Anthracene diphenyl ether compound with AIE characteristic, preparation method and application

Abstract

The invention discloses an anthracene diphenyl ether compound with AIE characteristics as well as a preparation method and application of the anthracene diphenyl ether compound. The anthracene diphenyl ether compound with the AIE characteristic is 9, 10-bis (bis (4-phenyl ether) vinyl)-9, 10-dihydroanthracene, has an excellent AIE effect, has higher fluorescence emission intensity compared with an existing fluorescence detection substance, has fluorescence enhancement response to Trp different from other amino acids, can specifically recognize tryptophan, is not influenced by other amino acids, and can be applied to detection of Trp. According to the present invention, the detection luminescence effect is significant, the sensitivity is good, the detection limit is low, the response time is short, the transient response can be performed on the Trp, the working time of the Trp detection is substantially shortened, and the working efficiency is effectively improved. Experimental results show that the fluorescence emission intensity of the compound reaches 2490, the detection limit of specific recognition of tryptophan is 89.35 nM, and the sensitivity (Ksv) is 9300 M <-1 >.

Description

Technical Field

[0001] The present invention belongs to the field of aggregation-induced enhanced luminescent materials, and in particular relates to an anthracene diphenyl ether compound with AIE characteristics, a preparation method and an application thereof. Background Art

[0002] Most organic luminescent molecules can emit strong fluorescence in dilute solutions, but the fluorescence disappears in concentrated solutions. This aggregation-induced quenching (ACQ) effect will limit the application of luminescent molecules in the field of materials. In 2001, Tang Benzhong's research group discovered a molecule with a completely different property from the traditional aggregation-induced quenching (ACQ), that is, the luminogen does not emit light in solution, but exhibits strong fluorescence in the aggregated state, and then proposed the concept of aggregation-induced emission (AIE). Compounds with AIE properties have many attractive advantages such as good biocompatibility, high resolution, strong resistance to photobleaching, low cytotoxicity, and easy functionalization. They have been widely used in cell tracking, sensors, drug delivery, fluorescence imaging, and clinical medicine.

[0003] Tryptophan is an important amino acid required by the human body. It plays an important role in preventing pellagra, depression, improving sleep and regulating emotions. It is widely used in the fields of food, medicine and feed additives. The analysis and determination of tryptophan is of great significance in the fields of biology, medicine and chemistry. At present, the main methods for determining tryptophan are spectrophotometry, fluorescence, high performance liquid chromatography, capillary electrophoresis, atomic absorption spectrometry and electrochemical methods. However, each method has its shortcomings: such as the need for specific instruments or trained personnel for detection, inconvenient operation, high analysis cost, slow test speed and high detection limit. Summary of the invention

[0004] In order to solve the above technical problems existing in the prior art, the present invention provides an anthracene diphenyl ether compound with AIE characteristics, a preparation method and application thereof.

[0005] The technical solution of the present invention is: an anthracene diphenyl ether compound with AIE characteristics, which is 9,10-bis(di(4-phenylether)vinyl)-9,10-dihydroanthracene, and the structural formula is as follows: .

[0006] The preparation method of the anthracene diphenyl ether compound with AIE characteristics, the synthesis route is as follows: .

[0007] The preparation method of the anthracene diphenyl ether compound having AIE properties comprises the following specific steps: Step 1. Put 4-phenylether phenylboronic acid, 9,10-bis(dibromomethylene)-9,10-dihydroanthracene, anhydrous potassium carbonate and Pd(PPh3)4 into a two-necked flask containing a magnetic particle; weigh 1,4-dioxane and distilled water as reaction solvents, pour them into the two-necked flask, and stir magnetically until the reactants are completely dissolved; the dosage ratio of the 4-phenylether phenylboronic acid, compound 6, anhydrous potassium carbonate, Pd(PPh3)4, 1,4-dioxane and distilled water is 3.36 mmol:0.674 mmol:6.51mmol:0.174mmol:120 mL:24 mL; Step 2. Under nitrogen atmosphere, after evacuation three times, the oil bath was heated to 90 °C for 24 h, and then the heating was stopped to cool the reactant to room temperature, extracted with 90 mL of dichloromethane, and then washed with saturated brine; Step 3. Collect the lower oily mixture and add anhydrous sodium sulfate to dry for 1 h; Step 4. Filter and evaporate under reduced pressure to obtain a crude product, purify it by column chromatography, use petroleum ether / dichloromethane with a volume ratio of 5:1 as eluent, and then add ethanol for recrystallization to obtain a green powder.

