CN116606313A - Aza-BODIPY-based high-brightness NIR-II region photothermal agent and preparation method and application thereof - Google Patents
Aza-BODIPY-based high-brightness NIR-II region photothermal agent and preparation method and application thereof Download PDFInfo
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- 239000012221 photothermal agent Substances 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
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- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 70
- 238000006243 chemical reaction Methods 0.000 claims description 68
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 53
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 42
- 239000007787 solid Substances 0.000 claims description 38
- 239000000243 solution Substances 0.000 claims description 38
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-diisopropylethylamine Substances CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 37
- 239000013067 intermediate product Substances 0.000 claims description 35
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- IAZDPXIOMUYVGZ-UHFFFAOYSA-N DMSO Substances CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 1
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- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic System
- C07F5/02—Boron compounds
- C07F5/022—Boron compounds without C-boron linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The application provides an Aza-BODIPY-based high-brightness NIR-II region photothermal agent and a preparation method and application thereof, and belongs to the technical field of photothermal treatment. A dual mechanism of improving the brightness and photo-thermal performance of the molecule by side chains is disclosed, and the side chain engineering can be used as an effective method for regulating and controlling the radiation transition and non-radiation transition processes simultaneously. Specifically, the introduction of side chains promotes intramolecular movement in the aggregated state by further distorting the molecular conformation, extending the intermolecular distance, and providing a more loosely packed environment for intramolecular movement (too large side chains can cause greater steric hindrance in the aggregated state), thereby promoting non-radiative transitions to produce photothermal properties. Furthermore, while intramolecular movement is promoted with the aid of side chains, pi-pi stacking also decreases with prolonged intermolecular distance, thereby promoting fluorescence in an aggregated state. The introduction of alkyl chain enhances both fluorescence and photothermal, and can be used for photothermal treatment under fluorescence guidance.
Description
Technical Field
The application belongs to the technical field of photothermal therapy, and particularly relates to an Aza-BODIPY-based high-brightness NIR-II region photothermal agent, a preparation method and application thereof.
Background
Photodiagnosis integrates photoimaging diagnosis and light-triggered accurate treatment, and is an emerging biotechnology. Because of their minimal invasiveness, high spatial-temporal resolution, high biosafety, and high anti-tumor capabilities, they play a significant and increasingly important role in accurate tumor therapy. As the core of the photo-thermal system, the development of high-performance photo-thermal agents is critical to the development of photo-thermal research. In particular, photothermal agents having near infrared two-region (NIR-II, 1000-1700 nm) emissions have attracted extensive attention over the past few years, with higher signal-to-noise ratios, deeper penetration, and lower light absorption, scattering, and autofluorescence interference from biological tissues. In order to obtain efficient NIR-II photothermal agents, four strategies are commonly applied to the red-shifted absorption and emission wavelengths of fluorophores: 1) Extending the conjugated chain of the fluorophore; 2) Enhancing intramolecular donor-acceptor (D-a) interactions; 3) Modulating the intensity and number of donors/acceptors in the fluorophore; 4) J-aggregates were prepared.
Despite many efforts, the structure-activity relationship of most NIR-II photothermal agents to date has not been explored to a great extent. In addition, the above-mentioned photothermal agents mostly have low fluorescence Quantum Yield (QY) and Photothermal Conversion Efficiency (PCE), resulting in low imaging resolution and limited anticancer activity. Therefore, the simultaneous improvement of QY and PCE of the NIR-II photothermal agent has important significance for the photothermal anticancer research.
Unfortunately, the simultaneous enhancement of QY and PCE has long been considered a contradiction, which makes the implementation of NIR-II photothermal agents with both high QY and strong PCE very challenging. Currently, the method for improving the aggregation state QY mainly comprises the following steps: 1) The steric effect is utilized to reduce the intermolecular pi-pi interaction; 2) Inhibiting distorted intramolecular charge transfer of the excited state. The main strategy to improve PCE relies on the introduction of fast moving rotors to accelerate the radiationless relaxation process. Almost without exception, however, these methods result in a decrease in molecular planarity and molar extinction coefficient (. Epsilon.) and thus decrease the total excited state energy of the photothermal agent. Therefore, the development of NIR-II photothermal agents with good planarity, high QY and PCE is very critical, but quite challenging.
Disclosure of Invention
In order to overcome the defects of the prior art, the application aims to provide an Aza-BODIPY-based high-brightness NIR-II region photothermal agent, a preparation method and application thereof, which are used for solving the technical problems of reduction of molecular planeness and molar extinction coefficient (epsilon), and thus reduction of total excited state energy of the photothermal agent, low imaging resolution and side effects of photothermal treatment.
In order to achieve the above purpose, the application is realized by adopting the following technical scheme:
the application provides an Aza-BODIPY-based high-brightness NIR-II region photothermal agent, which has the following structural formula:
the application also provides a preparation method of the Aza-BODIPY-based high-brightness NIR-II region photothermal agent, which comprises the following steps:
step one: adding N, N-dimethylformamide into phosphorus oxychloride, and stirring under a protective gas to obtain a Vilsmeier-Haack reagent; mixing julolidine and N, N-dimethylformamide, adding the mixture into a Vilsmeier-Haack reagent, heating to perform a first reaction, cooling to room temperature after the first reaction is finished, pouring the mixture into ice water to quench the reaction, filtering and drying to obtain an intermediate product S1;
step two: uniformly mixing the intermediate product S1, 4-bromoacetophenone with sodium hydroxide aqueous solution and ethanol for a second reaction, pouring the mixed solution of the second reaction product into ice water for quenching reaction after the second reaction is finished, and then filtering and drying to obtain an intermediate product S2;
step three: uniformly mixing an intermediate product S2, nitromethane, N-diisopropylethylamine and methanol, then heating to perform a third reaction, pouring a solution of the third reaction product into a saturated sodium chloride aqueous solution to stop the reaction, and then extracting, drying, filtering and purifying to obtain an intermediate product S3;
step four: mixing the intermediate product S3, ammonium acetate and n-butanol uniformly, heating to perform a fourth reaction, then concentrating in vacuum, filtering, separating solids, and washing to obtain an intermediate product S4;
step five: mixing intermediate products S4, N-diisopropylethylamine with dried dichloromethane, adding boron trifluoride diethyl etherate, stirring at room temperature in a dark place for a fifth reaction, diluting the fifth reaction product with ice water, extracting, drying, filtering, concentrating and purifying to obtain an intermediate product S5;
step six: under the protection gas, heating a solution of an intermediate product S5, 4-pyridine phenylboronic acid, tetra (triphenylphosphine) palladium, potassium carbonate and 1,4 dioxane mixed with water for a sixth reaction, cooling to room temperature after the sixth reaction is finished, concentrating, filtering and purifying to obtain an intermediate product S6;
step seven: under the protection gas, mixing the intermediate product S6 with 1-bromododecane and chloroform, heating to perform a seventh reaction, cooling to room temperature after the seventh reaction is finished, concentrating, filtering and purifying to obtain WS5.
