A metal ruthenium complex with aggregation-induced emission properties and its preparation method and application

Abstract

The present invention discloses a metal ruthenium complex with aggregation-induced luminescence properties, as well as a preparation method and application thereof. By introducing an aggregation-induced luminescence molecule and a lysosome-targeting morpholino group into a metal ruthenium complex precursor, a complex with both cell fluorescence imaging and anti-cancer activity is constructed. The complex of the present invention can effectively inhibit the proliferation of human tumor cells and achieve good anti-tumor activity by inducing the production of reactive oxygen species and mitochondrial autophagy in tumor cells. In addition, the complex of the present invention can induce immunogenic death of tumor cells, further activating the body's immune response, achieving a synergistic effect of chemotherapy-immunotherapy, and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to a metal ruthenium complex with aggregation-induced luminescence properties, and also relates to a preparation method and application of the complex. Technical Background

[0002] In recent years, transition metal complexes have emerged as promising anticancer drug candidates due to their exceptional photostability, excellent photophysical properties, and good solubility in aqueous solutions. For example, ruthenium complexes exhibit rapid scavenging, high singlet oxygen generation, long luminescence lifetimes, and large two-photon absorption cross-sections. In 2001, Academician Tang Benzhong discovered a photophysical phenomenon in which molecular aggregates exhibit stronger emission than individual molecules, termed aggregation-induced emission (AIE). Unlike ACQ fluorophores, AIE luminogens (AIEgens) exhibit bright emission in the aggregated state. To date, many AIEgens have been used as photosensitizers (PSs) to promote disease phototherapy by generating high yields of cytotoxic reactive oxygen species (ROS). In cells, excessive ROS can lead to lipid peroxidation and damage to proteins and DNA, ultimately leading to cell death. In recent years, research on the combined therapeutic effects of transition metal complexes with AIE molecules with unique optical properties has attracted widespread attention.

[0003] Cancer immunotherapy is a new cancer treatment method following traditional surgery, chemotherapy, and radiotherapy. Among them, immunogenic cell death (ICD) is a special form of cell death that can activate the host immune system to a certain extent, thereby achieving a systemic anti-tumor effect. Immunogenic cell death (ICD) is a form of cell death accompanied by the release of damage-associated molecular patterns (DAMPs). These include calreticulin (CRT) exposed on the surface of tumor cells, high-mobility group protein B1 (HMGB1) released by cells, and adenosine triphosphate (ATP) secreted by cells. These DAMPs can effectively recruit and activate antigen-presenting cells (APCs) to take up tumor antigens, such as dendritic cells (DCs), and present tumor antigens to T lymphocytes, subsequently triggering cytotoxic T lymphocyte responses and inducing systemic anti-tumor immune responses. To date, most metal-based anticancer agents with AIE properties have failed to successfully induce ICD in cancer treatment. Summary of the Invention

[0004] Purpose of the invention: The purpose of the present invention is to provide a metal ruthenium complex with aggregation-induced emission properties. By introducing aggregation-induced emission molecules and lysosome-targeted morpholino groups into the metal ruthenium complex precursor, a complex with both cell fluorescence imaging and anti-cancer activity is constructed. At the same time, the complex of the present invention can also induce immunogenic death through the synergistic effect of multi-organelle damage, thereby achieving the purpose of combined chemical-immunotherapy; another purpose of the present invention is to provide a preparation method and application of the above-mentioned metal ruthenium complex with aggregation-induced emission properties.

[0005] Technical solution: The metal ruthenium complex with aggregation-induced emission properties described in the present invention has the following structural formula:

[0006]

[0007] The method for preparing the above-mentioned metal ruthenium complex having aggregation-induced emission properties comprises the following steps:

[0008] (1) Synthesis of morpholine-modified aggregation-induced emission ligand: The aggregation-induced emission molecule DP-OH (3-(9,9-dimethylacridin-10(9H)-yl)-5-(pyridin-4-yl)phenol), 4-(2-chloroethyl)morpholine and K2CO3 were dissolved in acetone; the mixture was stirred and refluxed under an argon atmosphere; after the reaction, the supernatant was obtained by centrifugation, and the crude product was purified by silica gel column chromatography using CH2Cl2 / MeOH as eluent to obtain a light yellow solid product, which was named DP-M;

[0009]

[0010] (2) Synthesis of ruthenium complex: DP-M, Ru(phen)2Cl2 (bisphenanthroline ruthenium dichloride) and EtOH / H2O were mixed and stirred at high temperature under an argon atmosphere. After the reaction was completed and cooled to room temperature, NH4PF6 was added to obtain a crude product, which was washed three times with ethyl acetate. The crude product was purified by silica gel column chromatography (CH2Cl2 / MeOH) to obtain the target product, named Ru-AM;

[0011]

[0012] Wherein, in step (1), the molar ratio of DP-OH to 4-(2-chloroethyl)morpholine is 1:1.3.

