ICD inducer Ru (II) complex with capacity of photodynamically breaking DNA as well as preparation method and application of ICD inducer Ru (II) complex

Abstract

The invention discloses an ICD inducer Ru (II) complex with photodynamic DNA breaking capacity and a preparation method and application thereof, and relates to the field of antitumor drugs, the structural formula of the Ru (II) complex is [Ru (dip) 2 (L)] (PF6) 2, and the Ru (II) complex can enter cell nucleus to break DNA and activate pyroptosis and cGAS-STING pathway mediated immunogenic death when applied to photodynamic therapy, so that the antitumor immune effect of a body is improved. In an in-vivo experiment, the vaccine group has a good effect on immune cells of a tumor microenvironment in a mouse, and the three-needle vaccine has a better anti-tumor immune effect; hamp, Hamp; the E staining result shows that the vaccine has no obvious pathological injury on the main organs of the mouse, and the vaccine is proved to have better biological safety.

Description

Technical Field

[0001] The present invention belongs to the technical field of anti-tumor drugs, and in particular relates to an ICD inducer Ru(II) complex with the ability to photodynamically break DNA, and a preparation method and application thereof. Background Art

[0002] Photodynamic therapy (PDT) is a non-invasive cancer treatment whose core mechanism is to activate photosensitizers through light energy to exert its therapeutic effects. The synergistic effect of light energy, oxygen, and photosensitizers (PSs) generates large amounts of reactive oxygen species (ROS) within tumor cells. These ROS interact with intracellular biomacromolecules such as proteins, acting on key targets such as mitochondria, the endoplasmic reticulum, and the nucleus, disrupting the normal function and organelle structure of tumor cells, thereby inducing cell death. Compared with traditional cancer treatments such as surgery, chemotherapy, and radiotherapy, PDT offers significant advantages, including less physical trauma, fewer toxic side effects, and greater spatiotemporal precision. These advantages have made PDT a promising approach to improving therapeutic efficacy and mitigating adverse reactions. In recent years, it has garnered increasing attention and has gradually become a cutting-edge field in oncology research.

[0003] Pyroptosis is a novel form of cell death that relies on the gasdermin family of proteins to form pores in the cell membrane and stimulate the formation of inflammasomes. Within the tumor cell environment, pyroptosis can alter the tumor microenvironment by releasing specific inflammatory mediators and cellular contents, making it easier for the immune system to recognize and eliminate these altered cancer cells. PDT can induce pyroptosis in HeLa cells, thereby increasing their susceptibility to the immune system. This process, through endoplasmic reticulum stress, leads to the release of immunostimulatory factors and damage-associated molecular patterns (DAMPs), enhancing the inflammatory response. It also helps activate the innate immune response and enhance immunogenic cell death (ICD).

[0004] ICD is a process of cancer cell death triggered by certain chemotherapeutic drugs, physical chemotherapy, photodynamic therapy, and radiotherapy, which exposes or releases specific molecules known as DAMPs. One key molecule is calreticulin (CRT), which is originally localized to the endoplasmic reticulum but, during ICD, translocates to the cell membrane. This shift is considered a crucial "signal," as its exposure to the cell's exterior helps attract antigen-presenting cells. High-mobility group protein B1 (HMGB1) is released to the cell's exterior, acting as a "danger signal" to stimulate immune cell responses. ATP is also released to the cell's exterior, acting as a chemokine to attract immune cells and activate inflammasomes. These molecules, considered "danger signals," can promote tumor-specific immune responses, thereby directly participating in the elimination of cancer cells and enhancing the long-term effects of anticancer therapy by activating the immune system.

[0005] The genetic information and signaling factors carried by the cell nucleus play a key role in cell growth and proliferation and are considered the primary targets of many anti-tumor drugs. Through photodynamic therapy, photosensitizers can enhance the effect of inducing tumor cell death, thereby improving the body's ability to fight tumors.

[0006] Ru(II) complexes are considered candidates for the next generation of photosensitizers due to their excellent photochemical properties, tunable cellular uptake, and low dark toxicity. However, existing ruthenium complexes still have the following limitations: (1) most remain at the apoptosis / necrosis induction stage; (2) they lack clear nuclear-targeted DNA breakage capabilities; (3) their activation mechanisms for immune pathways such as cGAS-STING are unclear; and (4) their in vivo anti-tumor immune effects lack systematic verification. Summary of the Invention

[0007] To address the above technical problems, the present invention proposes a Ru(II) complex, an ICD inducer with the ability to photodynamically fragment DNA, as well as its preparation method and application. This Ru(II) complex exhibits anti-tumor immune activity. Research into the anti-tumor activity of this Ru(II) complex has revealed that it exhibits strong anti-tumor activity against tumor cells and significant phototoxicity against HeLa cells. It also induces pyroptosis in HeLa cells and enhances their immunogenic death via the cGAS-STING pathway.