In a specific embodiment, in the first step, the ratio of the amounts of phosphorus oxychloride, N-dimethylformamide and julolidine is 7.62:22.87:6.93mmol; the temperature of the first reaction is 20-30 ℃; the time of the first reaction is 3-4 h.
In a specific embodiment, in the second step, the ratio of the amounts of the substances of the intermediate products S1, 4-bromoacetophenone is 1:1, a step of; the temperature of the second reaction is 25-30 ℃.
In a specific embodiment, in the third step, the ratio of the amounts of the substances of the intermediate product S2 to N, N-diisopropylethylamine is 5:5.5; the temperature of the third reaction is 60-70 ℃.
In a specific embodiment, in the fourth step, the ratio of the amount of the intermediate product S3 to the amount of the substance of ammonium acetate is 1:15; the temperature of the fourth reaction is 100-110 ℃.
In a specific embodiment, in the fifth step, the ratio of the amounts of the intermediate products S4, N-diisopropylethylamine and boron trifluoride etherate is 0.11:1.1:2.
in a specific embodiment, in the sixth step, the ratio of the amounts of the substances of the intermediate product S5, 4-pyridine phenylboronic acid, tetrakis (triphenylphosphine) palladium and potassium carbonate is 0.06:0.12:0.1:7.5; the temperature of the sixth reaction is 70-80 ℃; the volume ratio of the 1,4 dioxane to the water is 5:1.
in a specific embodiment, in the seventh step, the ratio of the amount of the intermediate product S6 to the amount of the 1-bromododecane is 1:1, a step of; the temperature of the seventh reaction is 80-90 ℃.
The application also provides an application of the high-brightness NIR-II region photothermal agent in preparation of photothermal therapeutic drugs.
Compared with the prior art, the application has the following beneficial effects:
the present application provides an Aza-BODIPY based high intensity NIR-II region photothermal agent (WS 5) wherein the introduction of alkyl chains promotes intra-molecular movement in an aggregated state by further distorting the molecular conformation, extending the intermolecular distance and providing a more loosely packed environment for intra-molecular movement to promote non-radiative transitions to produce highly efficient photothermal Properties (PCE). Furthermore, while intramolecular movement is promoted with the aid of alkyl chains, pi-pi stacking also decreases with prolonged intermolecular distance, thereby promoting fluorescence (QY) in an aggregated state. Thus, the introduction of alkyl chains enhances both fluorescence and photothermal, and can be used for photothermal therapy under fluorescence guidance.
The application further discloses a photo-thermal agent (WS 5) which has strong absorption in a near infrared two-region, high fluorescence quantum yield and photo-thermal conversion efficiency, and DSPE-PEG 5000 The coated material has good biocompatibility, small toxic and side effects on organisms, can be biologically metabolized, and has certain biological application potential. The double-improvement mechanism of the side chain structure for improving the brightness and the photo-thermal performance is explored, and a new thought is provided for solving the problem of energy distribution depending on two competitive photophysical processes. The deep tumor treatment effect of the high-brightness and high-photo-thermal performance double-optimized photo-thermal agent under the near infrared two-region imaging guidance is studied.
Drawings
FIG. 1 is a graph showing the photophysical properties of WS5 molecule; wherein, the graph (a) is a space configuration graph of WS5 molecules optimized by density functional theory; FIG. (b) is a graph showing the molar absorptivity of WS 5; FIG. (c) is an emission spectrum of WS5 molecules; FIG. d is a graph showing the photo-thermal properties of WS5 molecules;
FIG. 2 is a graph of the test of the underlying photophysical properties of WS5@NPs of this application; wherein, the graph (a) is a DLS and TEM characterization graph of WS5@NPs; FIG. (b) is a graph showing the molar absorptivity of WS5@NPs; FIG. (c) is an emission spectrum of WS5@NPs; the graph (d) is a test graph of the photo-thermal performance of WS5@NPs; FIG. (e) is a graph of the photostability of WS5@NPs; panel (f) is a test chart of the thermal stability of ws5@nps;
FIG. 3 is an experimental diagram of in vitro cells (143B cells) of WS5@NPs of this application; wherein, the graph (a) is a cell viability test graph (0.23W cm -2 ) The method comprises the steps of carrying out a first treatment on the surface of the Panel (b) is an IC50 value measurement plot of WS5@NPs; FIG. (c) is a schematic diagram of the uptake of WS5@NPs to 143B cells by cells; (d) Live-dead staining experimental plots of ws5@nps, phosphate buffered saline scale = 100 μm; (e) Flow cytometry detection 143B cell apoptosis;
FIG. 4 (a) is a whole body vascular imaging of mice injected with WS5@NPs in vivo with 980nm laser; wherein, the images (1), (2) and (3) are the whole body vascular imaging images of mice injected with WS5@NPs at 1100LP, 1200LP and 1300LP respectively;
FIG. 4 (b) is a graph showing the comparison of the diameters of blood vessels between the green, blue and red lines;
FIG. 5 is a schematic illustration of the photothermal treatment of mouse osteosarcoma by WS5@NPs; wherein, figure (a) is a schematic diagram of an in vivo experimental time axis; panel (b) is a photo-thermal image contrast plot for the PBS +1064nm group and ws5@nps +1064nm group; panel (c) is x-ray images of osteosarcoma mice at 0, 6, 10 days; panel (d) is a camera photograph of osteosarcoma mice at days 0, 6, 10; panel (e) is a graphical representation of the change in tumor size of mice after 10 days of photothermal treatment (n=5, < p < 0.05); FIG. (f) is a photograph of a tumor; FIG. (g) is a graph of tumor weight after dissection in mice; panel (h) is a panel of immunohistochemical assays for treatment groups; FIG. (i) is a histological examination; FIG. j is a schematic view of the skin histology of the laser irradiated region;
FIG. 6 is a schematic diagram showing the preparation method of NIR-II photothermal agent with high brightness and high photothermal performance according to example 1 of the present application;
FIG. 7 is a schematic diagram of a process for generating WS5@NPs by WS5 nanocrystallization;
fig. 8 is a structural formula of WS5.
