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

Abstract

The application discloses an aggregation-induced emission photosensitizer, a medicine composition constructed by the aggregation-induced emission photosensitizer, a preparation method and application of the aggregation-induced emission photositizer. The aggregation-induced emission photosensitizer MTCN-3 has excellent active oxygen generation capacity and photothermal effect, and plays a double effect of inducing pyroptosis and ferroptosis of tumor cells. The aggregation-induced emission photosensitizer can also play a light immune synergistic curative effect with an immune stimulant Poly(I:C), the co-assembled medicine composition has a near-infrared aggregation-induced emission effect, has a good subcellular distribution and has an active targeting selectivity to tumor cells; the tumor is eliminated and the tumor metastasis is prevented by preventing immune escape, the growth of a primary tumor and a distal tumor is inhibited, and a systemic anti-tumor immune effect is played; the aggregation-induced emission photosensitizer has good stability, low toxic and side effects, and has a wide clinical application prospect.

Description

Technical Field

[0001] The present invention relates to an aggregation-induced luminescence photosensitizer and a pharmaceutical composition, preparation method and application thereof, and in particular to an aggregation-induced luminescence photosensitizer capable of inducing cell pyroptosis and ferroptosis and a pharmaceutical composition, preparation method and application thereof with synergistic photoimmunological efficacy. Background Art

[0002] Immunogenic cell death (ICD) is an immune-stimulating response induced by necrosis or programmed cell death. During ICD, cells produce a series of signaling molecules called damage-associated molecular patterns (DAMPs). DAMPs primarily include calreticulin exposed on the cell surface, highly mobile proteins diffusing out of the nucleus, ATP released by cells, and heat shock proteins. These DAMPs act as natural immune agonists, binding to pattern recognition receptors on the surface of dendritic cells, promoting dendritic cell maturation and initiating a series of cellular responses, ultimately activating both innate and adaptive immune responses.

[0003] Ferroptosis and pyroptosis are two typical immunogenic cell death mechanisms that can activate or alter the immune system. The essence of ferroptosis is the depletion of glutathione, the decreased activity of glutathione peroxidase (GPX4), and the inability of lipid oxides to be metabolized through the glutathione reductase reaction catalyzed by GPX4. Pyroptosis is a new type of lytic cell death, activated by the gasdermin protein family (GSDM) (including GSDMD, GSDME, etc.). The cleaved GSDM will pore in the cell membrane, thereby destroying the cell membrane and releasing a large amount of pro-inflammatory factors and intracellular substances.

[0004] Pyroptosis- and ferroptosis-mediated immunotherapy primarily relies on chemotherapy drugs, but they suffer from severe toxic side effects. Poly(I:C), an artificial interferon inducer, can induce apoptosis and immunogenic cell death (ICD), but it requires high dosage, low immune response rates, and a narrow therapeutic window. Therefore, the clinical application of these immunotherapeutic drugs is limited and cannot meet the demand for highly effective, low-toxic treatments. Summary of the Invention

[0005] Purpose of the invention: The first purpose of the present invention is to provide an aggregation-induced luminescence photosensitizer that targets and induces pyroptosis and ferroptosis in tumor cells. The second purpose is to provide a pharmaceutical composition constructed with the aggregation-induced luminescence photosensitizer and having a synergistic photoimmunological therapeutic effect. The third purpose is to provide a preparation method thereof. The fourth purpose is to provide their pharmaceutical applications.

[0006] Technical solution: The aggregation-induced emission photosensitizer of the present invention has the following structure:

[0007]

[0008] Phototherapy is an interventional treatment method for tumors with adjustable light damage, non-invasiveness and low side effects. Photodynamic therapy (PDT) and photothermal therapy (PTT) are two reasonable ways to achieve phototherapy. In the photodynamic process, the photosensitizer produces cytotoxic reactive oxygen species (ROS) under light; while in the photothermal process, light energy is effectively converted into thermal energy by the photothermal agent. The reactive oxygen species and photothermal energy produced by the photosensitizer targeting lysosomes make the lysosomal membrane permeabilized and the lysosomal function abnormal, thereby further triggering ICD. The AIE photosensitizer designed in the present invention has shown great advantages in fluorescence imaging and photodynamic therapy due to its high brightness in the aggregated state, good photostability and large Stokes shift, and has the potential for the design and application of lysosome-targeted drugs.

