CN115385851A

CN115385851A - Near-infrared aggregation-induced emission type ultra-efficient photosensitizer with asymmetric diacetonitrile structure, and preparation method and application thereof

Info

Publication number: CN115385851A
Application number: CN202210862936.XA
Authority: CN
Inventors: 张莎莎; 梅菊; 田禾; 杨文芳; 张心仪; 刘涛
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2022-07-21
Filing date: 2022-07-21
Publication date: 2022-11-25

Abstract

The invention relates to the technical field of biological medical treatment, and discloses a near-infrared aggregation-induced emission type ultra-high efficiency photosensitizer with an asymmetric diacetonitrile-based structure, and a preparation method and application thereof. The photosensitizer has the following general structure:

the preparation method is that the p-aryl diacetonitrile and the aryl methylAnd carrying out Knoevenagel condensation reaction on aldehyde, carrying out one-step Knoevenagel condensation reaction on the generated intermediate product and N-substituted aryl formaldehyde, and finally carrying out salt forming reaction to obtain the photosensitizer. The photosensitizer has typical Aggregation Induced Emission (AIE) performance, the maximum emission wavelength is positioned in a near infrared region (more than 650 nm), and the photosensitizer can be used for near infrared fluorescence imaging. The photosensitizer specifically targets mitochondria in cells, and can efficiently generate singlet oxygen under illumination (A and B) ¹ O ₂ ) Which is ¹ O ₂ The yield is more than 6 times of that of a classical photosensitizer rose bengal B, and the photodynamic therapy effect is good. Has potential clinical application value in the photodynamic disease treatment guided by near infrared fluorescence imaging.

Description

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer with asymmetric diacetonitrile structure, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biological medicines, relates to a photosensitizer, and particularly relates to a near-infrared aggregation-induced emission type ultra-high-efficiency photosensitizer with an asymmetric diacetonitrile structure, a preparation method and application thereof in imaging-guided photodynamic therapy.

Background

Cancer has become a major health problem threatening social progress and human development. The rapid proliferation and spread of malignant cells makes cancer treatment exceptionally difficult. Conventional cancer treatment modalities, such as surgical resection, radiation therapy, and chemotherapy, have significant limitations, such as invasiveness or adverse side effects. Photodynamic therapy (PDT) as a non-invasive, spatio-temporally controllable and non-invasive treatment is becoming an effective strategy for achieving personalized and accurate treatment of cancer. The principle is that Photosensitizers (PSs) generate Reactive Oxygen Species (ROS) under specific light irradiation, wherein singlet oxygen (R) ((R)) ¹ O ₂ ) It can destroy genetic material in tumor cells, leading to apoptosis or necrosis. It follows that the properties of the photosensitizer play a key role in achieving efficient PDT.

Among the photosensitizers, small-molecule organic fluorophores with photosensitizing properties can be used for tumor imaging, tracking and visualization of photosensitizers inside tumors, and are distinguished by better biocompatibility, degradability, easily adjustable structure and optical properties. While the traditional photosensitizer has a planar rigid pi-conjugated structureAnd poor water solubility can lead to aggregate fluorescence quenching effect (ACQ), thereby reducing fluorescence emission intensity and singlet oxygen: ( ¹ O ₂ ) The efficiency of production limits its wide use in biomedical systems. Exciting, the focus-induced emission (AIE) concept was first proposed by the team of down loyal academies in 2001, successfully overcoming the drawbacks of ACQ fluorophores. More importantly, the aggregation-induced emission (AIEgens) is not only beneficial to fluorescence imaging, but also in an aggregation state, a distorted three-dimensional structure can block a non-radiative relaxation channel, so that the triplet quantum yield and stability can be increased, and the generation efficiency of ROS can be further improved.

To date, a number of aggregation-induced emission photosensitizers (AIE PSs) have been successfully developed and have made favorable progress, but the AIE PSs reported so far emit wavelengths mostly in the visible range, which is very disadvantageous for achieving high signal-to-noise ratio and deep tissue penetration. In the diagnostic technology, the near infrared fluorescence imaging has the obvious advantages of negligible biological substrate autofluorescence interference, strong tissue penetration capability, intuition and high contrast. However, the preparation of near-infrared AIE PSs generally involves multi-step reactions, harsh synthetic conditions and time-consuming purification or isolation. In addition, the proliferation of cancer cells is closely related to apoptosis and mitochondrial function. In recent years, many researchers have focused their research on improving the efficacy of photosensitizer therapy by virtue of mitochondrial targeting functions. The research result shows that the mitochondria are promoted ¹ O ₂ The generation of the active oxygen can effectively solve the problems of short diffusion distance and short service life of the active oxygen, thereby obviously improving the treatment effect. Although there are also a number of reports of mitochondrially targeted AIE systems with long wavelength emission, their clinical use is limited by the yield of reactive oxygen species (photosensitizing properties). Therefore, in order to further bring high-performance AIE PSs to clinical application, the development of a novel near-infrared aggregation-induced emission type ultra-high-efficiency photosensitizer with mitochondrial targeting performance is of great significance.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention synthesizes a near-infrared aggregation-induced emission type ultra-high-efficiency photosensitizer with an asymmetric diacetonitrile structure, provides a preparation method of the photosensitizer and realizes near-infrared fluorescence imaging-guided photodynamic therapy.

The invention provides a near-infrared aggregation-induced emission type ultra-efficient photosensitizer with an asymmetric diacetonitrile structure, which is an electron donor (D) -pi-electron acceptor (A) type compound designed by taking the asymmetric diacetonitrile structure as a core and has the following general formula structure:

x and Y comprise C, N, O or S;

m and n are natural numbers of 0-3 and are not 0 at the same time;

Ar ₁ selected from any of the following structures:

wherein p is a natural number of 1 to 5, R ₂ And R ₃ Each independently selected from one of H, hydroxy, amine, optionally substituted alkyl, alkoxy, alkylthio, alkenyl, alkynyl, cycloalkyl, cycloalkyloxy, cycloalkylthio, amido, aryl, heterocyclyl, heteroaryl, heterocycloalkyl, monoalkylamine, or dialkylamino;

Ar ₂ selected from any of the following structures:

z is independently selected from one of halide, acetate, trifluoroacetate, dihydrogen phosphate, hydrogen sulfate, tetrafluoroborate, hexafluorophosphate and sulfonate;

R ₁ is linear alkyl, cycloalkyl, carboxyl substituted alkyl, aldehyde substituted alkyl, sulfonic substituted alkyl, azide substituted alkyl, quaternary ammonium substituted alkyl, preferablyCH ₃ 、CH ₂ CH ₃ 、CH ₂ COOH、CH ₂ CHO、CH ₂ CH ₃ SO ₃ Or CH ₂ CH ₃ N ₃ ；R ₂ And R ₃ Each independently selected from H and NH ₃ 、OH、OCH ₃ 、OCH ₂ CH ₃ Or CONH ₂ One kind of (1).

