Adenosine triphosphate aggregation-induced luminescent compound and preparation method and application thereof

Abstract

The present invention discloses an adenosine triphosphate aggregation-induced luminescent compound and a preparation method and application thereof. The present invention uses 2-benzylbenzimidazole, p-bromobenzylamine, etc. as raw materials to synthesize an aggregation-induced luminescent compound for adenosine triphosphate (ATP) detection. The compound has an obvious fluorescence aggregation induction effect and belongs to an aggregation-induced luminescent material. As an aggregation-induced fluorescent probe, it shows extremely high sensitivity and selectivity to ATP, is not interfered by ADP, PPi and common ions, and responds quickly. It has good AIE characteristics, can specifically identify ATP, and is used to detect ATP molecules. The ATP aggregation-induced luminescent compound provided by the present invention is simple to synthesize and has high specificity, which solves the problem of poor specificity of ATP detection by conventional methods, can realize accurate detection of adenosine triphosphate, and provides more detection products and detection methods with high specificity, high sensitivity, simple synthesis, and easy operation for ATP.

Description

Technical Field

[0001] The present invention belongs to the technical field of small molecule fluorescent probes, and more specifically, relates to an adenosine triphosphate aggregation-induced luminescent compound and a preparation method and application thereof. Background Art

[0002] Adenosine triphosphate (ATP) is a high-energy phosphate compound and the main carrier of cellular energy. It releases energy by hydrolysis into ADP and inorganic phosphate, providing power for various cellular processes, including muscle contraction, nerve signal transmission, material transport and biosynthesis. The level of ATP in the body is closely related to the occurrence and development of some diseases, such as ischemia, hypoxia, hypoglycemia, cardiovascular disease, Parkinson's disease and malignant tumors. At the same time, ATP can also be used as an indicator to reflect food safety, quality control and environmental analysis, and is of great significance in biochemical analysis, clinical disease diagnosis and food hygiene and safety testing.

[0003] At present, the methods used to detect ATP mainly include high performance liquid chromatography, colorimetry, electrochemical method, fluorescence method, etc. These methods usually have the problems of complex operation, high cost, low sensitivity, poor selectivity, etc. The use of fluorescent probes for detection stands out among many methods due to its advantages such as simple operation, high sensitivity, and can be used for cell or living body imaging. The use of fluorescent probes for the detection of biological and environmental analytes has been widely developed, so it is necessary to develop a simple, rapid, and low-cost ATP detection method.

[0004] Aggregation-induced emission (AIE) is considered to be a photophysical phenomenon displayed by a class of fluorescent materials. This type of luminescent material does not emit light when dissolved in a good solvent, but emits light in the form of aggregates when in a poor solvent or solid state. Therefore, AIE active materials have high application value in the field of fluorescent sensors. In recent years, people have synthesized a variety of molecular compounds with AIE properties, such as hexaphenylsilane (HPS), 9,10-dibenzanthracene (DSA), quinoline-propionitrile (QM), tetraphenylpyrazine (TPP) and tetraphenylethylene (TPE), but the synthesis methods of these AIE probes are cumbersome and the synthesis conditions are harsh. Since the existing ATP detection method is easily affected by adenosine diphosphate (ADP), pyrophosphate (PPi), etc., the problem of poor specific recognition occurs. At present, there are few AIE molecular probes for specific recognition of ATP. In order to solve the problem of poor specific recognition of ATP in the existing conventional detection methods, it is urgent to develop more AIE molecular probes with high specificity, high sensitivity, simple synthesis and easy operation to achieve ATP detection through fluorescence aggregation induced effect. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the problem of poor specific recognition of ATP in existing conventional detection methods and the shortcomings of AIE molecular probes for specific recognition of ATP, and to provide an adenosine triphosphate aggregation-induced luminescent compound and a preparation method and application thereof.

[0006] The object of the present invention is to provide an aggregation-induced luminescent compound.

[0007] Another object of the present invention is to provide a method for preparing the aggregation-induced luminescent compound.

[0008] Another object of the present invention is to provide applications of aggregation-induced luminescent compounds.

[0009] Another object of the present invention is to provide a product for detecting adenosine triphosphate.