Detailed Description
So that those skilled in the art can appreciate the features and effects of the present application, a general description and definition of the terms and expressions set forth in the specification and claims follows. Unless otherwise defined, 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, and in the event of a conflict, the present specification shall control.
The theory or mechanism described and disclosed herein, whether right or wrong, is not meant to limit the scope of the application in any way, i.e., the present disclosure may be practiced without limitation to any particular theory or mechanism.
All features such as values, amounts, and concentrations that are defined herein in the numerical or percent ranges are for brevity and convenience only. Accordingly, the description of a numerical range or percentage range should be considered to cover and specifically disclose all possible sub-ranges and individual values (including integers and fractions) within the range.
Herein, unless otherwise indicated, "comprising," "including," "having," or similar terms encompass the meanings of "consisting of … …" and "consisting essentially of … …," e.g., "a includes a" encompasses the meanings of "a includes a and the other and" a includes a only.
In this context, not all possible combinations of the individual technical features in the individual embodiments or examples are described in order to simplify the description. Accordingly, as long as there is no contradiction between the combinations of these technical features, any combination of the technical features in the respective embodiments or examples is possible, and all possible combinations should be considered as being within the scope of the present specification.
The application provides an Aza-BODIPY-based high-brightness NIR-II region photothermal agent, a preparation method and application thereof.
The application will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present application and are not intended to limit the scope of the present application. Furthermore, it should be understood that various changes and modifications can be made by one skilled in the art after reading the teachings of the present application, and such equivalents are intended to fall within the scope of the application as defined in the appended claims.
The following examples use instrumentation conventional in the art. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. The following examples used various starting materials, unless otherwise indicated, were conventional commercial products, the specifications of which are conventional in the art. In the description of the present application and the following examples, "%" means weight percent, and "parts" means parts by weight, and ratios means weight ratio, unless otherwise specified.
The application provides a preparation method of an Aza-BODIPY-based high-brightness NIR-II region photothermal agent, which comprises the following specific steps:
step one: synthesis of Compound S1
Under ice salt bath condition, phosphorus oxychloride (POCl) 3 7.62 mmol) of N, N-dimethylformamide containing dry matter was slowly dropped into the reactorDMF,22.87 mmol) in a 100mL round bottom flask. At N 2 After stirring for 2h under protection of (2) the Vilsmeier-Haack reagent was obtained. Julolidine (6.93 mmol) and dry DMF (10 mL) were then slowly added to the flask containing Vilsmeier-Haack reagent. Then, stirring is carried out for 3 to 4 hours at the temperature of 20 to 30 ℃. After cooling to room temperature, the mixture was poured into ice water (100 mL) to quench the reaction. The precipitate was then filtered to give a pale yellow solid. Finally, the solid was dried in a vacuum oven overnight to give compound S1. Namely, the ratio of the amounts of phosphorus oxychloride, N-dimethylformamide and julolidine was 7.62:22.87:6.93mmol.
Step two: synthesis of Compound S2
Compound S1 (10 mmol), 4-bromoacetophenone (10 mmol) and aqueous sodium hydroxide (NaOH, 20%,10 mL) were slowly added to a 100mL round bottom flask containing ethanol (20 mL). The mixture was stirred at room temperature 25-30 ℃ for 24 hours, then the reaction was quenched by pouring the mixed solution after the reaction into ice water (100 mL), and after stirring for 2 hours, the precipitate was filtered to give a red solid. Subsequently, the red solid product was dried in a vacuum oven overnight to give compound S2. Namely, the ratio of the amounts of the substances of the compounds S1, 4-bromoacetophenone is 1:1
Step three: synthesis of Compound S3
Compound S2 (5 mmol), nitromethane (1.00 mL) and N, N-diisopropylethylamine (DIPEA, 5.5 mmol) were slowly added to a solution containing methanol (CH) 3 OH,20 mL). After stirring at 60-70℃for 24h, the reaction solution was poured into a beaker containing a saturated aqueous sodium chloride solution (10 mL) to stop the reaction, followed by extraction with ethyl acetate (30 mL). The resulting organic solution was dried over anhydrous sodium sulfate, filtered to give a crude product, which was purified by column chromatography (silica gel, ethyl acetate/petroleum ether=1/10) to give a yellow solid. Namely, the ratio of the amounts of the substances of the compound S2 to N, N-diisopropylethylamine is 5:5.5.
step four: synthesis of Compound S4
Compound S3 (1.0 mmol) and ammonium acetate (NH) 4 OAc,15.0 mmol) was added to a 100mL round bottom flask containing n-butanol (n-BuOH, 20 mL) and stirred at 100-110 ℃After 24h, it was cooled to room temperature, then concentrated to 5mL in vacuo and filtered off with suction. The solid obtained by separation was washed with ethanol (2X 5 mL) to finally obtain a blue-black solid. The ratio of the amount of compound S3 to the amount of ammonium acetate is 1:15.
step five: synthesis of Compound S5
At N 2 Under the protection of (1) compound S4 (0.11 mmol) and DIPEA (1.1 mmol) were added to a 100mL round bottom flask containing dried dichloromethane (DCM, 20 mL). Then BF is carried out 3 ·Et 2 O (2.0 mmol) was slowly added dropwise to the solution, stirred at room temperature in the dark for 24h, then diluted with ice water (20 mL) and extracted with DCM (3X 20 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (silica gel, dichloromethane/petroleum ether=1/1) to give a solid with metallic luster. Namely, the ratio of the amounts of the compounds S4, N-diisopropylethylamine and boron trifluoride etherate was 0.11:1.1:2.