In a second aspect of the present invention, there is provided a process for preparing the above photosensitizer, the process is as follows:

the preparation method comprises the following steps:

s1, performing Knoevenagel condensation reaction on p-aryldiacetonitrile (II) and aryl formaldehyde (III) to obtain an intermediate product IV;

s2, carrying out one-step Knoevenagel condensation reaction on the intermediate product (IV) and N-substituted aryl formaldehyde (V) to obtain an asymmetric diacetonitrile-based intermediate product (VI);

and S3, carrying out substitution or salt formation reaction on the intermediate product (VI) and halogenated alkane, sultone, sulfonic group substituted alkane or other various substituted alkanes to obtain a target product in the structural formula shown in the I.

Preferred conditions for each preparation step are as follows:

the Knoevenagel condensation reaction in the step S1 comprises the following specific processes: under the protection of inert gas, adding p-aryldiacetonitrile (II) and arylformaldehyde (III) into an organic solvent, adding a small amount of alkaline substance as a catalyst, and reacting at room temperature for 6-12 h.

Preferably, the mass ratio of II to III is 1; the molar weight of the alkaline substance is 0.1-0.5 equivalent, and the alkaline substance is any one or a mixture of more of pyridine, piperidine, potassium carbonate, sodium carbonate, potassium hydroxide, sodium alkoxide and potassium alkoxide; the inert gas is nitrogen; the organic solvent is selected from one or more of ethanol, isopropanol, tetrahydrofuran and acetonitrile.

The Knoevenagel condensation reaction in the step S2 comprises the following specific processes: under the protection of inert gas, adding the intermediate product (IV) and N-substituted aryl formaldehyde (V) into an organic solvent, adding a small amount of alkaline substance as a catalyst, heating to the reflux reaction temperature, and reacting for 6-12 h.

Preferably, the molar ratio of the intermediate product (IV) to the aryl formaldehyde (V) is 1 to 2, the proportion and the amount of the basic catalyst, the organic solvent and the inert gas are in the step S1, the reflux reaction temperature is 70 to 120 ℃, and the reaction time is 6 to 12 hours.

The product separation and purification method comprises the following steps: the resulting mixture is cooled to room temperature, the crude product obtained is concentrated under reduced pressure, washed several times with additional hot organic solvent (e.g. ethanol, methanol) and filtered to give intermediate vi.

And S3, in the salt forming reaction process, carrying out substitution or salt forming reaction under the protection of inert gas, dissolving the reactant in an organic solvent, heating to the reflux reaction temperature, and reacting for 12-24h.

Preferably, the molar ratio of the intermediate product VI to halogenated alkane, sultone, sulfonic substituted alkane or other various substituted alkanes is 1-5; the temperature of the reflux reaction is 90-150 ℃, and the inert gas is nitrogen; the organic solvent is one or more of 1, 4-dioxane, acetonitrile, N' -dimethylformamide, dimethyl sulfoxide, acetone or toluene.

The method for separating and purifying the product in the step comprises the following steps: the mixture obtained by the reaction is cooled to room temperature, the crude product obtained by the reduced pressure concentration is added with ethyl acetate for washing for a plurality of times in a small amount, and the target compound is obtained by filtration.

The invention provides a near-infrared aggregation-induced emission type ultra-high-efficiency photosensitizer with an asymmetric diacetonitrile structure, which has aggregation-induced emission performance, has the maximum emission wavelength in a near-infrared region, can be used for near-infrared fluorescence imaging, can specifically target mitochondria, and is an excellent contrast agent for mitochondrial imaging.

In addition, the photosensitizer can generate singlet oxygen in cells with ultra-high efficiency under the irradiation of a light source, and effectively kill tumor cells.

Therefore, in a third aspect of the invention, there is provided the use of the photosensitizer in the preparation of a photodynamic therapeutic agent, in particular for use in near infrared fluorescence imaging guided photodynamic therapeutic agents.

In terms of photosensitizer structure, the invention takes para-aryldiacetonitrile as a structural element and changes the molecular structure Ar ₁ Electron donating groups (A) and Ar in position ₂ With electron-withdrawing groups in place (D) and R as auxiliary ₁ The flexible ion groups of the sites are synthesized into a series of super-efficient photosensitizers with aggregation-induced emission performance. The electron-withdrawing group (A) and the electron-donating group (D) are bridged through a conjugated pi system, the D-pi-A effect can be enhanced, the probe has near infrared luminescence, and the electron cloud distribution of HOMO (highest occupied molecular orbital) and LUMO (lowest unoccupied molecular orbital) of the compound can be obviously separated, so that the probe is favorable for the near infrared luminescence ¹ O ₂ The compound can be used as a photosensitizer to realize high-efficiency photodynamic therapy guided by near infrared imaging. In addition, the introduction of the flexible cationic chain not only enables the photosensitizer to have the capability of specifically targeting mitochondria, but also enhances the biocompatibility of the photosensitizer, and is more beneficial to biological application. From the aspect of probe performance, the compounds show near infrared AIE characteristics, have the advantages of strong penetrability, low background interference, small photodamage and the like, and have great potential in the field of photodynamic therapy.

Compared with the prior art, the photosensitizer with the asymmetric diacetonitrile structure provided by the invention has the following beneficial effects that: (1) The synthesis method of the photosensitizer is simple, the raw materials are convenient to obtain, and the photosensitizer has high yield; (2) The photosensitizer has AIE characteristics, the maximum absorption peak is in a visible light region, and the AIE photosensitizer can be activated by common white light; (3) The photosensitizer has near infrared emission (more than 650 nm), has strong near infrared fluorescence signal penetrating property when being used for fluorescence imaging, and is beneficial to eliminating interference of autofluorescence of biological tissues during imaging; (4) The photosensitizer has ultrahigh singlet oxygen yield which can be more than 6 times of rose bengal B (a common reference compound for determining the singlet oxygen yield); (5) The photosensitizer with near infrared AIE performance provided by the invention can specifically mark mitochondria in living cells,produced in mitochondria ¹ O ₂ Cancer cells can be killed in a short time, and PDT efficiency is improved; (6) The photosensitizer can also realize living body imaging of animal level, can gradually enrich in a tumor part after being injected into a body through tumor, and has good fluorescence imaging capability; (7) The photosensitizer has better light stability and biocompatibility and lower dark toxicity, and can be used for photodynamic therapy guided by tumor-bearing mouse near-infrared fluorescence imaging after the tumor-bearing mouse is constructed.

