阅读说明：本技术 一种喹啉衍生物及其制备方法与应用 (Quinoline derivative and preparation method and application thereof ) 是由 王战辉 温凯 沈建忠 张会艳 于雪芝 余文博 史为民 张素霞 于 2019-11-21 设计创作，主要内容包括：本发明涉及化工技术领域,具体公开了一种喹啉衍生物及其制备方法与应用。所述喹啉衍生物具有如下结构：该喹啉衍生物表面具有一个高活性的羧基可直接用于标记,且该喹啉衍生物由于具有蓝色的发射波长,填补了目前传统的有机染料荧光素在低波长段的空白,拓宽了多种标记模式的荧光素选择范围。此外,本发明的喹啉衍生物光稳定性好,在有机试剂中稳定、能保持良好的荧光特性,适于有机溶剂中的标记反应。所述喹啉衍生物的制备方法为：将4-苯甲酰基丁酸经酰胺化、肟化、还原反应后,首先合成该喹啉衍生物的侧链,然后再与6-溴喹啉发生缩合反应合成最终的喹啉衍生物。方法简便、温和。(The invention relates to the technical field of chemical industry, and particularly discloses a quinoline derivative and a preparation method and application thereof. The quinoline derivative has the following structure:)

1. A quinoline derivative having the structure:

2. a process for preparing the quinoline derivative of claim 1, comprising:

(1) 4-benzoyl butyric acid is subjected to amidation reaction to obtain a compound I,

(2) oximation reaction is carried out on the compound I to obtain a compound II,

(3) the compound II is subjected to reduction reaction to obtain a compound I,

(4) reacting the compound I with 6-bromoquinoline to obtain a compound IV,

(5) hydrolyzing the compound IV.

3. The method according to claim 2, wherein in step (1), the 4-benzoylbutyric acid is reacted with dimethylamine hydrochloride, N-diisopropylethylamine, tetramethylurea hexafluorophosphate in a first organic solvent to obtain the compound i;

and/or the molar ratio of the 4-benzoylbutyric acid to the dimethylamine hydrochloride is 1: (1-1.5); the molar ratio of the 4-benzoylbutyric acid to the N, N-diisopropylethylamine is 1: (4-8); the molar ratio of the 4-benzoylbutyric acid to the tetramethylurea hexafluorophosphate ester is 1: (1-2);

and/or, the reaction temperature of the step (1) is 25 +/-1 ℃, and the reaction time is 12-18 hours;

and/or the first organic solvent is dichloromethane.

4. The method according to claim 2 or 3, wherein in the step (2), the compound I is reacted with hydroxylamine hydrochloride and potassium carbonate in a second organic solvent to obtain the compound II;

and/or the molar ratio of the compound I to the hydroxylamine hydrochloride is 1: (1-1.5); the molar ratio of the compound I to the potassium carbonate is 1: (1-2);

and/or, the reaction temperature of the step (2) is 100 +/-10 ℃, and the reaction time is 5-7 hours;

and/or the second organic solvent is ethanol.

5. The method according to any one of claims 2 to 4, wherein in step (3), the compound II is reacted with hydrogen in a third organic solvent in the presence of palladium on carbon as a catalyst to obtain the compound II;

and/or, the reaction temperature of the step (3) is 25 +/-1 ℃, and the reaction time is 48-55 hours;

and/or the third organic solvent is ethanol.

6. The method according to any one of claims 2 to 5, wherein in the step (4), the compound II and the 6-bromoquinoline are reacted in a fourth organic solvent to obtain the compound IV by using palladium acetate, (±) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl and cesium carbonate as catalysts;

and/or the molar ratio of the compound II to the 6-bromoquinoline is 1: (1-2); the molar ratio of the compound II to the palladium acetate is 1: (0.2-0.6); the molar ratio of the compound II to the (+/-) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl is 1: (0.3-1); the molar ratio of said compound ii I to said cesium carbonate is 1: (2-3);

and/or, the reaction temperature of the step (4) is 100 ℃ +/-5 ℃, and the reaction time is 24-28 hours;

and/or, the fourth organic solvent is toluene.

7. The process according to any one of claims 2 to 6, wherein in step (5), the compound IV is reacted with an aqueous sodium hydroxide solution in a fifth organic solvent;

and/or, the reaction temperature of the step (5) is 50 +/-10 ℃, and the reaction time is 24-30 hours;

and/or the fifth organic solvent is ethanol.

