A binuclear ruthenium photocatalyst, and a preparation method and application thereof

Abstract

The application discloses a binuclear ruthenium photocatalyst and a preparation method and application thereof, and relates to a compound with the structure shown in the following formula: The compound with the structure of the application scheme can realize full-spectrum absorption, has good photocatalytic proliferation inhibition capacity (IC 50 The compound has a high light treatment index (PI=143.43) and can generate superoxide anions, singlet oxygen and active oxygen components under light irradiation, which has important significance for researching binuclear metal ruthenium complexes as antitumor drugs and provides a new idea for developing a treatment method for triple-negative breast cancer in a clinic.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a binuclear ruthenium photocatalyst, its preparation method, and its application. Background Technology

[0002] Cancer poses a significant threat to human health worldwide, and remarkable achievements have been made in cancer treatment thanks to continuous advancements in medical technology. However, traditional treatments such as chemotherapy and radiotherapy, while achieving good results in many cases, also have limitations and side effects. These include damage to healthy tissues, the development of treatment tolerance, and recurrence after treatment. Therefore, we urgently need new treatment strategies to overcome these challenges.

[0003] Photodynamic therapy (PDT), based on photosensitizers, offers a promising new treatment paradigm. Combining optical and pharmacological principles, it utilizes specific wavelengths of light to activate photosensitizers, thereby achieving highly selective killing of cancer cells while maximizing the protection of surrounding healthy tissue. Compared to traditional treatments, PDT offers several significant advantages. First, it enables precise treatment at a localized level, reducing systemic toxicity and improving treatment safety. Second, PDT can be precisely tuned to suit different types and stages of cancer. This provides clinicians with a highly customized treatment option, contributing to improved treatment effectiveness. Furthermore, PDT demonstrates potential for multimodal efficacy, meaning it can be combined with other treatment methods to create synergistic effects and further enhance therapeutic outcomes.

[0004] Therefore, developing new photosensitizers is an urgent need in the current pharmaceutical field. This will provide us with a powerful tool to fight cancer more effectively and safely, and to offer patients better treatment options. Summary of the Invention

[0005] This invention aims to at least solve one of the technical problems existing in the prior art. To this end, this invention proposes a binuclear ruthenium photocatalyst, which shows promising application prospects in the treatment of triple-negative breast cancer.

[0006] The present invention also proposes a method for preparing the above-mentioned binuclear ruthenium photocatalyst.

[0007] This invention also proposes the application of the aforementioned binuclear ruthenium photocatalyst.

[0008] According to one aspect of the present invention, a binuclear ruthenium photocatalyst is provided, comprising compounds with the following structures:

[0009]

[0010] According to a preferred embodiment of the present invention, at least the following beneficial effects are achieved: the compound with the structure of the present invention can achieve full-spectrum absorption and has good photocatalytic proliferation inhibition ability (IC50) against mouse triple-negative breast cancer cells (4T1). 50 It has a phototherapy index (PI = 143.43) of 0.08 μM and can generate superoxide anions, singlet oxygen and reactive oxygen species under light irradiation. This is of great significance for the research of binuclear ruthenium complex anti-tumor drugs and provides a new approach for the clinical development of triple-negative breast cancer treatment methods.

[0011] According to another aspect of the present invention, a method for preparing the above-mentioned catalyst is provided, comprising the following steps:

[0012] S1. 1,3-cyclohexadiene is reacted with a ruthenium(III) salt to generate precursor compound I;

[0013] S2. The precursor compound is reacted with 3-bromo-1,10-phenanthroline to generate precursor compound II;

[0014] S3. React the precursor compound II with 2,2'-bipyridine and then with hexafluorophosphate to generate precursor compound III;

[0015] S4. The precursor compound III is reacted with 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrolo-1,4-(2H,5H)-dione to obtain the product.

[0016] The preparation method according to a preferred embodiment of the present invention has at least the following beneficial effects: the preparation process of the present invention is simple, easy to operate, and has good prospects for industrial application.

[0017] In some embodiments of the present invention, the reaction conditions of step S1 include at least one of the following conditions:

[0018] (1) The reaction is carried out in an alcohol solvent system; preferably a lower alcohol; more preferably, the lower alcohol is a C1-C6 alcohol; more preferably ethanol;

[0019] (2) The reaction is carried out under reflux at 70-90°C; preferably the reaction temperature is 75-85°C; more preferably 80°C;

[0020] (3) The reaction time is 4-8 hours; more preferably 6 hours.