[0009] The anti-tumor pharmaceutical composition of the present invention is obtained by co-assembling an aggregation-induced luminescence photosensitizer selected from any of the following structures and an immune agonist Poly(I:C) through electrostatic interaction and modifying it with an amphiphilic PEG modifier,

[0010]

[0011] The pharmaceutical composition designed by the present invention can target tumors and be distributed in the lysosomes within the cells. Under illumination, the photosensitizer in the pharmaceutical composition can generate a large amount of ROS and heat, thereby causing lysosomal dysfunction in the cells, increasing lysosomal membrane permeability, and releasing substances within the lysosomes into the cytoplasm, inducing pyroptosis and ferroptosis in the cells. At the same time, the photosensitizer promotes the release of cell contents and inflammatory cytokines by inducing cell pyroptosis, and can synergize with the immune agonist Poly (I: C), thereby activating anti-tumor immune responses, 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 by the present invention an effective, low-side effect, targeted dual-induction drug for tumor cell pyroptosis and ferroptosis.

[0012] Preferably, the average particle size of the pharmaceutical composition of the present invention is 100 to 200 nm.

[0013] More preferably, the average particle size of the pharmaceutical composition of the present invention is 103.4 nm.

[0014] Preferably, the amphiphilic PEG modifier is selected from one or both of DSPE-PEG and DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 200 to 20,000.

[0015] More preferably, the amphiphilic PEG modifier is DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 2000.

[0016] Preferably, the mass ratio of the aggregation-induced luminescence photosensitizer to the immune agonist Poly (I:C) is (1-5):1.

[0017] More preferably, the mass ratio of the aggregation-induced emission photosensitizer to the immune agonist Poly (I:C) is 3:1.

[0018] Preferably, the mass ratio of the sum of the amounts of the aggregation-induced luminescence photosensitizer and the immune agonist Poly (I:C) to the amphiphilic PEG modifier is 1: (3-10).

[0019] More preferably, the mass ratio of the sum of the amounts of the aggregation-induced luminescence photosensitizer and the immune agonist Poly (I:C) to the amphiphilic PEG modifier is 1:3.

[0020] The aggregation-induced luminescence photosensitizer of the present invention is prepared by reacting 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridin-1-ium and 5-(4-(di-p-toluylamino)phenyl)thiophene-2-carboxaldehyde in an inert gas via piperidine catalysis.

[0021] Preferably, the molar ratio of 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridin-1-ium, 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carbaldehyde and piperidine catalyst is 1:1:0.05.

[0022] Preferably, the reaction solvent of the reaction is anhydrous ethanol.

[0023] Preferably, the reaction is a reflux reaction.

[0024] The preparation method of the pharmaceutical composition of the present invention comprises the following steps:

[0025] (1) The aggregation-induced luminescence photosensitizer and the immune agonist Poly(I:C) were dissolved separately, and both solutions were added to water, and co-assembled nanoparticles were spontaneously formed under stirring;

[0026] (2) dissolving the amphiphilic PEG modifier and adding it to the co-assembled nanoparticles prepared in step (1), and removing the solvent to obtain the pharmaceutical composition.

[0027] More preferably, in step (1), the aggregation-induced luminescence photosensitizer is dissolved in DMSO, and the immune agonist Poly(I:C) is dissolved in water.

[0028] More preferably, in step (1), the solution of the aggregation-induced luminescence photosensitizer and the immune agonist Poly (I:C) spontaneously forms co-assembled nanoparticles at a stirring speed of 1200 rpm.

