Aggregation-induced emission photosensitizer and pharmaceutical composition constructed therefrom, preparation method therefor and use thereof

Abstract

Disclosed are an aggregation-induced emission photosensitizer and a pharmaceutical composition constructed therefrom, a preparation method therefor and the use thereof. The aggregation-induced emission photosensitizer MTCN-3 has a great reactive oxygen species generation capacity and a photothermal effect, thus exerting the dual effect of inducing pyroptosis and ferroptosis in tumor cells. The aggregation-induced emission photosensitizer is also capable of exerting a synergistic photoimmunotherapeutic effect with an immune agonist Poly(I:C). A co-assembled pharmaceutical composition has a near-infrared aggregation-induced emission effect, exhibits good subcellular distribution, and has active targeting selectivity for tumor cells. The pharmaceutical composition eliminates tumors and prevents tumor metastasis by means of inhibiting immune evasion, and inhibits the growth of both primary and distal tumors, thereby exerting a systemic anti-tumor immune effect. The pharmaceutical composition exhibits high stability and low toxic side effects, and has broad prospects for clinical application.

Description

An aggregation-induced emission photosensitizer, a pharmaceutical composition constructed by the same, a preparation method and application thereof TECHNICAL FIELD

[0001] The present application relates to an aggregation-induced emission photosensitizer, a pharmaceutical composition constructed by the same, a preparation method and application thereof, in particular to an aggregation-induced emission photosensitizer capable of inducing pyroptosis and ferroptosis, a pharmaceutical composition constructed by the same with a synergistic effect of photoimmunity, a preparation method and application thereof. BACKGROUND

[0002] Immune cell death (ICD) is an immune stimulating reaction induced by necrosis or programmed cell death. During the process of ICD, a series of signal molecules, damage-associated molecular patterns (DAMPs), are produced by cells. DAMPs mainly include calreticulin exposed on the cell surface, high mobility group protein diffused outside the nucleus, ATP released by cells, and heat shock proteins, etc. They can act as natural immune agonists to bind to the pattern recognition receptors on the surface of dendritic cells, promote the maturation of dendritic cells, initiate a series of cellular responses, and ultimately activate the innate and acquired immune responses.

[0003] Ferroptosis and pyroptosis, as two typical immune cell deaths, can activate or change the immune system. The essence of ferroptosis is the depletion of glutathione, the decrease of glutathione peroxidase (GPX4) activity, and the inability of lipid oxidants to be metabolized by GPX4-catalyzed glutathione reductase reaction. Pyroptosis is a new type of lytic cell death, which is activated by the gasdermin protein family (GSDM) including GSDMD, GSDME, etc. The lysed GSDM will punch holes in the cell membrane, thereby destroying the cell membrane and releasing a large amount of pro-inflammatory factors and intracellular substances.

[0004] Ferroptosis and pyroptosis-mediated immunotherapy mainly relies on chemotherapeutic drugs, but their toxic side effects are serious. Poly (I:C), as an artificial interferon inducer, can induce apoptosis and immune cell death (ICD), but its use dose is high, the immune response rate is low, and the treatment window is narrow. Therefore, the clinical application of the above immunotherapeutic drugs has limitations and cannot meet the needs of efficient and low-toxic treatment. SUMMARY

[0005] The first object of the present application is to provide an aggregation-induced emission photosensitizer capable of targeting inducing pyroptosis and ferroptosis of tumor cells, the second object is to provide a pharmaceutical composition constructed by the aggregation-induced emission photosensitizer with a synergistic effect of photoimmunity, the third object is to provide a preparation method thereof, and the fourth object is to provide a pharmaceutical application thereof.

[0006] Technical solution: The aggregation-induced emission photosensitizer provided by the present application has the following structure:

[0007] Phototherapy is a tumor intervention treatment method with adjustable light damage, non-invasive and small side effects. Photodynamic therapy (PDT) and photothermal therapy (PTT) are two reasonable ways to achieve phototherapy. In the photodynamic process, photosensitizers produce cytotoxic reactive oxygen species (ROS) under light; while in the photothermal process, light energy is effectively converted into heat energy through photothermal agents. The active oxygen and photothermal generated by lysosome-targeted photosensitizers make the lysosome membrane permeable, and the lysosome function abnormal, thereby further triggering ICD. The AIE photosensitizer designed in the present application has great advantages in fluorescence imaging and photodynamic therapy due to its high brightness in aggregated state, good light stability and large Stokes shift, and has application potential as a lysosome-targeted drug.