[0008] To achieve the above objectives, the present invention provides an ICD inducer Ru(II) complex having the ability to photodynamically break DNA, having the following structure: .

[0009] The present invention also provides a method for preparing the above-mentioned ICD inducer Ru(II) complex having the ability to photodynamically break DNA, comprising the following steps: Under argon protection, Ru(dip)2Cl2 and ligand L are refluxed in an organic solvent to obtain a Ru(II) complex; The ligand L is 2-(2-(4-((4-chlorophenyl)(phenyl)methyl)piperazine-1-yl)ethoxy)-N-(1,10-phenanthroline-5-yl)acetamide; dip is 4,7-diphenyl-1,10-phenanthroline.

[0010] Furthermore, the organic solvent is anhydrous ethanol.

[0011] Furthermore, the reflux reaction temperature is 70-80° C., and the reaction time is 1-3 hours.

[0012] Furthermore, after the reflux reaction is completed, the method further comprises: adding an excess of saturated NH4PF6 solution, filtering and drying the precipitate, and purifying.

[0013] The present invention also provides an anti-tumor pharmaceutical composition comprising the aforementioned ICD inducer Ru(II) complex having the ability to photodynamically break DNA and a pharmaceutically acceptable carrier.

[0014] Furthermore, the anti-tumor drug composition is a photodynamic therapy agent, and the anti-tumor drug is used to induce immunogenic death of tumor cells.

[0015] The present invention also provides a use of the above-mentioned ICD inducer Ru(II) complex having the ability to photodynamically break DNA in the preparation of anti-tumor drugs.

[0016] Furthermore, the anti-tumor drug is used to induce immunogenic death of tumor cells; and the tumor is an in situ solid tumor.

[0017] Furthermore, the tumor cells are mouse breast cancer cells or human cervical cancer cells.

[0018] The present invention also provides a use of the above-mentioned ICD inducer Ru(II) complex having the ability to photodynamically cleave DNA in the preparation of an anti-tumor vaccine, wherein the anti-tumor vaccine is prepared by the following steps: (a) incubating tumor cells with the ICD inducer Ru(II) complex; (b) Irradiation with a light source of 400-500 nm wavelength; (c) washing and resuspending the cells to obtain the anti-tumor vaccine; The tumor cells are mouse breast cancer cells or human cervical cancer cells.

[0019] Compared with the prior art, the present invention has the following advantages and technical effects: This invention provides a ruthenium(II) complex with an o-phenanthroline derivative as the primary ligand, exhibiting excellent anti-tumor immune activity. Because it contains metal ions and carries an inherent charge, it enhances in vivo penetration and retention compared to traditional organic small molecules. Furthermore, the metal complex possesses a multi-coordinate configuration, allowing for modification with different ligands to achieve superior biological activity.