8. The method as claimed in any one of claims 2 to 7, wherein the reaction of step (1) is finished and further comprises a step of separating the compound I by thin layer chromatography, the developing solvent in step (1) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (45-55): 1, preferably 50: 1;

and/or, after the reaction in the step (2) is finished, the step of separating the compound II by adopting thin layer chromatography is further included, the developing solvent in the step (2) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (45-55): 1, preferably 50: 1;

and/or, after the reaction in the step (3) is finished, the method further comprises a step of separating the compound III by adopting thin layer chromatography, wherein the developing solvent in the step (3) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (25-35): 1, preferably 30: 1;

and/or, after the reaction in the step (4) is finished, the step of separating the compound IV by adopting thin layer chromatography is further included, the developing solvent in the step (4) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (100-30): 1, preferably 50: 1.

9. the method according to claim 7 or 8, wherein the step (5) further comprises, after the reaction is finished: removing the fifth organic solvent, adjusting the pH value to 2.8-3.2, precipitating, filtering, and recrystallizing in methanol.

10. Use of a quinoline derivative according to claim 1 or a method according to any one of claims 2-9 for labelling a small molecule or a protein molecule comprising an amino group.

Technical Field

The invention relates to the technical field of chemical industry, in particular to a quinoline derivative and a preparation method and application thereof.

Background

The parent structure of the quinoline derivative is quinoline, which was first isolated from coal tar. Quinoline derivatives are a very important heterocyclic compound, and have obvious pharmacological activity and biological activity, so that the quinoline derivatives are widely applied to the fields of drug screening, dye industry and chemical analysis. A great deal of research shows that many compounds with quinoline rings have obvious biological activity and pharmacological activity such as antibiosis, sterilization, pain relief, antianaphylaxis, antimalarial, antitumor, anticancer, antihypertensive, antidepressant, memory enhancement and the like, and in recent years, quinoline derivatives are also used for developing drugs for treating AIDS. In addition, due to the excellent optical properties of quinoline derivatives, quinoline heterocyclic derivatives play an important role in the fields of chemical sensors, analytical detection, synthesis of fluorescent sensor molecules, biological analysis imaging and the like. As a class of high-performance organic fluorescein, the most common quinoline derivative fluorescein at present is quinine sulfate, the fluorescein plays an important role in treating malaria and measuring the relative quantum yield of novel fluorescein with similar emission bands, and the quinoline derivative has structure specificity capable of forming chelate with certain metal ions, and the fluorescein is specially used for identifying and detecting metal ions such as zinc, copper, mercury and the like. However, the quinoline derivative fluorescein can not be used for labeling common small molecular compounds or biological macromolecules such as proteins, so that the quinoline derivative fluorescein has no universality and has great limitation in the fields of immunoassay, small molecule detection and the like.

Therefore, it is desirable to provide a new quinoline derivative, a preparation method and an application thereof to solve the problems in the prior art.

Disclosure of Invention

In order to solve the problems in the prior art, the invention aims to provide a quinoline derivative which is beneficial to labeling of small molecular compounds or biological macromolecules, has good water solubility and is suitable for labeling in an organic solvent.

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

a quinoline derivative (fluorescein) having the structure:

the surface of the quinoline derivative (4- (6-imine quinoline) -5-phenyl-pentanoic acid) has a high-activity carboxyl which can be directly used for marking, and provides a foundation for the application of the quinoline derivative in cell marking, immune reaction and the like. And the quinoline derivative has blue emission wavelength, so that the blank of the traditional organic dye fluorescein in a low wavelength band at present is filled, and the fluorescein selection range of various marking modes is widened. In addition, the quinoline derivative has good light stability, is stable in organic reagents such as ethanol and methanol, can keep good fluorescence characteristics, and is suitable for labeling reaction in organic solvents.

It is another object of the present invention to provide a simple and mild process for preparing the quinoline derivatives.

The method for preparing the quinoline derivative comprises the following steps:

(1) 4-benzoyl butyric acid is subjected to amidation reaction to obtain a compound I,

(2) oximation reaction is carried out on the compound I to obtain a compound II,

(3) the compound II is subjected to reduction reaction to obtain a compound I,

(4) reacting the compound I with 6-bromoquinoline to obtain a compound IV,

(5) hydrolyzing the compound IV.

