阅读说明：本技术 一种吲哚啉异色满类衍生物及其酶催化合成方法和应用 (Indoline isochroman derivative and enzymatic synthesis method and application thereof ) 是由 钟芳锐 郭欢 戴志锋 于 2021-10-08 设计创作，主要内容包括：本发明属于精细化工合成技术领域,具体涉及一种吲哚啉异色满类衍生物及其酶催化合成方法和应用,以2,3-二羟基苯甲酸和吲哚类化合物为底物,多铜氧化酶为催化剂,反应得到吲哚啉异色满类衍生物。本发明从廉价易得的化学原料起,通过多铜氧化酶为催化剂催化的氧化偶联反应得到高附加值的吲哚啉异色满类化合物,反应条件温和,操作简单,成本低廉,绿色环保。(The invention belongs to the technical field of fine chemical synthesis, and particularly relates to an indoline isochroman derivative, an enzymatic synthesis method and application thereof. The indoline isochroman compound with high added value is obtained by the oxidation coupling reaction catalyzed by the multi-copper oxidase serving as the catalyst from cheap and easily available chemical raw materials, and the method is mild in reaction condition, simple to operate, low in cost, green and environment-friendly.)

1. An indoline isochroman derivative, which is characterized in that: the structure is as follows:

the R is¹Is any one of hydrogen, alkyl and heteroaromatic radical; r²Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano and acyl; r³Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano, acyl, amino and aryl.

2. A preparation method of indoline isochroman derivatives is characterized by comprising the following steps: 2, 3-dihydroxy benzoic acid and indole compounds are taken as substrates, copper-rich oxidase is taken as a catalyst, and the indoline isochroman derivatives are obtained through reaction, wherein the reaction route is as follows:

the R is¹Is any one of hydrogen, alkyl and heteroaromatic radical; r²Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano and acyl; r³Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano, acyl, amino and aryl.

3. The method for producing an indoline isochroman derivative according to claim 2, characterized in that: the indole compound is one of a formula (1) to a formula (6), and the structures of the formula (1) to the formula (6) are as follows:

4. the method for preparing indoline isochroman derivatives according to claim 2, comprising the steps of:

(1) sequentially adding 2, 3-dihydroxybenzoic acid, indole compounds and copper-rich oxidase into a mixed system of an organic solvent, water and acid for catalytic reaction;

(2) and after the reaction is finished, performing extraction separation, and drying to obtain the indoline isochroman derivative.

5. The method for preparing indoline isochroman derivative according to claim 4, wherein the organic solvent is one of dimethyl sulfoxide, N-dimethylformamide and acetonitrile; the acid is one of hydrochloric acid, formic acid and acetic acid; the volume of the organic solvent in the mixed system accounts for 10-50%.

6. The method of claim 4, wherein the multicopper oxidase is at least one of a bacterial manganese oxidase, a copper efflux oxidase, and a Bacillus licheniformis laccase.

7. The method for producing an indoline isochroman derivative according to claim 4, wherein the mass ratio of the multicopper oxidase to the indole compound is 0.01-0.1:1, and the mass ratio of the 2, 3-dihydroxybenzoic acid to the indole compound is 1: 1-1.2.

8. The method for producing an indoline isochroman derivative according to claim 4, further comprising an auxiliary oxidizing agent, wherein the auxiliary oxidizing agent is added simultaneously with the copper-rich oxidase, and the auxiliary oxidizing agent is at least one of air, 30% aqueous hydrogen peroxide, and TBHP.

9. The method for preparing indoline isochroman derivative according to claim 4, wherein the temperature of the catalytic reaction is 4-37 ℃.

10. Use of the indoline isochroman derivative according to any one of claims 1-9 in the manufacture of a medicament or natural product.

Technical Field

The invention belongs to the field of fine chemical engineering, and particularly relates to an indoline isochroman derivative, and an enzymatic synthesis method and application thereof.