[0021] (4) The molar ratio of 1,3-cyclohexadiene to ruthenium(III) salt is 2-2.2:1; preferably 2:1.

[0022] In some preferred embodiments of the present invention, step S1 specifically involves dissolving 1,3-cyclohexadiene and ruthenium(III) trichloride hydrate in an ethanol solution, then refluxing the reaction at 70-90°C for 4-8 hours, and cooling to room temperature. The reaction formula for this step is as follows:

[0023]

[0024] In some embodiments of the present invention, the reaction conditions of step S2 include at least one of the following conditions:

[0025] (1) The reaction is carried out in an alcohol solvent system; preferably, the alcohol solvent is ethylene glycol;

[0026] (2) The reaction is carried out under reflux at 70-90°C; preferably the reaction temperature is 75-85°C; more preferably 80°C;

[0027] (3) The reaction time is 3-5 hours; more preferably 4 hours.

[0028] (4) The molar ratio of the precursor compound I to 3-bromo-1,10-phenanthroline is 1:1-1.2; more preferably 1:1.

[0029] In some preferred embodiments of the present invention, step S2 specifically involves dissolving precursor compound I and 3-bromo-1,10-phenanthroline in an ethanol solution, then refluxing at 70-90°C for 3-5 hours and cooling to room temperature. The reaction formula for this step is as follows:

[0030]

[0031] In some embodiments of the present invention, the reaction conditions of step S3 include at least one of the following conditions:

[0032] (1) The reaction is carried out in an alcohol solvent system; preferably, the alcohol solvent is ethylene glycol;

[0033] (2) The reaction is carried out under reflux at 180-220°C; preferably, the reaction temperature is 190-210°C; more preferably 200°C.

[0034] (3) The reaction time is 20-40 min; preferably 30 min;

[0035] (4) The molar ratio of the precursor compound II to 2,2'-bipyridine is 1:2-2.2; more preferably 1:2.

[0036] In some preferred embodiments of the present invention, step S3 specifically involves: dissolving precursor compound II and 2,2'-bipyridine in ethylene glycol solution, refluxing at 180-220°C for 20-40 min, cooling to room temperature, then adding ammonium hexafluorophosphate for ion exchange, followed by filtration, washing, and drying for purification. The reaction formula for this step is as follows:

[0037]

[0038] In some embodiments of the present invention, the reaction conditions of step S4 include at least one of the following conditions:

[0039] (1) The reaction is carried out under a protective atmosphere; preferably, the protective atmosphere is an argon atmosphere;

[0040] (2) The reaction is carried out in a mixture of toluene and N,N-dimethylformamide; preferably, the volume ratio of toluene to N,N-dimethylformamide is 1:1;

[0041] (3) The reaction temperature is 100-135℃; preferably 110-120℃; more preferably 115℃;

[0042] (4) The reaction time is 18-24 h; preferably 18-22 h; more preferably 20 h;

[0043] (5) The molar ratio of the precursor compound III to 1,3-bis(2-ethylhexyl)-5,7-bis(5-(trimethyltinyl)thiophen-2-yl)-4H,8H-benzo[1,2-c:4,5-c']dithiophene-4,8-dione is 1:(1.8-2.2); preferably, the molar ratio of the ruthenium complex to 1,3-bis(2-ethylhexyl)-5,7-bis(5-(trimethyltinyl)thiophen-2-yl)-4H,8H-benzo[1,2-c:4,5-c']dithiophene-4,8-dione is 1:1.9-2.1; preferably 1:2;

[0044] (6) The reaction is carried out under the catalysis of a catalyst, the catalyst comprising tetra(triphenylphosphine)palladium; preferably, the molar ratio of the precursor compound III to the catalyst is 1:0.08 to 0.12; more preferably 1:0.1.

[0045] In some preferred embodiments of the present invention, step S4 specifically involves: under argon protection, [(bpy)2Ru(3-bromo-1,10-phenanthroline)] 2+(PF6)2,2,5-Di(2-ethylhexyl)-3,6-Di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrolo-1,4-(2H,5H)-dione and tetra(triphenylphosphine)palladium were dissolved in dry N,N-dimethylformamide and stirred at 100-135°C for 18-24 h. After the reaction was completed, the mixture was cooled to room temperature, saturated brine was added, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate, and the solvent was evaporated to obtain the final product. The reaction formula for this step is as follows:

[0046]

[0047] According to another aspect of the present invention, the application of the above-described catalyst in the preparation of antitumor photocatalytic drugs is proposed.