[0013] Wherein, in step (1), the reflux reaction temperature is 65° C., and the reflux reaction time is 12 h.

[0014] Wherein, in step (2), the molar ratio of DP-M to Ru(phen)2Cl2 is 0.9:1.

[0015] Wherein, in step (2), the reaction temperature is 65° C. and the stirring time is 12 h.

[0016] Application of the metal ruthenium complex with aggregation-induced emission properties in the preparation of anti-tumor drugs or anti-tumor drug components.

[0017] Beneficial effects: Compared with the prior art, the present invention has the following significant effects: (1) The complex Ru-AM of the present invention can solve the problems of low efficacy, large dosage, inability to image cells and induction of immunogenic death when a single metal complex precursor or AIE molecule is used as an anti-tumor drug; (2) The complex Ru-AM of the present invention exhibits high cytotoxicity to a variety of tumor cells, and its IC value for human lung cancer cells (A549) is 50 The value is 3.8uM, which can effectively inhibit the proliferation of tumor cells; (3) After entering the cell, the complex Ru-AM of the present invention is distributed in multiple subcellular organelles, and achieves good anti-tumor activity by inducing the production of reactive oxygen species and cell autophagy in cancer cells; (4) The complex Ru-AM of the present invention can also induce immunogenic death of tumor cells, activate the host's immune system, and induce tumor-specific immune response, thereby achieving a combined effect of chemo-immunotherapy; (5) The fluorescence emission wavelength of the complex Ru-AM of the present invention is in the near-infrared region (725nm), which can effectively image the tumor area of ​​mice and effectively inhibit tumor growth in tumor-bearing mice, and has a wide range of application scenarios. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Figures 1 and 2 are fluorescence spectra of the Ru-AM complex of the present invention in a water / DMSO mixed solvent; (a) is a fluorescence emission spectrum of the Ru-AM complex in a water / DMSO mixed solvent with different water components; (b) is a graph of the relative fluorescence intensity of Ru-AM at different water contents, where I0 represents the fluorescence emission intensity in DMSO;

[0019] Figure 2 These are laser confocal images of the co-localization of the complex Ru-AM of the present invention with mitochondria, lysosomes and endoplasmic reticulum;

[0020] Figure 3 This is a flow cytometric graph showing that the complex Ru-AM of the present invention induces A549 cells to produce reactive oxygen species;

[0021] Figure 4 Schematic diagram of the immunogenic death of A549 cells induced by the complex Ru-AM of the present invention; (a) is a confocal fluorescence image of calreticulin CRT; (b) is a confocal fluorescence image of HMGB1; (c) is a flow cytometric image of calreticulin CRT; (d) is a schematic diagram of the extracellular ATP content;

[0022] Figure 5 Schematic diagram of the complex Ru-AM of the present invention inducing autophagy in A549 cells;

[0023] Figure 6 The fluorescence distribution images of the complex Ru-AM of the present invention in the mouse body at different times after intratumoral injection;

[0024] Figure 7 Figure 2 shows the changes in tumor tissue volume, weight, and body weight of tumor-bearing mice after treatment with the Ru-AM complex of the present invention; (a) is a physical picture of the mouse tumor dissection after treatment; (b) is the change in body weight of the mouse tumor; (c) is the change in tumor volume of the mouse; and (d) is the change in body weight of the mouse. DETAILED DESCRIPTION

[0025] Example 1

[0026] The method for preparing a ruthenium metal complex (Ru-AM) having aggregation-induced emission properties of the present invention comprises the following steps:

