁺318.0692Found 318.0695.

Experimental testing

Compounds A, B, C, D, E and F were tested for anti-cancer activity according to the MTT method described above and the results are given in table one below. The results show that the compound D has obvious inhibition effect on the growth of human kidney cancer cells, the compounds A, E and F have anticancer activity on the human kidney cancer cells, and the compounds B and C have potential anticancer activity on the human kidney cancer cells.

Activity test of epipolysubstituted Isobenzofuran compounds on human renal carcinoma cells

In light of the foregoing description of the preferred embodiment of the present invention, many modifications and variations will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.