[0008] The anti-tumor pharmaceutical composition described in the present application is obtained by electrostatic interaction co-assembly of an aggregation-induced emission photosensitizer selected from any one of the following structures and an immune agonist Poly (I:C), and modification by an amphiphilic PEG modifier,

[0009] The pharmaceutical composition designed in the present application can target tumors and distribute in lysosomes in cells. Under light, the photosensitizer in the pharmaceutical composition can generate a large amount of ROS and heat, thereby causing lysosome dysfunction in cells, increasing lysosome membrane permeability, releasing lysosome contents into the cytoplasm, and inducing pyroptosis and ferroptosis of cells. At the same time, the photosensitizer promotes the release of cell contents and inflammatory cytokines by inducing cell pyroptosis, and can play a synergistic role with the immune agonist Poly (I:C), thereby activating the anti-tumor immune response, promoting the production of tumor-specific antigens and the maturation of dendritic cells, promoting T cell activation and proliferation, and providing systemic anti-tumor immunity. This makes the pharmaceutical composition designed in the present application an effective, low-side-effect, targeted dual-inducing drug for tumor cell pyroptosis and ferroptosis.

[0010] Preferably, the average particle size of the pharmaceutical composition described in the present application is 100-200 nm.

[0011] Further preferably, the average particle size of the pharmaceutical composition described in the present application is 103.4 nm.

[0012] Preferably, the amphiphilic PEG modifier is selected from one or two of DSPE-PEG, DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 200-20000.

[0013] Further preferably, the amphiphilic PEG modifier is DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 2000.

[0014] Preferably, the mass ratio of the aggregation-induced emission photosensitizer to the immune stimulator Poly(I:C) is (1-5):1.

[0015] Further preferably, the mass ratio of the aggregation-induced emission photosensitizer to the immune stimulator Poly(I:C) is 3:1.

[0016] Preferably, the mass ratio of the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C) to the amphiphilic PEG modifier is 1:(3-10).

[0017] Further preferably, the mass ratio of the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C) to the amphiphilic PEG modifier is 1:3.

[0018] The aggregation-induced emission photosensitizer of the present application is prepared by the reaction of 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridine-1-ium and 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carboxaldehyde in an inert gas catalyzed by piperidine.

[0019] Preferably, the molar ratio of 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridine-1-ium, 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carboxaldehyde and the piperidine catalyst is 1:1:0.05.

[0020] Preferably, the reaction solvent of the reaction is anhydrous ethanol.

[0021] Preferably, the reaction is a reflux reaction.

[0022] The preparation method of the pharmaceutical composition of the present application comprises the following steps:

[0023] (1) separately dissolve the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C), and add both solutions to water to spontaneously form co-assembled nanoparticles under stirring;

[0024] (2) dissolve the amphiphilic PEG modifier and add it to the co-assembled nanoparticles prepared in step (1), and remove the solvent to obtain the pharmaceutical composition.

[0025] Further preferably, in step (1), the aggregation-induced emission photosensitizer is dissolved in DMSO, and the immune stimulator Poly(I:C) is dissolved in water.

[0026] Further preferably, in step (1), the solutions of the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C) spontaneously form co-assembled nanoparticles under stirring at a stirring speed of 1200 rpm.

[0027] It is further preferred that the amphiphilic PEG modifier in step (2) is dissolved in DMSO.

[0028] It is further preferred that the solvent is removed by lyophilization in step (2).

[0029] It is further preferred that the volume ratio of DMSO used in steps (1) and (2) is 1:1.

[0030] It is further preferred that the volume ratio of water used in steps (1) and (2) is 1:18.

[0031] It is further preferred that the total volume ratio of DMSO and water used in steps (1) and (2) is 1:19.

[0032] The aggregation-induced emission photosensitizer or the pharmaceutical composition constructed therefrom according to the present application is applied in the preparation of a drug for inducing pyroptosis, ferroptosis, lysosomal dysfunction of tumor cells.

[0033] The aggregation-induced emission photosensitizer or the pharmaceutical composition constructed therefrom according to the present application is applied in the preparation of a drug for activating anti-tumor immune response, inhibiting tumor growth, and inhibiting tumor metastasis.

[0034] Preferably, the tumor is a breast cancer tumor.

[0035] The aggregation-induced emission photosensitizer or the pharmaceutical composition constructed therefrom according to the present application eliminates tumors and prevents tumor metastasis by preventing immune escape, can effectively inhibit the growth of primary tumors and distant tumors, and plays a systemic anti-tumor immune role.