[0010] Another object of the present invention is to provide a method for specifically detecting adenosine triphosphate.

[0011] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0012] The present invention provides an aggregation-induced luminescent compound having the following molecular formula:

[0013] .

[0014] The present invention uses 2-benzylbenzimidazole, p-bromobenzylamine and the like as raw materials to synthesize an aggregation-induced luminescent compound for adenosine triphosphate (ATP) detection. The compound has an obvious fluorescence aggregation induction effect and belongs to an aggregation-induced luminescent material. As an aggregation-induced fluorescent probe, it exhibits extremely high sensitivity and selectivity to ATP, is not interfered by ADP, PPi and common ions, and responds quickly, has good AIE characteristics, can specifically identify ATP, and can be used to detect ATP molecules. The ATP aggregation-induced luminescent compound provided by the present invention is simple to synthesize and has high specificity, which overcomes the problem of poor specificity of conventional methods for detecting ATP, can realize accurate detection of adenosine triphosphate, and provides more detection products and methods for ATP with high specificity, high sensitivity, simple synthesis and easy operation.

[0015] The present invention provides a method for preparing an adenosine triphosphate aggregation-induced luminescent compound, comprising the following steps:

[0016] (1) Preparation of intermediate 3: 2-benzylbenzimidazole and cuprous chloride are dissolved in an organic solvent, and then p-bromobenzylamine and trimethylacetic acid are added, and stirred at 90°C-160°C for 2-36 hours. After the reaction is completed, the product is separated and purified by column chromatography to obtain intermediate 3, whose structural formula is:

[0017] ;

[0018] (2) Preparation of intermediate 5: In an inert gas atmosphere, intermediate 3, 4-pyridine boronic acid, tetrakis(triphenylphosphine)palladium and potassium carbonate are mixed and added to an organic solvent for dissolution, stirred at 60-100° C. for 6-12 hours, extracted, and separated and purified by column chromatography to obtain intermediate 5, whose structural formula is:

[0019] ;

[0020] (3) Preparation of aggregation-induced emission compound S-2: Dissolve the intermediate 5 and benzyl bromide in an organic solvent, stir at 60-100°C for 16-36 hours, and filter under reduced pressure after the reaction is completed. Collect the filtered product and wash it to obtain aggregation-induced emission compound S-2.

[0021] The synthetic route of ATP aggregation-induced luminescent compounds is:

[0022] .

[0023] Preferably, in (1), the molar ratio of 2-benzylbenzimidazole, cuprous chloride, p-bromobenzylamine and trimethylacetic acid is (0.1-1): (0.05-2): (1-10): (0.5-8).

[0024] Preferably, the organic solvent in (1) is o-dichlorobenzene.

[0025] More preferably, the molar ratio of 2-benzylbenzimidazole, cuprous chloride, p-bromobenzylamine and trimethylacetic acid is 1:(0.05-2):(1-10):(0.5-8), and the amount of the organic solvent o-dichlorobenzene is (0.5-4 mL).

[0026] Preferably, in the column chromatography method (1), a silica gel column is used for gradient elution, first eluting with petroleum ether: ethyl acetate = 10:1, and then eluting and separating with petroleum ether: ethyl acetate = 8:1.

[0027] Preferably, in (2), the molar ratio of the intermediate 3, 4-pyridineboronic acid, tetrakis(triphenylphosphine)palladium, and potassium carbonate is (0.1-1):(1-8):(0.05-1):(2-8).

[0028] Preferably, the organic solvent in (2) is a mixed solvent of 1,4-dioxane and water, with a volume ratio of 20:(1-5).

[0029] More preferably, the molar ratio of intermediate 3, 4-pyridineboric acid, tetrakis(triphenylphosphine)palladium and potassium carbonate is 1:(1-8):(0.05-1):(2-8); and the amount of the mixed solvent is (2-20 mL).

[0030] Preferably, in (2), dichloromethane and water are used as solvents for extraction, and the product is dissolved in dichloromethane; the column chromatography method uses a silica gel column for gradient elution, first eluting with petroleum ether: ethyl acetate = 1:1, and then eluting and separating with 100% ethyl acetate.