[0027] (1) Synthesis of compound DP-M: DP-OH (37.8 mg, 0.1 mmol), 4-(2-chloroethyl)morpholine M (19.45 mg, 0.13 mmol), K2CO3 (41.5 mg, 0.3 mmol) and acetone (15 mL) were added to a 50 mL round-bottom flask; under an argon atmosphere, the mixture was stirred and refluxed at 65°C for 12 hours; after the reaction was completed and cooled to room temperature, the supernatant was obtained by centrifugation; the crude product was purified by silica gel column chromatography using CH2Cl2 / MeOH as the eluent; the light yellow solid product was collected, namely DP-M (40 mg, yield 79%). Characterized by NMR and mass spectrometry, 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.68-8.59 (m, 2H), 7.87-7.81 (m, 2H), 7.61 (t, J = 2. 0Hz,1H),7.50(dd,J=7.8,1.6Hz,2H),7.41(t,J=1.6Hz,1H),7.07(t,J=2.1Hz,1H) ,7.00(ddd,J=8.4,7.2,1.6Hz,2H),6.91(td,J=7.5,1.3Hz,2H),6.27(dd,J=8.2,1 .3Hz, 2H), 4.25 (t, J = 5.6Hz, 2H), 3.57 (t, J = 4.7Hz, 4H), 2.73 (s, 2H), 1.64 (s, 6H). ESI-MS (in CH3OH): theoretical value: 494.26, experimental value: 491.64;

[0028]

[0029] (2) Synthesis of complex Ru-AM: DP-M (22.12 mg, 0.045 mmol), Ru(phen)2Cl2 (26.6 mg, 0.05 mmol) and EtOH / H2O (ethanol and water volume ratio of 3:1, 15 mL) were mixed and stirred at 65°C under argon atmosphere for 12 h. After the reaction was completed and cooled to room temperature, NH4PF6 (30 mg, 0.18 mmol) was added to obtain a crude product, which was washed three times with ethyl acetate. The crude product was purified by silica gel column chromatography (CH2Cl2 / MeOH) to obtain the target product, named Ru-AM. Yield: 31.5 mg, 62%. Characterized by NMR and mass spectrometry, 1 H NMR(400MHz,DMSO-d6)δ(ppm):10.21(dd,J=5.3,1.3Hz,1H),9.07(dd,J=5.3,1.3Hz,1H),8.84(dt,J=8.2,1.4Hz,2H),8.48(ddd,J=11.9,8 .2,1.2Hz,2H),8.38(d,J=8.9Hz,1H),8.32(dt,J=8.2,2.3Hz,2H),8.26(d,J=8.9Hz,1H),8.21(d,J=8.9Hz,1H),8.15-8.10(m,2H),7.80-7. 73(m,3H),7.59-7.43(m,6H),7.35(s,1H),7.05(s,1H),6.91(dtd,J=21.0,7.3,1.5Hz,5H),6.17(dd,J=8.0,1.4Hz,2H),4.18(t,J=5.7Hz, 3H), 3.53 (t, J=4.6Hz, 5H), 2.68 (t, J=5.7Hz, 2H), 2.45 (d, J=4.8Hz, 4H), 1.61 (s, 6H). ESI-MS (CH3OH): calculated: 1132.24, found: 984.3.

[0030]

[0031] Comparative Example 1

[0032] A method for preparing a metal ruthenium complex (Ru-A) having aggregation-induced emission properties, specifically comprising:

[0033] DP-OH (17 mg, 0.045 mmol), Ru(phen)2Cl2 (26.6 mg, 0.05 mmol), and EtOH / H2O (3:1, 15 mL) were mixed and stirred at 65°C under an argon atmosphere for 12 h. After the reaction was completed and cooled to room temperature, NH4PF6 (30 mg, 0.18 mmol) was added to obtain a crude product, which was washed three times with ethyl acetate. The crude product was purified by silica gel column chromatography (CH2Cl2 / MeOH) to obtain the target product, named Ru-A; yield: 31.5 mg, 61%. Characterization by NMR and mass spectrometry, 1 HNMR (400MHz, DMSO-d6) δ (ppm): 10.20 (s, 1H), 9.07 (d, J = 5.3Hz, 1H), 8.89-8.78 (m, 3H), 8.46 (dd, J = 12.9,7.8Hz,3H),8.37(d,J=9.1Hz,1H),8.34-8.29(m,2H),8.25(d,J=8.9Hz,1H),8.22-8.18(m,1H) ,8.16-8.08(m,2H),7.75(d,J=5.3Hz,1H),7.68(d,J=5.9Hz,2H),7.61-7.41(m,5H),7.31(s,1H),7.17(s,1H),6.91(dt,J=23.7,7.3Hz,4H),6.78(s,1H),6.20(d,J=8.0Hz,2H),1.59(s,6H).ESI-MS(in CH3OH):Theoretical value: 1020.15, experimental value: 875.2.

