Cobalt-iridium metal complex, nano particle, preparation method and application

Abstract

The invention provides a cobalt-iridium metal complex, nanoparticles, a preparation method and application, and belongs to the technical field of photodynamic therapy drugs. The structural formula of the cobalt-iridium metal complex is shown as a formula (I). The invention also provides a preparation method of the cobalt-iridium metal complex. The invention also provides nanoparticles, the cobalt-iridium metal complex prepared by the invention has a chemiluminescence property, so that the cobalt-iridium metal complex can simultaneously realize photo-thermal, photodynamic and ultrasonic synergistic thrombolysis treatment effects, and the oxidation resistance of the cobalt-iridium metal complex has a remarkable curative effect on a vascular injury part. The complex provided by the invention has good photo-thermal, photodynamic and ultrasonic response capabilities, and can overcome the problems of short excitation wavelength of exciting light and limited light penetration depth required by the existing metal iridium complex. Experiments show that the material has a good treatment effect on thrombus.

Description

Technical Field

[0001] This invention belongs to the field of phototherapy and acoustic therapy drug technology, specifically relating to a cobalt-iridium metal complex, nanoparticles, preparation method, and application in multifunctional combined thrombolysis using photothermal, photodynamic, and acoustic therapies. Background Technology

[0002] Phototherapy, including photodynamic therapy (PDT) and photothermal therapy (PTT), is a new type of non-invasive, real-time controllable light stimulation with broad application prospects in disease treatment. As a non-invasive technique, low-intensity ultrasound is widely used in clinical diagnosis because it can penetrate the human body to a greater depth than light. Recently, ultrasonic dynamic therapy (SDT) has been explored as a promising alternative to thrombolytic therapy. Compared with traditional drug and surgical methods, phototherapy and sound therapy have inherent advantages such as being non-invasive, having low toxicity, being simple to operate, and having rapid recovery, and have attracted much attention in the medical field.

[0003] Surgical removal of the embolus or thrombolysis are the main clinical treatments for ischemic stroke. However, current thrombolytic drugs often exhibit only limited therapeutic effects due to their low selectivity, short half-life, and significant bleeding risk. In recent years, the non-invasive application of photosensitizers in the diagnosis and treatment of cardiovascular and cerebrovascular diseases has attracted widespread attention, especially in thrombolysis, and has the potential to become a next-generation treatment method.

[0004] While photothermal torsion (PTT) has shown potential in thrombolytic therapy, its non-specific thermal damage to nearby tissues is a limiting factor. Furthermore, high cost, insufficient photothermal effect, low biodegradability, and potential toxicity also restrict its clinical translation in thrombolytic therapy. PDT, due to its rapid onset of action, mild adverse reactions, and ability to be used in combination with other therapies, has been used in thrombolytic therapeutic research. However, the clinical application of PDT is also limited by its oxygen dependence, insufficient ROS generation efficiency of the photosensitizer, and, like PDT, the limited penetration depth due to insufficient light source wavelength.

[0005] As research progresses, the integration of PTT and PDT into a single system for synergistic treatment demonstrates promising application prospects for high efficiency and safety. The addition of sonic therapy can effectively compensate for the insufficient penetration depth of phototherapy, thereby enhancing the thrombolytic effect.

[0006] Transition metal materials are highly favored due to their rich excited-state properties, such as high luminescence quantum efficiency, ultra-long luminescence lifetime, high stable luminescence, and multicolor luminescence. Compared with other transition metal complexes, cyclic iridium complexes have long triplet excitation lifetimes, large Stokes shifts, and outstanding photothermal generation capabilities, making them the most widely used class of transition metal complex materials. In 2020, Professor Gou Shaohua's team at Southeast University, for the first time, rationally designed a metal iridium complex with donor-acceptor-donor (DAD) based on a simple framework structure, achieving a highly efficient antithrombotic therapeutic effect by utilizing the synergistic effect of photodynamics and photothermality. However, the synthesis of this photosensitizer is relatively complex, limiting its widespread application. (AnIridium (III) Complex Bearing a Donor–Acceptor–Donor Type Ligand for NIR‐Triggered Dual Phototherapy[J].Advanced Functional Materials, 2020.) In 2021, the inventors rationally designed mononuclear and tetranuclear metal iridium complexes based on the porphyrin framework structure and achieved excellent photothermal properties. However, the fluorescence photosensitizers would affect the imaging effect due to their own background (Zhang Liping. Design, synthesis and optical therapy application of deep red / near-infrared photosensitizers based on metal iridium complexes and small organic molecules [D]. Northeast Normal University, 2021.).