[0031] Preferably, in (3), the molar ratio of intermediate 5 to benzyl bromide is (0.05-2):(1-10).

[0032] Preferably, the organic solvent in (3) is acetonitrile.

[0033] More preferably, the molar ratio of intermediate 5 to benzyl bromide is 1:(1-10), and the amount of the organic solvent acetonitrile used is (0.5-4 mL).

[0034] Preferably, in (3), the washing is first performed with petroleum ether for 2 to 4 times, and then with dichloromethane for 2 to 4 times.

[0035] The present invention provides application of an aggregation-induced emission compound as an aggregation-induced emission material.

[0036] The invention provides application of an aggregation-induced luminescence compound as an adenosine triphosphate aggregation-induced fluorescent probe.

[0037] The invention provides application of an aggregation-induced luminescence compound in detecting adenosine triphosphate or in preparing a product for detecting adenosine triphosphate.

[0038] The invention provides a product for detecting adenosine triphosphate, which contains an aggregation-induced luminescence compound.

[0039] At the same time, the present invention provides a method for specifically detecting adenosine triphosphate, which uses the above-mentioned aggregation-induced luminescence compound to detect ATP through the fluorescence aggregation-induced effect. Compared with existing conventional methods, this method has the advantages of good specificity, high sensitivity, and rapid response.

[0040] The present invention has the following beneficial effects:

[0041] The aggregation-induced luminescent compound S-2 synthesized by the present invention using 2-benzylbenzimidazole, p-bromobenzylamine and the like as raw materials has good AIE characteristics and belongs to an aggregation-induced luminescent material. As an aggregation-induced fluorescent probe, it exhibits extremely high sensitivity and selectivity to ATP and is not interfered by ADP, PPi and common ions. The aggregation-induced luminescent compound provided by the present invention is simple to synthesize and has high specificity. As an adenosine triphosphate-induced aggregation-type fluorescent probe, it can specifically identify ATP and detect ATP through the fluorescent aggregation-induced effect, which can overcome the problem of poor ATP specificity in existing detection methods. At the same time, the present invention provides a method for specifically detecting adenosine triphosphate, and the detection of ATP is realized through the fluorescent aggregation-induced effect of the aggregation-induced luminescent compound S-2. Compared with existing conventional methods, this method has the advantages of simple synthesis, high sensitivity, good specificity, and rapid response. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 is the fluorescence value of probe S-2 in different solvents.

[0043] Figure 2 The emission spectra of probe S-2 in toluene / DMSO solutions with different toluene ratios.

[0044] Figure 3 is the fluorescence value of probe S-2 in different ion solutions.

[0045] Figure 4 (a) is the fluorescence value curve of probe S-2 under ATP gradient concentration (0-10 μM); Figure 4 (b) is a straight line fitting graph of the fluorescence values ​​of probe S-2 under ATP gradient concentration (0-50μM).

[0046] Figure 5 This is the curve of the fluorescence value of probe S-2 changing with time in HEPES buffer with the addition of ATP. DETAILED DESCRIPTION

[0047] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0048] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0049] The chemical reagents and solvents used in the following examples were purchased from reagent companies, and the fluorescence spectra were recorded using a fluorescence spectrophotometer.

[0050] Example 1 Synthesis of fluorescent probe S-2

[0051] 1. Preparation of intermediate 3:

[0052] 2-Benzylbenzimidazole (84 mg, 0.4 mmol) and cuprous chloride (15.84 mg, 0.16 mmol) were added to a 10 mL glass test tube, 2 mL of o-dichlorobenzene was added to dissolve, and then p-bromobenzylamine (303 mL, 2.4 mmol) and trimethylacetic acid (44 μL, 0.4 mmol) were added in sequence, and stirred at 110 ° C in air for 10 hours. After the reaction solution was cooled, 98 mg of intermediate 3 (yield: 45%, yellow solid) was obtained by column chromatography (petroleum ether / ethyl acetate = 10:1 → petroleum ether / ethyl acetate = 8:1).