[0020] The ruthenium (II) complex provided by the present invention is not only a metal ruthenium (II) complex with good anti-tumor activity, but also, under the action of photodynamic therapy, Ru1 induces mitochondrial damage (such as decreased membrane potential, ROS burst) and endoplasmic reticulum stress (calcium ion disorder) through a cascade delivery mechanism from mitochondria and endoplasmic reticulum to the cell nucleus, ultimately entering the cell nucleus to break DNA, activating pyroptosis and immunogenic death mediated by the cGAS-STING pathway, thereby enhancing the body's anti-tumor immune effect. In in vivo experiments, the vaccine group had a good effect on the immune cells in the tumor microenvironment in mice, and the three-dose vaccine had a better anti-tumor immune effect. The results of H&E staining showed that the vaccine did not cause significant pathological damage to the major organs of mice, confirming its good biosafety. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a diagram showing the effect of the complex Ru1 entering the HeLa cell nucleus in Example 3; Figure 2 Graphs showing the photocleavage effect of the complex Ru1 on DNA in Example 4; (A) DNA gel electrophoresis of Ru1 under dark conditions; (B) DNA gel electrophoresis of Ru1 under light conditions; Figure 3 The effect diagram of the effect of the complex Ru1 on the HeLa cell cycle in Example 5; wherein, (A) flow cytometric analysis results of the complex Ru1 on the HeLa cell cycle distribution; (B) histogram of flow cytometric analysis data processing; (C) Western Blot analysis of the complex Ru1 on HeLa cell cycle-related proteins; Figure 4 Graphs showing the effect of the complex Ru1 in Example 6 on inducing γ-H2AX expression in HeLa cells; (A) Confocal imaging of Ru1 without the addition of the antioxidant NAC; (B) Confocal imaging of Ru1 with the addition of the antioxidant NAC; Figure 5 This is a diagram showing the effect of the complex Ru1 in Example 7 on the expression of cGAS-STING pathway-related proteins in HeLa cells under light irradiation; Figure 6 Figures depict the immune effects of 4T1 cells treated with the complex Ru1 on subcutaneously implanted tumors in female BALB / c mice inoculated with the complex Ru1 in Example 8. (A) Statistical graph showing changes in mouse weight; (B) Statistical graph showing changes in mouse tumor volume; (C) Schematic diagram showing tumor size on day 30 after 4T1 cell injection; (D) H&E staining of sections of major mouse organs. Note: Cisplatin was used as a control drug. DETAILED DESCRIPTION

[0022] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0023] All raw materials of the present invention are not particularly limited in their sources and can be purchased from the market or prepared according to conventional methods well known to those skilled in the art.

[0024] There is no particular limitation on the purity of all raw materials in the present invention. The present invention preferably uses analytically pure raw materials or raw materials with a purity commonly used in the field of chemical synthesis.

[0025] Example 1 Preparation of Ru(II) Complex Ru1, an ICD Inducer with Photodynamic DNA Fragmentation Capability Under argon, Ru(dip)2Cl2·2H2O (0.33 g, 0.4 mmol) and ligand L (0.22 g, 0.4 mmol) were placed in anhydrous ethanol and refluxed at 78 °C for 2 hours. After completion of the reaction, the solvent was mostly removed by rotary evaporation under reduced pressure to obtain a crude product. The crude product was isolated and purified by silica gel column chromatography using dichloromethane and methanol (10:1, v / v) as the eluent. The main yellow band was collected and dried to obtain 0.24 g of yellow powder Ru1, with a yield of 36.9%.

[0026] The reaction equation is as follows:

[0027] The structural formula of Ru(II) complex Ru1 is as follows:

[0028] The above complex can be abbreviated as [Ru(dip)2(L)](PF6)2, where ligand L = 2-(2-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-1-yl)ethoxy)-N-(1,10-phenanthroline-5-yl)acetamide, and dip structure = 4,7-diphenyl-1,10-phenanthroline.

[0029] Elemental analysis C 81 H 64 ClF 12 N9O2P2Ru (molecular weight 1621.32), theoretical value: C, 59.98%; H, 3.98%; N, 7.77%; experimental value: C, 59.89%; H, 3.88%; N, 7.89%.

[0030] ESI-MS: Theoretical value: [M-2PF6 - ] 2+ m / z = 665.7, experimental value: [M-2PF6 - ] 2+ m / z = 666.0.

[0031] Example 2 In vitro antitumor activity test of Ru(II) complexes The MTT method was used to evaluate the cytotoxicity of ruthenium(II) complexes.

[0032] (1) Cell culture: Human cervical cancer cells (HeLa) and mouse breast cancer cells (4T1) were seeded in 96-well plates (density 5×10 3 / well) and cultured at 37°C, 5% CO2 for 24 hours.

[0033] (2) Medication treatment: Dark toxicity group: 100 μL of culture medium containing different concentrations of Ru1 (0.001–10 μM) was added and cultured in the dark for 48 h.

[0034] Phototoxicity group: Ru1 was added and cultured for 4 hours, and then a 450 nm LED light source (40 mW / cm 2 ) for 10 minutes (total light energy 36 J / cm 2 ) and continue culturing for 44 hours.

[0035] (3) Detection: Add MTT solution (10 μL, 5 mg / mL) to each well. After incubation for 4 hours, add 150 μL of DMSO to dissolve the formazan, and measure the absorbance (OD) at 595 nm with a microplate reader.

[0036] The phototoxicity index (PI) reflects the median lethal concentration (IC50) of a drug under dark and light conditions. 50 The cell survival rate of the reference group without the addition of the metal ruthenium (II) complex was set to 100%, and the cell survival rate was calculated according to the following formula.