step six: synthesis of Compound S6
At N 2 Under protection, the compound S5 (0.06 mmol), 4-pyridine phenylboronic acid (0.12 mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh) 3 ) 4 0.1 mmol), potassium carbonate (K) 2 CO 3 7.5 mmol) and 1,4 dioxane/water (10 mL, 5/1) were added separately to a 50mL round bottom flask. The mixture was stirred at 70-80 ℃ for 24 hours, then the reaction was cooled to room temperature, the solvent was concentrated to 5mL in vacuo, and filtered to give the crude product. The crude product was purified by column chromatography (silica gel, methanol/dichloromethane=1/50) to give a blue-black solid. Namely, the mass ratio of the compounds S5, 4-pyridine phenylboronic acid, tetrakis (triphenylphosphine) palladium and potassium carbonate was 0.06:0.12:0.1:7.5.
step seven: synthesis of Compound WS5
At N 2 Under protection, S6 (0.06 mmol) and 1-bromododecane (0.06 mmol) were added to a solution containing chloroform (CHCl) 3 10 mL) was stirred at 80-90 ℃ for 24h, the reaction cooled to room temperature and the solvent concentrated in vacuo. Finally, purification by column chromatography (silica gel, methanol/dichloromethane=1/25) afforded WS5 as a blue-green solid. Namely, the ratio of the amount of the compound S6 to the amount of the 1-bromododecane is 1:1。
step eight: synthesis of the Compound WS5@NPs
WS5 and DSPE-mPEG 5000 Dissolving in tetrahydrofuran, and ultrasonic mixing to obtain stock solution. And then the stock solution is injected into deionized water by a needle tube and dispersed under a cell disruption instrument. The solution is then treated with N 2 Agitation in a fume hood was protected to evaporate the tetrahydrofuran. Finally, the resulting nanoparticles (fixed volume to 1 mL) were collected by filtration and purification using a 0.45 μm ultrafiltration filter (Millipore). Finally, WS5@NPs of 1mg/mL are obtained.
Wherein, the introduction of the side chain structure enhances the electron pushing-pulling capability of the donor and the acceptor, and the absorption and emission main peak of WS5 molecules are more than 1000nm. In addition, WS5 molecules exhibit absorption/wavelength red shift with increasing polarity of the solvent in solvents of different polarity, indicating the presence of significant intramolecular charge transfer states in the molecule. Meanwhile, the ultraviolet absorption spectrum at different concentrations shows that the molar absorptivity of WS5 molecules is higher than 10000M -1 cm -1 I.e. WS5 molecules exhibit a higher light absorption capacity. Furthermore, WS5 was prepared using amphiphilic polymer (DSPE-mPEG) 5000 ) Coated into nano particles (WS5@NPs).
The absorption spectrum of ws5@nps was significantly changed compared to WS5 molecules. The main causes of these changes are usually coulomb coupling effects and intermolecular interactions in the nanoparticles. From the photophysical test results, the absorption spectrum at ws5@nps was significantly broadened compared to that of WS5, indicating that multiple aggregation states coexist. Compared with the absorption spectrum of WS5, the absorption spectrum of WS5@NPs has slight blue shift, which indicates that coulomb coupling effect exists in molecules in the coated nano particles.
In summary, the introduction of the side chains lengthens the intermolecular distance under the nanoparticle, which is manifested by a decrease in intermolecular pi-pi interactions and an increase in fluorescence. And the prolonged intermolecular distance provides a more loose movement space for molecules, and photo-thermal enhancement. Thus, a NIR-II photothermal agent of high brightness and high photothermal performance is obtained.
Example 1
As shown in fig. 6, 7 and 8, compound S1 was prepared: on iceUnder salt bath condition, phosphorus oxychloride (POCl) 3 1.17g,7.62 mmol) was slowly added dropwise to a 100mL round bottom flask containing dry N, N-dimethylformamide (DMF, 1.67g,22.87 mmol). At N 2 After stirring for 2h under protection of (2) the Vilsmeier-Haack reagent was obtained. Julolidine (1.20 g,6.93 mmol) and dry DMF (10 mL) were then slowly added to the flask containing Vilsmeier-Haack reagent. Thereafter, the mixture was stirred at 30℃for 4 hours. After cooling to room temperature, the mixture was poured into ice water (100 mL) to quench the reaction. The precipitate was then filtered to give a pale yellow solid. Finally, the solid was dried in a vacuum oven overnight to give compound S1 (yield 78%). 1 H NMR(500MHz,CDCl 3 )δ/ppm 9.60(s,1H),7.29(s,2H),3.29(t,J＝5.0,4H),2.77(t,J＝10.0,4H),1.97(m,4H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 190.1,147.9,129.5,124.1,120.4,50.0,27.6,21.3.
Preparation of compound S2: compound S1 (2.00 g,10 mmol), 4-bromoacetophenone (2.00 g,10 mmol) and aqueous sodium hydroxide (NaOH, 20%,10 mL) were slowly added to a 100mL round bottom flask containing ethanol (20 mL). The mixture was stirred at room temperature for 24 hours, then the reaction was quenched by pouring the reacted mixture solution into ice water (100 mL), and after stirring for 2 hours, the precipitate was filtered to give a red solid. Subsequently, the red solid product was dried in a vacuum oven overnight to give compound S2 (yield 80%). 1 H NMR(500MHz,CDCl 3 )δ/ppm 7.85(d,J＝10.0Hz,2H),7.69(d,J＝20.0Hz,1H),7.59(d,J＝10.0Hz,2H),7.18(d,J＝20.0Hz,1H),7.11(s,2H),3.26(t,J＝5.0Hz,4H),2.76(t,J＝5.0Hz,4H),1.97(m,4H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 189.3,147.0,145.5,138.1,131.6,129.9,128.4,126.8,121.3,121.0,114.9,108.1,105.9,50.0,27.7,21.5.IT-TOF/MS:[M+H] + calcd:382.0728,found:382.0804.
