Water-soluble near-infrared two-region aggregation-induced emission material as well as preparation method and application thereof

Abstract

The invention discloses a water-soluble near-infrared two-region aggregation-induced emission material and a preparation method and application thereof, and belongs to the technical field of aggregation-induced emission materials.Benzobithiadiazole is used as an electron acceptor group, dithieno-cyclopentadiene and derivatives thereof are used as electron donor units, and benzobithiadiazole and dithieno-cyclopentadiene and derivatives thereof are coupled to form the water-soluble near-infrared two-region aggregation-induced emission material. The organic near-infrared second-region molecule with the intramolecular charge transfer characteristic is constructed. According to the invention, the positively charged quaternary ammonium salt group is introduced into the electron donor group, and the positively charged quaternary ammonium salt group has good hydrophilicity, so that the water solubility of molecules can be effectively improved. Moreover, the water-soluble near-infrared two-region aggregation-induced emission material has excellent photodynamic and photothermal activity, and realizes efficient killing of bacteria (especially propionibacterium acnes) through the synergistic effect of photodynamic and photothermal effects.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of aggregation-induced emission materials, and particularly relates to a water-soluble near-infrared two-region aggregation-induced emission material and a preparation method and application thereof. BACKGROUND

[0002] Bacterial infection is one of the main threats to human health. Studies have shown that many diseases are closely related to bacteria. Among them, Propionibacterium acnes is considered to be closely related to the occurrence and development of acne, and its excessive growth is an important factor in the pathogenesis of acne. At present, the main treatment method for acne is to use antibiotics to kill bacteria. However, the abuse of antibiotics can easily lead to bacterial drug resistance, which seriously affects the treatment effect. In recent years, phototherapy has attracted widespread attention from researchers due to its high spatial and temporal resolution, minimal invasion, low toxicity, and repeatable treatment. It has been applied to the treatment of acne. However, it has been found that Propionibacterium acnes is an anaerobic bacterium that grows in the deep part of the hair follicle, which leads to the poor effect of the current photodynamic therapy mainly excited by blue light and requiring a large amount of oxygen.

[0003] The luminescent material in the second near-infrared window (NIR-II, 1000-1700nm) has attracted much attention due to its high spatial and temporal resolution, strong penetration, and other unique advantages. These outstanding characteristics make NIR-II phototherapy materials an ideal choice for deep treatment in vivo. Therefore, developing new NIR-II phototherapy materials can help to enhance the killing effect of Propionibacterium acnes, thereby improving the treatment effect of acne. SUMMARY

[0004] To solve the above technical problems, the application provides a water-soluble near-infrared two-region aggregation-induced emission material and a preparation method and application thereof. The application develops a new NIR-II phototherapy material. The material has the absorption of NIR-I and the luminescence performance of NIR-II, and exhibits excellent type I photodynamic and photothermal activity, realizes the effective killing of Propionibacterium acnes, and provides a new method for the treatment of acne.

[0005] To achieve the above purpose, the application provides the following technical solutions.

[0006] The application provides a water-soluble near-infrared two-region aggregation-induced emission material, denoted as SBBTN, and the structural formula is as follows:

[0007]

[0008] The application selects benzobisthiazole with strong electron-withdrawing properties as an electron acceptor group, and constructs an organic fluorescent material with intramolecular charge transfer characteristics by taking dithienocyclopenta-diene and its derivatives as electron donor units.

[0009] The application further provides a preparation method of the water-soluble near-infrared two-region aggregation-induced emission material.

[0010] The benzobisthiazole derivative has a structural formula as shown in the following formula (I). The dithienocyclopenta-diene derivative has a structural formula as shown in the following formula (II).

[0011] Further, the molar ratio of the benzobisthiazole derivative and the dithienocyclopenta-diene derivative is 0.5:1.

[0012] The application further provides application of the water-soluble near-infrared two-region aggregation-induced emission material in preparation of a fluorescent contrast agent.

[0013] The application further provides application of the water-soluble near-infrared two-region aggregation-induced emission material in near-infrared two-region fluorescence imaging for non-therapeutic purposes.

[0014] The application further provides application of the water-soluble near-infrared two-region aggregation-induced emission material in preparation of an antibacterial drug.

[0015] Further, the antibacterial drug is a drug for resisting propionic acid acne bacillus.