[0029] More preferably, in step (2), the amphiphilic PEG modifier is dissolved in DMSO.

[0030] More preferably, the solvent is removed by freeze-drying in step (2).

[0031] More preferably, the volume ratio of DMSO used in steps (1) and (2) is 1:1.

[0032] More preferably, the volume ratio of water used in steps (1) and (2) is 1:18.

[0033] More preferably, the total volume ratio of DMSO to water used in steps (1) and (2) is 1:19.

[0034] The aggregation-induced luminescence photosensitizer of the present invention or the pharmaceutical composition constructed therefrom is used in the preparation of drugs for inducing pyroptosis, ferroptosis, and lysosomal dysfunction in tumor cells.

[0035] The aggregation-induced luminescence photosensitizer of the present invention or the pharmaceutical composition constructed therefrom is used in the preparation of drugs for activating anti-tumor immune responses, inhibiting tumor growth, and inhibiting tumor metastasis.

[0036] Preferably, the tumor is a breast cancer tumor.

[0037] The aggregation-induced luminescence photosensitizer of the present invention or the pharmaceutical composition constructed therefrom eliminates tumors and prevents tumor metastasis by preventing immune escape, can effectively inhibit the growth of primary tumors and distal tumors, and exert systemic anti-tumor immune effects.

[0038] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0039] The aggregation-induced luminescence photosensitizer designed in this invention has excellent reactive oxygen species generation capacity and photothermal effect, exerting the dual effects of inducing pyroptosis and ferroptosis in tumor cells. This aggregation-induced luminescence photosensitizer can also exert synergistic photoimmunological therapeutic effects with the immune agonist Poly(I:C). The co-assembled pharmaceutical composition has a near-infrared induced aggregation emission effect, good subcellular distribution, and active targeting selectivity for tumor cells. It eliminates tumors and prevents tumor metastasis by preventing immune escape, inhibits the growth of primary and distal tumors, and exerts systemic anti-tumor immunity. It has good stability, low toxicity and side effects, and has broad clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The synthetic route of aggregation-induced emission photosensitizer MTCN-3;

[0041] Figure 2UV spectrum (a) and fluorescence spectrum (b) of MTCN-3 (10 μM) in DMSO solution at an excitation wavelength of 480 nm;

[0042] Figure 3 MTCN-3 at different water fractions (f t ) in a DMSO / toluene mixture;

[0043] Figure 4 This is the result of MTCN-3 producing reactive oxygen species (ROS) in aqueous solution;

[0044] Figure 5 For MTCN-3, laser (0.5W cm -2 ) Temperature rise results under irradiation;

[0045] Figure 6 Particle size distribution of M@P (a) and stability results in water and PBS (b);

[0046] Figure 7 Confocal images of lysosome stain Lysosome Green and M@P (scale bar: 50 nm);

[0047] Figure 8 Under light (520nm, 0.5W cm -2 , 1 min) and dark conditions, the cytotoxicity results of 4T1 cells after treatment with different concentrations of M@P (M@P for dark conditions, M@P+L for light conditions);

[0048] Figure 9 Confocal images of AO staining (scale bar, 50 nm);

[0049] Figure 10 Cell membrane expansion and content release before and after M@P treatment (scale bar: 20 nm);

[0050] Figure 11 The results show the changes of intracellular GSH (a) and MDA (b) before and after M@P treatment;

[0051] Figure 12 In vivo fluorescence images of 4T1 tumor-bearing mice after intravenous injection of M@P;

[0052] Figure 13 The primary tumor and distal tumor of 4T1 tumor-bearing mice after laser irradiation (n=3). DETAILED DESCRIPTION

[0053] The technical solution of the present invention will be further described below in conjunction with embodiments.