[0007] Currently, there are still limited metal complexes suitable for integrated thrombosis therapy, and there are few reports on drug design targeting the thrombotic microenvironment. Providing more metal complexes that can be used in integrated therapy is of great significance for clinical selection of thrombosis treatment. Summary of the Invention

[0008] The purpose of this invention is to provide a cobalt-iridium metal complex, nanoparticles, preparation method, and application. The cobalt-iridium metal complex of this invention has chemiluminescent properties, enabling visual monitoring in the early stages of disease development. Its PDT / PTT / SDT performance achieves effective ablation of thrombi, while its excellent antioxidant properties provide excellent therapeutic effects for vascular damage repair.

[0009] This invention first provides a cobalt-iridium metal complex, the structural formula of which is shown in formula (Ⅰ): Equation (I) The present invention also provides a method for preparing the above-mentioned cobalt-iridium metal iridium complex, comprising the following steps: S1. Under nitrogen protection, IrCl3·3H2O and phenylpyridine ligand were heated and refluxed to obtain phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2; S2. Under nitrogen protection, the phenylpyridine iridium dichlorobridge and porphyrin Schiff base ligand prepared in S1 were placed in the dark and refluxed in the presence of a solvent. After the reaction was completed, potassium hexafluorophosphate was added and stirring was continued to obtain the binuclear metal iridium complex 2Ir-NN. The metal iridium complex 2Ir-NN was acidified in hydrochloric acid to obtain the complex 2Ir. S3. Under nitrogen protection, the 2Ir obtained in S2 is mixed with cobalt chloride and reacted to obtain the cobalt-iridium metal complex Co2Ir.

[0010] Preferably, in step S2, the molar ratio of the phenylpyridine iridium dichlorobridge and the porphyrin Schiff base ligand is 1:1.

[0011] Preferably, in step S2, the reflux reaction temperature is 75~80℃ and the reflux reaction time is 6~8 h.

[0012] Preferably, in step S3, the molar ratio of the binuclear metallic iridium complex 2Ir to cobalt chloride is 1:(5~7).

[0013] Preferably, in step S3, the reaction temperature is 150℃-170℃ and the reaction time is 9-12 h.

[0014] The present invention also provides a nanoparticle prepared using the above-mentioned metal iridium complex conjugate, the structural formula of which is shown in formula (II): (II) In equation (II), the value of n ranges from 44 to 46.

[0015] This invention also provides a method for preparing nanoparticles, comprising: polyethylene glycol 2000 -TK-COOH and N-hydroxysuccinimide were dissolved in dimethyl sulfoxide and stirred in the dark. Then, the cobalt-iridium metal complex Co2Ir was added and stirred. After dialyzing, Co2Ir nanoparticles NPs were obtained.

[0016] The present invention also provides the application of the above-mentioned nanoparticles as phototherapy and sound therapy materials.

[0017] The present invention also provides the application of the above-mentioned nanoparticles in the preparation of drugs for treating thrombotic diseases.

[0018] Beneficial effects of the present invention This invention provides a cobalt-iridium metal complex, nanoparticles, preparation method, and application. This invention designs and synthesizes a cobalt-iridium metal complex Co2Ir NPs in a simple and convenient manner. The synthesized cobalt-iridium metal complex has chemiluminescent properties, enabling visual monitoring in the early stages of disease development. At the same time, its excellent antioxidant properties have a superior therapeutic effect on vascular damage repair.