[0034]

[0035] Example 2: Fluorescence spectrum of the complex Ru-AM in a water / DMSO mixed solvent

[0036] (1) Prepare a DMSO solution of the complex Ru-AM at a concentration of 5 mM; (2) Prepare a water / DMSO mixed solvent with different water contents, with the water content ranging from 0 to 98%; (3) Add the same volume of Ru-AM at a concentration of 50 uM to the above mixed solvents and measure its fluorescence emission spectrum.

[0037] The fluorescence emission spectrum of the complex in water / DMSO mixed solvent ( Figure 1) shows that when the water content is less than 80%, the fluorescence intensity of Ru-AM gradually decreases with increasing water content, presumably due to the solvatochromic effect caused by twisted intramolecular charge transfer (TICT). When the water content in the mixed solvent exceeds 80%, the fluorescence intensity of Ru-AM significantly increases, indicating that Ru-AM has significant AIE properties.

[0038] Example 3: In vitro cytotoxicity test of the complex Ru-AM on tumor cells

[0039] The cytotoxic activity of the complex Ru-AM in in vitro cells was studied. The study used the standard MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide] method, and selected human lung cancer cell line A549, human breast cancer cell line MCF-7 and normal cell HLF cells for testing. The cells were seeded in 96-well plates (about 5000 cells per well) and cultured at 37°C, 5% CO2 for 24 hours. After the cells adhered, they were replaced with DMEM culture medium containing different concentration gradients of Ru-AM and cultured for 48 hours. 20μL MTT was added to each well, and the cells were incubated in a 37°C incubator for another 4 hours. The supernatant was then aspirated and 150μL dimethyl sulfoxide (DMSO) was added to each well. A was detected using an enzyme-linked immunosorbent assay (ELISA). 570nm , calculate the cell proliferation inhibition rate and find IC 50 value.

[0040] Table 1 shows the IC values ​​of the complexes Ru-AM, Ru-A, compounds DP-OH, DP-M and cisplatin CDDP. 50 (μM) value

[0041]

[0042] The MTT test results of the Ru-AM complex are shown in Table 1. Compared with the compounds DP-OH and DP-M, the cytotoxicity of the Ru-A and Ru-AM complexes was significantly enhanced, indicating that the introduction of the cyclometallated ruthenium significantly enhanced the drug's toxicity against tumor cells. Importantly, the IC50 value of Ru-AM against A549 was 3.8 μM, 3.5-fold lower than that of Ru-A and 2.3-fold lower than that of cisplatin, indicating that the modification with the morpholino group significantly enhanced its cytotoxic activity against tumor cells. These results demonstrate that the target complex Ru-AM possesses superior ability to inhibit tumor cell growth and is a potential cancer therapeutic.

[0043] Example 4: Co-localization of Ru-AM complex in mitochondria, lysosomes and endoplasmic reticulum by laser confocal microscopy

[0044] (1) A549 cells were incubated with 10 μM Ru-AM for 6 h in the dark; (2) the cell culture medium containing the complex was discarded and commercial probes (Mito-Green, ER-Green, Lyso-Green) of corresponding working concentrations were added, and then the experimental requirements of each commercial probe were incubated for about 30 min; (3) the culture medium of different commercial probes was discarded and the cells were washed twice with PBS, and then the colocalization laser confocal imaging experiment was performed.

[0045] The co-localization laser confocal microscopy images of the Ru-AM complex with mitochondria, lysosomes and endoplasmic reticulum are shown in Figure 2. Figure 2 As shown in the figure, the red fluorescence of Ru-AM colocalizes with the green fluorescence of commercial dyes such as Lyso-Green, Mito-Green, and ER-Green, with colocalization coefficients of 0.33, 0.35, and 0.42, respectively, indicating that Ru-AM is mainly distributed in the cytoplasm and mitochondria.

