Aggregation-induced emission material as well as preparation method and application thereof

Abstract

The invention discloses an aggregation-induced emission material as well as a preparation method and application thereof, and belongs to the technical field of aggregation-induced emission materials. The aggregation-induced emission material provided by the invention can be used for long-time high-resolution imaging of cancer cell membranes, and can be used for cancer cell radiography. The aggregation-induced emission material provided by the invention also can destroy cancer cells through photodynamic therapy to realize efficient tumor resistance.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of aggregation-induced emission materials, and particularly relates to an aggregation-induced emission material and a preparation method and application thereof. BACKGROUND

[0002] Cancer is one of the major causes of death worldwide, which is usually caused by gene mutation, leading to uncontrolled cell growth and serious complications. Traditional cancer treatment methods such as surgery, chemotherapy and radiotherapy have problems such as drug resistance, gene mutation and side effects. In contrast, photodynamic therapy as a non-invasive, low side effect and simple operation treatment method has become a promising treatment method. Photodynamic therapy injects photosensitizers into lesions, and uses light of a specific wavelength to activate the photosensitizers to generate active oxygen, thereby killing cancer cells. In addition to the treatment effect, the photodynamic agent also has imaging characteristics, which can realize image-guided cancer treatment. At present, fluorescence imaging has become an imaging technology that attracts much attention due to its high sensitivity, biocompatibility and simple operation. However, traditional cyanine dye fluorescent molecules have the problem of quenching caused by aggregation. Unlike this, aggregation-induced emission fluorescent molecules have strong luminescent properties in the aggregated state, and become a promising alternative for developing multifunctional imaging-guided photodynamic therapy systems. However, the fluorescent dyes currently used for cancer cell membrane targeting still have problems such as short imaging time and low resolution. SUMMARY

[0003] The application aims to provide an aggregation-induced emission material and a preparation method and application thereof. The provided aggregation-induced emission material can image cancer cell membranes for a long time with high resolution, and can destroy cancer cells through photodynamic therapy.

[0004] To achieve the above-mentioned purpose, the application provides the following technical solutions.

[0005] One of the technical solutions of the application is to provide an aggregation-induced emission material, which has the following structural formula:

[0006]

[0007] The second technical solution of the application is to provide a preparation method of the above-mentioned aggregation-induced emission material, which comprises the following steps:

[0008] Compound c is prepared by cross-coupling reaction of compound a and compound b; compound e is prepared by Witting reaction of compound c and compound d; and the aggregation-induced emission material is prepared by reacting compound e with 1,3-propane sulfonic acid lactone;

[0009] The structural formula of the compound a is:

[0010] The structural formula of the compound b is:

[0011] The structural formula of the compound c is:

[0012] The structural formula of the compound d is:

[0013] The structural formula of the compound e is:

[0014] Preferably, in the cross-coupling reaction, the solvent used is N,N-dimethylformamide (DMF), the catalyst used is palladium acetate (Pd(AcO)2), the phosphine ligand used is tris(o-tolyl)phosphine ((o-tol)3P), the acid binding agent used is triethylamine (TEA), the reaction is carried out under an inert atmosphere, the reaction temperature is 80-100 DEG C, and the reaction time is 12-16 h.

[0015] Preferably, in the Wittig reaction, the solvent used is ethanol (EtOH), the base catalyst used is sodium ethoxide (EtONa), the reaction is carried out under an inert atmosphere, the reaction temperature is 80-90 DEG C, and the reaction time is 12-16 h.

[0016] Preferably, in the reaction of the compound e with the 1,3-propane sulfonic acid lactone, the solvent used is acetonitrile, the reaction is carried out under an inert atmosphere, the reaction temperature is 85-90 DEG C, and the reaction time is 12-16 h.

[0017] The third technical solution of the present application provides an application of the above-mentioned aggregation-induced emission material in preparation of a cancer cell contrast agent.

[0018] The fourth technical solution of the present application provides an application of the above-mentioned aggregation-induced emission material in preparation of a photodynamic therapy reagent.

[0019] The present application has the following beneficial technical effects:

[0020] The aggregation-induced emission material provided by the present application can image cancer cell membranes for a long time with high resolution, and can be used for cancer cell contrast.

[0021] The aggregation-induced emission material provided by the present application can destroy cancer cells through photodynamic therapy, and achieve high-efficiency anti-tumor effect. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 The cell dark toxicity test results of different concentrations of compound f in Example 2.