[0054] Example 1: Preparation of aggregation-induced emission photosensitizer MTCN-3

[0055] (1) Preparation of 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carboxaldehyde

[0056] 4-Bromo-N,N-di-p-tolylaniline (3.38 g, 9.6 mmol), 5-formyl-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) mixed solution and all air was excluded. After the reaction mixture was cooled to room temperature, it was 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 obtain 1.53 g of a yellow solid in a yield of 62.5%. The nuclear magnetic resonance 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).

[0057] (2) Preparation of the compound 3-(cyanomethyl)-1-(3-(trimethylamino)propyl)pyridin-1-ium

[0058] (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 was removed by rotary evaporation, washed with ether, and the precipitate was filtered to obtain compound d as a white solid in a 74% yield. Its 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).

[0059] (3) Preparation of aggregation-induced luminescence photosensitizer MTCN-3

[0060] like Figure 1 As shown, the compound 5-(4-(di-p-tolylamino)phenyl)thiophene-2-carbaldehyde (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), piperidine (4.26 mg, 0.05 mmol) was added as a catalyst, and the reaction was carried out in a nitrogen atmosphere (80° C.) for 7 h. It was then cooled to room temperature and the solvent was removed by evaporation under reduced pressure. The crude product was purified by silica gel column chromatography using DCM / MeOH (10 / 1, v / v) as the eluent to obtain a purple solid product in a yield of 50.23%. The structural characterization is as follows: 1 H NMR(600MHz,DMSO-d6)δ9.66(s,1H),9.13(d,J=5.9Hz,1H),8.89(s,1H),8.83(d,J=8.4H z,1H),8.29(dd,J=8.0,6.2Hz,1H),7.93(d,J=4.0Hz,1H),7.65(t,J=6.1Hz,3H),7.18(d ,J=8.2Hz,4H),7.01(d,J=8.2Hz,4H),6.92(d,J=8.7Hz,2H),4.79(dd,J=13.2,6.0Hz,2H ),3.48(dd,J=10.6,5.5Hz,2H),3.13(s,9H),2.57(dt,J=15.3,7.7Hz,2H),2.29(s,6H). 13 C NMR (151MHz, DMSO) δ152.27,149.39,144.32,143.79,142.37,141.58,141.54,140.16,135.13,134.82,133.98,130.75,12 8.65,127.72,125.73,124.97,124.10,120.93,117.35,98.08,62.25,58.59,53.06,24.87,20.93.HR-ESI-MS:calcd.forC 38 H 40 Br2N4S:m / z:1 / 2[M-2Br] 2+ :292.1482,found:m / z 292.14551.

[0061] Example 2: Evaluation of the photophysical properties of the aggregation-induced emission photosensitizer MTCN-3

[0062] like Figure 2As shown in Figure 2, the DMSO solution of MTCN-3 (10 μM) showed a main absorption peak at 499 nm and a fluorescence emission peak at 760 nm. Figure 3 As shown, MTCN-3 exhibits significant aggregation-induced emission properties in a mixed solution of DMSO and toluene.

[0063] like Figure 4 As shown, dichlorofluorescein (DCFH) was used as ROS indicator and 10 mW cm -2 The spectra were recorded immediately after irradiating a mixture of photosensitizer MTCN-3 (10 μM) and DCFH (10 μM) with white light (500-600 nm) for different periods of time. The fluorescence of DCFH is excited at 488 nm, demonstrating the excellent ROS generation capacity of MTCN-3.

[0064] like Figure 5 As shown, the photothermal behavior of MTCN-3 was evaluated using a 520 nm laser. -2 When the MTCN-3 solution (100 μM) was irradiated with light, the temperature gradually increased from 25°C to a maximum of 93.4°C. The photothermal conversion efficiency (η) of MTCN-3 was 24.02%, indicating that MTCN-3 has a photothermal effect in vitro.