[0019] This invention also provides a cobalt-iridium metal complex Co2Ir bio-encapsulated in polyethylene glycol. 2000 The nanoparticles Co2Ir NPs formed in -TK-COOH exhibit excellent bioluminescent properties, specifically recognizing the location of thrombi. The Co2Ir NPs provided by this invention can chemically react with excess ROS at the site of inflammation without the need for an external excitation light source, generating bioluminescence. Experiments show that they have a high penetration depth of 10 mm, overcoming the limitations of existing iridium complexes which require short excitation wavelengths and have limited light penetration depth. Test results show that Co2Ir NPs have good antioxidant capacity, indicating that this material has a good therapeutic effect on thrombi and holds promise for development as a phototherapy and sound therapy material, and for use as a thrombus treatment drug. Attached Figure Description

[0020] Figure 1 Mass spectrometry data of Co2Ir prepared in Example 1 of this invention; Figure 2 The particle size potential diagram of Co2Ir NPs prepared in this invention; Figure 3 The UV absorption spectra of 2Ir NPs and Co2Ir NPs prepared in this invention in aqueous solution; Figure 4 AIE performance test graphs of 2Ir NPs and Co2Ir NPs prepared for this invention under the condition of DMSO as a good solvent and water as a poor solvent; Figure 5 shows the first-order kinetic curves of the aqueous solutions of 2Ir NPs and Co2Ir NPs prepared in this invention under ultrasonic conditions; Figure 6 shows the first-order kinetic curves of the aqueous solutions of 2Ir NPs and Co2Ir NPs prepared in this invention under light irradiation conditions; Figure 7 Photothermal data of 2Ir NPs and Co2Ir NPs prepared in this invention in aqueous solution; Figure 8 The graph shows the chemiluminescence performance test data of Co2Ir NPs prepared in this invention; Figure 9 The image shows thrombolytic data of Co2Ir NPs prepared in this invention under different conditions. Figure 10 The graph shows the blood compatibility test data of 2Ir NPs and Co2Ir NPs prepared in this invention. Figure 11 This is a graph showing the antioxidant capacity of Co2Ir NPs prepared according to the present invention. Figure 12 This is a cell migration assay diagram of Co2Ir NPs prepared in this invention; Figure 13 This is a monitoring test image of Co2Ir NPs cells forming tubes prepared according to the present invention; Figure 14 This is a test image of Co2Ir NPs in mice prepared according to the present invention for thrombosis imaging monitoring. Figure 15 The graph shows the thrombolysis data of mice under different conditions using the Co2Ir NPs prepared in this invention. Figure 16 The images show H&E slice data of blood vessels and mouse organs of Co2Ir NPs prepared in this invention. Detailed Implementation

[0021] This invention first provides a cobalt-iridium metal complex, the structural formula of which is shown in formula (Ⅰ): Equation (Ⅰ).

[0022] The present invention also provides a method for preparing the above-mentioned cobalt-iridium metal complex, comprising the following steps: S1. IrCl3·3H2O and phenylpyridine ligand are added to a reaction vessel containing solvent and water. The mixture is heated under nitrogen protection and refluxed, preferably at 120~130℃ for 24~30 h. After the reaction is cooled to room temperature, a large amount of undesirable solvent water is added to precipitate the precipitate, which is then filtered. The solvent is washed away with a large amount of water and ethanol. The resulting solid is dried to obtain phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2. The molar ratio of IrCl3·3H2O to phenylpyridine is preferably 1:(2.5~3), and the solvent is preferably 2-ethylene glycol ethyl ether. S2. Add the obtained phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2 and porphyrin Schiff base ligand (L4) to the reaction vessel, then add a solvent, preferably a mixture of dichloromethane and methanol (v:v=1:1). Under the condition of sufficient purging with inert gas N2, place the reaction vessel in the dark and reflux it. The reflux temperature is preferably 75-80℃, and the reaction time is preferably 6-8 hours. After the reaction is completed and cooled to room temperature, add potassium hexafluorophosphate solid to the solution in the flask and continue stirring at room temperature. The stirring time is preferably 45-60 seconds. The solvent in the system was removed using a rotary evaporator. Then, the system was extracted with dichloromethane and water to remove excess potassium hexafluorophosphate solid. The obtained substance was washed with petroleum ether and dried, and purified by column chromatography to obtain a purple-red solid, namely the binuclear metallic iridium complex 2Ir-NN. The molar ratio of the phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2 to the porphyrin Schiff base ligand was 1:1; the preferred molar ratio of the phenylpyridine iridium dichlorobridge to potassium hexafluorophosphate was 1:5. 2Ir-NN was dissolved in acetone, acidified with hydrochloric acid, and extracted with dichloromethane and water to obtain the product 2Ir. In the acidification process, 10 mL of concentrated hydrochloric acid was added, and the acidification time was preferably 30 min.