[0048] According to another aspect of the present invention, an antitumor photocatalytic drug is provided, wherein the active ingredient of the drug comprises the above-mentioned binuclear ruthenium photocatalyst.

[0049] According to another aspect of the present invention, an antitumor metal photosensitizer is provided, wherein the active ingredient of the metal photosensitizer comprises the aforementioned binuclear ruthenium photocatalyst.

[0050] In some embodiments of the present invention, the tumor is breast cancer.

[0051] In some embodiments of the present invention, the tumor is triple-negative breast cancer.

[0052] In some embodiments of the present invention, the tumor is a mouse triple-negative breast cancer.

[0053] In some embodiments of the present invention, the cancer cells of the mouse triple-negative breast cancer are 4T1 cancer cells.

[0054] In some embodiments of the present invention, the drug (or photosensitizer) further includes a pharmaceutically acceptable carrier and / or excipient. That is, the drug or photosensitizer uses a binuclear ruthenium complex as the main active ingredient, is mixed with a pharmaceutically acceptable carrier and / or excipient to prepare a composition, and is then formulated into a clinically acceptable dosage form.

[0055] In some embodiments of the present invention, the excipient refers to a diluent, binder, lubricant, disintegrant, solubilizer, stabilizer, and other pharmaceutical matrix that can be used in the pharmaceutical field.

[0056] In some embodiments of the invention, the different pharmaceutical excipients used for the drug dosage form (or photosensitizer) may vary depending on the specific medical application. Pharmaceutical excipients can be used to adjust the solubility and bioavailability of the photocatalyst, increase its stability, modulate the host's immune response, and act as emulsifiers, antioxidants, aerosol propellants, tablet binders, and tablet disintegrants. Preferred pharmaceutical excipients include, but are not limited to, binders / fillers, coating agents, disintegrants, lubricants, and sweeteners adapted to the use of photosensitizers.

[0057] In some embodiments of the present invention, the carrier is a functional pharmaceutical excipient acceptable in the pharmaceutical field, including surfactants, suspending agents, emulsifiers, and some novel pharmaceutical polymers, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.

[0058] In some embodiments of the present invention, there are no particular limitations on the dosage form of the aforementioned drug (or photosensitizer). The active substance can be administered together with an assimilated edible carrier, an inert diluent, or directly combined with food. Drug dosage forms include, but are not limited to, hard-shell or soft-shell gelatin capsules, tablets, pills, powder for injection, solutions, suspensions, elixirs, syrups, wafers, gels, buccal or sublingual tablets, films, suppositories, and enemas.

[0059] In other embodiments of the invention, the photocatalyst of the invention can be formulated without any formulation adjuvants or using other drug delivery systems known in the prior art, such as forming components with liposomes, vectored and non-vectored proteins, organic and inorganic nanoparticles, nanoemulsions and microemulsions, nanocrystals, individual solvents or suitable solvent mixtures, with components such as lactose, polyvinylpyrrolidone (PVP), etc.

[0060] In some embodiments of the present invention, the prepared drug (or photosensitizer) may be administered orally, via any part of the gastrointestinal tract (e.g., mouth, pharynx, esophagus, stomach, small intestine (duodenum, jejunum, ileum), large intestine (cecum, colon, rectum) and anus), or via parenteral route (e.g., intravenous, subcutaneous, intraperitoneal or local). If certain drugs are unstable under gastric conditions, they may be prepared as enteric-coated tablets.

[0061] The application of a preferred embodiment of the present invention has at least the following beneficial effects: the present invention has good application prospects in the field of preparation of anti-tumor drugs such as anti-triple-negative breast cancer drugs.

[0062] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0063] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0064] Figure 1 The ultraviolet absorption spectrum of the binuclear ruthenium photocatalyst prepared in this invention is shown.

[0065] Figure 2 The fluorescence excitation and emission spectra of the binuclear ruthenium photocatalyst prepared in this invention are shown.

[0066] Figure 3 The ability of the binuclear ruthenium photocatalyst prepared in this invention to photocatalyze the generation of superoxide anions;

[0067] Figure 4 This invention relates to the ability of the binuclear ruthenium photocatalyst to photocatalyze the generation of singlet oxygen.