[0016] The application further provides a pharmaceutical composition for treating propionic acid acne bacillus, and the effective component of the pharmaceutical composition is the water-soluble near-infrared two-region aggregation-induced emission material.

[0017] Compared with the prior art, the application has the following advantages and technical effects:

[0018] The application provides an aggregation-induced emission phototherapy material capable of emitting light in the NIR-II (1000 nm to 1500 nm) region and having photodynamic and photothermal activities, and the water solubility of the material is further improved by introducing a positively charged quaternary ammonium salt group into an electron donor group. BRIEF DESCRIPTION OF DRAWINGS

[0019] The accompanying drawings, which form a part of this application, are included to provide a further understanding of the application and are incorporated in and constitute a part of this application. The drawings illustrate embodiments of the present application and, together with the description, serve to explain the application. In the drawings:

[0020] Figure 1 The test results of water-solubility of SBBTN, a is the photo of SBBT and SBBTN in 1-octanol and water mixed solution (v / v = 1:1), b is the particle size distribution of SBBTN in phosphate buffer (concentration: 10 μM).

[0021] Figure 2 The characterization results of photophysical properties of SBBTN, a is the UV-Vis absorption spectrum and fluorescence emission spectrum of SBBTN, b is the fluorescence emission spectrum of SBBTN in aqueous solution with different contents of tetrahydrofuran, c is the curve of the fluorescence intensity peak of SBBTN with the change of the content of tetrahydrofuran.

[0022] Figure 3 The characterization results of photodynamic activity of SBBTN, a is the characterization results of total active oxygen production capacity, b is the characterization results of singlet oxygen production capacity, c is the characterization results of superoxide radical production capacity, d is the characterization results of superoxide radical production capacity under normoxic and anoxic conditions, e is the characterization results of hydroxyl radical production capacity, f is the characterization results of hydroxyl radical production capacity under normoxic and anoxic conditions.

[0023] Figure 4 The characterization results of photothermal activity of SBBTN, a is the temperature change curve of SBBTN aqueous solution (concentration of 50 μM) under laser irradiation (irradiation intensity of 1 W / cm 2 ), b is the temperature rise curve of SBBTN aqueous solution (concentration of 50 μM) under different power intensities (0.4, 0.6, 0.8 and 1 W / cm 2 ), c is the temperature rise curve of SBBTN aqueous solution with different concentrations (10, 20, 40 and 80 μM) under laser irradiation (irradiation intensity of 1 W / cm 2 ), d is the characterization results of photothermal stability of SBBTN and indocyanine green.

[0024] Figure 5 The characterization results of antibacterial activity of SBBTN, a is the photo of bacterial colonies on agar plates, b is the characterization results of bactericidal activity of SBBTN under laser irradiation and in the dark under different concentrations (0, 2.5 μM, 5 μM, 10 μM and 20 μM).

[0025] Figure 6 The characterization results of in vitro biocompatibility of SBBTN. DETAILED DESCRIPTION

[0026] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0027] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0028] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0029] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0030] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0031] An embodiment of the present invention provides a water-soluble near-infrared II aggregation-induced emission material, denoted as SBBTN, with the following structural formula:

[0032]

[0033] The SBBTN of this invention exhibits AIE (aggregation-induced emission) properties, emitting fluorescence in the near-infrared II region from 1000 nm to 1500 nm. This invention selects benzobisthiadiazole, with its strong electron-withdrawing properties, as the electron acceptor group, and uses dithienylcyclopentadiene and its derivatives as electron donor units to construct an organic fluorescent material with intramolecular charge transfer characteristics. This invention introduces a positively charged quaternary ammonium salt group into the electron donor group; due to the good hydrophilicity of the positively charged quaternary ammonium salt group, the water solubility of the molecule can be effectively improved.

[0034] The embodiment of the present application also provides a preparation method of the water-soluble near-infrared two-region aggregation-induced emission material, and the preparation method is as follows: using benzodithiazole derivatives and dithienocyclopenta derivative as raw materials, a water-soluble near-infrared two-region aggregation-induced emission material is prepared by Stille coupling reaction and quaternization reaction.