[0023] S3. Under nitrogen protection, the 2Ir prepared in step S2 is mixed with cobalt chloride at a molar ratio of 1:(5~7), and DMF is added as a reaction solvent. The reaction is preferably carried out at 150℃-170℃ for 9-12 h, and more preferably at 160℃ with stirring for 11 h. The product is extracted with dichloromethane and water, and the solvent in the system is removed by rotary evaporation. After drying, a purplish-black solid product, namely the cobalt-iridium metal complex, is obtained.

[0024] The synthetic route of this preparation method is shown below:

[0025]

[0026]

[0027] The present invention also provides a nanoparticle prepared using the above-mentioned bimetallic iridium complex coupling compound, wherein the nanoparticle has a particle size of 100-200 nm and a structural formula as shown in formula (II): (II).

[0028] In equation (II), the value of n ranges from 44 to 46.

[0029] This invention also provides a method for preparing nanoparticles, comprising: polyethylene glycol2000 -TK-COOH and N-hydroxysuccinimide were dissolved in dimethyl sulfoxide and stirred in the dark for two hours. Then, the cobalt-iridium metal complex Co2Ir was added and stirred at room temperature for 48 hours. After dialysis, Co2Ir nanoparticles (NPs) were obtained. Polyethylene glycol was also present. 2000 The preferred molar ratio of -TK-COOH, N-hydroxysuccinimide, and the cobalt-iridium metal complex Co2Ir is 2:1:1, and the reaction process is as follows:

[0030] The present invention also provides the application of the above-mentioned nanoparticles as phototherapy materials, wherein the phototherapy and acoustic therapy materials are preferably photosensitive and acoustic initiators.

[0031] The present invention also provides the application of the above-mentioned nanoparticles in the preparation of drugs for treating thrombotic diseases.

[0032] Unless otherwise stated, the terms used herein have the meanings conventionally understood by those skilled in the art. The technical solutions of the present invention will be described in more detail below with reference to embodiments, wherein the raw materials involved in the embodiments are all commercially available.

[0033] Example 1 S1. IrCl3·3H2O (0.1 mmol, 0.0352 g) and phenylpyridine ligand (0.3 mmol, 0.0471 g) were added to a round-bottom flask containing 30 mL of 2-ethylene glycol ethyl ether and 10 mL of water. The mixture was heated to reflux at 120 °C for 24 h under a nitrogen-protected atmosphere. After the reaction was cooled to room temperature, a large amount of water, a poor solvent, was added to precipitate the solid. The precipitate was then filtered and washed repeatedly with a large amount of water and ethanol. The resulting solid, when dried, was the phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2.