According to the invention, 4-benzoyl butyric acid is subjected to amidation, oximation and reduction reactions, and then the side chain of the quinoline derivative is firstly synthesized, and then the side chain and 6-bromoquinoline undergo a condensation reaction to synthesize the final quinoline derivative.

In the step (1), the 4-benzoyl butyric acid reacts with dimethylamine hydrochloride, N-diisopropylethylamine and tetramethylurea hexafluorophosphate in a first organic solvent to obtain a compound I;

and/or the molar ratio of the 4-benzoylbutyric acid to the dimethylamine hydrochloride is 1: (1-1.5); the molar ratio of the 4-benzoylbutyric acid to the N, N-diisopropylethylamine is 1: (4-8); the molar ratio of the 4-benzoylbutyric acid to the tetramethylurea hexafluorophosphate ester is 1: (1-2);

and/or, the reaction temperature of the step (1) is 25 +/-1 ℃, and the reaction time is 12-18 hours;

and/or the first organic solvent is dichloromethane.

In the step, the compound I is generated by amidation reaction of secondary amine in dimethylamine hydrochloride and carboxyl in 4-benzoyl butyric acid.

Preferably, after the reaction in step (1) is finished, the reaction product is washed by water, then dried by anhydrous sodium sulfate, and the precipitate is taken out by centrifugation for the subsequent separation step.

In the step (2), reacting the compound I with hydroxylamine hydrochloride and potassium carbonate in a second organic solvent to obtain a compound II;

and/or the molar ratio of the compound I to the hydroxylamine hydrochloride is 1: (1-1.5); the molar ratio of the compound I to the potassium carbonate is 1: (1-2);

and/or, the reaction temperature of the step (2) is 100 +/-10 ℃, and the reaction time is 5-7 hours;

and/or the second organic solvent is ethanol.

In the step, the amide group in the compound I is changed into oxime by utilizing the oximation reaction of hydroxylamine hydrochloride on carbonyl in the compound I, so that the compound I is generated.

Preferably, after the reaction in step (2) is completed, the second organic solvent is evaporated, water is added, and then, after extraction with ethyl acetate, the organic layer is dried over anhydrous sodium sulfate and concentrated for use in the subsequent separation step.

In the step (3), palladium carbon is used as a catalyst, and the compound II and hydrogen react in a third organic solvent to obtain the compound II;

and/or, the reaction temperature of the step (3) is 25 +/-1 ℃, and the reaction time is 48-55 hours;

and/or the third organic solvent is ethanol.

Preferably, the palladium content in the palladium on carbon is 5% (mass fraction).

In the step (3), the whole reaction is carried out in a hydrogen environment, and the addition of hydrogen and palladium carbon is preferably carried out in order to ensure that the reaction is fully carried out.

In the step, oxime is reduced to generate amino by using the catalytic action of palladium-carbon to obtain a compound II.

Preferably, after the reaction in step (3) is completed, the catalyst is removed by filtration, and the third organic solvent is evaporated for the subsequent separation step.

In the step (4), palladium acetate, (+ -) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl and cesium carbonate are used as catalysts, and the compound II and the 6-bromoquinoline react in a fourth organic solvent to obtain a compound IV;

and/or the molar ratio of the compound II to the 6-bromoquinoline is 1: (1-2); the molar ratio of the compound II to the palladium acetate is 1: (0.2-0.6); the molar ratio of the compound II to the (+/-) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl is 1: (0.3-1); the molar ratio of said compound ii I to said cesium carbonate is 1: (2-3).

And/or, the reaction temperature of the step (4) is 100 ℃ +/-5 ℃, and the reaction time is 24-28 hours;

and/or, the fourth organic solvent is toluene.

In the step, a compound II and 6-bromoquinoline are subjected to condensation reaction (reaction under the protection of nitrogen) under the catalytic action of palladium acetate, (+/-) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl and cesium carbonate to generate a compound IV.

Preferably, after the reaction of step (4) is completed, dichloromethane is added after the fourth organic solvent is removed by concentration, and after extraction with water, the organic layer is dried over anhydrous sodium sulfate and used in the subsequent separation step.

In step (5) of the present invention, reacting the compound IV with an aqueous solution of sodium hydroxide in a fifth organic solvent;

and/or, the reaction temperature of the step (5) is 50 +/-10 ℃, and the reaction time is 24-30 hours;

and/or the fifth organic solvent is ethanol.

In the present invention, the molar ratio of the compound IV to the sodium hydroxide is 1: (1-3).