[0053] The synthetic route of intermediate 3 is:

[0054]

[0055] Intermediate 3 compound 1 H NMR and 13 The characterization results of C NMR are as follows:

[0056] 1 H NMR (400 MHz, CDCl3) δ 7.85 (d, J = 8.2 Hz, 1H), 7.71 (d, J = 8.4Hz, 2H), 7.61 (d, J = 8.4 Hz, 2H), 7.47 – 7.23 (m, 10H), 7.06 (t, J = 7.8 Hz,1H), 6.87 (d, J = 8.3 Hz, 1H).

[0057] 13 C NMR (101 MHz, CDCl3) δ 150.13, 148.38, 147.55, 145.82, 136.81,133.57, 132.76, 132.41, 131.85, 131.07, 130.78, 130.16, 128.76, 128.49,127.52, 126.45, 125.69, 122.85, 122.04, 121.48, 120.53, 114.47.

[0058] 2. Preparation of intermediate 5:

[0059] Under nitrogen, intermediate 3 (221.6 mg, 0.4 mmol), 4-pyridineboronic acid (196.672, 1.6 mmol), tetrakis(triphenylphosphine)palladium (36.97 mg, 0.032 mmol), potassium carbonate (331.704 mg, 2.4 mmol) were added to a 50 mL round-bottom flask, and 8 mL of 1,4-dioxane and 2 mL of water were added to dissolve. Stir at 80 ° C for 8 hours under nitrogen. After the reaction solution was cooled, 30 mL of water was added to dissolve the reaction system, extracted with dichloromethane (30 mL×3), the organic layers were combined, dried with anhydrous sodium sulfate, filtered, and 95.6 mg of intermediate 5 (yield: 43%, yellow solid) was obtained by column chromatography (petroleum ether / ethyl acetate = 1:1→100% ethyl acetate).

[0060] The synthetic route of intermediate 5 is:

[0061]

[0062] Intermediate 5 1 H NMR and 13 The characterization results of C NMR are as follows:

[0063] 1 H NMR (400 MHz, CDCl3) δ 9.02 (s, 1H), 8.83 (s, 1H), 8.70 (d, J =4.9 Hz, 1H), 8.57 (d, J = 6.5 Hz, 1H), 8.05 (dt, J = 7.9, 2.0 Hz, 1H), 8.02 – 7.94 (m, 3H), 7.94 – 7.82 (m, 3H), 7.67 – 7.57 (m, 4H), 7.52 – 7.39 (m, 7H), 7.35 (dd, J = 8.3, 4.5 Hz, 1H), 7.15 (t, J = 8.4 Hz, 1H), 7.05 (d, J = 8.4Hz, 1H).

[0064] 13C NMR (101 MHz, CDCl3) δ 150.43, 149.34, 149.02, 148.62, 148.39,148.21, 145.95, 140.66, 137.97, 137.58, 134.59, 134.28, 133.95, 133.78,131.12, 130.99, 129.47, 128.81, 128.51, 127.83, 126.61, 126.47, 123.83,123.58, 122.00, 121.55, 120.56, 114.69.

[0065] 3. Preparation of probe S-2:

[0066] Intermediate 5 (55.12 mg, 0.1 mmol), benzyl bromide (68.41 mg, 0.4 mmol), and 2 mL of acetonitrile were added to a 25 mL pressure tube and stirred at 80°C for 24 hours. After the reaction system was cooled, it was filtered under reduced pressure, and the solid was washed with petroleum ether (25 mL×3) and then with dichloromethane (25 mL×3). 73 mg of probe S-2 was obtained (yield: 82%, yellow solid).

[0067] The synthetic route of probe S-2 is:

[0068]

[0069] Probe S-2 1 H NMR and 13 The characterization results of C NMR are as follows:

[0070] 1 H NMR (400 MHz, CDCl3) δ 8.53 (d, J = 7.1 Hz, 2H), 8.43 (d, J = 8.4Hz, 2H), 8.20 (s, 2H), 8.02 (d, J = 8.7 Hz, 2H), 7.88 (d, J = 8.2 Hz, 1H),7.67 (dd, J = 18.0, 7.4 Hz, 4H), 7.62 – 7.39 (m, 14H), 7.18 (t, J = 8.3 Hz,1H), 6.85 (d, J = 8.4 Hz, 1H), 5.95 (s, 2H), 5.86 (s, 2H).