[0034] S2. Add the phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2 (0.1 mmol, 0.1072 g) and porphyrin Schiff base (0.1 mmol, 0.1030 g) obtained above to a 100 mL single-necked flask, and then add 60 mL of dichloromethane and methanol (v:v=1:1) as the solvent for the reaction system. Under the condition of fully purging with inert gas N2, the reaction was carried out in the dark under reflux at 80°C for 8 h. After the reaction was completed and cooled to room temperature, potassium hexafluorophosphate solid (1 mmol, 0.1842 g) was added to the solution in the flask, and the mixture was stirred at room temperature for 45 min. The solvent in the system was removed by rotary evaporation. Excess potassium hexafluorophosphate solid was removed by extraction with dichloromethane and water. The obtained substance was washed with petroleum ether and dried to obtain a purple-red solid, namely the binuclear metallic iridium complex 2Ir-NN. 2Ir-NN was dissolved in acetone, acidified with 10 mL of hydrochloric acid for 30 min, and extracted with dichloromethane and water to obtain the product 2Ir.

[0035] S3. Add 2Ir (0.1 mmol, 0.1854 g) and cobalt chloride (0.5 mmol, 0.064 g) to a 100 mL beaker, using DMF as the reaction solvent. Stir the reaction at 160 °C for 10 h. Extract the product with dichloromethane and water, remove the solvent from the system using a rotary evaporator, and dry to obtain a purplish-black solid product, namely the cobalt-iridium complex Co2Ir, with a yield of 85% and a molecular formula of C. 100 H 70 CoIr2N 14 Its relative molecular mass is 1911.44 g / mol. Figure 1 The mass spectrometry data for Co₂Ir prepared in Example 1 shows a molecular weight of 955.67 g / mol, compared to the fitted molecular weight (molecular ion peak [M]). 2+ The fact that the same structure was obtained proves that the structure was successfully prepared.

[0036] Example 2 The experimental conditions and procedures were the same as in Example 1, except that in step S3, 2Ir (0.1 mmol, 0.1854 g) and cobalt chloride (0.6 mmol, 0.0768 g) were added.

[0037] Example 3 The experimental conditions and procedures were the same as in Example 1, except that in step S3, 2Ir (0.1 mmol, 0.1854 g) and cobalt chloride (0.7 mmol, 0.0896 g) were added.

[0038] Example 4 The experimental conditions and steps were the same as in Example 1, except that in step S3, the reaction was stirred at 150°C for 10 h.

[0039] Example 5 The experimental conditions and steps were the same as in Example 1, except that in step S3, the reaction was stirred at 170°C for 10 h.

[0040] Example 6 The experimental conditions and steps were the same as in Example 1, except that in step S3, the reaction was stirred at 160°C for 8 hours.

[0041] The properties of the cobalt-iridium metal complex Co2Ir prepared in Example 1 were characterized as follows: 1. Preparation of nanoparticles polyethylene glycol 2000 -TK-COOH (0.2 mmol) and N-hydroxysuccinimide (0.1 mmol) were dissolved in dimethyl sulfoxide and stirred in the dark for two hours. Then, the cobalt-iridium metal complex Co2Ir (0.1 mmol 0.19 g) prepared in step 2 of Example 1 was added and stirred at room temperature for 48 h. After dialysis, Co2Ir nanoparticles NPs were obtained.

[0042] (0.1 mmol 0.2 g) polyethylene glycol 2000 -TK-COOH and N-hydroxysuccinimide (0.1 mmol, 0.0115 g) were dissolved in dimethyl sulfoxide and stirred in the dark for two hours. Then, the prepared metal complex 2Ir (0.1 mmol, 0.18 g) was added, and the mixture was stirred at room temperature for 48 h. After dialysis, 2Ir nanoparticles (NPs) were obtained. The structural formula of 2Ir is as follows:

[0043] 2. Physical properties of Co2Ir NPs Figure 1 The mass spectrometry data for Co2Ir is shown, which matches the theoretical values ​​to demonstrate the structure. Figure 2 The figure shows the particle size of Co2Ir NPs (measured by a particle size potential analyzer; the figure shows the particle size of 2Ir NPs and Co2Ir NPs, respectively, and the potential). As can be seen from the figure, the particle size of Co2Ir NPs in aqueous solution is 105.33 nm, and the particle size increases to 878.84 nm under ROS response, which means that ROS can effectively release drugs.