[0068] Figure 5 This invention demonstrates the photocatalytic ability of the binuclear ruthenium photocatalyst to generate reactive oxygen species.

[0069] Figure 6 The dark toxicity and phototoxicity of the binuclear ruthenium photocatalyst prepared in this invention on mouse triple-negative breast cancer cells (4T1). Detailed Implementation

[0070] The following will clearly and completely describe the concept and technical effects of the present invention in conjunction with embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods; the materials and reagents used, unless otherwise specified, are commercially available. Unless otherwise specified, the same parameter value is the same in all embodiments. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0071] The term "room temperature" as used in this invention refers to any temperature between 25 and 5°C, and specifically 25°C in the embodiments.

[0072] Example

[0073] This embodiment prepares a dual-core ruthenium photocatalyst, the structural formula of which is as follows:

[0074]

[0075] The specific process is as follows:

[0076] (1) [Ru(π-C6H6)Cl2]2 is generated by the reaction of 1,3-cyclohexadiene with ruthenium(III) trichloride hydrate.

[0077] 1,3-cyclohexadiene (161 mg, 2 mmol) and ruthenium(III) trichloride hydrate (207.4 mg, 1 mmol) were dissolved in 15 mL of ethanol solution. The mixture was refluxed at 80 °C for 6 h and then cooled to room temperature. The reaction solution was filtered, and the filter cake was washed with ethanol and n-hexane and then dried under vacuum to obtain a brown solid powder [Ru(π-C6H6)Cl2]2 (204.3 mg, 82%).

[0078] The chemical reaction equations for the above reactions are shown below:

[0079]

[0080] (2) [Ru(π-C6H6)Cl2]2 reacts with 3-bromo-1,10-phenanthroline to form [Ru(π-C6H6)(3-bromo-1,10-phenanthroline)Cl] + Cl -

[0081] The precursor [Ru(π-C6H6)Cl2]2 (248.9 mg, 1 mmol) and 3-bromo-1,10-phenanthroline (259 mg, 1 mmol) were dissolved in 15 mL of ethanol solution. The mixture was refluxed at 80 °C for 4 h and then cooled to room temperature. The reaction solution was filtered, and the filter cake was washed with ethanol and n-hexane and then dried under vacuum to obtain the ruthenium complex [Ru(π-C6H6)(3-bromo-1,10-phenanthroline)Cl]. + Cl - Brown powder (447.2 mg, 88%).

[0082] The chemical reaction equations for the above reactions are shown below:

[0083]

[0084] (3)[Ru(π-C6H6)(3-bromo-1,10-phenanthroline)Cl] + Cl - It reacts with 2,2'-bipyridine to form [(bpy)2Ru(3-bromo-1,10-phenanthroline)] 2+ (PF6)2

[0085] Precursor [Ru(π-C6H6)(3-bromo-1,10-phenanthroline)Cl] + Cl - 2,2'-Bipyridine (254 mg, 0.5 mmol) was dissolved in 20 mL of ethylene glycol solution. The mixture was stirred at 200 °C for 30 min, then cooled to room temperature. A saturated aqueous solution of NH4PF6 was added, and the mixture was stirred at room temperature for 1 h. The mixture was then filtered, and the filter cake was washed with water and ethanol and dried under vacuum to obtain the ruthenium complex [(bpy)2Ru(3-bromo-1,10-phenanthroline)]. 2+ (PF6)2 orange-red powder (264 mg, 51%).

[0086] The mass spectrum of the product is: ESI-MS [CH3OH, m / z]: 335.97 [M-2PF6- ]2+ ;

[0087] The 1H NMR spectrum of the product is as follows: 1 HNMR (400MHz, acetone-d6) δ (ppm) = 9.07 (d, 1H, J = 1.6Hz), 8.86-8.80 (m, 5H), 8.48-8.44 (m, 3H), 8.37-8.35 (d, 1H, J = 9.2Hz), 8.28-8.24 (t, 2H, J = 8. 0Hz), 8.21-8.11(m, 4H), 8.06-8.04(d, 1H, J=5.2Hz), 7.97-7.93(q, 1H, J= 3.2Hz), 7.87-7.86 (d, 1H, J=5.6Hz), 7.65-7.58 (m, 2H), 7.41-7.37 (m, 2H).