[0037] Cell survival rate % = (average OD of the treatment group) / (average OD of the reference group) × 100% The test results are shown in Table 1 below: Table 1 IC of Ru(II) complexes 50 value a (μΜ)

[0038] Note: IC 50 a Refers to the concentration of complex required to kill 50% of tumor cells. The drug action time is 48 hours. The data were obtained by at least three repeated experiments and are expressed as mean ± standard deviation. Example 3 Nuclear localization of Ru1 (confocal imaging) (1) HeLa cells were seeded in confocal culture dishes (density 3×10 4 / dish), and Ru1 (0.5 μM) was added after adhesion.

[0039] (2) Light treatment: 450 nm LED irradiation for 10 minutes and incubation in the dark for 3 hours.

[0040] (3) Staining: Wash three times with PBS and stain the nuclei with DAPI (5 μg / mL) for 15 minutes in the dark.

[0041] (4) Imaging: Laser confocal microscopy observation (Ru1:λ ex =448 nm, λ em =530±20 nm; DAPI: λ ex =405 nm, λ em =460±20 nm).

[0042] like Figure 1 As shown, laser confocal imaging shows that the blue fluorescence representing DAPI is highly consistent with the red fluorescence produced by Ru1. After illumination, the complex Ru1 can enter the nucleus of HeLa cells and be enriched in the nucleolus.

[0043] Example 4 Photodynamic DNA fragmentation of Ru1 (gel electrophoresis) (1) pBR322 DNA (0.1 μg) was mixed with different concentrations of Ru1 (0–0.5 μM) and the mixture was placed in the dark or under 450 nm light (40 mW / cm 2 , 10 minutes) processing.

[0044] (2) Loading: Add the DNA-loading buffer mixture to the wells of 1% agarose gel.

[0045] (3) Electrophoresis: TBE buffer, 50 V, 45 min.

[0046] (4) Staining: The gel was stained with GoldView and imaged by UV.

[0047] like Figure 2 As shown, the experiment revealed that Ru1 can cause DNA breakage under light conditions (Form II relaxed ring, Form III linear), and the degree of breakage increases with the increase of Ru1 concentration (there is almost no breakage in the dark treatment group).

[0048] Example 5 Effect of Ru1 on Cell Cycle (Flow Cytometry + Western Blot) (1) HeLa cells were seeded in 60 mm culture dishes (Corning). When the cells were 70% adherent, Ru1 (0-0.5 μM) was added and then treated with light for 24 hours.

[0049] (2) Cell fixation: Trypsinize and collect cells. Wash cells three times with PBS, add 500 μL PBS to resuspend the cells, and fix them in 70% cold ethanol at 4°C overnight.

[0050] (3) Staining: Stain with RNase (50 μg / mL) and propidium iodide (PI) (50 μg / mL) for 30 minutes in the dark.

[0051] (4) Detection: Flow cytometric analysis of cycle distribution (ModFit software).

[0052] (5) Western blot: Extract total cell protein and detect Cyclin D1 / CDK4 expression.

[0053] like Figure 3 As shown, Figure 3 Specific analysis of A-3B showed that the proportion of G0 / G1 phase cells increased significantly from 62.3% in the blank control group to 79.1%. It should be noted that the proportion of G2 / M phase cells decreased from 31.6% in the blank control group to 14.8%. At the same time, the proportion of S phase cells did not change significantly. The inhibitory effect of the drug on the cell cycle after illumination is directly related to the drug concentration. This observation is consistent with the Western Blot analysis of cycle-related proteins ( Figure 3 The results in Figure 3 (C) are consistent with those in Figure 3. Based on cell cycle distribution analysis, it can be inferred that light exposure causes DNA damage, resulting in a decrease in the number of cells in the S phase. Furthermore, the number of cells in the G2 / M phase, which is associated with DNA repair, decreased with treatment with different drug concentrations. This suggests that Ru1 interferes with the repair process of damaged DNA.

