Quinoxalinone boride with high-efficiency double-state luminescence as well as preparation and application of quinoxalinone boride

Abstract

The invention discloses a quinoxalinone boride with efficient bi-state luminescence and preparation and application thereof.The material mainly takes a quinoxalinone-BF2 structure as a core, the luminescent material has the advantages that synthesis steps are simple, the preparation method is convenient, the luminescent property is excellent, the efficient bi-state photoluminescence property of a solution state and a solid state is achieved, and the luminescent material is suitable for large-scale production. The biotoxicity is low, the biocompatibility is high, and the long-time cell imaging function is realized. And the probe can become a novel biological probe molecular platform.

Description

Technical Field

[0001] The invention relates to the field of biological imaging, and in particular to a quinoxalinone boride with high-efficiency dual-state luminescence, and its preparation and application. Background Art

[0002] Traditional organic light-emitting molecules usually have bright fluorescence emission in the single-molecule state of solution, but in the aggregated state, the probability of non-radiative transition of excited state electrons increases due to molecular stacking, resulting in fluorescence quenching in the solid state. Although molecules can emit bright fluorescence in the solid or aggregated state through aggregation-induced emission (AIE), they have the characteristic of non-luminescence in solution. These factors make organic molecular light-emitting molecules only show bright fluorescence in a single state (solution or solid state). In recent years, the development of organic dual-state light-emitting materials has received increasing attention because they show strong fluorescence emission in both solution and solid state. However, there are relatively few organic molecules with dual-state light-emitting molecular structures. Therefore, the development of new organic molecular structures with solid-liquid dual-state light-emitting is of great significance to the wider application of organic light-emitting materials.

[0003] On the other hand, relatively few of the fluorescent dyes currently used can be used for long-term imaging in cells. The reason is that factors such as the photostability of the probe molecules and cell metabolism cause the probe molecules to stay in cells for a short time. Summary of the invention

[0004] The purpose of the present invention is to provide a quinoxalinone boride with high efficiency dual-state luminescence and its preparation and application. A quinoxalinone boride with high efficiency dual-state luminescence has a structure shown in Formula VI or Formula VII:

[0005]

[0006] Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring;

[0007] R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group (including a cycloalkyl group), an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

[0008] A method for preparing a quinoxalinone boride with high-efficiency dual-state luminescence comprises the following steps:

[0009]

[0010] Step 1: Add the compound represented by Formula I and the compound represented by Formula II or Formula III to a solvent, and heat to reflux for reaction; after the reaction, recrystallize with ethanol / ether to obtain a ligand containing a quinazolinone-quinoline or pyridine structure represented by Formula IV or Formula V;

[0011]

[0012] Step 2: adding a ligand containing a quinazolinone-quinoline or pyridine structure shown in Formula IV or Formula V to a non-polar solvent, adding a boron trifluoride ether solution and triethylamine, and performing a reflux reaction; cooling the reaction liquid, adding saturated sodium bicarbonate to quench the reaction, standing, layering, extracting with ethyl acetate, concentrating the organic phase, and separating the mixture by column chromatography to finally obtain a quinoxalinone boron compound shown in Formula VI or Formula VII;

[0013] Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring;

[0014] R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group (including a cycloalkyl group), an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

[0015] Preferably, the solvent in step 1 is ethanol, ethyl acetate, tetrahydrofuran, or 1,2-dichloroethane.

[0016] Preferably, the reaction time in step 1 is 16-48 hours, and the reaction temperature is 70°C to 90°C.

[0017] Preferably, the solvent in step 1 is ethanol, and the reaction time is 24 hours.

[0018] Preferably, the non-polar solvent in step 2 is ethyl acetate, tetrahydrofuran, 1,2-dichloroethane, or dichloromethane; preferably dichloromethane. The reflux reaction temperature in step 2 is 70 to 90° C., and the reflux reaction time is 2 to 4 hours.

[0019] A method for preparing a quinoxalinone boride with high-efficiency dual-state luminescence comprises the following steps:

[0020]

[0021] The compound of formula I and the compound of formula II or formula III are added to a solvent, and heated under reflux overnight; after the reaction liquid is cooled, a boron trifluoride ether solution and triethylamine are added, and the reflux reaction is continued; after the reaction liquid is cooled, saturated sodium bicarbonate is added, the mixture is allowed to stand, separated, extracted with ethyl acetate, and the organic phase is concentrated and separated by column chromatography to obtain a quinoxalinone boron compound of formula VI or formula VII;

[0022] Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring;

[0023] R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group (including a cycloalkyl group), an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

[0024] Preferably, the solvent is ethyl acetate, tetrahydrofuran, 1,2-dichloroethane, or dichloromethane.