Background

Indoline isochroman compounds are a class of compounds with monoterpene alkaloid structures and widely exist in a plurality of natural products and drug molecules, such as bisphylline and rhynchophylline, and the prior researches prove that the indoline isochroman compounds have potential biological activities, including antibacterial, anti-inflammatory and anti-tumor effects. Within the organic chemistry community, synthesis of bisphylline is considered a climbing-worthy mountain. The existing literature reports that the synthesis of indoline isochroman compounds often needs the application of transition metals, which brings huge influence on the residual quantity of metals in the following pharmaceutical research. Therefore, the traditional chemical synthesis of indoline isochroman compounds has the problems of impracticality, uneconomical and environmental pollution in chemical production, and the development of a green and efficient synthesis of indoline isochroman compounds is urgently needed.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides an indoline isochroman derivative and a preparation method thereof, which solve the defects of complex operation, high production cost and the like in the prior art, and the indoline isochroman compound with high added value is obtained by the oxidation coupling reaction catalyzed by taking copper-rich oxidase as a catalyst from cheap and easily available chemical raw materials, and the indoline isochroman derivative is mild in reaction condition, simple to operate, low in cost and environment-friendly.

In order to achieve the technical purpose, the technical scheme of the invention is as follows:

an indoline isochroman derivative has the following structure:

the R is¹Is any one of hydrogen, alkyl and heteroaromatic radical; r²Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano and acyl; r³Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano, acyl, amino and aryl.

A preparation method of indoline isochroman derivatives comprises the following steps of reacting 2, 3-dihydroxy benzoic acid and indole compounds serving as substrates and a copper-rich oxidase serving as a catalyst to obtain the indoline isochroman derivatives, wherein the reaction route is as follows:

the R is¹Is any one of hydrogen, alkyl and heteroaromatic radical; r²Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano and acyl; r³Is any one of hydrogen, halogen, alkyl, alkoxy, nitro, ester group, cyano, acyl, amino and aryl.

The indole compound is one of a formula (1) to a formula (6), and the structures of the formula (1) to the formula (6) are as follows:

the preparation method comprises the following steps:

(1) sequentially adding 2, 3-dihydroxybenzoic acid, indole compounds and copper-rich oxidase into a mixed system of an organic solvent, water and acid for catalytic reaction;

(2) and after the reaction is finished, performing extraction separation, and drying to obtain the indoline isochroman derivative.

Wherein, the organic solvent is one of dimethyl sulfoxide, N-dimethylformamide and acetonitrile, and acetonitrile is preferred.

The acid is one of hydrochloric acid, formic acid and acetic acid, and is preferably hydrochloric acid.

The volume ratio of the organic solvent in the mixed system is 10-50%, preferably 50%.

The multicopper oxidase is at least one of bacterial manganese oxidase, copper efflux oxidase and bacillus licheniformis laccase; further, the copper-rich oxidase is bacillus licheniformis laccase, and the bacillus licheniformis laccase is obtained by fermentation of escherichia coli.

The mass ratio of the copper-rich oxidase to the indole compound is 0.01-0.1:1, and the mass ratio of the 2, 3-dihydroxybenzoic acid to the indole compound is 1: 1-1.2.

The preparation method also comprises an auxiliary oxidizing agent, and the auxiliary oxidizing agent is added simultaneously with the copper-rich oxidase, and the auxiliary oxidizing agent is at least one of air, 30% aqueous hydrogen peroxide solution and tert-butyl aqueous hydrogen peroxide solution TBHP, wherein 30% aqueous hydrogen peroxide solution is preferred.

The temperature of the catalytic reaction is 4-37 ℃.

The application of the indoline isochroman derivative in preparing medicines or natural products.

From the above description, it can be seen that the present invention has the following advantages:

1. the invention solves the defects of complex operation, high production cost and the like in the prior art, and obtains the indoline isochroman compound with high added value through the oxidative coupling reaction catalyzed by taking copper-rich oxidase as a catalyst from cheap and easily-obtained chemical raw materials. The reaction mechanism of the invention is to form a benzoquinone intermediate by oxidizing 2, 3-dihydroxy benzoic acid substrate by bacillus licheniformis laccase CotA, then perform nucleophilic addition of indole compounds to the intermediate, and finally cyclize to obtain the indoline isochroman compounds. Multicopper oxidase is an oxidoreductase containing a metallic copper center, widely present in bacteria, fungi and higher plants. Most studied laccases, for example, are generally composed of four copper atoms and are classified according to three active sites, type 1 or blue, type 2 or normal, type 3 or binuclear. The T1 Cu active site accepts 4 electrons of substrate oxidation and transfers them to the T2/T3 Cu cluster, and in the T2/T3 Cu cluster, the molecular oxygen is completely reduced to 2 water molecules by accepting 4 electrons.