[0065] Example 3: Preparation of anti-tumor drug composition M@P for photoimmunotherapy

[0066] (1) Preparation of the anti-tumor drug composition M@P for photoimmunotherapy constructed with the aggregation-induced luminescence photosensitizer MTCN-3

[0067] Dissolve 3 mg of MTCN-3 in 0.5 mL of DMSO and 1 mg of Poly(I:C) in 1 mL of deionized water. Add both solutions simultaneously to 18 mL of water and stir at 1200 rpm. Dissolve 12 mg of DSPE-Hyd-PEG-Folate (PEG with a molecular weight of 2000) in 0.5 mL of DMSO and sonicate until completely dissolved. Then, add the mixture to the stirring solution and stir at 1200 rpm overnight.

[0068] (2) Preparation of anti-tumor drug composition for photoimmunotherapy constructed with aggregation-induced luminescence photosensitizers M1 and D1

[0069] Dissolve 3 mg of photosensitizer M1 or D1 in 0.5 mL of DMSO and 1 mg of Poly(I:C) in 1 mL of deionized water. Add both solutions simultaneously to 18 mL of water and stir at 1200 rpm. Dissolve 12 mg of DSPE-Hyd-PEG-Folate (PEG with a molecular weight of 2000) in 0.5 mL of DMSO and sonicate until completely dissolved. Then, add the solution to the stirring mixture and stir at 1200 rpm overnight.

[0070] The particle size and particle size distribution of the prepared pharmaceutical composition M@P were detected by dynamic light scattering. Figure 6 As shown, the average particle size of the pharmaceutical composition M@P was approximately 103.4 nm, and its zeta potential was approximately 18.85 ± 1.66 mV, indicating good nanoassembly capability. Furthermore, transmission electron microscopy (TEM) revealed that the pharmaceutical composition was uniformly spherical. The prepared pharmaceutical composition maintained good stability in water and PBS (pH 7.4), with no significant change in particle size over 7 days.

[0071] Example 4: Targeting evaluation of the pharmaceutical composition M@P

[0072] The cellular imaging and subcellular organelle distribution of the drug composition M@P were studied by colocalization analysis. MDA-MB-231 cells were cultured with the drug composition M@P (80 μg / mL) in a confocal culture dish for 4 hours, and then the culture medium was removed and the cells were washed with PBS. A commercial lysosomal green stain was then added and incubated for 10 minutes. Finally, the cells were washed 3 times with PBS and analyzed under a confocal laser scanning microscope (CLSM). Figure 7 As shown, when the pharmaceutical composition M@P is co-stained with a lysosomal stain, the red fluorescence of the pharmaceutical composition M@P overlaps well with the green fluorescence of the lysosomal stain, indicating that the pharmaceutical composition M@P has the ability to target lysosomes; the Pearson correlation coefficient between the pharmaceutical composition M@P and the lysosomal stain is 0.84, further indicating that the pharmaceutical composition M@P has good lysosomal targeting ability.

[0073] Example 5: Evaluation of the phototoxicity of the pharmaceutical composition M@P on cancer cells

[0074] The killing effect of the drug composition M@P on MDA-MB-231 tumor cells was detected by the tetrazolium nitro blue colorimetric method (MTT method). -2 , 1 min) (M@P+L group) and dark (M@P group), MDA-MB-231 cells were treated with different concentrations (0, 5, 10, 20, 40, 60, 80 μg / mL) of the pharmaceutical composition M@P, and their cytotoxicity was determined ( Figure 8 Each experiment was repeated at least three times. Under dark conditions, even at concentrations as high as 80 μg / mL, the survival rate of both cancer cells and normal cells remained above 80%. However, after laser irradiation, cell viability decreased significantly with increasing concentrations of the M@P composition, with the mortality rate exceeding 80%, indicating significant phototoxicity of the M@P composition to MDA-MB-231 cells.

[0075] Example 6: Effect of the pharmaceutical composition M@P on cellular lysosomal function

[0076] MDA-MB-231 cells were inoculated into confocal culture dishes and cultured for 24 h. The cells were divided into PBS (control), M@P (dark group), M@P+L (light group, 520 nm, 0.5 W cm -2 The cells were treated with AO (2 μM) for 15 min and washed with PBS three times. The fluorescence emission of AO in the cells was observed using a confocal laser scanning microscope. Figure 9 As shown, the red fluorescence of lysosomes disappeared in the M@P+L group, confirming that the pharmaceutical composition M@P can cause lysosomal dysfunction under light.