[0071] 13C NMR (101 MHz, DMSO) δ 154.57, 151.57, 149.28, 145.55, 145.28,136.59, 135.03, 135.00, 133.78, 131.65, 131.25, 130.37, 129.86, 129.78,129.74, 129.69, 129.63, 129.33, 129.28, 129.23, 128.33, 126.06, 125.38,123.27, 121.66, 119.36, 115.20, 63.00, 62.78.

[0072] Example 2 Fluorescence values ​​of probe S-2 in different solvents

[0073] The probe S-2 prepared in Example 1 was incubated in methanol, ethanol, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), dichloromethane and toluene ion solution (ion concentration of 10 μM) for 20 min, and the emission spectrum was tested by fluorescence spectrophotometer. The excitation wavelength was set to 420 nm, the detection band was 430-700 nm, the emission start wavelength was 430 nm, the emission stop wavelength was 700 nm, the excitation bandwidth was 5 nm, the emission bandwidth was 5 nm, and the gain (PMT) was 605 V.

[0074] The results of the test are as follows Figure 1 As shown in the figure, we can learn the basic information of probe S-2 in different solvents, such as the maximum emission wavelength and maximum fluorescence value. The ultraviolet absorption of probe S-2 was detected to be around 365 nm. When the excitation wavelength was 420 nm, there was a large Stokes shift and a small background light, and the performance was superior. In subsequent experiments, the excitation wavelength was 420 nm.

[0075] In addition, it was found that the maximum fluorescence value of probe S-2 in easily soluble solvents was smaller, and the maximum fluorescence value in poorly soluble solvents was larger, which preliminarily reflects that probe S-2 has certain AIE characteristics, and the Stokes shift of probe S-2 is about 50 nm.

[0076] Example 3 AIE properties of probe S-2

[0077] According to the measurement in Example 2, toluene is used as a sparingly soluble solvent and DMSO is used as a readily soluble solvent. The volume fractions of toluene are set to 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100%, respectively, for a total of 11 volume fractions. The concentration of probe S-2 used during detection was 10 μM, and the fluorescence value was measured. The instrument settings are: excitation wavelength: 420 nm, detection band: 430-700 nm, emission start wavelength: 430 nm, emission stop wavelength: 700 nm, excitation bandwidth: 5 nm, emission bandwidth: 5 nm, gain (PMT): 650 V. In theory, the fluorescence value of probe S-2 in the solvent should continue to increase as the ratio of toluene to toluene / DMSO increases.

[0078] The results of the test are as follows Figure 2 As shown, the probe S-2 exhibits good AIE properties, and the Stokes shift of the probe S-2 is about 50 nm. Specifically, the fluorescence value of the probe S-2 increases with the increase of the ratio of toluene to toluene / DMSO. When the toluene content is 100%, the fluorescence property of the probe S-2 is the strongest; when the toluene content is less than 90%, the fluorescence property of the probe S-2 is not strong. When the toluene content is less than 80%, the fluorescence value of the probe S-2 does not change significantly with the increase of the toluene content toluene / DMSO ratio, indicating that the probe S-2 has not begun to aggregate in large quantities. When the toluene content increases from 80% to 90% and from 90% to 100%, the fluorescence value of the probe S-2 changes significantly, indicating that the probe S-2 begins to aggregate in large quantities when the toluene content is 80% and is completely aggregated when the toluene content is 100%. The Stokes shift of probe S-2 increases with the increase of toluene content. When the toluene content is 100%, the maximum absorption peak changes from 620 nm to 570 nm, and the Stokes shift reaches the maximum value, with a shift value of about 50 nm. This shows that probe S-2 has a significant fluorescence aggregation induction effect and belongs to an aggregation-induced luminescence material, as an aggregation-induced fluorescent probe.