[0035] The benzodithiazole derivative has a structural formula as follows: The dithienocyclopenta derivative has a structural formula as follows:

[0036] In the embodiment of the present application, the molar ratio of the benzodithiazole derivative to the dithienocyclopenta derivative is 0.5:1.

[0037] The present application also provides application of the water-soluble near-infrared two-region aggregation-induced emission material in preparation of a fluorescent contrast agent.

[0038] The present application also provides application of the water-soluble near-infrared two-region aggregation-induced emission material in near-infrared two-region fluorescence imaging for non-therapeutic purposes.

[0039] The present application also provides application of the water-soluble near-infrared two-region aggregation-induced emission material in preparation of an antibacterial drug.

[0040] In the embodiment of the present application, the antibacterial drug is a drug against propionibacterium acnes.

[0041] The present application also provides a pharmaceutical composition for treating propionibacterium acnes, and the effective component of the pharmaceutical composition is the water-soluble near-infrared two-region aggregation-induced emission material.

[0042] In the present application, benzodithiazole is used as an electron acceptor group, dithienocyclopenta and its derivative are used as an electron donor unit, and the two are coupled to construct an organic near-infrared two-region molecule with intramolecular charge transfer characteristics. By introducing a positively charged quaternary ammonium salt group into the electron donor group, the water solubility of the molecule can be effectively improved because the positively charged quaternary ammonium salt group has good hydrophilicity. Moreover, the water-soluble near-infrared two-region aggregation-induced emission material has excellent photodynamic and photothermal activity, and through the synergistic effect of photodynamic and photothermal effect, efficient killing of bacteria (especially propionibacterium acnes) is realized.

[0043] Unless otherwise specified, the room temperature in the present application is 25±2℃.

[0044] Each raw material used in the embodiment of the present application is obtained by commercial purchase. As an example, NIH-3T3 cells are purchased from the China Academy of Sciences Cell Bank; propionibacterium acnes is purchased from Beina Biological Technology Co., Ltd. - Henan Industrial Microbial Strain Engineering Technology Research Center.

[0045] It should be noted that the invention is not described in detail, which is the conventional operation means in the art, and is not the focus of the invention, for example, MTT detection of cell toxicity, plate coating method to calculate the survival rate of bacteria, etc. The specific method is completed by using the conventional method.

[0046] The technical solutions of the present application are further illustrated by the following examples.

[0047] Synthesis of compound SBBT

[0048] The synthesis route of compound SBBT is as follows:

[0049]

[0050] Under the atmosphere of nitrogen, compound 1 (50.4 mg, 0.1 mmol) was dissolved in tetrahydrofuran (THF, 5 mL), and n-butyl lithium (0.11 mmol) was added dropwise at -78°C. After 1 h of reaction, tributyltin chloride (35.8 mg, 0.11 mmol) was added, and the temperature was restored to room temperature. After overnight reaction, the reaction solution was quenched with potassium fluoride, extracted with dichloromethane three times, and then the organic phase was dried with sodium sulfate. The obtained crude product 2 was distilled under reduced pressure. Under the atmosphere of nitrogen, the crude product 2 (79.3 mg, 0.1 mmol), compound 3 (34.3 mg, 0.05 mmol), tris(dibenzylideneacetone)dipalladium (Pd2(dba)3, 9.2 mg, 0.01 mmol) and tri(o-tolyl)phosphine (P(o-tol)3, 30.4 mg, 0.1 mmol) were dissolved in toluene, and refluxed overnight. After the reaction was completed, the reaction solution was cooled to room temperature, quenched with potassium fluoride, extracted with dichloromethane three times, and then the organic phase was dried with sodium sulfate. The obtained crude product was distilled under reduced pressure, and the product SBBT was separated by column chromatography (the yield was 58%). 1 H NMR (500 MHz, CDCI3) δ 7.31 (s, 2H), 7.22 (d, J = 4.5 Hz, 2H), 7.13 (s, 2H), 6.94 (d, J = 4.5 Hz, 2H), 3.35 (t, J1= 13.5 Hz, J2= 6.5 Hz, 8H), 2.58 (t, J1= 15.5 Hz, J2= 7.5 Hz, 4H), 1.87 (t, J1= 16.5 Hz, J2= 8.5 Hz, 8H), 1.78-1.72 (m, 8H), 1.67-1.61 (m, 4H), 1.34-1.28 (m, 8H), 1.20-1.08 (m, 20H), 0.99-0.95 (m, 8H), 0.75 (t, J1= 14 Hz, J2= 7 Hz, 6H). 13CNMR (126 MHz, CDC13) δ 158.35, 157.83, 153.19, 145.81, 141.23, 136.99, 136.58, 136.48, 127.72, 125.61, 125.18, 121.52, 118.71, 115.74, 53.63, 37.87, 34.08, 32.72, 31.54, 30.50, 30.25, 29.17, 29.10, 27.95, 24.30, 22.52, 14.05.