2. The preparation method disclosed by the invention is mild in reaction, the organic solvent with a certain proportion is beneficial to dissolving a reaction substrate, the acid can improve the conversion rate of the reaction, the cost is low, the operation is simple, and the preparation method is green and environment-friendly.

3. The preparation method has high conversion rate and yield, the adopted bacillus licheniformis laccase CotA is obtained by fermenting escherichia coli, the production cost is lower than that of the prior art, and the industrial production is facilitated.

Drawings

FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of the objective product prepared in example 1.

Detailed Description

With reference to fig. 1, a specific embodiment of the present invention is described in detail, but the present invention is not limited in any way by the claims.

The 2, 3-dihydroxybenzoic acid, hydrogen peroxide, indole compounds, hydrochloric acid in the examples were all purchased from Shanghai, and the Bacillus licheniformis laccase CotA (NCBI accession No. YP-077905) was obtained by fermentation of E.coli and purchased from Jinzhi Biotech, Inc., Suzhou.

Example 1

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg), 3-methylindole (0.15mmol,19.7mg), CotA (0.1 mol%), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added to a 10 mL reaction flask in this order, and magnetic stirring was carried out at room temperature for 48 hours, after the substrate conversion was completed, ethyl acetate was added to the reaction system for extraction (3mL 4), the organic phases were combined and dried over anhydrous sodium sulfate, and after filtration, vacuum concentration was carried out to obtain a crude product. The obtained crude product is purified and separated by column chromatography to obtain a yellow solid product, and the separation yield is 87%.

The synthetic route of example 1 is as follows:

FIG. 1 shows the NMR spectrum of the objective product obtained in example 1.

¹H NMR(400MHz,CDCl₃):11.52(s,1H),7.28(s,1H),7.11(q,J＝15.6Hz,8.0Hz,2H),6.93(d,J＝7.2Hz,1H),6.79(q,J＝17.6Hz,8.0Hz,2H),5.81(s,1H),5.69(s,1H),4.96(s,1H),1.64(s,3H)。

Example 2

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg), 6-fluoro-3-methylindole (0.15mmol,22.4mg), CotA (0.1 mol%), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added to a 10 mL reaction flask in this order, and magnetic stirring was carried out at room temperature for 48 hours, after the substrate conversion was completed, ethyl acetate was added to the reaction system for extraction (3mL 4), the organic phases were combined and dried over anhydrous sodium sulfate, and the crude product was obtained by vacuum concentration after filtration. The obtained crude product is purified and separated by column chromatography to obtain a yellow solid product, and the separation yield is 85%.

The synthetic route of example 2 is shown below:

the nuclear magnetic resonance hydrogen spectrum of the obtained target product is as follows:

¹H NMR(400MHz,CDCl₃):11.50(s,1H),7.30(s,1H),7.08(d,J＝8.0Hz,1H),6.85(q,J＝5.6Hz,8.8Hz,1H),6.50(d,J＝5.2Hz,2H),5.83(s,1H),5.08(s,1H),1.65(s,3H)。

example 3

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg), 5-chloro-3-methylindole (0.15mmol,24.8mg), CotA (0.1 mol%), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added to a 10 mL reaction flask in this order, and magnetic stirring was carried out at room temperature for 48 hours, after the substrate conversion was completed, ethyl acetate was added to the reaction system for extraction (3mL 4), the organic phases were combined and dried over anhydrous sodium sulfate, and the crude product was obtained by vacuum concentration after filtration. The obtained crude product is purified and separated by column chromatography to obtain a yellow solid product, and the separation yield is 83 percent.