Drawings

FIG. 1 shows TPA-Py-PF prepared in example 1 ₆ The nuclear magnetic resonance hydrogen spectrum of (1) characterizes the result;

FIG. 2 shows TPA-Py-PF prepared in example 1 ₆ The nuclear magnetic resonance carbon spectrum characterization result is obtained;

FIG. 3 shows TPA-Py-PF prepared in example 1 ₆ The high-resolution mass spectrum characterization result is obtained;

FIG. 4 shows TPE-Py-PF prepared in example 2 ₆ The result is characterized by the nuclear magnetic resonance hydrogen spectrum;

FIG. 5 shows TPE-Py-PF prepared as the compound in example 2 ₆ The nuclear magnetic resonance carbon spectrum characterization result is obtained;

FIG. 6 shows TPE-Py-PF prepared as the compound in example 2 ₆ The high-resolution mass spectrum characterization result is obtained;

FIG. 7 shows DEA-Py-PF which is a compound prepared in example 3 ₆ The result is characterized by the nuclear magnetic resonance hydrogen spectrum;

FIG. 8 shows DEA-Py-PF which is a compound prepared in example 3 ₆ The nuclear magnetic resonance carbon spectrum of (3) represents the result;

FIG. 9 shows DEA-Py-PF which is a compound prepared in example 3 ₆ The high resolution mass spectrum characterization result is obtained;

FIG. 10 shows TPA-Py-PF prepared in examples 1, 2 and 3 ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ UV-visible absorption spectrum in dimethyl sulfoxide (concentration 10. Mu.M);

FIG. 11 shows TPA-Py-PF prepared in examples 1, 2 and 3 ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ In a mixed solvent (volume) of dimethyl sulfoxide and tolueneFluorescence emission spectrum in a ratio of 1;

FIG. 12 shows TPA-Py-PF prepared in example 1 ₆ A fluorescence emission spectrum and a relative fluorescence intensity chart in a DMSO and toluene mixed system;

FIG. 13 shows TPE-Py-PF prepared in example 2 ₆ A fluorescence emission spectrum and a relative fluorescence intensity chart in a DMSO and toluene mixed system;

FIG. 14 shows DEA-Py-PF prepared in example 3 ₆ A fluorescence emission spectrum and a relative fluorescence intensity chart in a DMSO and toluene mixed system;

FIG. 15 shows TPA-Py-PF prepared in example 1 ₆ Singlet oxygen detection map of;

FIG. 16 shows TPE-Py-PF prepared in example 2 ₆ A singlet oxygen detection map of;

FIG. 17 shows DEA-Py-PF prepared in example 3 ₆ Singlet oxygen detection map of;

FIG. 18 is a TPA-Py-SO prepared in example 7 ₃ Singlet oxygen detection map of;

FIG. 19 is a graph of the absorption decay of ABDA in the presence of different photosensitizers, where A ₀ The initial absorbance is obtained, and A is the final absorbance after illumination;

FIG. 20 shows TPA-Py-PF of example 10 of this invention ₆ Production in mouse 4T1 cells ¹ O ₂ The performance test chart of (a) and (b) are laser confocal images of a fluorescence field and a bright field respectively;

FIG. 21 shows TPE-Py-PF in example 10 of the present invention ₆ Production in mouse 4T1 cells ¹ O ₂ The performance test chart of (a) and (b) are laser confocal images of a fluorescence field and a bright field respectively;

FIG. 22 shows DEA-Py-PF in example 10 of the present invention ₆ Production in mouse 4T1 cells ¹ O ₂ The (a) and (b) are laser confocal images of a fluorescence field and a bright field respectively;

FIG. 23 shows TPA-Py-PF of example 11 of the present invention ₆ Mitochondrial co-localization test plots in mouse 4T1 cells; wherein (a) is TPA-Py-PF ₆ The fluorescent signal of (a); (b) Is the fluorescence signal of a commercial mitochondrial dye; (c) is the mixed field of (a) and (b); (d) is the overlap factor of (a) and (b);

FIG. 24 shows TPE-Py-PF of example 11 of the present invention ₆ Mitochondrial co-localization test plots in mouse 4T1 cells; wherein (a) is TPE-Py-PF ₆ The fluorescent signal of (a); (b) is the fluorescence signal of a commercial mitochondrial dye; (c) is the mixed field of (a) and (b); (d) is the overlap factor of (a) and (b);

FIG. 25 shows that in example 11 of the present invention, DEA-Py-PF ₆ Mitochondrial co-localization test plots in mouse 4T1 cells; wherein (a) is DEA-Py-PF ₆ The fluorescent signal of (a); (b) is the fluorescence signal of a commercial mitochondrial dye; (c) is the mixed field of (a) and (b); (d) is the overlap factor of (a) and (b);

FIG. 26 shows TPA-Py-PF concentrations (0, 1, 2, 5, 10, 20, 50, 100 μmol/L) in light (white light) or dark (dark) according to example 12 of the present invention ₆ Survival of 4T1 cells in the coexistence;

FIG. 27 shows TPE-Py-PF at different concentrations (0, 1, 2, 5, 10, 20, 50, 100 μmol/L) under light (white light) or dark (dark) in example 12 of the present invention ₆ Survival of 4T1 cells in coexistence;

FIG. 28 is a graph showing that DEA-Py-PF is added at different concentrations (0, 1, 2, 5, 10, 20, 50, 100. Mu. Mol/L) under light (white light) or dark (dark) in example 12 of the present invention ₆ Survival of 4T1 cells in coexistence;

FIG. 29 shows TPA-Py-PF in example 13 of the present invention ₆ A near-infrared fluorescence imaging performance test chart of a tumor part of a 4T1 tumor-bearing mouse, wherein the chart (a) is a photosensitizer TPA-Py-PF prepared in example 1 ₆ After intratumoral injection into 4T1 tumor-bearing mice, TPA-Py-PF was administered to the mice in different treatment groups ₆ A graph of the change of the fluorescence signal with time; (b) The figure is a near infrared fluorescence imaging contrast diagram of isolated tissues (tumor, heart, liver, spleen, lung and kidney) obtained by dissecting a 4T1 tumor-bearing mouse 24h after the 4T1 tumor-bearing mouse is treated by PBS or a photosensitizer;

FIG. 30 is a graph showing the comparison of the treatment effects of photodynamic tumors on 4T1 tumor-bearing mice by different treatment groups in example 14, wherein (a) the tumor volume and (b) the body weight of the mice are varied with the number of days;

FIG. 31 is a graph comparing tumor size after 12 days of treatment for each group in example 14 of the present invention.