[0051] Synthesis of compound SBBTN of Example 1

[0052] The synthesis route of compound SBBTN is as follows:

[0053]

[0054] SBBT (100 mg, 0.07 mmol) prepared in Comparative Example 1 was dissolved in 2 mL of THF, trimethylamine (Me3N, 25 wt% THF solution, 1 mL) was added dropwise at a temperature of -78°C, after stirring for 1 hour, the reaction temperature was raised to 60°C, and stirred for 48 hours, during which 1 mL of methanol was added every 12 hours, after the reaction was completed, the reaction solution was cooled to room temperature, and the product SBBTN was obtained by distillation under reduced pressure (yield 90%). 1 H NMR (500 MHz, MeOD) δ 7.36-7.23 (m, 6H), 7.03-7.03 (m, 2H), 3.22-3.18 (m, 8H), 3.02 (s, 36H), 2.56 (t, J1= 16.0 Hz, J2= 8.5 Hz, 2H), 1.99-1.96 (m, 8H), 1.59-1.51 (m, 12H), 1.20 (s, 18H), 1.07-1.03 (m, 4H), 0.92-0.85 (m, 16H), 0.74-0.71 (m, 2H), 0.65-0.62 (m, 4H). Figure S4.4: 13C NMR(126MHz,MeOD)δ158.57,158.33,157.96,152.96,145.59,140.48,136.82,136.44,136.28,135.72,128.10,126.00,124.75,121.42 ,118.47,115.40,66.33,53.75,52.12,37.35,31.38,31.12,29.87,28.66,28.51,25.37,23.71,22.33,22.15,13.15,13.06,13.00.ESI HRMS calcd.for C80H118N8S8 4+ [m / z]:361.6806,found:361.6768.

[0055] Characterization of the water solubility of SBBT and SBBTN in Test Example 1

[0056] Figure 1 In Figure a, there is a photograph of the materials obtained from Comparative Example 1 and Example 1 in a mixed solution of 1-octanol and water (volume ratio = 1:1). It can be seen that SBBT is mainly distributed in 1-octanol, while SBBTN is mainly distributed in water, indicating that SBBTN has good water solubility. Figure 1 Figure b shows the dynamic light scattering (DLS) results of SBBTN in water, which indicates that SBBTN can be dispersed in water and self-assemble into nanoparticles.

[0057] Test Example 2: Photophysical Properties of SBBTN

[0058] Figure 2 The photophysical properties of SBBTN are characterized as follows: a) shows the UV-Vis absorption and fluorescence emission spectra of SBBTN; b) shows the fluorescence emission spectra of SBBTN in aqueous solutions with different tetrahydrofuran contents; c) shows the curve of SBBTN fluorescence intensity peak changing with tetrahydrofuran content. Figure 2 From a, we can see that SBBTN exhibits NIR-I absorption and NIR-II luminescence. From... Figure 2 As shown in b and c, SBBTN exhibits weak luminescence in the good solvent water, but its fluorescence gradually increases with the addition of the unsuitable solvent tetrahydrofuran, which leads to aggregate formation. This indicates that these materials all possess AIE properties.