[0036] Beneficial effects: Compared with the prior art, the present application has the following remarkable advantages:

[0037] The aggregation-induced emission photosensitizer designed in the present application has excellent active oxygen generation ability and photothermal effect, and plays a dual effect of inducing pyroptosis and ferroptosis of tumor cells. The aggregation-induced emission photosensitizer can also play a synergistic phototherapy effect with the immune stimulant Poly(I:C), and the co-assembled pharmaceutical composition has a near-infrared induced aggregation emission effect, good subcellular distribution, and active targeting selectivity for tumor cells; eliminates tumors and prevents tumor metastasis by preventing immune escape, inhibits the growth of primary tumors and distant tumors, and plays a systemic anti-tumor immune role; has good stability, low toxicity and side effects, and has a wide clinical application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0038] FIG. 1 is a synthesis route of the aggregation-induced emission photosensitizer MTCN-3.

[0039] Figure 2 is the UV spectrum (a) and fluorescence spectrum (b) of MTCN-3 (10 μM) in DMSO solution at an excitation wavelength of 480 nm;

[0040] Figure 3 is the fluorescence spectrum of MTCN-3 in DMSO / toluene mixture with different water fractions (f t ) at an excitation wavelength of 480 nm;

[0041] Figure 4 is the result of the generation of reactive oxygen species (ROS) by MTCN-3 in aqueous solution;

[0042] Figure 5 is the result of the temperature rise of MTCN-3 under laser (0.5 W cm -2 ) irradiation;

[0043] Figure 6 is the particle size distribution (a) and the stability results in water and PBS respectively (b) of M@P;

[0044] Figure 7 is the confocal image of the lysosome colorant Lysosome Green and M@P (scale bar: 50 nm);

[0045] Figure 8 is the result of the cytotoxicity of 4T1 cells treated with different concentrations of M@P under light (520 nm, 0.5 W cm -2 , 1 min) and dark conditions (M@P for dark conditions, M@P+L for light conditions);

[0046] Figure 9 is the confocal image of AO staining (scale bar: 50 nm);

[0047] Figure 10 is the cell membrane expansion and content release before and after M@P treatment (scale bar: 20 nm);

[0048] Figure 11 is the result of the changes in intracellular GSH (a) and MDA (b) before and after M@P treatment;

[0049] Figure 12 is the in vivo fluorescence image of 4T1 tumor-bearing mice after intravenous injection of M@P;

[0050] Figure 13 is the primary tumor and distal tumor of 4T1 tumor-bearing mice after laser irradiation (n = 3). DETAILED DESCRIPTION

[0051] The technical solutions of the present application are further described below in conjunction with examples.

[0052] Example 1: Preparation of aggregation-induced emission photosensitizer MTCN-3

[0053] (1) Preparation of 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carboxaldehyde

[0054] 4-Bromo-N,N-di-p-tolylaniline (3.38 g, 9.6 mmol), 5-aldehyde-2-thiopheneboronic acid (1 g, 6.4 mmol), potassium carbonate (35.3 g, 25.6 mmol), and tetrakis(triphenylphosphine)palladium (0.37 g, 0.32 mmol) were dissolved in a toluene (Tol, 90 mL) / methanol (MeOH, 90 mL) mixture, and all air was removed. The reaction mixture was cooled to room temperature and extracted with dichloromethane (DCM) and water, and dried over anhydrous Na₂SO₄. After evaporation of the solvent, the crude product was purified by silica gel column chromatography using petroleum ether (PE) / ethyl acetate (EA) (10 / 1, v / v) as eluent to give 1.53 g of a yellow solid, in 62.5% yield. NMR characterization is as follows: 1 H NMR (600MHz, CDCl3) δ9.84(s,1H),7.69(d,J=3.8Hz,1H),7.47(d,J=8.6Hz,2H),7.27(d,J=3 .9Hz,1H),7.10(d,J=8.0Hz,4H),7.03(d,J=8.1Hz,4H),6.99(d,J=8.6Hz,2H),2.33(s,6H).