Detailed Description

Objects, advantages and features of the invention will be illustrated and explained by the following non-limiting description of preferred embodiments, which is provided by way of example only and not to limit the scope of the invention. The technical features mentioned in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

It will be apparent to those skilled in the chemical, biochemical or related arts that many modifications and variations may be made without departing from the spirit of the invention and the scope of the appended claims.

And declaring: the raw materials used in the invention are all commonly available.

The invention adopts the following synthetic route to prepare the near-infrared aggregation-induced emission type ultra-high-efficiency photosensitizer with an asymmetric diacetonitrile structure:

firstly, under the protection of nitrogen, dissolving para-aryl diacetonitrile (II) and aryl formaldehyde (III) in an organic solvent according to the molar ratio of 1 (1-4), adding an alkaline substance as a catalyst, and reacting at room temperature for 6-12 h to obtain an intermediate product (IV).

And secondly, under the protection of nitrogen, dissolving the intermediate product (IV) obtained in the first step and N-substituted aryl formaldehyde (V) in an organic solvent, wherein the molar ratio of IV to V is 1 (1-2), adding an alkaline substance as a catalyst, and performing reflux reaction at 70-120 ℃ to obtain the intermediate product with the structural formula shown in VI.

Thirdly, under the protection of nitrogen, the intermediate product (VI) of the second step reaction and halogenated alkane, sultone, sulfonic group substituted alkane or other various substituted alkanes are mixed according to the proportion of 1: (1-5) dissolving in an organic solvent according to the molar ratio, and carrying out reflux reaction at 90-150 ℃ to obtain the target compound.

Example 1

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer TPA-Py-PF with asymmetric diacetonitrile structure ₆ The preparation method of (I-1) comprises the following steps:

(1) 468.2mg of terephthalonitrile (II, 3.0 mmol) were added to a three-necked flask, and 7.5mL of THF and 2.5mL of EtOH were added. 237.1mg of 4- (diphenylamine) benzaldehyde (III-1, 1.0 mmol) was dissolved in 15mL of THF, 20mg of sodium hydroxide was dissolved in 5mL of EtOH, and the two were mixed well and then charged into a dropping funnel having a constant pressure. After vacuumizing and nitrogen filling for three times, the constant-pressure dropping funnel is slowly opened, and the reaction is stirred at room temperature for 12 hours. The reaction mixture was extracted three times with 50mL of methylene chloride, and the organic phases were combined, washed three times with saturated brine and dried over anhydrous sodium sulfate. Distilling under reduced pressure to remove the organic solvent, separating and purifying the crude product by silica gel column chromatography, wherein the eluent is petroleum ether: ethyl acetate =10 (volume ratio), yielding 255mg of a yellow-green solid as (Z-2- (4- (cyanomethyl) phenyl) 3- (4- (diphenylamine) phenyl) acrylonitrile (iv-1) in 51% yield.

The reaction formula of this step is as follows:

(2) 411.2mg of IV-1, 128.4mg of 4-pyridine benzaldehyde (V-1, 1.2mmol) and 42.6. Mu.L of piperidine are added into 40mL of ethanol, and the temperature is raised to reflux reaction for 12 hours under the protection of nitrogen. The reaction solution was cooled to room temperature, extracted three times with 50mL of methylene chloride, and the organic phases were combined, washed three times with saturated brine and dried over anhydrous sodium sulfate. Distilling under reduced pressure to remove the organic solvent, separating and purifying the crude product by silica gel column chromatography, wherein the eluent is petroleum ether: ethyl acetate =1 (volume ratio), yielding 480.3mg of an orange-yellow solid as (Z) -2- (4- ((Z) 1-cyano-2- (4- (diphenylamine) phenyl) vinyl) phenyl) -3- (pyridine-4-acrylonitrile) (vi-1), yield: 85 percent.

The reaction formula of the step is as follows:

(3) 250.2mg of VI-1 (0.5 mmol) and 58. Mu.L of iodoethane (0.6 mmol) are added to 15mL of acetonitrile solvent, and the mixture is reacted for 6h after heating to reflux temperature under nitrogen. The reaction solution is cooled to room temperature, the organic solvent is removed by reduced pressure distillation, the crude product is dissolved by 15mL of acetone, and 5mL of KPF is added ₆ Saturated aqueous solution, and the mixture is heated to reflux temperature under the protection of nitrogen and then reacted for 3 hours. After cooling to room temperature, it was extracted three times with DCM and water, the organic phases were collected, the solvent was spin-dried and the mixed residue was washed with ethyl acetate, and 631.8mg of black solid obtained by filtration was TPA-Py-PF ₆ (I-1), yield: 91 percent.

Nuclear magnetic resonance spectral characterization and high resolution mass spectral characterization of compound I-1 see fig. 1-3, data as follows: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ9.20(d,J＝6.8Hz,2H),8.48(d,J＝6.7Hz,2H),8.41(s,1H),8.10(s,1H),7.95(dt,J＝16.7,8.9Hz,6H),7.42(t,J＝7.8Hz,4H),7.20(dd,J＝16.3,7.9Hz,6H),6.97(d,J＝8.9Hz,2H),4.65(q,J＝7.3Hz,2H),1.59(t,J＝7.3Hz,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ149.97,145.80,145.26,135.20,131.22,129.93,128.39,127.49,127.11,126.19,125.82,124.96,119.48,83.89,24.61.HRMS for C ₃₇ H ₂₉ N ₄ ⁺ [M–PF ₆ ] ⁺ ,calculated:529.2392；found:529.2392。

the reaction formula of this step is as follows:

example 2

Near-infrared aggregation-induced emission type super-efficient photosensitizer TPE-Py-PF with asymmetric diacetonitrile structure ₆ (I-2) production method of example 1 was repeated except that 4- (diphenylamine) benzaldehyde (III-1) in the above-mentioned step (1) was changed to 4- (1)2, 2-triphenylethylene) benzaldehyde (III-2), the other conditions being unchanged. Finally obtaining the target compound TPE-Py-PF ₆ Red solid of (I-2), yield: 83 percent.