Preparation of compound S3: compound S2 (1.00 g,5 mmol), nitromethane (1.00 mL) and N, N-diisopropylethylamine (DIPEA, 1.0mL,5.5 mmol) were slowly added to a solution containing methanol (CH) 3 OH,20 mL). After stirring at 70℃for 24 hours, the reaction solution was poured into a beaker containing a saturated aqueous sodium chloride solution (10 mL) to stop the reaction, followed by ethyl acetate(30 mL) extraction was performed. The resulting organic solution was dried over anhydrous sodium sulfate, filtered to give a crude product, which was purified by column chromatography (silica gel, ethyl acetate/petroleum ether=1/10) to give a yellow solid (yield 67%). 1 H NMR(500MHz,CDCl 3 )δ/ppm 7.76(d,J＝15.0Hz,2H),7.59(d,J＝10.0Hz,2H),6.63(s,2H),4,72(t,J＝5.0Hz,1H),4.60(t,J＝5.0Hz,1H),3.98(t,J＝10.0Hz,1H).3.38(m,2H).3.10(t,J＝5.0Hz,4H),2.70(t,J＝5.0Hz,4H),1.97(m,4H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 196.5,142.5,135.3,132.0,129.6,128.6,125.8,125.4,121.8,80.0,49.9,41.9,38.6,27.7,21.9,14.8.IT-TOF/MS:[M+H] + calcd:443.0892,found:443.0965.
Preparation of compound S4: compound S3 (1.00 g,1.0 mmol) and ammonium acetate (NH) 4 OAc,1.16g,15.0 mmol) was added to a 100mL round bottom flask containing n-butanol (n-BuOH, 20 mL), stirred at 110℃for 24h, cooled to room temperature, then concentrated to 5mL in vacuo and filtered off with suction. The solid obtained was separated and washed with ethanol (2×5 mL) to finally give a blue-black solid (yield 37%). 1 H NMR(500MHz,CDCl 3 )δ/ppm 7.77(d,J＝15.0Hz,4H),7.63(d,J＝10.0Hz,4H),7.56(s,4H),6.94(s,2H),5.30(s,1H),3.22(t,J＝5.0Hz,8H),2.76(t,J＝10.0Hz,8H),2.01(m,8H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 156.1,150.5,143.9,138.8,131.8,128.7,125.5,124.5,123.3,121.7,115.8,49.9,27.9,21.8,1.0.
Preparation of compound S5: at N 2 Under the protection of (1), compound S4 (80 mg,0.11 mmol) and DIPEA (0.2 mL,1.1 mmol) were added to a 100mL round bottom flask containing dried dichloromethane (DCM, 20 mL). Then BF is carried out 3 ·Et 2 O (0.28 mL,2.0 mmol) was slowly added dropwise to the solution, stirred at room temperature in the dark for 24h, then diluted with ice water (20 mL) and extracted with DCM (3X 20 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (silica gel, dichloromethane/petroleum ether=1/1) to give a solid with metallic luster (yield 84%). 1 H NMR(500MHz,CDCl 3 )δ/ppm 7.86(d,J＝10.0Hz,4H),7.60(s,4H),7.56(d,J＝10.0Hz,4H),6.69(s,2H),3.30(t,J＝5.0Hz,8H),2.77(t,J＝5.0Hz,8H),2.00(m,8H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 156.1,150.5,143.9,138.8,131.8,128.7,125.5,124.5,123.3,121.7,115.8,49.9,30.9,29.7,27.9,24.4,21.8,11.2,1.4.MALDI-TOF/MS:[M] + calcd:845.1530,found:845.1960.
Preparation of compound S6: at N 2 Under protection, compound S5 (50 mg,0.06 mmol), 4-pyridine phenylboronic acid (54 mg,0.12 mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh) 3 ) 4 115mg,0.1 mmol), potassium carbonate (K) 2 CO 3 1.03g,7.5 mmol) and 1,4 dioxane/water (10 mL, 5/1) were added separately to a 50mL round bottom flask. The mixture was stirred at 80 ℃ for 24 hours, then the reaction was cooled to room temperature, the solvent was concentrated to 5mL in vacuo, and filtered to give the crude product. The crude product was purified by column chromatography (silica gel, methanol/dichloromethane=1/50) to give a blue-black solid in 61% yield. 1H NMR (500 MHz, CDCl) 3 )δ/ppm 8.67(d,J＝5.0Hz,4H),8.13(d,J＝5.0Hz,4H),7.70(s,4H),7.64(d,J＝5.0Hz,4H),7.55(d,J＝5.0Hz,4H),6.78(s,2H)3.31(t,J＝5.0Hz,8H),2.78(t,J＝5.0Hz,8H),2.01(m,8H). 13 C NMR(126MHz,CDCl 3 )δ/ppm153.1,150.2,148.4,148.2,147.6,143.5,141.0,138.1,135.9,135.0,134.9,131.6,130.3,128.0,127.5,126.9,121.8,121.5,121.3,121.1,111.4,51.4,31.9,30.2,29.7,27.9,22.7,22.1,13.4.MALDI-TOF/MS:[M] + calcd:841.3876,found:841.7387.
Preparation of compound WS5: at N 2 Under protection, S6 (50 mg,0.06 mmol) and 1-bromododecane (9.9 mg,0.06 mmol) were added to a solution containing chloroform (CHCl) 3 10 mL) was stirred at 90 ℃ for 24h, the reaction cooled to room temperature and the solvent concentrated in vacuo. Finally, purification by column chromatography (silica gel, methanol/dichloromethane=1/25) gave WS5 as a blue-green solid (36% yield). WS5: 1 H NMR(500MHz,d 6 -DMSO),δ/ppm 9.13(d,J＝10.0,2H),8.68(m,4H),8.27(m,6H),7.96(d,J＝10.0,2H),7.83(s,2H),7.70(d,J＝25.0,4H),7.23(d,J＝15.0,2H),4.58(t,J＝10.0,2H),3.51(s,8H),2.71(s,8H),1.93(s,10H),1.27(d,J＝30.0,18H),0.84(t,J＝5.0,3H). 13 C NMR(126MHz,CDCl 3 )δ/ppm 145.0,130.6,130.2,129.3,127.9,127.1,124.8,121.4,120.7,114.6,77.2,70.7,50.4,32.0,31.6,29.8,29.7,29.6,29.5,29.2,28.2,26.3,22.8,21.8,14.3.MALDI-TOF/MS:[M-2Br] + calcd:1011.149,found:1011.055.
preparation of ws5@nps: WS5 (1 mg) and DSPE-mPEG 5000 (5 mg) was dissolved in tetrahydrofuran (1 mL), and the mixture was sonicated to prepare a stock solution. The stock solution was then injected into deionized water (9 mL) through a syringe and dispersed under a cell disrupter. The solution is then treated with N 2 Agitation in a fume hood was protected to evaporate the tetrahydrofuran. Finally, the resulting nanoparticles (fixed volume to 1 mL) were collected by filtration and purification using a 0.45 μm ultrafiltration filter (Millipore). Finally, WS5@NPs of 1mg/mL are obtained.