[0079] Example 4 Ion selectivity of probe S-2

[0080] Preparation of various ion solutions: Weigh ATP, ADP, and PPi separately and dissolve them in HEPES buffer to prepare a 10 mM stock solution for use; then weigh sodium chloride, potassium acetate, ferric chloride, calcium chloride, copper chloride, potassium dihydrogen phosphate, and potassium hydrogen phosphate separately and dissolve them in deionized water to prepare a 10 mM stock solution for use. The detection concentration of probe S-2 is 10 μM, the ion detection concentration is 10 μM, and the emission spectrum is tested after incubation for 20 min. Set the excitation wavelength to: 420 nm, detection band: 430-700 nm, emission start wavelength: 430 nm, emission stop wavelength: 700 nm, excitation bandwidth: 5 nm, emission bandwidth: 5 nm, gain (PMT): 650 V.

[0081] The results of the test are as follows Figure 3 As shown, in the selective detection of 10 ions, probe S-2 has the best recognition effect on ATP, and the fluorescence value is about 40 times that of probe S-2 in HEPES buffer; it has no recognition ability for the other 9 ions, and shows good specific recognition effect on ATP.

[0082] Example 5 Sensitivity test of probe S-2

[0083] The 10 mM ATP stock solution was diluted to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50 μM according to the concentration gradient, and HEPES buffer was used for dilution. The probe S-2 detection concentration was 10 μM, and the fluorescence value was detected after incubation for 20 minutes. The excitation wavelength was set to 420 nm, the detection band was 430-700 nm, the emission start wavelength was 430 nm, the emission stop wavelength was 700 nm, the excitation bandwidth was 5 nm, the emission bandwidth was 5 nm, and the gain (PMT) was 650 V.

[0084] The results of the test are as follows Figure 4 As shown in the figure, the fluorescence value of the probe S-2 molecule in the HEPES buffer is very low. As the concentration of ATP increases, the fluorescence value of the probe S-2 molecule solution continues to increase ( Figure 4 In (a) above, the maximum fluorescence value showed a good linear relationship with the ATP concentration. In the concentration range of 1-10 mM, a linear fit was performed between the maximum fluorescence value and the corresponding ATP concentration to obtain R 2 =0.98707, y= 139.99427x -54.00227. According to the linear fitting formula, it can be calculated that the minimum detection limit of ATP is 0.847 μM.

[0085] To further explore the linear relationship between the maximum fluorescence value and ATP concentration, we set a higher ATP concentration gradient of 10, 20, 30, 40, and 50 μM for detection and analysis. The results showed that as the concentration of ATP increased, the fluorescence value of the S-2 solution continued to increase and the maximum fluorescence value showed a good linear relationship with the ATP concentration ( Figure 4 In the concentration range of 10-50 mM, the maximum fluorescence value was linearly fitted with the corresponding ATP concentration, and R 2 =0.99634, y= 78.98647 x+578.19183, the minimum detection limit of ATP is 0.386 μM. In addition, according to the two fitting results, the linear relationship between probe S-2 and ATP can be adjusted by changing the ATP concentration used in the detection process. When the ATP concentration is as low as 386 nM, probe S-2 can still detect ATP.

[0086] Example 6 Detection of temporal changes of probe S-2

[0087] The detection concentration of probe S-2 was 10 μM, and the detection concentration of ATP was 10 μM. The fluorescence values ​​of the detection system were 30 s, 60 s, 90 s, 120 s, 150 s, 180 s, 210 s, 240 s, 270 s, 300 s, 600 s, 900 s, 1200 s, and 1500 s. The excitation wavelength was set to 420 nm, the detection band was 430-700 nm, the emission start wavelength was 430 nm, the emission stop wavelength was 700 nm, the excitation bandwidth was 5 nm, the emission bandwidth was 5 nm, and the gain (PMT) was 650 V.

[0088] The results of the test are as follows Figure 5 As shown, the fluorescence value of probe S-2 in HEPES buffer with ATP added increases with time. In the first 210 s, the fluorescence value of probe S-2 in HEPES buffer with ATP added increases with time at a large rate, and after 210 s, the fluorescence value of probe S-2 in HEPES buffer with ATP added increases with time at a slow rate. This may be because there are a large number of free ATP molecules in the solution before 210 s, which makes the probe S-2 bind to ATP more quickly, forms aggregates faster, and increases the fluorescence value faster; after 210 s, there are fewer free ATP molecules in the solution, which makes the probe S-2 bind to ATP slower, and finally shows a slow increase in fluorescence value.