[0025] Preferably, the reaction temperature for heating and reflux overnight is 70°C-90°C, the reaction time is 12-24h, and the temperature for continued reflux reaction is 40-60°C, the reaction time is 3-5h.

[0026] The quinoxalinone boronate is applied to long-term living cell imaging.

[0027] The long-term cell imaging of quinoxalinone boride mainly comprises the following steps:

[0028] The cells were divided into confocal culture dishes and grown overnight. The original culture medium was removed and replaced with a culture medium containing 5 μM probe compound and incubated for another 3 h. The confocal dish was taken out of the incubator and washed 3 times with PBS to remove the compound in the dish. The confocal dish was then imaged and detected by fluorescence confocal microscopy. After the cells were cultured for 72 h, the original culture medium was removed, the cells were washed 3 times with PBS, and the cells were imaged and observed by fluorescence confocal microscopy.

[0029] The long-term cell viability detection of a quinoxalinone boron compound mainly comprises the following steps:

[0030] Place the cells in RPMI 1640 medium containing 10% FBS and 1% penicillin-streptomycin. Then the cells were inoculated in a 96-well plate and waited for the cells to adhere. The compound was diluted in RPMI 1640 medium containing 10% FBS to a final concentration of 1-50 μM. The original medium in the 96-well plate was aspirated, and the medium containing the probe compound was added. The wells without probes were used as controls. Three replicate wells were set for each concentration and incubated in a 37°C incubator for 2-4 hours. After the compound entered the cells, the probe in the medium was washed away with PBS three times, and fresh medium was added. The cells were continued to be cultured in the incubator for 72 hours. 10 μL MTT solution (5 mg / mL) was added to each well and cultured for 4 hours. The culture was terminated, the culture supernatant in the air was aspirated, and 150 μL DMSO solution was added to each well. The wells were shaken for 10 minutes to fully dissolve the crystals. The absorbance value of each well at 490 nm was measured by an ELISA reader.

[0031] Beneficial effects of the invention: The luminescent material of the invention has simple synthesis steps, convenient preparation method, excellent luminescent performance, high-efficiency solution and solid dual-state photoluminescent properties, low biological toxicity and high biocompatibility, and long-term cell imaging function. It can become a new type of biological probe molecular platform. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The quinoxalinone boride prepared in Example 1 1 H NMR spectrum.

[0033] Figure 2 The crystal structure diagram of the quinoxalinone boride prepared in Example 1.

[0034] Figure 3 The ultraviolet absorption spectrum of the quinoxalinone boride prepared in Example 1 in an organic solvent.

[0035] Figure 4 The fluorescence emission spectrum of the quinoxalinone boride prepared in Example 1 in an organic solvent.

[0036] Figure 5 This is the fluorescence emission spectrum of the quinoxalinone boride prepared in Example 1 in the solid state.

[0037] Figure 6 (a) is a 3-hour fluorescence imaging experiment diagram of the quinoxalinone boride prepared in Example 1, and (b) is a 72-hour fluorescence imaging experiment diagram of the quinoxalinone boride.

[0038] Figure 7 This is a graph showing the cytotoxicity of the quinoxalinone boronate prepared in Example 1. DETAILED DESCRIPTION

[0039] The present invention is further described below by way of examples.

[0040] Example 1.

[0041]

[0042] The compound was synthesized in one step:

[0043] Add 4-bromo-2-aminodiphenylamine (2.62 g, 10 mmol) and ethyl 2-oxo-3-quinoline-2-propionate (2.43 g, 10 mmol) and dissolve in 300 ml of 1,2-dichloroethane, heat to 85°C and reflux for 24 h. Cool the reaction to room temperature, add 10 ml of triethylamine and 10 ml of boron trifluoride ether solution, heat to 50 degrees and react for 4 h. After the reaction is completed, add saturated sodium bicarbonate to quench the reaction, let stand, separate the layers, extract with ethyl acetate, concentrate the organic phase, and purify the mixture by column chromatography to obtain the product.

[0044] Example 2.

[0045]

[0046] The compound was synthesized in two steps:

[0047] Add 2-aminodiphenylamine (1.84 g, 10 mmol) and ethyl 2-oxo-3-pyridine-2-propionate (1.93 g, 10 mmol) and dissolve in 200 ml of ethanol, heat at 90°C and reflux for 24 h. After the reaction is completed, add ether for crystallization, filter the solid and recrystallize it with ethanol / ether to obtain the ligand.