[0008] The anthracene diphenyl ether compounds having AIE properties are used as fluorescent probes for identifying small molecules, and specifically as fluorescent probes for identifying tryptophan.

[0009] The anthracene diphenyl ether compounds prepared by the present invention have excellent AIE characteristic effects, and the fluorescence emission intensity is improved compared with the existing fluorescence detection substances. It has a fluorescence enhancement response to Trp that is different from other amino acids. It can specifically identify tryptophan and is not affected by other amino acids. The detection luminescence effect is obvious and has good sensitivity and a low detection limit. At the same time, the response time is short, and Trp can be responded to instantly, which greatly shortens the working time of Trp detection and effectively improves work efficiency. The experimental results show that the fluorescence emission intensity of the compound of the present invention reaches 2490, the detection limit of specific recognition of tryptophan is 89.35 nM, and the sensitivity (Ksv) is 9300M -1 . BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 is the compound prepared in Example 1 of the present invention 1 H NMR spectrum.

[0011] Figure 2 It is the fluorescence spectrum of the compound prepared in Example 1 of the present invention when different amino acids are added.

[0012] Figure 3It is a three-dimensional diagram of the interaction between the compound prepared in Example 1 of the present invention as a fluorescent probe and different types of amino acids at different concentration gradients.

[0013] Figure 4 It is the fluorescence spectrum and ultraviolet absorption spectrum of the compound prepared in Example 1 of the present invention as a fluorescent probe for titration experiment of different concentrations of Trp.

[0014] Figure 5 1 is a Stern-Volmer curve and a linear relationship diagram of the compound prepared in Example 1 of the present invention at different Trp concentrations.

[0015] Figure 6 This is a fluorescence spectrum diagram of the compound prepared in Example 1 of the present invention used as a fluorescent probe in interference experiments with other amino acids and Trp.

[0016] Figure 7 It is the fluorescence spectrum of the compound prepared in Experimental Example 1 with different response times to Trp.

[0017] Figure 8 It is a Job's curve diagram of the compound prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0018] The present invention is described in detail below by specific examples, but the protection scope of the present invention is not limited. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the experimental equipment, materials, reagents, etc. used can be obtained from commercial channels.

[0019] The preparation method of the anthracene diphenyl ether compound having AIE properties of the present invention is carried out in the following steps: Step 1. Weigh 4-phenylether phenylboronic acid (0.72 g, 3.36 mmol), 9,10-bis(dibromomethylene)-9,10-dihydroanthracene (0.3 g, 0.674 mmol), anhydrous potassium carbonate (0.9 g, 6.51 mmol) and Pd(PPh3)4 (0.12 g, 0.174 mmol), and put them into a two-necked flask containing a magnetic rod; weigh 1,4-dioxane (120 ml) and distilled water (24 ml) as reaction solvents using a measuring cylinder, pour them into the two-necked flask, and stir with a magnetic rod until all the reactants are dissolved; Step 2. Under nitrogen atmosphere, after three times of vacuuming, the oil bath was heated to 90 °C, and the Suzuki coupling reaction was started. The reaction was fully reacted for 24 h, and then the heating was stopped and the reactant was cooled to room temperature. It was extracted with 90 mL of dichloromethane three times, and then washed with saturated brine three times; Step 3. Collect the lower oily mixture, add appropriate amount of anhydrous sodium sulfate and dry for 1 h; Step 4. The crude product was filtered and evaporated under reduced pressure, and then purified by column chromatography using petroleum ether / dichloromethane (5 / 1, v / v) as eluent. Then, an appropriate amount of ethanol was added for recrystallization to obtain 0.4 g of green powder with a yield of 74.8%.