In the step, the amide bond is hydrolyzed by utilizing the hydrolysis action of sodium hydroxide to obtain the quinoline derivative with the carboxyl.

As an embodiment of the present invention, the reaction scheme of the method is as follows:

wherein Me is₂NHHCL as dimethylamine hydrochloride, DCM as dichloromethane, HATU as tetramethylurea hexafluorophosphate, DIPEA as N, N-diisopropylethylamine, EtOH as ethanol, Pd (OAc)₂Being palladium acetate, BINAP is (±) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl, toluene is toluene.

The method also comprises a step of separating the compound I by adopting a thin-layer chromatography after the reaction in the step (1) is finished, wherein a developing solvent in the step (1) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (45-55): 1, preferably 50: 1;

and/or, after the reaction in the step (2) is finished, the step of separating the compound II by adopting thin layer chromatography is further included, the developing solvent in the step (2) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (45-55): 1, preferably 50: 1;

and/or, after the reaction in the step (3) is finished, the method further comprises a step of separating the compound III by adopting thin layer chromatography, wherein the developing solvent in the step (3) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (25-35): 1, preferably 30: 1;

and/or, after the reaction in the step (4) is finished, the step of separating the compound IV by adopting thin layer chromatography is further included, the developing solvent in the step (4) is a mixed solution of dichloromethane and methanol, and the volume ratio of the dichloromethane to the methanol is (100-30): 1, preferably 50: 1.

the method also comprises the following steps after the step (5) of the invention is finished: removing the fifth organic solvent, adjusting the pH value to 2.8-3.2 (preferably 3), precipitating, filtering, and recrystallizing in methanol. Preferably, after the reaction of step (5) is completed, the fifth organic solvent is removed in a concentrated manner.

The invention also discloses application of the quinoline derivative or the method in marking small molecules or protein molecules containing amino groups.

The invention has the beneficial effects that:

the quinoline derivative can be used for directly marking micromolecules or protein molecules containing amino groups, has good water solubility and stable fluorescence property of aqueous solution, can keep good fluorescence property in organic solvents such as methanol, ethanol and the like, is suitable for marking reaction in the organic solvents, fills the blank of the traditional organic dye fluorescein in a low wavelength band at present, widens the selection range of probes in fluorescence marking, and can powerfully promote the application of the quinoline fluorescein in the aspects of immunoassay, molecular marking and the like. The synthesis method of the invention has mild reaction conditions, is simple and convenient, is easy to operate and is suitable for popularization.

Drawings

FIG. 1 is an absorption spectrum of a quinoline derivative according to the present invention;

FIG. 2 is a diagram of the excitation spectrum of a quinoline derivative according to the present invention;

FIG. 3 is a graph showing an emission spectrum of a quinoline derivative of the present invention;

FIG. 4 is a nuclear magnetic resonance C spectrum of a quinoline derivative according to the present invention;

FIG. 5 is a nuclear magnetic resonance H spectrum of a quinoline derivative of the present invention;

FIG. 6 is a mass spectrum of the quinoline derivative of the present invention.

Detailed Description

Preferred embodiments of the present invention will be described in detail with reference to the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1

This example provides a quinoline derivative of the present invention and a method for preparing the same.

The preparation method comprises the following specific steps:

(1) carrying out amidation reaction on 4-benzoyl butyric acid to obtain a compound I:

119g (100mmol) of 4-benzoylbutyric acid was dissolved in 500mL of Dichloromethane (DCM), and then 8.97g (110mmol) of dimethylamine hydrochloride, 50mL of N, N-Diisopropylethylamine (DIPEA), 41.80g (110mmol) of tetramethyluronium Hexafluorophosphate (HATU) was added, and the mixture was stirred at room temperature and 25 ℃ for 16 hours, after completion of the reaction, washed with 500mL of water each time, repeated twice, and then dried over anhydrous sodium sulfate, and the precipitate was centrifuged to obtain 14.25g of compound I, which was isolated by thin layer chromatography (dichloromethane/methanol (v/v,50:1) as a developing solvent, in 65% yield.