The synthetic route for example 3 is shown below:

the nuclear magnetic resonance hydrogen spectrum of the obtained target product is as follows:

¹H NMR(400MHz,CDCl₃):11.51(s,1H),7.31(d,J＝8.4Hz,1H),7.11–7.08(m,2H),6.88(d,J＝2.0Hz,1H),6.71(d,J＝8.4Hz,1H),5.82(s,1H),4.99(s,1H),1.66(s,3H)。

example 4

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg), 5-bromo-3-methylindole (0.15mmol,31.5mg), CotA (0.1 mol%), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added to a 10 mL reaction flask in this order, and magnetic stirring was carried out at room temperature for 48 hours, after the substrate conversion was completed, ethyl acetate was added to the reaction system for extraction (3mL 4), the organic phases were combined and dried over anhydrous sodium sulfate, and the crude product was obtained by vacuum concentration after filtration. The obtained crude product is purified and separated by column chromatography to obtain a yellow solid product, and the separation yield is 64%.

The synthetic route of example 4 is as follows:

the nuclear magnetic resonance hydrogen spectrum of the obtained target product is as follows:

¹H NMR(400MHz,CDCl₃):11.50(s,1H),7.31(d,J＝8.4Hz,1H),,7.25(dd,J＝6.0Hz,8.0Hz,1H),7.08(d,J＝8.0Hz,1H),7.01(d,J＝2.0Hz,1H),6.67(d,J＝8.4Hz,1H),5.80(s,1H),5.00(s,1H),1.66(s,3H)。

example 5

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg), 3-ethyl indole (0.15mmol,21.8mg), CotA (0.1 mol%)), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added in sequence into a 10 mL reaction flask, magnetic stirring was carried out at room temperature for 48 hours, ethyl acetate was added into the reaction system for extraction (3mL 4) after substrate conversion was completed, the organic phases were combined and dried over anhydrous sodium sulfate, and vacuum concentration was carried out after filtration to obtain a crude product. The obtained crude product is purified and separated by column chromatography to obtain a yellow solid product, and the separation yield is 76%.

The synthetic route of example 5 is as follows:

the nuclear magnetic resonance hydrogen spectrum of the obtained target product is as follows:

¹H NMR(400MHz,CDCl₃):11.56(s,1H),7.29(d,J＝6.4Hz,1H),7.15-7.11(m,1H),7.02(t,J＝8.4Hz,2H),6.85–6.81(m,1H),6.76(d,J＝8.0Hz,1H),5.99(s,1H),4.97(s,1H),2.51-2.15(m,1H),1.97-1.88(m,1H),0.93(t,J＝7.6Hz,3H)。

example 6

2, 3-dihydroxybenzoic acid (0.3mmol,46.2mg),2, 3-dimethyloindole (0.15mmol,21.8mg), CotA (0.1 mol%), hydrogen peroxide (0.75mmol,75uL), deionized water (1mL), acetonitrile (1mL), hydrochloric acid (0.3mmol,13uL) were added to a 10 mL reaction flask in this order, and magnetic stirring was carried out at room temperature for 48 hours, and ethyl acetate was added to the reaction system for extraction (3mL 4) after the substrate conversion was completed, and the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. The crude product was purified and separated by column chromatography to give a yellow solid product with a separation yield of 73%.

The synthetic route for example 6 is shown below:

the nuclear magnetic resonance hydrogen spectrum of the obtained target product is as follows:

¹H NMR(400MHz,CDCl₃):11.57(s,1H),7.29(d,J＝5.6Hz,1H),7.12(d,J＝8.4Hz,2H),6.81(d,J＝4.0Hz,2H),6.76(d,J＝7.6Hz,1H),5.71(s,1H),4.93(s1H),1.76(s,3H),1.59(s,3H)。

it should be understood that the detailed description of the invention is merely illustrative of the invention and is not intended to limit the invention to the specific embodiments described. It will be appreciated by those skilled in the art that the present invention may be modified or substituted equally as well to achieve the same technical result; as long as the use requirements are met, the method is within the protection scope of the invention.