[0055] (2) Preparation of compound 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridine-1-onium

[0056] (3-Bromopropyl)trimethylammonium bromide (a) (2.61 g, 10 mmol) and 3-pyridylacetonitrile (1.18 g, 10 mmol) were dissolved in anhydrous acetonitrile (ACN, 20 mL) in a 50 mL round flask. The mixture was refluxed at 80 °C for 4 h. After cooling to room temperature, the solvent in the reaction system was removed by rotary evaporation. The mixture was washed with diethyl ether, and the precipitate was filtered to give a white solid compound d in 74% yield. NMR characterization is as follows: 1 H NMR (600MHz, DMSO-d6) δ9.21(d,J=11.1Hz,1H),9.07(s,1H),8.51(d,J=7.6Hz,1H),8.12( t,J=6.6Hz,1H),4.69(d,J=4.7Hz,2H),3.50(d,J=4.9Hz,2H),3.14(s,11H),2.51(s,2H).

[0057] (3) Preparation of aggregation-induced emission photosensitizer MTCN-3

[0058] As shown in Figure 1, the compound 5-(4-(di-p-tolylamino)phenyl)thiophene-2- carboxaldehyde (0.38 g, 1 mmol) prepared in step (1) and the compound 3-(cyanomethyl)- 1-(3-(trimethylamino)propyl)pyridin-1-ium (0.38 g, 1 mmol) prepared in step (2) were added to anhydrous ethanol (30 mL), and piperidine (4.26 mg, 0.05 mmol) was added as a catalyst, and the reaction was carried out under a nitrogen atmosphere (80 °C) for 7 h. Then it was cooled to room temperature, and the solvent was removed by evaporation under reduced pressure. The crude product was purified by silica gel column chromatography with DCM / MeOH (10 / 1, v / v) as the eluent to obtain the product as a purple solid in a yield of 50.23%. The structural characterization is as follows: 1 H NMR (600 MHz, DMSO-d6) δ 9.66 (s, 1H), 9.13 (d, J = 5.9 Hz, 1H), 8.89 (s, 1H), 8.83 (d, J = 8.4 Hz, 1H), 8.29 (dd, J = 8.0, 6.2 Hz, 1H), 7.93 (d, J = 4.0 Hz, 1H), 7.65 (t, J = 6.1 Hz, 3H), 7.18 (d, J = 8.2 Hz, 4H), 7.01 (d, J = 8.2 Hz, 4H), 6.92 (d, J = 8.7 Hz, 2H), 4.79 (dd, J = 13.2, 6.0 Hz, 2H), 3.48 (dd, J = 10.6, 5.5 Hz, 2H), 3.13 (s, 9H), 2.57 (dt, J = 15.3, 7.7 Hz, 2H), 2.29 (s, 6H). 13 C NMR (151 MHz, DMSO) δ 152.27, 149.39, 144.32, 143.79, 142.37, 141.58, 141.54, 140.16, 135.13, 134.82, 133.98, 130.75, 128.65, 127.72, 125.73, 124.97, 124.10, 120.93, 117.35, 98.08, 62.25, 58.59, 53.06, 24.87, 20.93. HR-ESI-MS: calcd. for C 38 H 40 Br2N4S: m / z: 1 / 2 [M-2Br] 2+ : 292.1482, found: m / z 292.14551.

[0059] Example 2: Evaluation of the photophysical properties of the aggregation-induced emission photosensitizer MTCN-3

[0060] As shown in Figure 2, the DMSO solution (10 μΜ) of MTCN-3 showed a main absorption peak at 499 nm and a fluorescence emission peak at 760 nm. In addition, as shown in Figure 3, MTCN-3 showed significant aggregation-induced emission properties in the mixed solution of DMSO and toluene.

[0061] As shown in Figure 4, the ROS indicator DCFH was used to evaluate the photothermal behavior of MTCN-3. After irradiating the mixture of photosensitizer MTCN-3 (10 μΜ) and DCFH (10 μΜ) with 10 mW cm -2 of white light (500-600 nm) for different times, the spectrum was recorded immediately. The fluorescence of DCFH was excited at 488 nm, which showed that MTCN-3 had excellent active oxygen generation ability.

[0062] As shown in Figure 5, the photothermal behavior of MTCN-3 was evaluated using a 520 nm laser. The solution of MTCN-3 (100 μΜ) was irradiated with 520 nm, 0.5 W cm -2 of light, and the temperature gradually increased from 25 °C to a maximum temperature of 93.4 °C. The photothermal conversion efficiency (η) of MTCN-3 was 24.02%, which indicated that MTCN-3 had a photothermal effect in vitro.