[0044] 3. Photophysical properties of Co2Ir NPs The UV absorbance of Co2Ir NPs in aqueous solution was measured using a UV-Vis spectrophotometer, and the results are as follows: Figure 3 As shown. Among them. Figure 3 The image shows the UV absorption spectrum. In the UV absorption, the two original Q bands of the porphyrin disappear because the Co metal enters the porphyrin nucleus.

[0045] Figure 4 For the present invention 2Ir ( Figure 4 (a)) and Co2Ir ( Figure 4 (c) Both exhibited weak emission in pure acetonitrile solution. With increasing water content, 2Ir ( Figure 4 (b) and Co2Ir ( Figure 4 The luminescence intensity of (d) was enhanced, demonstrating that it possesses typical characteristics of AIE. This will facilitate the formation of aggregated nanoparticles for subsequent diagnostic and therapeutic applications.

[0046] Figure 5 For the Co2Ir NPs prepared in this invention, in vitro singlet oxygen production experiments and related kinetic curves were measured using a UV spectrophotometer under illumination and DPBF conditions (in the figure, B is the logarithmic curve of A; the larger the slope, the more singlet oxygen is produced). Figure 5 (a) is a line graph showing the decrease in DPBF of NPs at a specific nanometer. Figure 5 (b) is the first-order kinetic curve corresponding to 5 (a). The degradation of DPBF under ultrasound proves that Co2Ir NPs have good singlet oxygen generation ability.

[0047] Figure 6 For the Co2Ir NPs prepared in this invention, in vitro singlet oxygen production experiments and related kinetic curves were measured using a UV spectrophotometer under illumination and DPBF conditions (in the figure, B is the logarithmic curve of A; the larger the slope, the more singlet oxygen is produced). Figure 6 (a) is a line graph showing the decrease in DPBF of NPs at a specific nanometer. Figure 6 (b) is the first-order kinetic curve corresponding to (a). The degradation of DPBF under light proves that Co2Ir NPs have a good ability to generate singlet oxygen.

[0048] 4. Photothermal properties of Co2Ir NP Figure 7 Photothermal data of 2Ir NPs and Co2Ir NPs prepared in this invention in aqueous solution, including a control group, temperature changes at different concentrations, temperature changes at different powers, and the photothermal cycle of the drug. Figure 7 (a) is a graph showing the temperature variation of different NPs. Figure 7 (b) shows the temperature variation at different concentrations of NPs, where Figure 7 (c) shows the temperature variation of NPs with different power levels. Figure 7 (d) is a diagram of five heating and cooling cycles of Co2Ir NP. Figure 7 Figure (a) shows that the drug group (2Ir NPs, Co2Ir NPs) had better photothermal generation capacity compared to the control group H2O; Figure 7 Figure (b) shows that the higher the concentration of Co2Ir NPs, the higher the temperature under the same light exposure time, indicating that it is concentration-dependent. Figure 7 Figure (c) shows that Co2Ir NPs exhibit power dependence, with higher power resulting in higher heat generation and temperature rise. Figure 7 Figure (d) shows the temperature changes of Co2IrNPs after five heating and cooling cycles, demonstrating that the drug has good photothermal cycling ability.

[0049] 5. Chemiluminescence experiment of Co2Ir NPs Figure 8 The table below shows the relevant data on the chemiluminescence of Co2Ir NPs. Chemiluminescence assays were performed using an IVIS mouse in vivo imaging system. See also... Figure 8 (a) Under different RONS responses, different types of RONS were co-incubated with Co2Ir NPs in a 96-well plate, ONOO - The strongest luminescence intensity was observed at the inflammatory sites in mice. - As a marker substance, it can respond to Co2IrNPs to produce chemiluminescence, illuminating inflamed areas. See also Figure 8 (b) The dotted-line plots of chemiluminescence intensity at different nanometers show that the bioluminescence emission peak can reach as long as 660 nm. See also Figure 8 (c), ONOO - The luminescence decay of Co2Ir NPs can reach 60 minutes. See also: Figure 8 (d) By covering the drug with chicken breast of varying thicknesses, its chemiluminescence can penetrate to a depth of up to 10 mm. This high penetration depth is beneficial for imaging testing of the drug at deep inflammatory sites. This has significant advantages over photosensitizers that depend on short-wavelength excitation light.