[0054] Example 6 Immunofluorescence staining detection of Ru1-induced γ-H2AX expression in HeLa cells The cell density was 3 × 10 4HeLa cells were seeded / dish in a confocal microplate. After the cells adhered, different concentrations of Ru1 were added and the cells were exposed to light for 10 minutes. The incubation was continued for 24 hours. After following the instructions for the γ-H2AX antibody, rabbit anti-IgG (H+L) antibody was added and incubated in the dark for 1 hour at room temperature. The cells were washed (repeated three times with immunostaining wash buffer) and then DAPI was added for nuclear staining in the dark for 15 minutes. The cells were washed three times with wash buffer and observed under a laser confocal microscope. (DAPI: λ ex = 405 nm, λ em = 460 ± 20 nm; rabbit anti-IgG (H+L): λ ex = 488 nm, λ em = 520 ± 20 nm) The generation of γ-H2AX is one of the earliest responses to DNA damage, and this protein is often used as a key marker for diagnosing DNA damage. When HeLa cells were exposed to different concentrations of Ru1, the intensity of the green γ-H2AX fluorescence signal correlated with the concentration of Ru1 administered, indicating the occurrence of DNA damage within the cells. Importantly, the green fluorescence brightness of γ-H2AX was not completely lost in cells treated with the antioxidant NAC.

[0055] like Figure 4 As shown in the figure, the number of γ-H2AX foci increases with increasing Ru1 concentration, confirming DNA double-strand breaks. The experimental results also reveal that the main mechanism of DNA damage induced by Ru1 may not be solely induced by the production of large amounts of ROS.

[0056] Example 7 Western blot analysis of cGAS-STING pathway-related proteins HeLa cells were seeded in a six-well plate. When the cell density reached 70%, different concentrations of Ru1 were added and cultured in the dark for 4 hours. Then, a 450 nm wavelength LED light source (40 mW·cm -2 ) for 10 minutes (36 J· cm -2 After treating cells with Ru1 complex, perform the following steps according to the product manual: extract total protein, prepare gel and load sample, perform electrophoresis, transfer to membrane, block, incubate with primary antibody, incubate with secondary antibody, and develop.

[0057] The classic signaling pathway for pyroptosis is based on the activation of caspase-1. When tumor cells are stimulated by damage-associated molecular patterns (DAMPs), the inflammasome is activated, recruiting caspase-1 to cleave GSDMD, generating an active N-terminal fragment (GSDMD-NT) and a C-terminal fragment. In the classic caspase-1-dependent pyroptosis mechanism, cells exhibit swelling, membrane disruption, and the release of their contents. Compared to traditional cell death mechanisms, nuclear structure is often preserved during pyroptosis, and the process is accompanied by DNA damage. The cGAS-STING pathway plays a crucial role in mobilizing anti-tumor immune responses. In the field of tumor immunotherapy, cyclic guanosine monophosphate synthase (cGAS) has shown great potential, effectively promoting STING expression. In this process, STING activates nuclear factor κB (NF-κB) through ubiquitination, further enhancing anti-tumor immunity.

[0058] like Figure 5 As shown, Western blot analysis of cell-related protein expression revealed that Caspase-1 was activated, and N-GSDMD expression also increased. As Ru1 concentration increased, cGAS expression also increased, leading to increased p-STING levels and ultimately enhanced p-NF-κB expression. Therefore, it can be concluded that Ru1 in HeLa cells not only triggers Caspase-1-mediated pyroptosis but also activates the cGAS-STING signaling pathway, enhancing anti-tumor immune responses.

[0059] Example 8 In vivo experiments to evaluate the tumor immunity effect in mice (1) Preparation of tumor cell vaccine: Mouse breast cancer 4T1 cells in the logarithmic growth phase were inoculated into a cell culture dish and the cell concentration was adjusted to 1×10 6 Each dish was cultured overnight in a cell culture incubator. Different drug treatments were administered according to the groups: 1) 4T1 cells were co-cultured with CDDP (20 μM) for 36 hours; 2) 4T1 (mouse breast cancer) cells were treated with Ru1 and then illuminated for an additional 36 hours. After drug treatment, the cell suspension was collected into a centrifuge tube, washed three times with PBS, and the cell pellet was collected by centrifugation and resuspended in PBS to obtain a single-cell suspension, which served as the tumor cell vaccine.