Example 2
Step one: synthesis of Compound S1
Under ice salt bath condition, phosphorus oxychloride (POCl) 3 1.17g,7.62 mmol) was slowly added dropwise to a 100mL round bottom flask containing dry N, N-dimethylformamide (DMF, 1.67g,22.87 mmol). At N 2 After stirring for 2h under protection of (2) the Vilsmeier-Haack reagent was obtained. Julolidine (1.20 g,6.93 mmol) and dry DMF (10 mL) were then slowly added to the flask containing Vilsmeier-Haack reagent. Thereafter, the mixture was stirred at 30℃for 3 hours. After cooling to room temperature, the mixture was poured into ice water (100 mL) to quench the reaction. The precipitate was then filtered to give a pale yellow solid. Finally, the solid was dried in a vacuum oven overnight to give compound S1.
Step two: synthesis of Compound S2
Compound S1 (2.00 g,10 mmol), 4-bromoacetophenone (2.00 g,10 mmol) and aqueous sodium hydroxide (NaOH, 20%,10 mL) were slowly added to a 100mL round bottom flask containing ethanol (20 mL). The mixture was stirred at room temperature 25 ℃ for 24 hours, then the reaction was quenched by pouring the reacted mixed solution into ice water (100 mL), and after stirring for 2 hours, the precipitate was filtered to give a red solid. Subsequently, the red solid product was dried in a vacuum oven overnight to give compound S2.
Step three: synthesis of Compound S3
Compound S2 (1.00 g,5 mmol), nitromethane (1.00 mL) and N, N-diisopropylethylamine (DIPEA, 1.0mL,5.5 mmol) were slowly added to a solution containing methanol (CH) 3 OH,20 mL)In a round bottom flask. After stirring at 60℃for 24 hours, the reaction solution was poured into a beaker containing a saturated aqueous sodium chloride solution (10 mL) to stop the reaction, followed by extraction with ethyl acetate (30 mL). The resulting organic solution was dried over anhydrous sodium sulfate, filtered to give a crude product, which was purified by column chromatography (silica gel, ethyl acetate/petroleum ether=1/10) to give a yellow solid.
Step four: synthesis of Compound S4
Compound S3 (1.00 g,1.0 mmol) and ammonium acetate (NH) 4 OAc,1.16g,15.0 mmol) was added to a 100mL round bottom flask containing n-butanol (n-BuOH, 20 mL), stirred at 100deg.C for 24h, cooled to room temperature, then concentrated to 5mL under vacuum and filtered with suction. The solid obtained by separation was washed with ethanol (2X 5 mL) to finally obtain a blue-black solid.
Step five: synthesis of Compound S5
At N 2 Under the protection of (1), compound S4 (80 mg,0.11 mmol) and DIPEA (0.2 mL,1.1 mmol) were added to a 100mL round bottom flask containing dried dichloromethane (DCM, 20 mL). Then BF is carried out 3 ·Et 2 O (0.28 mL,2.0 mmol) was slowly added dropwise to the solution, stirred at room temperature in the dark for 24h, then diluted with ice water (20 mL) and extracted with DCM (3X 20 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (silica gel, dichloromethane/petroleum ether=1/1) to give a solid with metallic luster.
Step six: synthesis of Compound S6
At N 2 Under protection, compound S5 (50 mg,0.06 mmol), 4-pyridine phenylboronic acid (54 mg,0.12 mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh) 3 ) 4 115mg,0.1 mmol), potassium carbonate (K) 2 CO 3 1.03g,7.5 mmol) and 1,4 dioxane/water (10 mL, 5/1) were added separately to a 50mL round bottom flask. The mixture was stirred at 70 ℃ for 24 hours, then the reaction was cooled to room temperature, the solvent was concentrated to 5mL in vacuo, and filtered to give the crude product. The crude product was purified by column chromatography (silica gel, methanol/dichloromethane=1/50) to give a blue-black solid.
Step seven: synthesis of Compound WS5
At N 2 Under protection, S6 (50 mg,0.06 mmol) and 1-bromododecane (9.9 mg,0.06 mmol) were added to a solution containing chloroform (CHCl) 3 10 mL) was stirred at 80 ℃ for 24h, the reaction cooled to room temperature and the solvent concentrated in vacuo. Finally, purification by column chromatography (silica gel, methanol/dichloromethane=1/25) afforded WS5 as a blue-green solid.
Step eight: synthesis of the Compound WS5@NPs
WS5 (1 mg) and DSPE-mPEG 5000 (5 mg) was dissolved in tetrahydrofuran (1 mL), and the mixture was sonicated to prepare a stock solution. The stock solution was then injected into deionized water (9 mL) through a syringe and dispersed under a cell disrupter. The solution is then treated with N 2 Agitation in a fume hood was protected to evaporate the tetrahydrofuran. Finally, the resulting nanoparticles (fixed volume to 1 mL) were collected by filtration and purification using a 0.45 μm ultrafiltration filter (Millipore). Finally, WS5@NPs of 1mg/mL are obtained.
Example 3
Step one: synthesis of Compound S1
Under ice salt bath condition, phosphorus oxychloride (POCl) 3 1.17g,7.62 mmol) was slowly added dropwise to a 100mL round bottom flask containing dry N, N-dimethylformamide (DMF, 1.67g,22.87 mmol). At N 2 After stirring for 2h under protection of (2) the Vilsmeier-Haack reagent was obtained. Julolidine (1.20 g,6.93 mmol) and dry DMF (10 mL) were then slowly added to the flask containing Vilsmeier-Haack reagent. Thereafter, the mixture was stirred at 25℃for 4 hours. After cooling to room temperature, the mixture was poured into ice water (100 mL) to quench the reaction. The precipitate was then filtered to give a pale yellow solid. Finally, the solid was dried in a vacuum oven overnight to give compound S1.
Step two: synthesis of Compound S2
Compound S1 (2.00 g,10 mmol), 4-bromoacetophenone (2.00 g,10 mmol) and aqueous sodium hydroxide (NaOH, 20%,10 mL) were slowly added to a 100mL round bottom flask containing ethanol (20 mL). The mixture was stirred at room temperature for 24 hours, then the reaction was quenched by pouring the reacted mixture solution into ice water (100 mL), and after stirring for 2 hours, the precipitate was filtered to give a red solid. Subsequently, the red solid product was dried in a vacuum oven overnight to give compound S2.