[0089] The system was allowed to stand still, and its fluorescence values ​​were measured at 600 s, 900 s, 1200 s, and 1500 s, respectively. It can be clearly seen from the figure that when the time increased from 300 s to 600 s, the fluorescence value of the system still increased significantly; the increase in the fluorescence value of the system decreased from 600 s to 1200 s, and the fluorescence value of the system remained basically unchanged from 1200 s to 1500 s, indicating that the probe S-2 reacted almost completely with the ATP molecule at 1200 s, and the fluorescence value of the system reached a maximum.

[0090] In summary, the aggregation-induced luminescent compound S-2 synthesized by the present invention using 2-benzylbenzimidazole, p-bromobenzylamine, etc. as raw materials has good AIE characteristics, belongs to aggregation-induced luminescent materials, and as an aggregation-induced fluorescent probe, it shows extremely high sensitivity and selectivity to ATP, and is not interfered by ADP, PPi and common ions. The ATP aggregation-induced luminescent compound provided by the present invention is simple to synthesize and has high specificity. As an adenosine triphosphate-induced aggregation-type fluorescent probe, it can specifically identify ATP and detect ATP molecules. The problem of poor ATP specificity in existing detection methods can be overcome. At the same time, the present invention provides a method for specifically identifying and detecting adenosine triphosphate, and the detection of ATP is realized by the fluorescent aggregation-induced effect of the fluorescent probe S-2. Compared with existing conventional methods, this method has the advantages of simple synthesis, high sensitivity, good specificity, and rapid response.

[0091] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An aggregation-induced emission compound, characterized in that: The structure of the compound is shown below: 。 2. The method for preparing the aggregation-induced emission compound according to claim 1, characterized in that: The following steps are involved: (1) Preparation of intermediate 3: 2-benzylbenzimidazole and cuprous chloride are dissolved in an organic solvent, and then p-bromobenzylamine and trimethylacetic acid are added, and stirred at 90°C-160°C for 2-36 hours. After the reaction is completed, the product is separated and purified by column chromatography to obtain intermediate 3, whose structural formula is: ； (2) Preparation of intermediate 5: In an inert gas atmosphere, intermediate 3, 4-pyridine boronic acid, tetrakis(triphenylphosphine)palladium and potassium carbonate are mixed and dissolved in an organic solvent, stirred at 60-100° C. for 6-12 hours, extracted, and separated and purified by column chromatography to obtain intermediate 5, whose structural formula is: ； (3) Preparation of aggregation-induced emission compounds: The intermediate 5 and benzyl bromide are dissolved in an organic solvent, stirred at 60-100°C for 16-36 hours, and filtered under reduced pressure after the reaction is completed. The filtered product is collected and washed to obtain an aggregation-induced emission compound.

3. The preparation method according to claim 2, characterized in that: In (1), the molar ratio of 2-benzylbenzimidazole, cuprous chloride, p-bromobenzylamine and trimethylacetic acid is (0.1-1): (0.05-2): (1-10): (0.5-8).

4. The preparation method according to claim 2, characterized in that: In (2), the molar ratio of intermediate 3, 4-pyridineboronic acid, tetrakis(triphenylphosphine)palladium and potassium carbonate is (0.1-1):(1-8):(0.05-1):(2-8).

5. The preparation method according to claim 2, characterized in that: In (3), the molar ratio of intermediate 5 and benzyl bromide is (0.05-2):(1-10).

6. Use of the aggregation-induced emission compound according to claim 1 as an aggregation-induced emission material.

7. Use of the aggregation-induced emission compound according to claim 1 as an adenosine triphosphate aggregation-induced fluorescent probe.

8. Use of the aggregation-induced emission compound of claim 1 in detecting adenosine triphosphate for purposes other than disease diagnosis and treatment, or in preparing products for detecting adenosine triphosphate.

9. A product for detecting adenosine triphosphate, characterized in that: Containing the aggregation-induced emission compound according to claim 1.

10. A method for specifically detecting adenosine triphosphate for non-disease diagnosis and treatment purposes, characterized in that: The detection is performed using the aggregation-induced emission compound described in claim 1 or the product described in claim 9.

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