[0088] The chemical reaction equations for the above reactions are shown below:

[0089]

[0090] (4)[(bpy)2Ru(3-bromo-1,10-phenanthroline)] 2+ (PF6)2 reacts with 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrolo-1,4-(2H,5H)-dione to generate a novel binuclear ruthenium metal complex.

[0091] [(bpy)2Ru(3-bromo-1,10-phenanthroline)] 2+(PF6)2,2,5-Di(2-ethylhexyl)-3,6-Di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrolo-1,4-(2H,5H)-dione (85 mg, 0.1 mmol) and tetra(triphenylphosphine)palladium (12 mg, 0.01 mmol) were dissolved in 15 mL of N,N-dimethylformamide and stirred at 115 °C for 20 h under argon protection. After the reaction was completed, the mixture was cooled to room temperature, saturated brine was added, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous magnesium sulfate, and the solvent was evaporated to obtain 207 mg (85%) of a dark green solid powder.

[0092] The chemical reaction equations for the above reactions are shown below:

[0093]

[0094] The mass spectrum of the product is: ESI-MS [CH3OH, m / z]: 426.8 [M-4PF6] - ] 4+ ;

[0095] The 1H NMR spectrum of the product is as follows: 1 HNMR (500MHz, Acetonitrile-d3) δ (ppm) = 8.85 (t, J = 2.0Hz, 2H), 8.73 (t, J = 4.3Hz, 2H), 8.62 (dd, J = 8.3, 1.2Hz, 2H) , 8.54 (d, J=8.2Hz, 4H), 8.49 (t, J=7.5Hz, 5H), 8.27 (d, J=1.2Hz, 5H), 8.19–8.06 (m, 9H), 8.00 (tdd, J=8.0, 3.8, 1.5H z, 5H), 7.92 (d, J=5.8Hz, 2H), 7.85–7.80 (m, 2H), 7.77–7.68 (m, 5H), 7.57 (dt, J=5.6, 1.1Hz, 2H), 7.55–7.43 (m, 7H) , 7.24 (tdd, J=7.8, 5.7, 1.3Hz, 5H), 1.34 (t, J=7.1Hz, 6H), 1.28 (s, 6H), 1.19 (s, 5H), 0.84 (dt, J=40.7, 7.3Hz, 15H).

[0096] Application examples

[0097] The performance of the binuclear ruthenium photocatalyst prepared in the examples was tested, as follows:

[0098] 1. Determination of absorbance and fluorescence spectra of binuclear ruthenium photocatalysts (binuclear ruthenium metal complexes) in different solvents

[0099] (1) Absorbance of binuclear ruthenium complex in different solvents

[0100] The binuclear ruthenium complexes prepared in the examples were dissolved in 10 μM solution using phosphate-buffered saline (PBS), dichloromethane (DCM), and acetonitrile (MeCN), respectively. The UV absorption spectra of the binuclear ruthenium complexes were then recorded using a double-beam UV-Vis spectrophotometer. The absorbance characterization results in different solvents are shown below. Figure 1 As shown. From Figure 1 As can be seen, the binuclear ruthenium complex has good light absorption in organic solvents, and its absorption wavelength in PBS shows a significant red shift.

[0101] (2) Fluorescence excitation and emission spectra of binuclear ruthenium complexes

[0102] Using dichloromethane as a solvent, 10 μM sample solutions of the novel binuclear ruthenium complex from the examples were prepared. The fluorescence emission spectra of the ruthenium complexes were recorded using a fluorescence spectrophotometer at a fixed excitation wavelength of 598 nm. Similarly, the fluorescence excitation spectra of the ruthenium complexes were recorded at a fixed excitation wavelength of 688 nm. Figure 2 As shown. From Figure 2 The excitation and emission spectra of the compound in dichloromethane show that its optimal excitation wavelength in dichloromethane is 598 nm and its optimal emission wavelength is 688 nm.

[0103] 2. Determination of the ability of binuclear ruthenium complexes to generate superoxide anions

[0104] To test the photocatalytic ability of the binuclear ruthenium complex prepared in the examples to generate superoxide anions, dihydrorhodamine 123 (DHR123) was used to determine the superoxide anion generation capability of the novel binuclear ruthenium complex. When superoxide anions are generated in the solution, DHR123 immediately captures them and is oxidized to generate the fluorescent derivative rhodamine 123, which emits bright green fluorescence (Ex / Em = 500 / 536 nm). An increase in fluorescence intensity indicates the generation of superoxide anions in the solution. The superoxide anion generation capability can be reflected by monitoring the changes in the fluorescence spectra of the test sample and the DHR123 mixture under different illumination times using a fluorescence spectrophotometer.