[0048] The ligand (1.56 g, 5 mmol) was dissolved in 200 ml of 1,2-dichloroethane, and then 5 ml of triethylamine and 5 ml of boron trifluoride ether solution were added. The mixture was refluxed at 80°C for 3 h. After the reaction was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was allowed to stand, separated and extracted with ethyl acetate. The organic phase was concentrated and purified by column chromatography to obtain the product.

[0049] Example 3.

[0050]

[0051] The compound was synthesized in two steps:

[0052] (4-methoxy-2-aminoaniline)pyridine (2.15 g, 10 mmol) and ethyl 2-oxo-3-pyridine-2-propionate (1.93 g, 10 mmol) were added and dissolved in 200 ml of ethyl acetate, and heated to 80° C. and refluxed for 24 h. After the reaction was completed, the solvent was removed and the ligand was recrystallized from ethanol / ether to obtain the ligand.

[0053] The ligand (1.97 g, 5 mmol) was dissolved in 200 ml of 1,2-dichloroethane, and then 5 ml of triethylamine and 5 ml of boron trifluoride ether solution were added. The mixture was refluxed at 80°C for 3 h. After the reaction was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was allowed to stand, separated and extracted with ethyl acetate. The organic phase was concentrated and the mixture was purified by column chromatography to obtain the product.

[0054] Example 4.

[0055]

[0056] The compound was synthesized in one step:

[0057] Add diamine raw material (0.31g, 1mmol) and ethyl 2-oxo-3-pyridine-2-propionate (0.25g, 1mmol) dissolved in 50ml of 1,2-dichloroethane, heat to 85℃ and reflux for 24h. Cool the reaction to room temperature, add 1.5ml of triethylamine and 1.5ml of boron trifluoride ether solution, heat to 50℃ and react for 4h. After the reaction is completed, add saturated sodium bicarbonate to quench the reaction, let stand, separate, extract with ethyl acetate, concentrate the organic phase, and purify by column chromatography to obtain the product.

[0058] Example 5.

[0059]

[0060] The compound was synthesized in one step:

[0061] Add nitro-containing quinoline substituted diamine (0.28 g, 1 mmol) and ethyl 2-oxo-3-pyridine-2-propionate (0.25 g, 1 mmol) dissolved in 50 ml of 1,2-dichloroethane, and heat to 85 ° C for 24 hours. The reaction was cooled to room temperature, 1 ml of triethylamine and 1 ml of boron trifluoride ether solution were added, and the temperature was raised to 50 degrees for 4 hours. After the reaction was completed, saturated sodium bicarbonate was added to quench the reaction, stand, separate, extract with ethyl acetate, and concentrate the organic phase to obtain a mixture, and purify by column chromatography to obtain the product.

[0062] Example 6.

[0063]

[0064] The compound was synthesized in two steps:

[0065] (4-carbazole-2-aminoaniline)pyridine (0.35 g, 1 mmol) and ethyl 2-oxo-3-quinoline-2-propionate (0.19 g, 1 mmol) were added and dissolved in 50 ml of ethyl acetate, and heated to 80° C. and refluxed for 24 h. After the reaction was completed, the solvent was removed and the ligand was recrystallized from ethanol / ether to obtain the ligand.

[0066] The ligand (0.27 g, 0.5 mmol) was dissolved in 50 ml of 1,2-dichloroethane, and then 1 ml of triethylamine and 1 ml of boron trifluoride ether solution were added. The mixture was refluxed at 80°C for 3 h. After the reaction was completed, saturated sodium bicarbonate was added to quench the reaction. The mixture was allowed to stand, separated and extracted with ethyl acetate. The organic phase was concentrated and the obtained mixture was purified by column chromatography to obtain the product.

[0067] The long-term cell imaging of quinoxalinone boride mainly comprises the following steps:

[0068] The cells were divided into confocal culture dishes and grown overnight. The original culture medium was removed and replaced with a culture medium containing 5 μM probe compound and incubated for another 3 h. The confocal dish was taken out of the incubator and washed 3 times with PBS to remove the compound in the dish. The confocal dish was then imaged and detected by fluorescence confocal microscopy. After the cells were cultured for 72 h, the original culture medium was removed, the cells were washed 3 times with PBS, and the cells were imaged and observed by fluorescence confocal microscopy.