[0063] Example 3: Preparation of the anti-tumor drug composition M@P for photo-immune synergistic therapy

[0064] (1) Preparation of the anti-tumor drug composition M@P for photo-immune synergistic therapy constructed by the aggregation-induced emission photosensitizer MTCN-3

[0065] 3 mg of MTCN-3 was dissolved in 0.5 mL of DMSO, and 1 mg of Poly(I:C) was dissolved in 1 mL of deionized water. Both solutions were added to 18 mL of water at the same time and stirred at 1200 rpm. 12 mg of DSPE-Hyd-PEG-Folate (wherein the molecular weight of PEG is 2000) was dissolved in 0.5 mL of DMSO and ultrasonicated until completely dissolved. Then, it was added to the stirring mixed solution and stirred at 1200 rpm overnight.

[0066] (2) Preparation of the anti-tumor drug composition for photo-immune synergistic therapy constructed by the aggregation-induced emission photosensitizers M1, D1

[0067] 3mg photosensitizer M1 or D1 was dissolved in 0.5mL DMSO, 1mg Poly(I:C) was dissolved in 1mL deionized water. The two solutions were added into 18mL water at the same time and stirred at 1200rpm. 12mg DSPE-Hyd-PEG-Folate (wherein the molecular weight of PEG is 2000) was dissolved in 0.5mL DMSO and ultrasonicated until completely dissolved. Then, it was added into the stirring mixed solution and stirred at 1200rpm overnight.

[0068] The particle size and size distribution of the prepared drug composition M@P were detected by dynamic light scattering method. As shown in Figure 6, the average particle size of the drug composition M@P was about 103.4nm, and its Zeta potential was about 18.85±1.66mV, indicating that it had good nano-assembly ability. In addition, transmission electron microscopy (TEM) showed that the drug composition was uniform spherical. The prepared drug composition maintained good stability in water and PBS (pH 7.4), and the particle size did not change significantly within 7 days.

[0069] Example 4: Evaluation of the targeting property of the drug composition M@P

[0070] Through co-localization analysis, the cell imaging and subcellular organelle distribution of the drug composition M@P were studied. MDA-MB-231 cells were cultured with the drug composition M@P (80μg / mL) in a confocal dish for 4h, and then the culture medium was removed and the cells were washed with PBS. Commercial lysosome green colorant was added and cultured for 10min. Finally, the cells were washed with PBS for 3 times, and analyzed under confocal laser scanning microscope (CLSM). As shown in Figure 7, when the drug composition M@P was co-stained with lysosome colorant, the red fluorescence of the drug composition M@P had good overlap with the green fluorescence of the lysosome colorant, which indicated that the drug composition M@P had the ability to target lysosomes; the Pearson correlation coefficient between the drug composition M@P and the lysosome colorant was 0.84, which further indicated that the drug composition M@P had good lysosome targeting ability.

[0071] Example 5: Evaluation of the phototoxicity of the drug composition M@P on cancer cells

[0072] The killing effect of the drug composition M@P on MDA-MB-231 tumor cells was detected by tetrazolium blue colorimetry (MTT method). The cells were irradiated (520nm, 0.5W cm-2) for 20min, and then the culture medium was removed and 100μL MTT solution (5mg / mL) was added. After 4h, the MTT solution was removed and 100μL DMSO was added. After 12h, the absorbance of the sample was detected at 570nm. As shown in Figure 8, the drug composition M@P had a good killing effect on MDA-MB-231 tumor cells, and the killing effect was better than that of the photosensitizer M1 alone. -2MDA-MB-231 cells were treated with different concentrations (0, 5, 10, 20, 40, 60, 80 pg / mL) of the pharmaceutical composition M@P under dark (M@P group) and light (M@P+L group) conditions for 1 min, and the cytotoxicity was determined (Figure 8). Each group of experiments should be repeated at least 3 times. Under dark conditions, the survival rate of cancer cells and normal cells can still reach more than 80% even if the concentration of the pharmaceutical composition M@P is as high as 80 pg / mL. However, after laser irradiation, the cell viability decreased significantly with the increase of the concentration of the pharmaceutical composition M@P, and the mortality rate reached more than 80%, indicating that the pharmaceutical composition M@P has obvious phototoxicity to MDA-MB-231 cells.

[0073] Example 6: Effect of the pharmaceutical composition M@P on lysosome function

[0074] MDA-MB-231 cells were seeded in confocal culture dishes for 24 h and divided into 3 groups: PBS (control), M@P (dark group), and M@P+L (light group, 520 nm, 0.5 W cm -2 for 5 min), then stained with AO (2 mM) for 15 min, washed with PBS for 3 times, and observed the fluorescence emission of AO in cells by confocal laser scanning microscopy. As shown in Figure 9, the red fluorescence of lysosomes disappeared in the M@P+L group, confirming that the pharmaceutical composition M@P can cause lysosome dysfunction under light.