[0105] An aqueous solution containing a binuclear ruthenium complex (5 μM) and DHR123 reagent (5 μM) was placed in a cuvette, and its superoxide anion generation capacity was measured under dark and light conditions. The results are as follows: Figure 3 As shown in the figure, this binuclear ruthenium complex has the ability to generate superoxide anions after illumination.

[0106] 3. Determination of the ability of binuclear ruthenium complexes to generate singlet oxygen

[0107] To detect the photocatalytic ability of the synthesized binuclear ruthenium complex to generate singlet oxygen, the singlet oxygen probe 9,10-anthrayl-bis(methylene)dimalonic acid (ABDA) was used to determine the ability of the novel binuclear ruthenium complex to generate singlet oxygen. When singlet oxygen is generated in the solution, ABDA immediately captures the singlet oxygen in the solution and reacts to generate an endogenous oxidation product, causing the characteristic absorption peak of ABDA to decrease. The rate of decrease of the ABDA absorption peak is the singlet oxygen generation rate. The singlet oxygen generation ability can be reflected by monitoring the changes in the UV-Vis absorption spectra of the test sample and the ABDA mixture solution under different illumination times using a UV-Vis spectrophotometer.

[0108] Two aqueous solutions containing the same binuclear ruthenium complex (5 μM) and ABDA reagent (200 μM) were placed in cuvettes, and their singlet oxygen generation capacity was measured under light and dark conditions. The results are as follows: Figure 4 As shown in the figure, this binuclear ruthenium complex has the ability to generate singlet oxygen after illumination.

[0109] 4. Determination of the ability of binuclear ruthenium metal complexes to generate reactive oxygen species

[0110] To test the photocatalytic ability of the synthesized binuclear ruthenium complex to generate reactive oxygen species (ROS), hydroxyphenyl fluorescein (HPF) was used to determine this ability. When ROS are generated in the solution, HPF immediately captures them and is oxidized to produce fluorescein with high fluorescence intensity, emitting bright green fluorescence (Ex / Em = 540 / 590 nm). An increase in fluorescence intensity indicates the generation of ROS in the solution. The ability to generate ROS can be reflected by monitoring the changes in the fluorescence spectrum of the test sample and the HPF mixture under different illumination times using a fluorescence spectrophotometer.

[0111] An aqueous solution containing a binuclear ruthenium complex (5 μM) and HPF reagent (5 μM) was placed in a cuvette, and its reactive oxygen species (ROS) generation capacity was measured under both dark and light conditions. The results are as follows: Figure 5 As shown. From Figure 5 As can be seen from the data, this binuclear ruthenium complex has the ability to generate reactive oxygen species after being exposed to light.

[0112] 5. Photodynamic therapy effect of novel binuclear ruthenium metal complex on mouse triple-negative breast cancer cell lines

[0113] Resazurin solution is blue and is commonly used as an acid-base indicator (orange to deep purple at pH 3.8) and a redox indicator. In cell viability assays, resazurin can penetrate cells and be irreversibly reduced to pink by living cells, simultaneously producing the red fluorescence of resorufin. The absorbance or fluorescence intensity of resorufin is positively correlated with cell number and reducing capacity; therefore, cell proliferation can be analyzed using an enzyme-linked immunofluorescence assay (ELISA).

[0114] The experimental steps for the azure blade are as follows:

[0115] (1) First, revive one tube of 4T1 tumor cells and culture them in fresh complete culture medium (DMEM medium + 10 vol% fetal bovine serum + 1 vol% penicillin-streptomycin mixture). After passage twice, start the experiment.

[0116] (2) When the cells reach the logarithmic growth phase, seed them into two 96-well plates at a density of 5000 cells / well (each well is cultured with 100 μL of culture medium, one plate is the light group and the other is the dark control group), and incubate them in a 37°C, 5 vol% CO2 incubator.

[0117] (3) After the ruthenium complex adheres to the wall, remove the original culture medium and add 100 μL of ruthenium complex at 7 concentrations of 100, 50, 10, 1, 0.1, 0.01 and 0.001 μM to each well. Shake gently and incubate in a carbon dioxide incubator (37℃, 5 vol% CO2) in the dark.