[0046] Example 5: Effect of the complex Ru-AM on the production of reactive oxygen species in A549 cells

[0047] A549 cells were incubated with Ru-AM for 24 hours and then stained with 10 μM DCF-DA in serum-free medium at 37°C for 30 minutes in the dark. The cells were centrifuged and the supernatant discarded. The cells were washed three times with serum-free medium to remove any DCF-DA that had not entered the cells. Within half an hour of harvesting the cells, green fluorescence intensity was measured by flow cytometry using an excitation wavelength of 488 nm and an emission wavelength of 530 ± 20 nm. The mean green fluorescence intensity was analyzed using FlowJo 7.6 (Tree Star, OR, USA) software.

[0048] The effect of the complex Ru-AM on the production of reactive oxygen species in A549 cells is shown in Figure 3. Compared with the control group, treatment with the complex Ru-AM can effectively induce an increase in the content of reactive oxygen species in the cells, and show a dose-dependent manner. This indicates that after the complex Ru-AM enters the cancer cells, it induces the cancer cells to produce reactive oxygen species (ROS), thereby leading to the death of the cancer cells.

[0049] Example 6: Application of the complex Ru-AM to induce immunogenic death of A549 cells

[0050] Immunofluorescence detection of CRT and HMGB1 proteins: A549 cells were seeded into 35 mm confocal culture dishes and observed under a confocal microscope. After overnight culture, the cells were treated with the test drug at 37°C for 24 hours. The cells were then fixed with 4% paraformaldehyde / PBS for 30 minutes. After washing three times with PBS, the cells were stained with either calreticulin or HMGB1 antibodies at room temperature for 1 hour and incubated with a FITC-conjugated secondary antibody at room temperature for 1 hour. The cells were counterstained with 300 nM DAPI for 10 minutes and immediately observed under a confocal microscope. Extracellular ATP content: A549 cells were seeded into 6-well culture dishes and cultured overnight until cell attachment reached 60-70%. The medium was then replaced with medium containing Ru-A or Ru-AM, respectively. After 24 hours of culture, the supernatant was collected for extracellular ATP measurement. The cells were washed three times with PBS and lysed with 100 μL of lysis reagent. The cell lysate was centrifuged at 12,000 g and 4°C for 5 minutes. In a 96-well white plate, the supernatant of the cell lysate was added to the ATP working solution according to the manufacturer's instructions. Luminescence in each well was immediately detected using a multi-functional microplate reader.

[0051] The results of the complex Ru-AM inducing immunogenic death of A549 cells are as follows Figure 4 Compared with the control group, Ru-AM significantly enhanced the fluorescence intensity of CRT and HMGB1 proteins in A549 cells, and significantly increased the extracellular ATP content, indicating that the complex Ru-AM induced the release and exposure of intracellular CRT, the release of HMGB1, and the exocytosis of ATP, while no obvious phenomenon was seen in the control complex Ru-A treatment group, indicating that the complex Ru-AM of the present invention effectively induced the immunogenic death of tumor cells, while the complex Ru-A could not induce the immunogenic death of tumor cells.

[0052] Example 7: Effect of the complex Ru-AM on autophagy in A549 cells

[0053] A549 cells were cultured in 10 cm culture dishes and treated with Ru-A and Ru-AM for 48 h, respectively. The cells were collected and lysed in cell lysis buffer containing PMSF (1%) at 4 ° C for 30 minutes. The lysate was collected by centrifugation for 20 minutes. The proteins in the cell lysate were separated on SDS-PAGE and transferred to a polyvinylidene fluoride (PVDF) membrane. The membrane was blocked with PBST containing 5% skim milk powder for 1 hour and then incubated with specific antibodies overnight at 4 ° C with gentle shaking. Afterwards, the membrane was incubated with secondary antibodies at room temperature for 1 hour. Protein blots were detected with chemiluminescent reagents.

[0054] Effects of the complex Ru-AM on the expression of autophagy-related proteins in A549 cells Figure 5Compared with the untreated control group or the Ru-A drug group, the levels of autophagy marker proteins LC3-II / LC3-I increased in a concentration-dependent manner, and the level of autophagy adaptor protein P62 decreased in a concentration-dependent manner after Ru-AM treatment, indicating that the Ru-AM complex induced the death of A549 cells through autophagy.

[0055] Example 8: Fluorescence image distribution of the complex Ru-AM in mice

[0056] A 2 mM Ru-AM solution (120 μL) was injected intratumorally into the test site of the mouse. Fluorescence signals were collected at 0, 1, 3, 6, 12, 24, and 48 hours. Following the experiment, the tumors were dissected and major organs (heart, liver, spleen, lungs, and kidneys) were removed for fluorescence imaging.