[0023] Figure 2 The cell light toxicity test results of different concentrations of compound f in Example 2.

[0024] Figure 3 Results of membrane staining of cancer cells (Hela) for compound f in Example 3.

[0025] Figure 4 Results of membrane staining of normal cells (3T3) for compound f in Example 3. DETAILED DESCRIPTION

[0026] Various illustrative embodiments of the present application are described in detail below. The detailed description is presented in terms of specific embodiments which include particular components, materials, and dimensions. Those skilled in the art will recognize that the application can be practiced with

[0027] It should be noted that the present application does not describe in detail the conventional operations in the art, and is not the focus of the present application.

[0028] In addition, for numerical ranges that are expressly recited herein, it is understood that every intervening value between the upper and lower limits of that range is also specifically contemplated. In celling values, and ranges between any stated values or intervening values in stated ranges are also specifically encompassed. The upper and lower limits of these smaller ranges can independently be included or excluded in the range, and are also encompassed within the application, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of the limits are also included.

[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application.

[0030] As used herein, the terms "comprise", "comprising", "include", "including", "have", "having" and the like are open-ended and do not exclude additional elements or steps.

[0031] Example 1

[0032] (1) Synthesis of compound c:

[0033]

[0034] Compound a (243 mg), (o-tol)3P (304 mg), palladium acetate (224 mg) were dissolved in DMF under argon protection, then triethylamine (101 mg) and compound b (105 mg) were added. The mixture was stirred at 85°C under argon protection for 12 h. After that, the reaction mixture was extracted with dichloromethane, and the organic solvent was evaporated under reduced pressure to obtain the crude product. The purified compound c was obtained by column chromatography with a yield of 75%.

[0035] (2) Synthesis of compound e:

[0036]

[0037] Compound c (220 mg), compound d (273 mg) and sodium ethoxide (68 mg) were dissolved in ethanol under argon protection. The mixture was stirred at 90°C under argon protection for 12 h. After the reaction was completed, the mixture was filtered, and the filter residue was washed with ethanol, and finally the purified compound e was obtained with a yield of 67%.

[0038] (3) Synthesis of compound f:

[0039]

[0040] Compound e (475 mg) and 1,3-propanesultone (122 mg) were dissolved in acetonitrile under argon protection. The mixture was refluxed at 90°C under argon protection for 12 h. After the reaction was completed, the mixture was filtered, and the filter residue was washed with acetonitrile, and finally the purified compound f (solid powder) was obtained with a yield of 71%.

[0041] The nuclear magnetic detection result of compound f is:

[0042] 1 H NMR (500 MHz, DMSO-d6) δ (ppm): 9.00-8.94 (d, J = 6.0 Hz, 2H), 8.27-8.22 (s, 2H), 8.08-8.02 (m, 2H), 7.92-7.81 (m, 6H), 7.62-7.57 (m, 1H), 7.44-7.36 (m, 4H), 7.22-7.13 (m, 6H), 6.99-6.92 (m, 2H), 4.67-4.60 (m, 2H), 2.47-2.41 (m, 2H), 2.26-2.19 (m, 2H). 13 C NMR (125 MHz, DMSO-d6) δ (ppm): 152.61, 149.75, 145.91, 144.52, 142.72, 139.68, 135.92, 135.27, 131.07, 129.93, 128.84, 125.88, 125.74, 124.86, 123.94, 119.70, 118.33, 105.39, 58.64, 47.06, 27.25.

[0043] Example 2

[0044] Application of compound f in photodynamic therapy:

[0045] Dark cytotoxicity was assessed using the MTT assay. HeLa cells were seeded in 96-well plates and cultured at 37°C in an incubator containing 5% CO2 / 95% air at a cell density of 5 × 10⁶ cells / well. 4 Cells / mL. The original culture medium was discarded, and equal volumes of culture medium containing different concentrations of compound f (10, 20, 30, 40, 50 μg / mL) were added. Cells were then cultured in 96-well plates at 37°C for 6 h. The culture medium was discarded, and culture medium containing MTT (100 μL, 0.5 mg / mL) was added, and the cells were cultured for another 4 h. After removing the MTT-containing culture medium, 100 μL of DMSO was added to each well to lyse the cells. Finally, the absorbance at 490 nm was measured using a microplate reader, and cell viability was calculated. Results are as follows: Figure 1 As shown, when the concentration of compound f reaches 50 μM, the survival rate of HeLa cells is still above 75%, indicating that this aggregation-induced luminescence material has low cell dark toxicity and can be used for cell imaging studies.