[0060] (2) Evaluation of tumor immunity effect in mice: 40 healthy 4- to 5-week-old SPF-grade female BALB / c mice were selected and 4T1 cell suspension was inoculated into the right hind limb of the mice by subcutaneous injection to promote the formation of subcutaneous tumors, so as to evaluate whether the prepared vaccine can effectively stimulate anti-tumor immune response in vivo. The experimental mice were randomly divided into four groups: one group was the control group, another group was treated with CDDP, and there was also a group that received a single injection of Ru1 vaccine and a group that received three injections of Ru1 vaccine. The specific method of animal experiment is as follows: the right groin to the midpoint of the dorsal thigh of the mouse was disinfected and injected subcutaneously in the area of ​​the mouse with a syringe: Control group: only 100 μL PBS was injected; CDDP group: injection of cell suspension treated with CDDP; Ru1 one-shot vaccine group: injection of cell suspension treated with Ru1, and the steps of Ru1 three-shot vaccine group were to repeat the one-shot injection process three times. Seven days later, 30 mice were disinfected from the right groin to the midpoint of the dorsal thigh, and active 4T1 cells were injected subcutaneously in this area. Tumor size and body weight of female BALB / c mice were monitored and recorded every two days. The formula for calculating tumor volume (V) is V = W 2 × L / 2, where W is the width of the tumor and L is the length of the tumor. At the end of the in vivo mouse experiment, H&E staining was performed on the major organs of the remaining mice to observe possible histological changes.

[0061] like Figure 6 As shown, during the entire animal experiment period, the body weight of mice gradually increased without significant fluctuations ( Figure 6 A). The experiment found that the growth rate of tumors in the control group and mice treated with cisplatin (CDDP) was significantly higher than that in the mice vaccinated with the vaccine. On the 30th day after tumor inoculation, tumor samples from each group of mice were collected and observed ( Figure 6 B-6C). Analysis by H&E staining ( Figure 6 D) observed no significant pathological changes in the mice's major organs (including the heart, spleen, liver, lungs, and kidneys). The results indicate that the vaccine did not cause significant tissue damage or pathological changes in these organs, confirming its good biosafety. This study provides critical support for the vaccine's biosafety and effectiveness.

[0062] The Ru(II) complex provided by the present invention can induce HeLa cell pyroptosis by damaging DNA with a low dose of Ru1 photosensitizer and enhance immunogenic death through the cGAS-STING pathway.

[0063] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art shall fall within the scope of protection defined by the claims of the present invention.

Claims

1. An ICD inducer Ru(II) complex having the ability to photodynamically break DNA, characterized in that: Has the following structure: 。 2. A method for preparing the ICD inducer Ru(II) complex having the ability to photodynamically break DNA according to claim 1, characterized in that: The following steps are involved: Under argon protection, Ru(dip)2Cl2 and ligand L are refluxed in an organic solvent to obtain a Ru(II) complex; The ligand L is 2-(2-(4-((4-chlorophenyl)(phenyl)methyl)piperazine-1-yl)ethoxy)-N-(1,10-phenanthroline-5-yl)acetamide; dip is 4,7-diphenyl-1,10-phenanthroline.

3. The preparation method according to claim 2, characterized in that The organic solvent is anhydrous ethanol.

4. The preparation method according to claim 2, characterized in that The reflux reaction temperature is 70-80° C., and the reaction time is 1-3 hours.

5. The preparation method according to claim 2, characterized in that After the reflux reaction is completed, the method further comprises: adding an excess saturated NH4PF6 solution, filtering and drying the precipitate, and purifying.

6. An anti-tumor pharmaceutical composition, characterized in that: The invention comprises the ICD inducer Ru(II) complex having the ability of photodynamic DNA fragmentation according to claim 1 and a pharmaceutically acceptable carrier.

7. The antitumor pharmaceutical composition according to claim 6, characterized in that The anti-tumor drug composition is a photodynamic therapy agent, and the anti-tumor drug is used to induce immunogenic death of tumor cells.

8. Use of the ICD inducer Ru(II) complex having the ability to photodynamically cleave DNA according to claim 1 in the preparation of anti-tumor drugs.

9. The use according to claim 8, characterized in that The anti-tumor drug is used to induce immunogenic death of tumor cells; the tumor cells are mouse breast cancer cells or human cervical cancer cells.

10. Use of the Ru(II) complex, an ICD inducer with the ability to photodynamically cleave DNA, according to claim 1, in the preparation of an anti-tumor vaccine, characterized in that: The anti-tumor vaccine is prepared by the following steps: (a) incubating tumor cells with the ICD inducer Ru(II) complex; (b) Irradiation with a light source of 400-500 nm wavelength; (c) washing and resuspending the cells to obtain the anti-tumor vaccine; The tumor cells are mouse breast cancer cells or human cervical cancer cells.

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