(2) Carrying out oximation reaction on the compound I to obtain a compound II;

14.25g (65mmol) of Compound I, 4.80g (69mmol) of hydroxylamine hydrochloride and 17.94g (130mmol) of potassium carbonate were dissolved in 250mL of an ethanol solution and heated under reflux at 100 ℃ for 5 hours. After the reaction was complete, the reaction solvent, ethanol, was evaporated, 300mL of water was added, and the mixture was extracted twice with 250mL of ethyl acetate each time. The organic layer was dried over anhydrous sodium sulfate, concentrated and then separated by thin layer chromatography (dichloromethane/methanol (v/v,50:1) as a developing solvent) to obtain 11.20g of compound II as a white solid in 73% yield.

(3) Carrying out reduction reaction on the compound II to obtain a compound I;

11.20g (47.5mmol) of Compound II were dissolved in 100mL of ethanol, 4g of palladium on carbon (5%) catalyst was added, and the whole reaction system was exposed to pure hydrogen and reacted at 25 ℃ for 48 hours. After the reaction was completed, the catalyst was removed by filtration, the organic solvent was evaporated off, and the residue was separated by thin layer chromatography (dichloromethane/methanol (v/v, 30: 1) as a developing solvent) to obtain 6.5g of compound II as a colorless oil in 62% yield.

(4) Reacting the compound II I with 6-bromoquinoline to obtain a compound IV:

6.24g (30mmol) of the compound II, 6.6g (30mmol) of 6-bromoquinoline, 1.34g (6mmol) of palladium acetate, 7.47g (12mmol) (. + -.) -2,2 '-bis (diphenylphosphino) -1, 1' -binaphthyl, 19.55g (60mmol) of cesium carbonate were dissolved in 250mL of toluene, reacted at 95 ℃ for 24h under a pure nitrogen atmosphere, concentrated after the reaction to remove toluene, and the residue was extracted twice with 500mL of dichloromethane and 250mL of water. The organic layer was dried over anhydrous sodium sulfate and separated and purified by thin layer chromatography (dichloromethane/methanol (v/v,50:1) as a developing solvent to finally obtain 4.60g of compound IV with a yield of 44%.

(5) (ii) hydrolyzing the compound IV;

4.60g (21mmol) of Compound IV, 30mL (6M) of aqueous sodium hydroxide solution and 30mL of ethanol are heated to 42 ℃ and evaporated under reflux for 24 hours. After the reaction was completed, the mixture was concentrated to remove ethanol, the aqueous phase was adjusted to pH 3 with 2M hydrochloric acid, the precipitate was filtered and the solid was collected and recrystallized from methanol to finally obtain 1.90g of the quinoline derivative of the present invention as a white powder in 28% yield.

The quinoline derivative prepared in this example was subjected to an absorption spectrum test (spectrum shown in fig. 1, test condition 0.1mg/mL in aqueous solution), an excitation spectrum test (spectrum shown in fig. 2, test condition 0.1mg/mL in aqueous solution, excitation wavelength 450nm), an emission spectrum test (spectrum shown in fig. 3, test condition 0.1mg/mL in aqueous solution, emission wavelength 360nm), and it was found from each spectrum that the quinoline derivative had three absorption peaks, 230nm,270nm, and 360nm, respectively. The wavelength range of excitation is 220-430nm, where excitation emission peaks occur at 279nm and 368nm, respectively. The emission wavelength is in the range of 380-570nm, and the emission peak is 445 nm.

The quinoline derivatives prepared in this example were subjected to structural identification, and the nuclear magnetic and mass spectrum data thereof were as follows:

¹H NMR(300M Hz,DMSO-d₆)δ1.42-1.90(m,4H),2.24(t,J＝6.3Hz,2H),4.49-4.51(m,1H),6.50(s,1H),7.07(d,J＝7.2Hz,1H),7.14-7.25(m 3H),7.31(t,J＝7.5Hz,2H),7.43(t,J＝7.5Hz,2H),7.70(d,J＝8.7Hz,1H),8.11(d,J＝5.7Hz,1H),8.81(s,1H).¹³C NMR(125MHz,DMSO-d₆)δ21.78,33.61,36.99,56.14,100.58,118.18,119.42,121.90,126.33,126.61,128.15,128.22,137.53,142.73,143.85,149.29,150.58,174.39。

ESI-HRMS:m/z for[M+H]⁺C₂₀H₂₃N₃O₂,321.15975,found 321.16。

the nuclear magnetic resonance C spectrum, the nuclear magnetic resonance H spectrum and the mass spectrum of the quinoline derivative prepared in the example are shown in the figures 4, 5 and 6 respectively.

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. The person skilled in the art can obtain quinoline molecule derivatives with different substituent groups on the basis of the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.