Structural characterization of compound I-2 referring to fig. 4-6, the data are: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ9.22(d,J＝6.8Hz,2H),8.48(d,J＝6.7Hz,2H),8.44(s,1H),8.12(s,1H),7.99(q,J＝8.7Hz,4H),7.78(d,J＝8.4Hz,2H),7.17(dt,J ₁ ＝8.8Hz,J ₂ ＝4.2Hz,11H),7.07–6.95(m,6H),4.66(q,J＝7.3Hz,2H),1.59(t,J＝7.3Hz,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ193.41,151.09,150.47,149.58,141.29,140.76,136.61,131.92,126.77,125.44,122.72,122.67,122.64,122.13,119.19,114.02,100.15,43.83,12.41.HRMS for C ₄₅ H ₃₄ N ₃ ⁺ [M–PF ₆ ] ⁺ ,calculated:616.2753；found:616.2757。

the synthetic route is as follows:

example 3

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer DEA-Py-PF with asymmetric diacetonitrile structure ₆ (I-3) preparation method, example 1 was repeated except that 4- (diphenylamine) benzaldehyde (III-1) in the above-mentioned step (1) was changed to 4- (diethylamino) benzaldehyde (III-3), and other conditions were not changed. Finally obtaining a target compound DEA-Py-PF ₆ Black solid of (I-3), yield: and 90 percent.

Structural characterization maps for compound I-3 see fig. 7-9, data for: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ9.20(d,J＝6.8Hz,2H),8.47(d,J＝6.8Hz,2H),8.38(s,1H),8.10–7.80(m,7H),6.82(d,J＝9.1Hz,2H),4.65(q,J＝7.3Hz,2H),3.46(q,J＝6.9Hz,4H),1.58(t,J＝7.3Hz,3H),1.24–1.05(m,6H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ149.75,148.83,144.89,137.83,132.11,131.05,127.37,126.66,125.55,119.83,119.13,118.81,116.08,111.08,99.80,56.23,43.86,16.06,12.47.HRMS for C ₂₉ H ₂₉ N ₄ ⁺ [M–PF ₆ ] ⁺ ,calculated:433.2392；found:433.2384。

the synthetic route is as follows:

example 4

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer TPA-Qu-PF with asymmetric diacetonitrile structure ₆ (I-4) preparation method, repeat example 1, the difference lies in example 1 (2) step in 4-pyridine benzaldehyde (V-1) change 4-quinoline benzaldehyde (V-2), finally obtain target compound TPA-Qu-PF ₆ (I-4). The synthetic route is as follows:

the structural characterization data for compound I-4 is: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ9.01(s,1H),8.60(s,1H),8.42(s,1H),8.23(s,1H),8.10(s,1H),7.94(s,2H),7.89(s,2H),7.75(s,1H),7.33(s,2H),7.25(d,J＝10.0Hz,6H),7.18(s,2H),7.08(s,4H),7.00(s,2H),4.80(s,2H),1.57(s,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ146.93,145.86,143.71,143.27,143.14,142.60,142.43,134.49,132.57,131.93,130.39,129.80,129.27,128.36,126.82,124.67,124.23,124.18,123.15,122.99,121.32,121.16,112.55,108.47,54.30,12.57.HRMS for C ₄₁ H ₃₁ N ₄ ⁺ [M–PF ₆ ] ⁺ ,calculated:579.2543；found:433.2534。

example 5

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer MeO-TPA-Py-PF with asymmetric diacetonitrile structure ₆ The preparation method of (I-5) comprises the following steps:

(1) 500mg of aniline (VII, 5.5 mmol), 5.0g of 1-iodo-4-methoxybenzene (VIII, 22.0 mmol), 1.3g of copper (22.0 mmol), 300mg of 18-crown-6 (1.1 mmol) and6.0g K ₂ CO ₃ (44.0 mmol) was placed in a completely dry two-neck flask under nitrogen. 50mL of degassed o-dichlorobenzene was then charged to the flask, the mixture was stirred and warmed to reflux temperature for 48h. The reaction solution was cooled to room temperature, filtered, extracted three times with dichloromethane, the organic phases were combined and washed three times with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was distilled off under reduced pressure. And (3) separating and purifying the crude product by using silica gel column chromatography, wherein the eluent is petroleum ether: ethyl acetate =3 (volume ratio), yielding 1.3g of 4,4' -dimethoxytriphenylamine (IX) as a white solid, in yield: 81 percent.

The reaction formula of this step is as follows:

(2) 1.0g of 4,4' -dimethoxytriphenylamine (IX, 3.3 mmol) was dissolved in 20mL of DMF at 0 ℃ and 400. Mu.L of OCl ₃ (5.0 mmol) was added dropwise to the reaction. The mixture was stirred at 0 ℃ for 30 minutes and then at room temperature until the mixture turned red. After color change, the mixture was heated to reflux temperature and stirred for 3 hours. After the reaction, the reaction mixture was poured into a beaker filled with ice water, neutralized with 0.5M sodium bicarbonate under stirring, extracted with ethyl acetate, and the organic solvent was removed by evaporation under reduced pressure. This gave 4- (bis- (4-methoxyphenyl) amino) benzaldehyde as a yellow viscous liquid, 1.0g of (III-4), yield: 95 percent.

The reaction formula of the step is as follows:

the rest of the procedure in example 1 was repeated except that 4- (diphenylamino) benzaldehyde (III-1) in the step (1) in example 1 was changed to 4- (bis- (4-methoxyphenyl) amino) benzaldehyde (III-4). Finally obtaining the target product MeO-TPA-Py-PF ₆ (I-5), the synthetic route is as follows:

the structural characterization data for compound I-5 is: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ8.81(s,2H),7.91(d,J＝25.0Hz,4H),7.75(s,2H),7.33(s,2H),7.26(s,2H),7.22–7.18(m,3H),7.18–7.15(m,3H),6.79(s,4H),4.51(s,2H),3.79(s,6H),1.53(s,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ160.03,145.86,144.35,142.60,140.30,138.07,134.49,131.93,130.39,127.01,126.31,124.23,123.15,121.16,114.48,108.47,56.08,54.00,11.68.HRMS for C ₃₉ H ₃₃ N ₄ O ₂ ⁺ [M–PF ₆ ] ⁺ ,calculated:589.2598；found:589.2566。