[0050] 6. In vitro thrombolysis experiment of Co2Ir NPs Five mL of different groups of Co2Ir NPs were placed together with thrombus clots in a 10 mL vial for in vitro thrombolysis testing. The results were as follows: Figure 9 As shown. Among them. Figure 9(a) A graph showing the color changes of the thrombus and solution before and after thrombolysis, where (I represents PBS, II represents before urokinase (UK), III represents 2Ir NPs, VI represents Co2Ir NPs, V represents PBS + L (where L represents light exposure); VI represents after UK; VII represents 2Ir NPs + L; VIII represents Co2Ir NPs + L; IX represents 2Ir NPs + US; X represents Co2Ir NPs + US; XI represents 2Ir NPs + L + US; XII represents Co2Ir NPs + L + US;) Figure 9 (b) This is a graph showing the thrombus dissolution rate. Compared with the control group, the Co2Ir NPs group achieved a thrombus dissolution rate of 92% when both PDT / PTT / US dual-mode thrombolysis were activated simultaneously. The experimental results indicate that Co2Ir NPs have good in vitro thrombolytic ability. Figure 9 (c) is a graph showing the hemoglobin and fibrin content in the supernatant after thrombolysis.

[0051] 7. Blood compatibility test of Co2Ir NPs Figure 10 (a) and Figure 10 (b) After incubating blood cells with different concentrations of Co2Ir NPs and 2Ir NPs for 2 hours, the supernatant was collected for testing, and the hemolysis rate was calculated. As shown in the figure, the hemolysis rates of both 2Ir NPs and Co2Ir NPs were less than 5%, indicating that 2Ir NPs and Co2Ir NPs have good blood compatibility and can be used for in vivo testing. The inset plots in the figure correspond to the changes in the solution before and after the addition of blood cells, with the horizontal axis representing different concentrations.

[0052] 8. Antioxidant performance experiment of Co2Ir NPs Figure 11 The results show the antioxidant capacity of Co2Ir NPs, including total antioxidant capacity and hydrogen peroxide consumption. See also... Figure 11 ,in Figure 11 (a) is a DPPH degradation diagram. Figure 11 (b) This is a graph showing the degradation of hydrogen peroxide, detected using a kit. It can be seen that when the Co2Ir NPs concentration reaches 300 μg / mL, the DPPH consumption rate reaches approximately 68%. See also... Figure 11 Co-incubation with hydrogen peroxide revealed a consumption rate of nearly 68.2%, demonstrating that Co2Ir NPs possess excellent antioxidant capacity and show promising therapeutic effects against inflammation in vivo.

[0053] 9. Cellular experiments promoting angiogenesis with Co2Ir NPs Figure 12 and Figure 13 To investigate endothelial cell migration during angiogenesis, Co2Ir NPs were co-incubated with HUVEC cells. Figure 12 (a) and tube formation Figure 12 (b) The ability to achieve wound closure. Scratch closure was observed at 12 and 24 hours, and representative images of wound healing in all groups are shown below. Figure 12 As shown in (b). Statistical results show that after 12 hours of culture, the wound healing rate of the Co2Ir NPs treatment group was significantly higher than that of the 2Ir NPs treatment group. Figure 13 This is a monitoring test diagram of Co2Ir NPs cells prepared according to the present invention. Figure 13 (a) is a tube formation image of Co2Ir NPs after 6 h of co-incubation with cells. Figure 13 The quantitative data in (b) and 13(c) also show a significant increase in the number of tubes formed. This indicates that Co2Ir NPs can effectively promote angiogenesis.