[0046] Cell phototoxicity was assessed using the MTT assay. HeLa cells were seeded in 96-well plates and cultured at 37°C in an incubator containing 5% CO2 / 95% air at a cell density of 5 × 10⁶ cells / well. 4 Cells / mL. The original culture medium was discarded, and equal volumes of culture medium containing different concentrations of compound f (0.5, 1, 2, 5, 10, 15, 20 μg / mL) were added to each well. Cells were then cultured in 96-well plates at 37°C for 1 h. Subsequently, white light (10 mW) was applied for 30 min, and cells were cultured in 96-well plates at 37°C for another 6 h. The culture medium was discarded, and culture medium containing MTT (100 μL, 0.5 mg / mL) was added, followed by 4 h of culture. After removing the MTT-containing culture medium, 100 μL of DMSO was added to each well to lyse the cells. Finally, the absorbance at 490 nm was measured using a microplate reader, and cell viability was calculated. Results are as follows: Figure 2 As shown, when the concentration of compound f reaches 20 μM, the survival rate of HeLa cells is 15%, indicating that this aggregation-induced luminescent material has good cell phototoxicity and can be used for photodynamic therapy research of tumor cells.

[0047] Example 3

[0048] Applications of compound f cell imaging:

[0049] Cell imaging experiments were performed using compound f. HeLa cells or 3T3 cells were seeded in 35 mm confocal culture dishes (≥10 cells per well). 5Cells were cultured in 6-well plates (5 x 105cells / well) and incubated at 37 °C for 24 h. Subsequently, live cells were incubated with the dyes at the indicated concentrations and times. The procedure was as follows: 500 μM of compound f was added, the plates were gently shaken at room temperature for 30 s, and then immediately imaged using a confocal laser scanning microscope (CLSM) (excitation wavelength λex= 405 nm, laser power 5%; emission wavelength collection range 650-800 nm). The results are shown in Figure 3 (HeLa cells) and Figure 4 (3T3 cells).

[0050] Figure 3 and Figure 4 It is shown that compound f at 500 μM successfully achieved imaging of the membranes of cancer cells within an incubation time of 30 s, which indicates that the aggregation-induced emission material can achieve rapid staining of the membranes of cancer cells and has good anti-permeation ability. Normal cells were not stained, which indicates that compound f can distinguish between normal cells and cancer cells and selectively image the membranes of cancer cells.

[0051] The above-described embodiments are merely preferred modes of the present application and are not intended to limit the scope of the present application. Without departing from the spirit of the present application, various modifications and improvements to the technical solutions of the present application made by those of ordinary skill in the art should fall within the scope of protection of the present application as defined by the claims.

Claims

1. An aggregation-induced emission material, characterized in that, The structure of the aggregation-induced emission material is as follows:

2. A method for preparing the aggregation-induced emission material according to claim 1, characterized in that, The method comprises the following steps: Compound c is prepared by cross-coupling reaction of compound a and compound b; compound e is prepared by Wittig reaction of compound c and compound d; and the aggregation-induced emission material is prepared by reaction of compound e and 1,3-propane sultone; The structural formula of the compound a is: The structural formula of the compound b is: The structural formula of the compound c is: The structural formula of the compound d is: The structural formula of the compound e is:

3. The method for preparing the aggregation-induced emission material according to claim 2, characterized in that, In the cross-coupling reaction, the solvent is N,N-dimethylformamide, the catalyst is palladium acetate, the phosphine ligand is tri(o-methylphenyl)phosphine, the acid binding agent is triethylamine, the reaction is carried out in an inert atmosphere, the reaction temperature is 80-100 DEG C, and the reaction time is 12-16 h.

4. The method for preparing the aggregation-induced emission material according to claim 2, characterized in that, In the Wittig reaction, the solvent is ethanol, the base catalyst is sodium ethoxide, the reaction is carried out in an inert atmosphere, the reaction temperature is 80-90 DEG C, and the reaction time is 12-16 h.

5. The method for preparing the aggregation-induced emission material according to claim 2, characterized in that, In the reaction of compound e and 1,3-propane sultone, the solvent is acetonitrile, the reaction is carried out in an inert atmosphere, the reaction temperature is 85-90 DEG C, and the reaction time is 12-16 h.

6. The use of the aggregation-induced emission material in claim 1 in the preparation of a cancer cell contrast agent.

7. The use of the aggregation-induced emission material in claim 1 in the preparation of a photodynamic therapy reagent.

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