Step three: synthesis of Compound S3
Compound S2 (1.00 g,5 mmol), nitromethane (1.00 mL) and N, N-diisopropylethylamine (DIPEA, 1.0mL,5.5 mmol) were slowly added to a solution containing methanol (CH) 3 OH,20 mL). After stirring at 65℃for 24 hours, the reaction solution was poured into a beaker containing a saturated aqueous sodium chloride solution (10 mL) to stop the reaction, followed by extraction with ethyl acetate (30 mL). The resulting organic solution was dried over anhydrous sodium sulfate, filtered to give a crude product, which was purified by column chromatography (silica gel, ethyl acetate/petroleum ether=1/10) to give a yellow solid.
Step four: synthesis of Compound S4
Compound S3 (1.00 g,1.0 mmol) and ammonium acetate (NH) 4 OAc,1.16g,15.0 mmol) was added to a 100mL round bottom flask containing n-butanol (n-BuOH, 20 mL), stirred at 105℃for 24h, cooled to room temperature, concentrated in vacuo to 5mL and filtered with suction. The solid obtained by separation was washed with ethanol (2X 5 mL) to finally obtain a blue-black solid.
Step five: synthesis of Compound S5
At N 2 Under the protection of (1), compound S4 (80 mg,0.11 mmol) and DIPEA (0.2 mL,1.1 mmol) were added to a 100mL round bottom flask containing dried dichloromethane (DCM, 20 mL). Then BF is carried out 3 ·Et 2 O (0.28 mL,2.0 mmol) was slowly added dropwise to the solution, stirred at room temperature in the dark for 24h, then diluted with ice water (20 mL) and extracted with DCM (3X 20 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (silica gel, dichloromethane/petroleum ether=1/1) to give a solid with metallic luster.
Step six: synthesis of Compound S6
At N 2 Under protection, compound S5 (50 mg,0.06 mmol), 4-pyridine phenylboronic acid (54 mg,0.12 mmol), tetrakis (triphenylphosphine) palladium (Pd (PPh) 3 ) 4 115mg,0.1 mmol), potassium carbonate (K) 2 CO 3 1.03g,7.5 mmol) and 1,4 dioxane/water (10 mL, 5/1) were added separately to a 50mL round bottom flask. The mixture was stirred at 75℃for 24 hours, then reversedThe reaction mixture was cooled to room temperature, the solvent was concentrated to 5mL in vacuo, and the crude product was obtained by filtration. The crude product was purified by column chromatography (silica gel, methanol/dichloromethane=1/50) to give a blue-black solid.
Step seven: synthesis of Compound WS5
At N 2 Under protection, S6 (50 mg,0.06 mmol) and 1-bromododecane (9.9 mg,0.06 mmol) were added to a solution containing chloroform (CHCl) 3 10 mL) was stirred at 85 ℃ for 24h, the reaction cooled to room temperature and the solvent concentrated in vacuo. Finally, purification by column chromatography (silica gel, methanol/dichloromethane=1/25) afforded WS5 as a blue-green solid.
Step eight: synthesis of the Compound WS5@NPs
WS5 (1 mg) and DSPE-mPEG 5000 (5 mg) was dissolved in tetrahydrofuran (1 mL), and the mixture was sonicated to prepare a stock solution. The stock solution was then injected into deionized water (9 mL) through a syringe and dispersed under a cell disrupter. The solution is then treated with N 2 Agitation in a fume hood was protected to evaporate the tetrahydrofuran. Finally, the resulting nanoparticles (fixed volume to 1 mL) were collected by filtration and purification using a 0.45 μm ultrafiltration filter (Millipore). Finally, WS5@NPs of 1mg/mL are obtained.
FIG. 1 is a schematic diagram showing the measurement of the photophysical properties of WS5 molecule. Test of the photo-thermal Properties of WS5 molecule of (d) figures (a), (b), (c) as in 1 (0.2 mg mL) -1 ，0.23W cm -2 1064 nm). The photophysical properties of WS5 molecules indicate higher molar absorptivity, fluorescence brightness and photothermal properties. The electron withdrawing capability of the acceptor is enhanced due to the introduction of the side chain structure, so that the push-pull capability of electrons in molecules and the delocalization capability of electrons are enhanced, the absorption and emission main peak of WS5 molecules exceeds 1000nm, and the introduction of the flexible side chain is found to enhance the molecular motion and promote the non-radiative decay process.
As shown in fig. 2, the photophysical properties of the nanoparticles are greatly changed, especially fluorescence and photothermal properties, compared to the photophysical properties of the organic molecules. From the photophysical test results, the absorption spectrum of the nanoparticle is obviously widened compared with that of the organic molecule, which indicates that a plurality of aggregation states coexist. Compared with the absorption spectrum of organic molecules, the absorption spectrum of the nano-particles has slight blue shift, which indicates that the molecules in the coated nano-particles have coulomb coupling effect. Photophysical test results show that the introduction of a suitable side chain structure, H-aggregation and prolonged intermolecular distance can promote intramolecular movement to enhance photothermal heating, and pi-pi accumulation also decreases with prolonged intermolecular distance, thereby promoting fluorescence in an aggregated state. Thus, a NIR-II photothermal agent of high brightness and high photothermal performance is obtained.
As shown in fig. 3, in vitro cell experiments demonstrated that ws5@nps have a photo-induced thermo-effect tumor killing mechanism. The WS5@NPs with different concentrations are respectively selected for cell viability experiments, and after laser irradiation of 1064nm for 5 minutes (0.23W cm) -2 ) Tumor cells die rapidly. Especially at concentrations of 50. Mu.g/mL and 100. Mu.g/mL, rapid cell death can be seen. However, in the absence of laser irradiation, the activity of the tumor cells was hardly reduced, indicating that the material has excellent safety and a photothermal effect of inducing death of tumor cells. Meanwhile, WS5@NPs can effectively induce 143B cells to undergo apoptosis under the excitation of 1064 nm. In the laser group administered, the proportion of necrotic cells was only 5.85%, whereas the total of early and late apoptosis was as high as 76.02%. These results indicate that ws5@nps induce apoptosis in cells under laser irradiation, thus achieving autonomous programmed death.