[0054] 10. In vivo experiments of Co2Ir NPs in mice Figure 14 In vivo chemiluminescence imaging of Co2Ir NPs. It is evident that after tail vein administration, the drug circulates to the thrombus site over time, exhibiting significant chemiluminescence under illumination, reaching its maximum aggregation level at 60 minutes, at which point thrombolytic therapy can be initiated.

[0055] Figure 15 This is an in vivo thrombolysis assay of Co2Ir NPs. Mice were anesthetized and the right carotid artery was located via thoracotomy. The carotid artery was incubated with 10% FeCl3 solution for approximately 10 minutes, causing the vessel to turn black, indicating thrombolysis was complete. After drug administration, the drug circulated to the thrombus site. Ultrasound (1.0 W cm⁻¹) was used to visualize the thrombus. -2 1 MHz) and laser (635 nm, 0.8 W cm⁻¹) -2 Irradiating the thrombus site has a good thrombolytic effect.

[0056] Figure 16 This is a H&E section of blood vessels and mouse heart, liver, spleen, lungs and kidneys after thrombolysis. Figure 16 (a) The vascular section clearly shows the formation of embolic vessels, and after thrombolysis, the plaque in the vessels is significantly reduced, demonstrating the good thrombolytic effect of RuIr NPs. Figure 16 (b) The organ sections of the mice were all normal, which proves that Co2Ir NPs have good biosafety.

[0057] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A cobalt-iridium metal complex, the structural formula of which is shown in formula (Ⅰ): （Ⅰ）。 2. The method for preparing the above-mentioned cobalt-iridium metal complex according to claim 1, characterized in that, Includes the following steps: S1. Under nitrogen protection, IrCl3·3H2O and phenylpyridine ligand were heated and refluxed to obtain phenylpyridine iridium dichlorobridge [Ir(ppy)2Cl2]2; S2. Under nitrogen protection, the phenylpyridine iridium dichlorobridge and porphyrin Schiff base ligand prepared in S1 were placed in the dark and refluxed in the presence of a solvent. After the reaction was completed, potassium hexafluorophosphate was added and the mixture was stirred to obtain the binuclear metallic iridium complex 2Ir-NN. The metallic iridium complex 2Ir-NN was acidified in hydrochloric acid and extracted with dichloromethane and water to obtain the complex 2Ir. S3. Under nitrogen protection, the 2Ir obtained in S2 is mixed with cobalt chloride and reacted to obtain the cobalt-iridium metal complex Co2Ir.

3. The method for preparing the above-mentioned cobalt-iridium metal complex according to claim 2, characterized in that, In step S2, the molar ratio of the phenylpyridine iridium dichlorobridge and the porphyrin Schiff base ligand is 1:

1.

4. The method for preparing the above-mentioned cobalt-iridium metal complex according to claim 2, characterized in that, In step S2, the reflux reaction temperature is 75~80℃, and the reflux reaction time is 6 h.

5. The method for preparing the above-mentioned cobalt-iridium metal complex according to claim 2, characterized in that, In step S3, the molar ratio of 2Ir to cobalt chloride is 1:(5~7).

6. The method for preparing the above-mentioned cobalt-iridium metal complex according to claim 2, characterized in that, In step S3, the reaction temperature is 150℃-170℃, and the reaction time is 6-9 h.

7. A nanoparticle, characterized in that, The nanoparticles were prepared using the cobalt-iridium metal complex described in claim 1, and the structural formula is shown in formula (II): （Ⅱ）， In equation (II), the value of n ranges from 44 to 46.

8. The method for preparing nanoparticles according to claim 7, characterized in that, include: polyethylene glycol 2000 -TK-COOH and N-hydroxysuccinimide were dissolved in dimethyl sulfoxide and stirred in the dark for two hours. Then, the cobalt-iridium metal complex Co2Ir was added and stirred at room temperature for 48 hours. After dialysis, Co2Ir nanoparticles NPs were obtained.

9. The application of the nanoparticles according to claim 7 as phototherapy and sound therapy materials.

10. The use of the nanoparticles according to claim 7 in the preparation of drugs for treating thrombotic diseases.

Images