[0057] The biodistribution results of the complex Ru-AM in mice at different times after intratumoral injection are shown in the following figure: Figure 6 As shown in the figure, a clear fluorescence signal appeared at the tumor site after injection of the Ru-AM complex around 6 hours later. The fluorescence signal gradually increased between 6 and 48 hours, and strong fluorescence was still observed at 48 hours. Meanwhile, the fluorescence was less abundant or even very weak in other organs of the mouse (heart, liver, spleen, lungs, and kidneys). This indicates that Ru-AM has significant tumor retention and shows great potential in tumor imaging.

[0058] Example 9: Application of the complex Ru-AM in anti-tumor effect in mice in vitro

[0059] Tumor-bearing mouse models were established by subcutaneous injection of tumor cells in female BALB / c nude mice. Drug treatment was initiated once tumors reached a certain size, with intratumoral injections administered every other day. Tumor volume and mouse weight were recorded every two days during the course of treatment. After treatment, tumors were removed from the mice for observation and weight.

[0060] After Ru-AM treatment, the changes in tumor weight (a, b), volume (c), and body weight (d) of mice were shown in Figure 2. Figure 7 As shown. The tumor in the control group grew rapidly, while the tumor growth in the Ru-AM-treated group was significantly inhibited, with the tumor volume being 65.2% of that in the control group, indicating that Ru-AM has excellent anti-tumor efficacy in vivo. In addition, no significant changes in mouse body weight were observed in the three treatment groups, indicating that the complex Ru-AM had weak toxic side effects on mice at this dose and exhibited good biosafety. The above experimental results show that the metal ruthenium complex Ru-AM with AIE properties prepared in Example 1 has a good tumor inhibitory effect in vivo.

[0061] The complex of the present invention can effectively inhibit the proliferation of human tumor cells and achieve good anti-tumor activity by inducing the production of reactive oxygen species and mitochondrial autophagy in tumor cells. In addition, the complex of the present invention can induce immunogenic death of tumor cells, further activating the body's immune response, achieving a synergistic effect of chemotherapy-immunotherapy, and has broad application prospects.

Claims

1. A metal ruthenium complex having aggregation-induced emission properties, characterized in that: Its structural formula is shown below: 。 2. The method for preparing the metal ruthenium complex having aggregation-induced emission properties according to claim 1, characterized in that: The following steps are involved: (1) Synthesis of morpholine-modified aggregation-induced emission ligand: The aggregation-induced emission molecule DP-OH, 4-(2-chloroethyl)morpholine, and K2CO3 were dissolved in acetone; the mixture was stirred and refluxed under an argon atmosphere; after the reaction, the supernatant was obtained by centrifugation, and the crude product was purified by silica gel column chromatography using CH2Cl2 / MeOH as the eluent to collect a light yellow solid product, named DP-M; ； (2) Synthesis of ruthenium complex: DP-M, Ru(phen)2Cl2 and EtOH / H2O were mixed and stirred at 65°C under an argon atmosphere. After the reaction was completed and cooled to room temperature, NH4PF6 was added to obtain a crude product. The crude product was washed three times with ethyl acetate and purified by silica gel column chromatography using CH2Cl2 / MeOH as eluent to obtain the target product, named Ru-AM. 。 3. The method for preparing a metal ruthenium complex having aggregation-induced emission properties according to claim 2, wherein: In step (1), the molar ratio of DP-OH to 4-(2-chloroethyl)morpholine is 1:1.

3.

4. The method for preparing a metal ruthenium complex having aggregation-induced emission properties according to claim 2, wherein: In step (1), the reflux reaction temperature is 65° C. and the reflux reaction time is 12 h.

5. The method for preparing a metal ruthenium complex having aggregation-induced emission properties according to claim 2, wherein: In step (2), the molar ratio of DP-M to Ru(phen)2Cl2 is 0.9:

1.

6. The method for preparing a metal ruthenium complex having aggregation-induced emission properties according to claim 2, wherein: In step (2), the stirring time is 12 h.

7. Use of the metal ruthenium complex with aggregation-induced emission properties according to claim 1 in the preparation of anti-tumor drugs.

Images