[0020] The compound 1 H NMR spectrum Figure 1 As shown, the structural formula is as follows: .

[0021] Experiment 1: AIE performance experiment of anthracene diphenyl ether compounds.

[0022] In order to prove that the compound has AIE effect, it was dissolved in tetrahydrofuran (THF) to prepare a solution, using THF as a good solvent and water as a poor solvent. -4 mol / L). The excitation wavelength of the fluorescence spectrum was set to 300 nm, the slit width was set to 10:10, and the voltage was set to 550 V. By changing the volume fraction of water, (f w ) from 0% to 90% in THF / H2O mixture, and the intensity change of the solution (fw) from 0% to 90% was visually observed under UV light (365 nm).

[0023] Experiments show that the fluorescence intensity of the compound in pure THF solution is 20.97. When the water content increases to 60%, the fluorescence quantum yield is 41%, and the fluorescence intensity reaches a maximum value of 2490, which is 118.75 times higher than that of pure THF solution. Correspondingly, the emission wavelength blue-shifts from 474.4 nm to 472 nm, and the absorbance drops from 3.5 to 1.0.

[0024] The above data show that the compound has typical AIE characteristics and is an AIE compound.

[0025] Experiment 2: Selectivity of anthracene diphenyl ether compounds for amino acids In order to confirm the specific recognition of amino acids by the compound, the compound prepared in Example 1 of the present invention was used as a fluorescent probe to interact with different amino acids. Specifically, 2 ml of the compound was added to 100 μM of 12 amino acids (Val, Thr, Ccp, Ala, Leu, Met, Pro, Ser, Cys, Lys, Glu, Trp), and the fluorescence spectra were as follows: Figure 2 As shown. Figure 2It can be seen that only when 100 μM Trp is added, the compound undergoes a blue shift and a dramatic change in fluorescence intensity. When other equal amounts of amino acids are added under the same conditions, the fluorescence intensity changes only slightly and remains basically unchanged compared to when no amino acids are added. When tryptophan is added alone, the fluorescence intensity changes dramatically, indicating that the compound specifically recognizes tryptophan.

[0026] Experiment 3: Fluorescence titration test of amino acids The compound prepared in Example 1 of the present invention was used as a fluorescent probe to interact with different types of amino acids at different concentration gradients. The specific operation was to take 2 ml of the compound and add 0-100 μM of the above 12 amino acids in sequence for fluorescence titration test. Figure 3 As shown. Figure 3 It can be seen that with the increase of Trp concentration, the fluorescence intensity of the compound increased dramatically, and the fluorescence intensity of the compound hardly changed during the gradual increase of the concentration of other amino acids. Among them, Ccp, Ala, and Val concentrated amino acids only showed a slight upward trend when added.

[0027] Experiment 4: Tryptophan concentration titration The compound prepared in Example 1 of the present invention was used as a fluorescent probe to titrate different concentrations of Trp. The specific operation was to sequentially add 0-100 μM Trp to the compound to obtain the fluorescence emission spectrum and the ultraviolet absorption spectrum. Figure 4 As shown in the fluorescence emission spectrum, it can be seen that the emission wavelength of the compounds has undergone a significant blue shift and the fluorescence intensity has increased dramatically; the ultraviolet absorption spectrum shows that the absorbance of the compounds has decreased.

[0028] Experiment 5: Sensitivity and detection limit for Trp The detection limit of a compound is calculated using the following formula: detection limit = 3σ / k; Where σ is the standard deviation of blank compound measurement, and k is the slope value of the linear relationship graph between fluorescence intensity and Trp concentration. The linear relationship graph of the compound is shown in Figure 5 As shown, the correlation coefficient R 2 It is 0.98005, indicating that it has a good linear relationship. The k value is 61.88. The corresponding detection limit is calculated to be 89.35 nM through σ and.