[0075] Example 7: Effect of the pharmaceutical composition M@P on pyroptosis and ferroptosis

[0076] Confocal microscopy was used to observe the morphological changes of cells before and after treatment with the pharmaceutical composition M@P (520 nm, 0.5 W cm -2 ). As shown in Figure 10, when the cancer cells treated with the pharmaceutical composition M@P were irradiated with light for 3 min, the cells swelled, bubbles appeared on the cell membrane and gradually expanded, showing a clear process of pyroptosis. The markers of ferroptosis are the decrease of GSH content and the increase of LPO content in cells. GSH and MDA detection kits were used to detect the contents of the two in cells to verify whether ferroptosis occurred. MDA-MB-231 cells were treated with PBS, M@P (dark group), and M@P+L (light group, 520 nm, 0.5 W cm -2 for 5 min), then the cells in different groups were collected, repeatedly frozen and thawed at -80 °C and 25 °C for 3 times, centrifuged at 8000 rpm for 10 min, and the supernatant was taken for determination. As shown in Figure 11, the GSH content decreased and the LPO content increased in cells treated with the pharmaceutical composition M@P after light irradiation, verifying the occurrence of ferroptosis.

[0077] Example 8: Evaluation of therapeutic effect of pharmaceutical composition M@P

[0078] Intratumoral injection method was used to establish 4T1 tumor-bearing mouse model (mouse breast cancer tumor) to evaluate the therapeutic effect of pharmaceutical composition M@P. The pharmaceutical composition M@P solution (10 mg / kg; 10 mg per kg of mouse) was injected into 4T1 tumor-bearing BALB / c mice through intravenous injection. As shown in Figure 12, the fluorescence of pharmaceutical composition M@P still gathered at the tumor site 24 h after injection, showing a good tumor targeting effect. To further demonstrate the effect of synergistic photoimmunotherapy on systemic anti-tumor immune response, a bilateral axillary 4T1 tumor model was established in mice to evaluate whether photoimmunotherapy could effectively induce specific systemic anti-tumor effect. The growth of primary tumors (treated) and distant tumors (untreated) was monitored to evaluate the therapeutic effect. As shown in Figure 13, 9 days after photoimmunotherapy, the primary tumor was effectively inhibited, and the growth of distant tumor was also inhibited. Therefore, pharmaceutical composition M@P effectively inhibited distant tumor through photoimmunotherapy by triggering systemic immunity.

Claims

1. An aggregation-induced emission photosensitiser, characterised in that, has the following structure:

2. An antitumor pharmaceutical composition, characterized by, co-assembled by electrostatic interaction with an immune agonist Poly(I:C) and modified by an amphiphilic PEG modifier agent selected from the group consisting of any one of the following structures, 3. The pharmaceutical composition of claim 2, wherein, The average particle size is 100-200 nm.

4. The pharmaceutical composition of claim 2, wherein, The amphiphilic PEG modifier is selected from one or two of DSPE-PEG, DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 200-20000.

5. The pharmaceutical composition of claim 2, wherein, The mass ratio of the aggregation-induced emission photosensitizer to the immune stimulator Poly(I:C) is (1-5):

1.

6. The pharmaceutical composition of claim 2, wherein, The mass ratio of the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C) to the amphiphilic PEG modifier is 1:(3-10).

7. A method of preparing the aggregation-induced emission photosensitizers of claim 1, characterized in that, It is prepared by the reaction of 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridine-1-ium and 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carboxaldehyde in an inert gas catalyzed by piperidine.

8. A method of preparing the pharmaceutical composition of claim 2, characterized by, The method comprises the following steps: (1) respectively dissolve the aggregation-induced emission photosensitizer and the immune stimulator Poly(I:C), and add both solutions into water to spontaneously form co-assembled nanoparticles under stirring; (2) dissolve the amphiphilic PEG modifier and add it into the co-assembled nanoparticles prepared in step (1), and remove the solvent to obtain the pharmaceutical composition.

9. Use of the aggregation-induced emission photosensitizer of claim 1 or the pharmaceutical composition of claim 2 in the preparation of a drug for inducing pyroptosis, ferroptosis, lysosomal dysfunction of tumor cells.

10. Use of the aggregation-induced emission photosensitizer of claim 1 or the pharmaceutical composition of claim 2 in the preparation of a drug for activating anti-tumor immune response, inhibiting tumor growth, and inhibiting tumor metastasis.

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