[0118] (4) After incubation for 6 hours, the cell culture plates of the light-illuminated group were placed under a white light lamp for 20 minutes (light dose of 52.56 J / cm²). 2 Then, the cells were returned to the incubator and incubated in the dark for another 42 hours (the cells in the dark control group were kept in the incubator in the dark throughout the incubation).

[0119] (5) After incubation for 48 h, the culture medium was discarded from each well, and 80 μL of resazurin (100 μg / mL) was added to each well. The cells were then incubated at 37 °C for another 4 h. The EX540 / EM590 ratio was detected using the fluorescence plate of an ELISA reader, and the cell proliferation inhibition rate was calculated. The IC50 value was then determined. 50 Value (drug concentration when the inhibition rate is equal to 50%).

[0120] The cytotoxic effect of different concentrations of ruthenium complexes on mouse triple-negative breast cancer cells (4T1 cells) under dark and light treatment conditions was determined by the resazurite assay. Figure 6 As shown. From Figure 6 As can be seen, under no-light conditions, the IC50 of mouse triple-negative breast cancer cells (4T1 cells) was significantly reduced. 50=11.8 μM, IC50 of mouse triple-negative breast cancer cells under light conditions 50 The concentration was 0.08 μM, indicating that the ruthenium complex of the present invention has a strong photodynamic therapy effect.

[0121] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A dual-nuclear ruthenium photocatalyst, characterized in that: Compounds including those with the following structures: 。 2. The method for preparing the binuclear ruthenium photocatalyst according to claim 1, characterized in that: Includes the following steps: S1. 1,3-cyclohexadiene is reacted with ruthenium(III) salt to generate precursor compound I; S2. The precursor compound is reacted with 3-bromo-1,10-phenanthroline to generate precursor compound II; S3. React the precursor compound II with 2,2'-bipyridine and then with hexafluorophosphate to generate precursor compound III; S4. The precursor compound III is reacted with 2,5-di(2-ethylhexyl)-3,6-di(5-(trimethyltin)thiophen-2-yl)pyrrolo[3,4-c]pyrrolo-1,4-(2H,5H)-dione to obtain the product.

3. The preparation method according to claim 2, characterized in that: The reaction conditions for step S1 include at least one of the following conditions: (1) The reaction is carried out in an alcohol solvent system; (2) The reaction is carried out under reflux at 70-90℃; (3) The reaction time is 4-8 hours; (4) The molar ratio of 1,3-cyclohexadiene to ruthenium(III) salt is 2-2.2:

1.

4. The preparation method according to claim 3, characterized in that: The reaction conditions for step S1 include at least one of the following conditions: (1) The alcohol solvent is a lower alcohol; (2) The reaction temperature is 75-85℃; (3) The reaction time is 6 hours; (4) The molar ratio of 1,3-cyclohexadiene to ruthenium(III) salt is 2:

1.

5. The preparation method according to claim 4, characterized in that: The reaction conditions for step S1 include at least one of the following conditions: (1) The lower alcohol is a C1~C6 alcohol; (2) The reaction temperature is 80℃; (3) The reaction time is 6 hours.

6. The preparation method according to claim 5, characterized in that: The lower alcohol is ethanol.

7. The preparation method according to claim 2, characterized in that: The reaction conditions for step S2 include at least one of the following conditions: (1) The reaction is carried out in an alcohol solvent system; (2) The reaction is carried out under reflux at 70-90℃; (3) The reaction time is 3-5 hours; (4) The molar ratio of the precursor compound I to 3-bromo-1,10-phenanthroline is 1:1-1.

2.

8. The preparation method according to claim 7, characterized in that: The reaction conditions for step S2 include at least one of the following conditions: (1) The alcohol solvent is ethylene glycol; (2) The reaction temperature is 75-85℃; (3) The reaction time is 4 hours; (4) The molar ratio of the precursor compound I to 3-bromo-1,10-phenanthroline is 1:

1.

9. The preparation method according to claim 7, characterized in that: The reaction conditions for step S2 include: a reaction temperature of 80°C.

10. The preparation method according to claim 2, characterized in that: The reaction conditions for step S3 include at least one of the following conditions: (1) The reaction is carried out in an alcohol solvent system; (2) The reaction is carried out under reflux at 180-220℃; (3) The reaction time is 20-40 min; (4) The molar ratio of the precursor compound II to 2,2'-bipyridine is 1:2-2.

2.