As shown in FIG. 4, in vivo fluorescence imaging of NIR-IIa region shows that WS5@NPs can rapidly light the blood vessels and lymphatic system of mice within 5 minutes, and high imaging resolution is shown.
As shown in fig. 5, tumor-bearing mice were subjected to photothermal treatment of deep tumors 24 hours after intravenous administration under the guidance of NIR-II imaging, and the efficacy thereof was evaluated. The temperature of the administration group was significantly increased under irradiation of laser light compared with the control group, and the temperature was increased to 44℃for 3 minutes, with the highest temperature being 46 ℃. No significant recurrence of the tumor was seen in mice after cessation of treatment. In the whole treatment process, the skin at the irradiated part is not obviously burnt, and the material has the outstanding advantage of high-efficiency photo-thermal performance in deep tumor treatment.
The application not only discloses deep tumor treatment of a photothermal agent with high brightness and high photo-thermal performance under the guidance of NIR-II imaging, but also discloses a double-improvement mechanism of a side chain for improving molecular brightness and photo-thermal performance, and the side chain engineering can be used as an effective method for simultaneously regulating and controlling radiation transition (QY) and non-radiation transition (PCE). Specifically, the introduction of side chains promotes intramolecular movement in the aggregated state by further distorting the molecular conformation, extending the intermolecular distance, and providing a more loosely packed environment for intramolecular movement (too large side chains can cause greater steric hindrance in the aggregated state), thereby promoting non-radiative transitions to produce photothermal properties. Furthermore, while intramolecular movement is promoted with the aid of side chains, pi-pi stacking also decreases with prolonged intermolecular distance, thereby promoting fluorescence in an aggregated state. Therefore, the introduction of the alkyl chain enhances fluorescence and photothermal, can be used for photothermal treatment under the guidance of fluorescence, and provides a new idea for solving the problem of dependence on two competitive photophysical processes.
The above is only for illustrating the technical idea of the present application, and the protection scope of the present application is not limited by this, and any modification made on the basis of the technical scheme according to the technical idea of the present application falls within the protection scope of the claims of the present application.
Claims (10)
1. The Aza-BODIPY-based high-brightness NIR-II region photothermal agent is characterized by comprising the following structural formula:
2. a process for the preparation of an Aza-BODIPY-based high intensity NIR-II region photothermal agent according to claim 1, comprising the steps of:
step one: adding N, N-dimethylformamide into phosphorus oxychloride, and stirring under a protective gas to obtain a Vilsmeier-Haack reagent; mixing julolidine and N, N-dimethylformamide, adding the mixture into a Vilsmeier-Haack reagent, heating to perform a first reaction, cooling to room temperature after the first reaction is finished, pouring the mixture into ice water to quench the reaction, filtering and drying to obtain an intermediate product S1;
step two: uniformly mixing the intermediate product S1, 4-bromoacetophenone with sodium hydroxide aqueous solution and ethanol for a second reaction, pouring the mixed solution of the second reaction product into ice water for quenching reaction after the second reaction is finished, and then filtering and drying to obtain an intermediate product S2;
step three: uniformly mixing an intermediate product S2, nitromethane, N-diisopropylethylamine and methanol, then heating to perform a third reaction, pouring a solution of the third reaction product into a saturated sodium chloride aqueous solution to stop the reaction, and then extracting, drying, filtering and purifying to obtain an intermediate product S3;
step four: mixing the intermediate product S3, ammonium acetate and n-butanol uniformly, heating to perform a fourth reaction, then concentrating in vacuum, filtering, separating solids, and washing to obtain an intermediate product S4;
step five: mixing intermediate products S4, N-diisopropylethylamine with dried dichloromethane, adding boron trifluoride diethyl etherate, stirring at room temperature in a dark place for a fifth reaction, diluting the fifth reaction product with ice water, extracting, drying, filtering, concentrating and purifying to obtain an intermediate product S5;
step six: under the protection gas, heating a solution of an intermediate product S5, 4-pyridine phenylboronic acid, tetra (triphenylphosphine) palladium, potassium carbonate and 1,4 dioxane mixed with water for a sixth reaction, cooling to room temperature after the sixth reaction is finished, concentrating, filtering and purifying to obtain an intermediate product S6;
step seven: under the protection gas, mixing the intermediate product S6 with 1-bromododecane and chloroform, heating to perform a seventh reaction, cooling to room temperature after the seventh reaction is finished, concentrating, filtering and purifying to obtain WS5.
3. The method according to claim 2, wherein in the first step, the ratio of the amounts of phosphorus oxychloride, N-dimethylformamide and julolidine is 7.62:22.87:6.93mmol; the temperature of the first reaction is 20-30 ℃; the time of the first reaction is 3-4 h.
4. The preparation method according to claim 2, wherein in the second step, the ratio of the amounts of substances of the intermediate products S1, 4-bromoacetophenone is 1:1, a step of; the temperature of the second reaction is 25-30 ℃.
5. The method according to claim 2, wherein in the third step, the ratio of the amounts of the substances of the intermediate product S2 to N, N-diisopropylethylamine is 5:5.5; the temperature of the third reaction is 60-70 ℃.
6. The method according to claim 2, wherein in the fourth step, the ratio of the amounts of the intermediate product S3 to the ammonium acetate is 1:15; the temperature of the fourth reaction is 100-110 ℃.
7. The method according to claim 2, wherein in the fifth step, the ratio of the amounts of the intermediate products S4, N-diisopropylethylamine and boron trifluoride etherate is 0.11:1.1:2.
8. the method according to claim 2, wherein in the sixth step, the ratio of the amounts of the substances of the intermediate products S5, 4-pyridine phenylboronic acid, tetrakis (triphenylphosphine) palladium and potassium carbonate is 0.06:0.12:0.1:7.5; the temperature of the sixth reaction is 70-80 ℃; the volume ratio of the 1,4 dioxane to the water is 5:1.
9. the method according to claim 2, wherein in the seventh step, the ratio of the amounts of the intermediate S6 and the 1-bromododecane is 1:1, a step of; the temperature of the seventh reaction is 80-90 ℃.
10. Use of a high intensity NIR-II region photothermal agent according to claim 1 in the preparation of a photothermal therapeutic agent.
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