example 6

Near-infrared aggregation-induced emission type ultra-efficient photosensitizer MeO-TPA-Qu-PF with asymmetric diacetonitrile structure ₆ (I-6) preparation method the first three steps of example 5 were repeated to synthesize (Z) 3- (4- (bis (4-methoxyphenyl) amino) benzene) 2- (4- (cyanomethyl) phenyl) propenenitrile (IV-4). The last two steps of example 4 were then repeated, with the difference that IV-1 was replaced by IV-4, the synthetic route of which is as follows:

the structural characterization data for compound I-6 is: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ9.01(s,1H),8.60(s,1H),8.42(s,1H),8.23(s,1H),8.10(s,1H),7.92(d,J＝25.0Hz,4H),7.75(s,1H),7.33(s,2H),7.26(s,2H),7.18(s,6H),6.79(s,4H),4.80(s,2H),3.79(s,6H),1.57(s,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ160.03,145.86,143.71,143.27,143.14,142.60,142.43,138.07,134.49,132.57,131.93,130.39,129.80,128.36,127.01,126.82,124.23,124.18,123.15,121.32,121.16,114.48,112.55,108.47,56.08,54.30,12.57.HRMS for C ₄₃ H ₃₅ N ₄ O ₂ ⁺ [M–PF ₆ ] ⁺ ,calculated:639.2755；found:639.2759。

example 7:

near-infrared aggregation-induced emission type ultra-efficient photosensitizer TPA-Py-SO with asymmetric diacetonitrile structure ₃ (Compound I-7) A production method, except that the iodoethane in the (3) step of example 1 was changed to 1, 3-propane sultone, to finally obtain the objective compound TPA-Py-SO ₃ (I-7). The synthetic route of the step is as follows:

the structural characterization data for compound I-7 is: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ8.81(s,2H),7.92(d,J＝25.0Hz,4H),7.75(s,2H),7.33(s,2H),7.25(d,J＝10.0Hz,5H),7.18(s,3H),7.08(s,4H),7.00(s,2H),3.53(s,2H),2.57(d,J＝20.0Hz,4H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ146.93,145.86,145.24,142.60,140.47,134.49,131.93,130.39,129.27,126.54,124.67,124.23,123.15,122.99,121.16,108.47,59.27,47.83,23.84.HRMS for C ₃₈ H ₃₀ N ₄ O ₃ S ⁺ [M+Na] ⁺ ,calculated:645.1936；found:645.1930。

example 8:

near-infrared aggregation-induced emission type ultra-efficient photosensitizer MeO-TPA-Bt-PF with asymmetric diacetonitrile structure ₆ The difference in the preparation method of (I-8) is that in the step (2) of example 5, 4-pyridinebenzaldehyde (V-1) was changed to 6-benzothiazolecarbaldehyde (V-3), and finally the objective compound MeO-TPA-Bt-PF was obtained ₆ (I-8), the synthetic route is as follows:

the structural characterization data for compound I-8 is: ¹ H NMR(400MHz,DMSO-d ₆ )δ[ppm]:δ8.10(s,1H),7.89(s,2H),7.75(s,2H),7.46(s,1H),7.38(s,5H),7.18(s,6H),6.79(s,4H),5.89(s,1H),4.80(s,2H),3.79(s,6H),1.57(s,3H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ[ppm]:δ163.90,160.03,145.86,143.21,142.60,138.07,135.95,135.17,134.49,134.46,131.93,130.39,127.01,126.96,124.23,124.01,123.15,121.16,119.33,114.48,110.88,108.47,56.08,53.20,13.62.HRMS for C ₄₁ H ₃₃ N ₄ O ₂ S ⁺ [M–PF ₆ ] ⁺ ,calculated:645.2319；found:645.2324。

example 9: study of photophysical Properties and characteristics of singlet oxygen production

(1) Study of photophysical Properties

With the photosensitizer (TPA-Py-PF) produced in examples 1, 2 and 3 ₆ 、TPE-Py-PF ₆ 、DEA-Py-PF ₆ ) The examples are given for the purpose of illustration, and further description is omitted for the purpose of illustration.

Respectively dissolving the synthesized photosensitizer in DMSO solution to prepare the photosensitizer with the concentration of 10 ^–3 M mother liquor, diluting the mother liquor to 10 ^–5 M was dissolved to test the change of the ultraviolet absorption spectrum and the fluorescence emission spectrum in DMSO/toluene mixed solvents of different solvent volume ratios. TPA-Py-PF as shown by UV-VIS absorption spectrum in FIG. 10 ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ The absorption band of (2) is very wide, and the maximum absorption peaks are respectively positioned at 476nm, 419nm and 495 nm. As shown in FIG. 11, TPA-Py-PF ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ The emission spectrum of the fluorescent material is distributed at 500-800nm, the maximum emission wavelength is about 800nm, and the fluorescent material belongs to the near-infrared light-emitting category. The fluorescence emission peak in DMSO is weak, and after toluene is gradually added, the probe gradually forms an aggregate along with the increase of the proportion of the toluene, so that the intramolecular movement is limited, a non-radiative channel is effectively blocked, the radiative transition of the non-radiative channel is activated, and the fluorescence intensity of the photosensitizer is obviously enhanced. As shown in fig. 12-14, the maximum emission wavelengths of the photosensitizers were located at 660nm, 783nm, and 768nm, respectively, when the toluene content reached 90%. The results of the study show that all three fluorescent molecules exhibit typical AIE properties. In addition, the resulting fluorescent molecules have stokes shifts in excess of 200nm, avoiding interference of excitation light and self-absorption of emitted light during biomedical imaging.

(2) Ability to generate singlet oxygen

The photosensitizer (TPA-Py-PF) produced in examples 1, 2, 3 and 7 ₆ 、TPE-Py-PF ₆ 、DEA-Py-PF ₆ 、TPA-Py-SO ₃ ) Other products will not be described again by way of example.