[0069] The long-term cell viability detection of a quinoxalinone boron compound mainly comprises the following steps:

[0070] Place the cells in RPMI 1640 medium containing 10% FBS and 1% penicillin-streptomycin. Then the cells were inoculated in a 96-well plate and waited for the cells to adhere. The compound was diluted in RPMI 1640 medium containing 10% FBS to a final concentration of 1-50 μM. The original medium in the 96-well plate was aspirated, and the medium containing the probe compound was added. The wells without probes were used as controls. Three replicate wells were set for each concentration and incubated in a 37°C incubator for 2-4 hours. After the compound entered the cells, the probe in the medium was washed away with PBS three times, and fresh medium was added. The cells were continued to be cultured in the incubator for 72 hours. 10 μL MTT solution (5 mg / mL) was added to each well and cultured for 4 hours. The culture was terminated, the culture supernatant in the air was aspirated, and 150 μL DMSO solution was added to each well. The wells were shaken for 10 minutes to fully dissolve the crystals. The absorbance value of each well at 490 nm was measured by an ELISA reader.

Claims

1. A quinoxalinone boride having high-efficiency dual-state luminescence, characterized in that: The quinoxalinone boron compound with high-efficiency dual-state luminescence has a structure shown in Formula VI or Formula VII: Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring; R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group, an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

2. The method for preparing a quinoxalinone boride having high-efficiency dual-state luminescence according to claim 1, characterized in that: The following steps are involved: Step 1: Add the compound represented by Formula I and the compound represented by Formula II or Formula III to a solvent, and heat to reflux for reaction; after the reaction, recrystallize with ethanol / ether to obtain a ligand containing a quinazolinone-quinoline or pyridine structure represented by Formula IV or Formula V; Step 2: adding a ligand containing a quinazolinone-quinoline or pyridine structure shown in Formula IV or Formula V to a non-polar solvent, adding a boron trifluoride ether solution and triethylamine, and performing a reflux reaction; cooling the reaction liquid, adding saturated sodium bicarbonate to quench the reaction, standing, layering, extracting with ethyl acetate, concentrating the organic phase, and separating the mixture by column chromatography to finally obtain a quinoxalinone boron compound shown in Formula VI or Formula VII; Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring; R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group, an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

3. The method according to claim 2, characterized in that The solvent in step 1 is ethanol, ethyl acetate, tetrahydrofuran, and 1,2-dichloroethane.

4. The method according to claim 2, characterized in that: The reaction time in step 1 is 16-48 hours, and the reaction temperature is 70°C to 90°C.

5. The method according to claim 2, characterized in that: The solvent in step 1 is ethanol, and the reaction time is 24 hours.

6. The method according to claim 2, characterized in that The non-polar solvent described in step 2 is ethyl acetate, tetrahydrofuran, 1,2-dichloroethane, dichloromethane; preferably dichloromethane, the reflux reaction temperature in step 2 is 70-90° C., and the reflux reaction time is 2-4 hours.

7. The method for preparing a quinoxalinone boride with high-efficiency dual-state luminescence according to claim 1, characterized in that: The following steps are involved: The compound of formula I and the compound of formula II or formula III are added to a solvent, and heated under reflux overnight; after the reaction liquid is cooled, a boron trifluoride ether solution and triethylamine are added, and the reflux reaction is continued; after the reaction liquid is cooled, saturated sodium bicarbonate is added, the mixture is allowed to stand, separated, extracted with ethyl acetate, and the organic phase is concentrated and separated by column chromatography to obtain a quinoxalinone boron compound of formula VI or formula VII; Wherein, Ar is selected from: phenyl, substituted phenyl, naphthalene, and nitrogen heteroaromatic ring; R is selected from the group consisting of a hydrogen atom, a halogen atom, an alkyl group, an azaalkyl group, an oxaalkyl group, an alkyl heterocycle, an amino group, a substituted amino group, a diphenylamino group, an aromatic ring, and an aromatic heterocycle.

8. The method according to claim 7, characterized in that The solvent is ethyl acetate, tetrahydrofuran, 1,2-dichloroethane, and dichloromethane.

9. The method according to claim 7, characterized in that: The reaction temperature for heating and reflux overnight is 70°C-90°C, and the reaction time is 12-24h. The temperature for continued reflux reaction is 40-60°C, and the reaction time is 3-5h.

10. The use of a quinoxalinone boride with high-efficiency dual-state luminescence according to claim 1, characterized in that the quinoxalinone boride is used in long-term living cell imaging.

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