[0077] Example 7: Effects of the pharmaceutical composition M@P on cell pyroptosis and ferroptosis

[0078] Confocal microscopy was used to observe the morphological changes of cells before and after treatment with the drug composition M@P (520 nm, 0.5 W cm -2 ).like Figure 10 As shown in the figure, when the cancer cells treated with the pharmaceutical composition M@P were irradiated with light for 3 minutes, the cells swelled, bubbles appeared on the cell membrane and gradually expanded, showing an obvious process of cell pyroptosis. The hallmark of ferroptosis is a decrease in intracellular GSH content and an increase in LPO content. GSH and MDA detection kits were used to detect the content of both in the cells to verify whether ferroptosis occurred. PBS, M@P (dark group) and M@P+L (light group, 520nm, 0.5Wcm -2 , 5min) to treat MDA-MB-231 cells, and then collect cells from different groups, freeze-thaw three times at -80℃ and 25℃, centrifuge at 8000rpm for 10min, and take the supernatant for determination. Figure 11 As shown, the GSH content of cells treated with the drug composition M@P decreased and the LPO content increased after illumination, verifying the occurrence of ferroptosis.

[0079] Example 8: Evaluation of the therapeutic effect of the pharmaceutical composition M@P

[0080] The 4T1 tumor-bearing mouse model (mouse breast cancer tumor) was established by intratumoral injection to evaluate the therapeutic effect of the pharmaceutical composition M@P. The pharmaceutical composition M@P solution (10 mg / kg; 10 mg per kg mouse) was injected intravenously into 4T1 tumor-bearing BALB / c mice. Figure 12 As shown in the figure, the fluorescence of the pharmaceutical composition M@P still accumulated at the tumor site 24 hours 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 can effectively induce specific systemic anti-tumor effects. The growth of primary tumors (treated) and distant tumors (untreated) was monitored to evaluate the therapeutic effect. Figure 13 As shown, 9 days after photoimmunotherapy, the primary tumor was effectively suppressed, and the growth of distant tumors was also inhibited. Therefore, the pharmaceutical composition M@P effectively inhibits distant tumors by triggering systemic immunity through photoimmunotherapy.

Claims

1. An anti-tumor pharmaceutical composition, characterized in that: The aggregation-induced luminescence photosensitizer with the following structure and the immune agonist Poly(I:C) are co-assembled through electrostatic interaction and modified with an amphiphilic PEG modifier. MTCN-3 The amphiphilic PEG modifier is DSPE-Hyd-PEG-Folate, wherein the molecular weight of PEG is 2000; the mass ratio of the aggregation-induced luminescence photosensitizer to the immune agonist Poly(I:C) is (1-5):1; and the mass ratio of the sum of the amounts of the aggregation-induced luminescence photosensitizer and the immune agonist Poly(I:C) to the amphiphilic PEG modifier is 1:(3-10).

2. The pharmaceutical composition according to claim 1, characterized in that The average particle size is 100~200 nm.

3. A method for preparing the pharmaceutical composition according to claim 1, characterized in that: The following steps are involved: (1) Dissolve the aggregation-induced luminescence photosensitizer and the immune agonist Poly(I:C) separately, add both solutions into water, and spontaneously form co-assembled nanoparticles under stirring; (2) dissolving the amphiphilic PEG modifier and adding it to the co-assembled nanoparticles prepared in step (1), and removing the solvent to obtain the pharmaceutical composition.

4. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for inducing pyroptosis, ferroptosis, and lysosomal dysfunction in tumor cells.

5. Use of the pharmaceutical composition according to claim 1 in the preparation of a drug for activating anti-tumor immune response, inhibiting tumor growth, and inhibiting tumor metastasis.

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