To evaluate the generation of singlet oxygen by the photosensitizer under light: ( ¹ O ₂ ) The capacity of (A) was measured by using a commercially available 9, 10-anthracenediyl-bis (methylene) dipropionic acid (ABDA) as an indicator and rose bengal B (RB), a widely used photosensitizer, as a standard reference ¹ O ₂ The efficiency of generation of (a). As shown in FIGS. 15-18, when photosensitizer is present, 25mW/cm is used ² The ultraviolet absorption of ABDA at 378nm shows a rapid descending trend after the white light is irradiated for 2 minutes, and the faster the absorbance descending rate of ABDA is, the generation of the photosensitizer can be shown ¹ O ₂ The rate is faster. As shown in FIG. 19, it is evident from the attenuation plot of ABDA that TPA-Py-PF ₆ And TPE-Py-PF ₆ Photosensitizer generation ¹ O ₂ Much higher than the commercial photosensitizer rose bengal B. Calculated TPA-Py-PF of 10.00nmol per minute ₆ And TPE-Py-PF ₆ The amounts of degradable ABDA were 38.70nmol and 32.60nmol, respectively, whereas the amount of degradable ABDA of rose bengal B was only 8.75nmol under the same conditions. Further proves that the photosensitizer TPA-Py-PF ₆ And TPE-Py-PF ₆ Active oxygen can be efficiently generated. This means that such AIE fluorophores would have good therapeutic potential as photosensitizers in photodynamic therapy techniques.

Example 10: photosensitizer production in mouse breast cancer cells ¹ O ₂ Evaluation of Capacity

Inoculating mouse breast cancer cell (4T 1 cell) in confocal culture dish with green Singlet Oxygen Sensor (SOSG) as active oxygen indicator, and placing in CO with temperature of 37 deg.C and volume concentration of 5% ₂ The culture was carried out overnight in an incubator. Then the old medium was treated with a medium containing TPA-Py-PF prepared in example 1, 2 or 3 ₆ 、TPE-Py-PF ₆ Or DEA-Py-PF ₆ Replaced with fresh medium solution and incubation continued for 1h. White light (100 mW/cm) for removing the drug-containing petri dish ² ) Irradiating for different time (0, 30s, 1min, 2min, 5min, 10min, 15min or 30 min), and replacing the fresh culture medium containing SOSG, and culturing for 1h. Finally, the cells were washed three times with PBS, 1mL of fresh medium was added, and the green fluorescence intensity in 4T1 cells in each dish was observed using a confocal laser scanning microscope.

As shown in FIGS. 20a-22a, the fluorescence intensity of the cells in the green channel gradually increased with the increase of the illumination time. Furthermore, as shown in the bright field images of FIGS. 20b-22b, TPA-Py-PF was used after 15min of white light irradiation ₆ Or TPE-Py-PF ₆ Both groups of cells treated almost become round, cytosolic leakage, cytoskeleton collapse, and begin to slough off the plate. DEA-Py-PF ₆ Similar phenomena occurred in the treated cells after 20min of light exposure. The combination of the above results confirms that the photosensitizer synthesized according to the above technical route can be produced under illumination conditions ¹ O ₂ And (3) effectively killing tumor cells.

Example 11: assessment of mitochondrial co-localization capability

Inoculating mouse breast cancer cell (4T 1 cell) in confocal culture dish, and placing in CO at 37 deg.C and 5% volume concentration ₂ The culture was carried out overnight in an incubator. Then using a commercial mitochondrial bioprobe (MitoTracker Deep Red) and TPA-Py-PF ₆ 、TPE-Py-PF ₆ Or DEA-Py-PF ₆ The two groups of treated cells were co-stained to determine the location of photosensitizer enrichment in cancer cells. And observing the overlapping degree of the two fluorescence signals by using a confocal laser microscope.

The results from fig. 23-25 show that the red fluorescent signal from the photosensitizer blends well with the green signal (false color) from the mitochondrial dye. Indicating their ability to target and image cancer cell mitochondria, which is beneficial for improving PDT efficiency.

Example 12: TPA-Py-PF ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ Lethality evaluation on 4T1 cells

After trypsinizing the cells in logarithmic growth phase, the cells were diluted with complete medium to a cell suspension, which was subsequently diluted at 1X 10 ⁴ The density of individual/well was seeded in 96-well plates,placing at 37 deg.C and 5% CO ₂ Culturing in an incubator for 12h, adding TPA-Py-PF with different concentrations ₆ 、TPE-Py-PF ₆ Or DEA-Py-PF ₆ The samples were incubated for 1h so that the final concentrations were 0, 1, 2, 5, 10, 15, 20, 50 and 100. Mu.M, respectively, and then illuminated with white light (100 mW/cm) ² ) At the same time, the dark toxicity study was also performed in the group that was not illuminated under the same experimental conditions for 30 minutes. After culturing for 4h,12h and 24h, respectively, the photosensitizer-containing medium was removed, followed by culturing for 1h in the dark with fresh 10-CCK-8-containing medium (no FBS), and then measuring the absorbance value (OD value) at 450nm with a microplate reader, and the corresponding cell viability was calculated by the following equation: cell survival rate (%) = (OD) _Sample(s) –OD _Background )/(OD _{Control of} –OD _Background )×100％。

The killing of 4TI cells by different concentrations of photosensitizer is shown in figures 26-28. Under the condition of illumination, TPA-Py-PF ₆ 、TPE-Py-PF ₆ And DEA-Py-PF ₆ The killing of 4TI cells was concentration dependent. When the concentration of the photosensitizer is as low as 10 mu mol/L, the cell survival rate reaches below 20 percent, and the prepared photosensitizer shows a high-efficiency photodynamic therapy effect on 4TI cells.

Example 13: tumor imaging test

PBS and TPA-Py-PF prepared in example 1 ₆ (0.4 mg/mL, 50. Mu.L) was injected intratumorally into 4T1 tumor-bearing mice and photographs of the mice were taken at different time points after injection using a small animal in vivo imaging system.

Mouse in vivo TPA-Py-PF ₆ The time-dependent change of the near-infrared fluorescence imaging of (2) is shown in FIG. 29a, which shows that TPA-Py-PF ₆ The fluorescence intensity of the compound tends to be stable after 9 hours until the fluorescence intensity is not obviously weakened after 24 hours, and the compound has good drug retention and enrichment capacity at tumor parts; injection of TPA-Py-PF ₆ 24 After h, isolated tissues (tumor and heart, liver, spleen, lung, kidney) obtained by dissecting 4T1 tumor-bearing mice were subjected to near infrared fluorescence imaging contrast. As shown in FIG. 29b, only the tumor site showed fluorescence signals, further demonstrating that TPA-Py-PF ₆ Abundant enrichment at the tumor site.The above results show that TPA-Py-PF ₆ Near infrared fluorescence imaging can be realized in mice.