[0059] Characterization of the photodynamic activity of SBBTN in Test Example 3

[0060] The photodynamic activity of SBBTN was characterized using different indicators (2',7'-dichlorodihydrofluorescein diacetate, DCFH; 9,10-anthracenediyl-bis(methylene)dipropanoic acid, ABDA; dihydro rhodamine 123, DHR123; hydroxyphenyl fluorescein, HPF), and the results are shown in Table 1. Figure 3 , a is the characterization result of total active oxygen production capacity, b is the characterization result of singlet oxygen production capacity, c is the characterization result of superoxide radical production capacity, d is the characterization result of superoxide radical production capacity under normoxic and anoxic conditions, e is the characterization result of hydroxyl radical production capacity, and f is the characterization result of hydroxyl radical production capacity under normoxic and anoxic conditions. From the results in Table 1, it can be seen that under normoxic and anoxic conditions, SBBTN can effectively produce active oxygen under the irradiation of 808 nm laser, and the main types are free radicals (hydroxyl radicals, superoxide anion radicals), which is a type I photosensitizer. Figure 3

[0061] Test Example 4 Characterization of the photothermal activity of SBBTN

[0062] Figure 4 The characterization results of the photothermal activity of SBBTN are shown in Table 2, a is the temperature change curve of SBBTN aqueous solution (concentration of 50 μM) under laser irradiation (irradiation intensity of 1 W / cm 2 ), b is the temperature change curve of SBBTN aqueous solution (concentration of 50 μM) under different power intensities (0.4, 0.6, 0.8 and 1 W / cm 2 ), c is the temperature change curve of SBBTN aqueous solution with different concentrations (10, 20, 40 and 80 μM) under laser irradiation (irradiation intensity of 1 W / cm 2 ), and d is the characterization results of the photothermal stability of SBBTN and indocyanine green. The results show that SBBTN can effectively generate heat under the irradiation of 808 nm laser, and the photothermal conversion efficiency is 50.4%, which shows excellent photothermal properties and photothermal stability.

[0063] Test Example 5 Characterization of the antibacterial activity of SBBTN

[0064] SBBTN was incubated with P. acnes for 30 min, and then irradiated with 808 nm laser for 10 min. The survival rate of bacteria was calculated by plate coating method. Figure 5 The characterization results of the antibacterial activity of SBBTN are shown in Table 3, a is the colony photograph on agar plate, and b is the characterization results of the bactericidal activity of SBBTN under laser irradiation and in the dark under different concentrations (0, 2.5 μM, 5 μM, 10 μM and 20 μM). The results show that SBBTN can effectively kill P. acnes under the irradiation of 808 nm laser, and under the conditions of a concentration of 10 μM and an irradiation time of 10 min, the bactericidal rate is 99.99%.

[0065] ​Example 6: Evaluation of the biocompatibility of SBBTN at the cellular level

[0066] Evaluation of the cytotoxicity of SBBTN using NIH-3T3 cells: NIH-3T3 cells were plated in 96-well plates and incubated for 24 h, then co-incubated with SBBTN at different concentrations for 24 h. The cytotoxicity was detected by MTT. The results are shown in Table 1. Figure 6 SBBTN had no significant cytotoxicity.

[0067] The above merely describes the preferred embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can easily think of changes or replacements within the technical scope disclosed by the present application, which should be covered by the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A water-soluble near-infrared two-region aggregation-induced emission material, characterized in that, The structural formula is as follows:

2. A method for preparing the water-soluble near-infrared two-region aggregation-induced emission material according to claim 1, characterized in that, The water-soluble near-infrared two-region aggregation-induced emission material is prepared by using Stille coupling reaction and quaternization reaction with benzobisthiazole derivatives and dithienocyclopenta-diene derivatives as raw materials; The benzodithiazolyl derivative has a structural formula as shown in the following formula (I) or (II) The dithienocyclopentadiene derivative has a structural formula as shown in the following formula (III) or (IV) 3. The method for preparing the water-soluble near-infrared II aggregation-induced emission material according to claim 2, characterized in that, The molar ratio of the benzobisthiazole derivatives and the dithienocyclopenta-diene derivatives is 0.5:

1.

4. Application of the water-soluble near-infrared two-region aggregation-induced emission material in claim 1 in preparation of a fluorescent contrast agent.

5. Application of the water-soluble near-infrared two-region aggregation-induced emission material in claim 1 in near-infrared two-region fluorescence imaging for non-therapeutic purposes.

6. Application of the water-soluble near-infrared two-region aggregation-induced emission material in claim 1 in preparation of an antibacterial drug.

7. Use according to claim 6, characterized in that, The antibacterial drug is an anti-P. acnes drug.

8. A pharmaceutical composition for treating P. acnes, characterized by, The effective component is the water-soluble near-infrared two-region aggregation-induced emission material in claim 1.

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