11. The preparation method according to claim 10, characterized in that: The reaction conditions for step S3 include at least one of the following conditions: (1) The alcohol solvent is ethylene glycol; (2) The reaction temperature is 190-210℃; (3) The reaction time is 30 min; (4) The molar ratio of the precursor compound II to 2,2'-bipyridine is 1:

2.

12. The preparation method according to claim 10, characterized in that: The reaction conditions for step S3 include a reaction temperature of 200°C.

13. The preparation method according to claim 2, characterized in that: The reaction conditions for step S4 include at least one of the following conditions: (1) The reaction is carried out under a protective atmosphere; (2) The reaction was carried out in a mixture of toluene and N,N-dimethylformamide; (3) Reaction temperature: 100-135℃; (4) The reaction time is 18-24 hours; (5) The molar ratio of the precursor compound III to 1,3-bis(2-ethylhexyl)-5,7-bis(5-(trimethyltinyl)thiophen-2-yl)-4H,8H-benzo[1,2-c:4,5-c']dithiophene-4,8-dione is 1:(1.8-2.2). (6) The reaction is carried out under the catalysis of a catalyst, which includes tetra(triphenylphosphine)palladium.

14. The preparation method according to claim 13, characterized in that: The reaction conditions for step S4 include at least one of the following conditions: (1) The protective atmosphere is an argon atmosphere; (2) The volume ratio of toluene to N,N-dimethylformamide is 1:1; (3) The reaction temperature is 110-120℃; (4) The reaction time is 18-22 hours; (5) The molar ratio of the ruthenium complex to 1,3-bis(2-ethylhexyl)-5,7-bis(5-(trimethyltinyl)thiophen-2-yl)-4H,8H-benzo[1,2-c:4,5-c']dithiophene-4,8-dione is 1:1.9-2.1; (6) The molar ratio of the precursor compound III to the catalyst is 1:0.08~0.

12.

15. The preparation method according to claim 13, characterized in that: The reaction conditions for step S4 include at least one of the following conditions: (1) The volume ratio of toluene to N,N-dimethylformamide is 1:1; (2) The reaction temperature is 115℃; (3) The reaction time is 20 hours; (4) The molar ratio of the ruthenium complex to 1,3-bis(2-ethylhexyl)-5,7-bis(5-(trimethyltinyl)thiophen-2-yl)-4H,8H-benzo[1,2-c:4,5-c']dithiophene-4,8-dione is 1:2; (5) The molar ratio of the precursor compound III to the catalyst is 1:0.

1.

16. The application of the binuclear ruthenium photocatalyst as described in claim 1 in the preparation of antitumor photocatalytic drugs.

17. An antitumor drug, characterized in that: The active ingredient of the antitumor drug includes the binuclear ruthenium photocatalyst as described in claim 1.

18. The antitumor drug according to claim 17, characterized in that: The antitumor drug has at least one of the following characteristics: (1) The drug is a photosensitizer; (2) The tumor is breast cancer; (3) The drug also includes pharmaceutically acceptable carriers and / or excipients; (4) The dosage form of the drug includes at least one of hard-shell or soft-shell gelatin capsules, tablets, pills, powder injections, solutions, suspensions, elixirs, syrups, dry films, gels, buccal or sublingual tablets, films, suppositories and enemas; (5) The raw materials for preparing the drug also include at least one of the following: liposome forming components, carrierd or uncarrierized proteins, organic or inorganic nanoparticles, nanoemulsions, microemulsions, nanocrystals, and solvents; (6) The routes of administration of the drug include oral, gastrointestinal or parenteral.

19. The antitumor drug according to claim 18, characterized in that: The antitumor drug has at least one of the following characteristics: (1) The tumor is triple-negative breast cancer; (2) The excipient refers to at least one of the following: diluent, coating agent, binder, lubricant, disintegrant, solubilizer or stabilizer that can be used in the pharmaceutical field; the carrier is a functional pharmaceutical excipient acceptable in the pharmaceutical field, including at least one of surfactant, suspending agent or emulsifier.

20. The antitumor drug according to claim 18, characterized in that: The antitumor drug has at least one of the following characteristics: (1) The tumor is a triple-negative breast cancer in mice; (2) The carrier includes at least one of cyclodextrin, chitosan, polylactic acid, polyglycolic acid-polylactic acid copolymer or hyaluronic acid.

21. The antitumor drug according to claim 20, characterized in that: The cancer cells in the mouse triple-negative breast cancer were 4T1 cancer cells.

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