[0029] To explore compound detection Where I0 is the fluorescence intensity of the initial added compound, I is the corresponding fluorescence intensity when different concentrations of Trp are added, and [Q] is the molar concentration of Trp. The Stern-Volmer curve is shown in Figure 5 As shown, the correlation coefficient R2 The sensitivity (Ksv) is 9300M. -1 , the compound has a higher sensitivity and therefore responds better to tryptophan.

[0030] Experiment 6: Interference Experiment In order to explore whether the presence of other amino acids would affect the detection of tryptophan by the compound of the present invention, an interference experiment was conducted using the compound prepared in Example 1 of the invention as a fluorescent probe with other amino acids and Trp. Specifically, 10 μM of other amino acids and tryptophan were added alternately to measure their fluorescence intensity (twice each). The results are as follows Figure 6 As shown. The experimental results show that when other amino acids are added, the fluorescence intensity changes slightly, while when tryptophan is added, the fluorescence intensity rises rapidly. When 10 μM other amino acids are repeatedly added, the effect on the fluorescence intensity is not significant. It proves that the compound of the present invention is not interfered by other amino acids when detecting tryptophan, can specifically identify tryptophan, and is basically unaffected and has a similar change trend when equal amounts of other amino acids are added.

[0031] Experiment 7: Response Time Test To investigate the fluorescence intensity of the compound at different response times to Trp, this experiment selected a Trp solution with a maximum titration concentration of 100 μM and directly added it to the compound. The fluorescence spectrum is shown in the figure below: Figure 7 As shown. The results show that the fluorescence intensity of the solution rises rapidly when Trp is added to the compound, and then changes slightly with the increase of response time. This shows that the compound can respond to Trp instantaneously, greatly shortening the working time of Trp detection and improving work efficiency.

[0032] Experiment 8: Job's Curve Analysis In order to explore the optimal binding ratio of the compound to Trp, Job's curve was used to illustrate. By controlling the total concentration of the compound and Trp to 100 μM, the ratio of the compound to Trp was changed. Figure 8 Through linear fitting, it can be observed that the fluorescence intensity of the combination of the two is the highest at about 0.5 [compounds] / [compounds+Trp], indicating that the compounds are most effective when combined with Trp in a 1:1 ratio.

Claims

1. An anthracene diphenyl ether compound having AIE properties, characterized in that 9,10-Bis(di(4-phenylene oxide)vinyl)-9,10-dihydroanthracene, the structural formula is as follows: 。 2. The method for preparing anthracene diphenyl ether compounds having AIE properties according to claim 1, characterized in that The synthetic route is as follows: 。 3. The method for preparing anthracene diphenyl ether compounds having AIE properties according to claim 1, characterized in that Follow the steps below: Step 1. Put 4-phenylether phenylboronic acid, 9,10-bis(dibromomethylene)-9,10-dihydroanthracene, anhydrous potassium carbonate and Pd(PPh3)4 into a two-necked flask containing a magnetic particle; weigh 1,4-dioxane and distilled water as reaction solvents, pour them into the two-necked flask, and stir magnetically until the reactants are completely dissolved; the dosage ratio of the 4-phenylether phenylboronic acid, compound 6, anhydrous potassium carbonate, Pd(PPh3)4, 1,4-dioxane and distilled water is: 3.36mmol:0.674mmol:6.51mmol:0.174mmol:120mL:24mL; Step 2. Under nitrogen atmosphere, after evacuation three times, the oil bath was heated to 90 °C for 24 h, and then the heating was stopped to cool the reactant to room temperature, extracted with 90 mL of dichloromethane, and then washed with saturated brine; Step 3. Collect the lower oily mixture and add anhydrous sodium sulfate to dry for 1 h; Step 4. Filter and evaporate under reduced pressure to obtain a crude product, purify it by column chromatography, use petroleum ether / dichloromethane with a volume ratio of 5:1 as eluent, and then add ethanol for recrystallization to obtain a green powder.

4. Application of an anthracene diphenyl ether compound with AIE properties as a fluorescent probe for identifying small molecules, characterized in that Application as a fluorescent probe for identifying tryptophan.

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