Example 14: in vivo anti-tumor assessment

The 4T1 tumor-transplanted mice were randomly divided into 4 groups of 5 mice each, including PBS group, PBS + light group, TPA-Py-PF ₆ Combination of TPA-Py-PF ₆ + light group, 100 μ L TPA-Py-PF was injected by intratumoral injection ₆ (8 mg/kg) or a physiological saline solution. Thereafter, the light group was irradiated with white light for 20 minutes for treatment. After each treatment, tumor size and body weight of the mice were recorded daily. Tumor volume was measured with a vernier caliper according to the formula V = (tumor length. Times. Tumor width) ² ) And/2 calculation. To further evaluate the effect of light treatment, mice were sacrificed 12 days after treatment, tumor tissues of different treatment groups were removed, and tumor volumes were measured with a vernier caliper and weighed.

The photodynamic therapy results of tumors in 4 groups of 4T1 tumor-bearing mice are shown in FIG. 30a, and only the treatment group (TPA-Py-PF) ₆ + light group) had the effect of inhibiting tumor growth and even tumor ablation, as shown in fig. 30b, there was no significant change in body weight in mice of the different treatment groups, indicating TPA-Py-PF ₆ Has low dark toxicity. Mice were sacrificed 12 days after treatment, tumor volumes were collected and measured. As shown in fig. 31, the tumor volume in the treatment group was found to be significantly reduced, and there was a case of complete cure. Evidence of TPA-Py-PF ₆ Has the ability to perform PDT efficiently in complex biological systems.

Claims

1. A near-infrared aggregation-induced emission type ultra-efficient photosensitizer with an asymmetric diacetonitrile structure is characterized in that the structural general formula of the photosensitizer is shown as formula I:

wherein X and Y are any one of C, N, O and S;

m and n are natural numbers of 0-3 and are not 0 at the same time;

Ar ₁ selected from any of the following structural formulae:

wherein p is a natural number of 1 to 5, R ₂ And R ₃ Each independently selected from any one of H, hydroxyl, amino, substituted alkyl, alkoxy, alkylthio, alkenyl, alkynyl, cycloalkyl, cycloalkyloxy, cycloalkylthio, amido, aryl, heterocyclic group, heteroaryl, heterocycloalkyl, monoalkylamino or dialkylamino;

Ar ₂ selected from any of the following structural formulas:

z is selected from any one of halide, acetate, trifluoroacetate, dihydrogen phosphate, hydrogen sulfate, tetrafluoroborate, hexafluorophosphate and sulfonate;

R ₁ the alkyl is any one of linear alkyl, cycloalkyl, carboxyl substituted alkyl, aldehyde substituted alkyl, sulfonic group substituted alkyl, azide substituted alkyl and quaternary amine substituted alkyl.

2. The near-infrared aggregation-induced emission type ultra-high efficiency photosensitizer with an asymmetric diacetonitrile-based structure according to claim 1, wherein R is R ₁ Is CH ₃ 、CH ₂ CH ₃ 、CH ₂ COOH、CH ₂ CHO、CH ₂ CH ₃ SO ₃ And CH ₂ CH ₃ N ₃ Any one of the above; r ₂ And R ₃ Each independently selected from H, NH ₃ 、OH、OCH ₃ 、OCH ₂ CH ₃ Or CONH ₂ One kind of (1).

3. The preparation method of the near-infrared aggregation-induced emission type ultra-high efficiency photosensitizer with the asymmetric diacetonitrile structure as claimed in claim 1-2, characterized in that the reaction route is as follows:

the method comprises the following steps:

firstly, adding p-aryl diacetonitrile (II) and aryl formaldehyde (III) into an organic solvent under the protection of inert gas, adding a small amount of alkaline substances as a catalyst, and reacting at room temperature to obtain an intermediate product (IV);

secondly, dissolving the intermediate product (IV) and N-substituted aryl formaldehyde (V) in an organic solvent under the protection of inert gas, and then adding an alkaline catalyst to carry out reflux reaction to obtain an asymmetric diacetonitrile-based intermediate product (VI);

thirdly, under the protection of inert gas, carrying out salt forming reaction on the intermediate product (VI) and Z-substituted alkane to obtain a target product (I), wherein Z is selected from any one of halogen, acetic acid group, trifluoroacetic acid group, dihydrogen phosphate group, hydrogen sulfate group, tetrafluoroboric acid group, hexafluorophosphoric acid group and sulfonic group.

4. The production method according to claim 3, characterized in that: in the first Knoevenagel condensation reaction, the molar ratio of the aryl diacetonitrile (II) to the aryl formaldehyde (III) is 1-4;

in the second Knoevenagel condensation reaction, the molar ratio of the intermediate product (IV) to the N-substituted aryl formaldehyde (V) is 1-2.

5. The production method according to claim 3, characterized in that: in the first and second Knoevenagel condensation reactions, the molar weight of the alkaline substance is 0.1-0.5 equivalent, and the alkaline substance is any one or mixture of more of pyridine, piperidine, potassium carbonate, sodium carbonate, potassium hydroxide, sodium alkoxide and potassium alkoxide;

the organic solvent is selected from any one or a mixture of ethanol, isopropanol, tetrahydrofuran and acetonitrile;

the inert gas is any one of nitrogen or argon.

6. The preparation method according to claim 3, wherein in the first Knoevenagel condensation reaction, the reaction is carried out at room temperature for 6 to 12 hours; in the second step of Knoevenagel condensation reaction, the reflux reaction is carried out for 6 to 12 hours at the temperature of between 70 and 120 ℃.

7. The process according to claim 3, wherein in the salt-forming reaction of the third step, the molar ratio of the intermediate (VI) to the Z-substituted alkane is 1 to 5; the inert gas is nitrogen; the selected organic solvent is one or a mixture of more of 1, 4-dioxane, acetonitrile, N' -dimethylformamide, dimethyl sulfoxide, acetone or toluene,

during the reaction, the reflux reaction is carried out for 12 to 24 hours at the temperature of between 90 and 150 ℃.

8. The production method according to claim 3, characterized in that: the product separation method of the second step and the third step is as follows: cooling the product to room temperature, carrying out reduced pressure concentration to obtain a crude product, adding low-carbon alcohol or ester thereof, washing for a few times, and filtering to obtain the target compound.

9. Use of the near-infrared aggregation-induced emission type ultra-efficient photosensitizer having an asymmetric diacetonitrile-based structure according to claim 1 or 2 for the preparation of a photodynamic therapeutic agent or a fluorescence developer or an image-guided photodynamic therapeutic agent.

10. Use according to claim 9, characterized in that: the photodynamic therapeutic agent is a living tumor photodynamic therapeutic agent or a fluorescence imaging agent or an imaging-guided photodynamic therapeutic agent.