Metal iridium complex with porphyrin part for multi-mode thrombolysis, preparation method, nanoparticles and application

Abstract

The invention provides a metal iridium complex with a porphyrin part for multi-mode thrombolysis, a preparation method, nanoparticles and application, and belongs to the technical field of chemical synthesis and biomedicine. The structural formula of the metal iridium complex is shown as a formula 1 or a formula 2. The invention also provides a preparation method of the metal iridium complex, and also provides a nano particle which is prepared from the metal iridium complex with a porphyrin part for multi-mode thrombolysis as shown in a formula 1 or a formula 2. The invention also provides application of the nanoparticles as a medicine in preparation of medicines for treating thrombosis diseases. The metal iridium complex disclosed by the invention not only can realize PTT / SDT dual-mode combined treatment of thrombus, but also can be used as a chemiluminescent substrate to act with active oxygen for imaging, so that integration of diagnosis and treatment is realized.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical synthesis and biomedicine, and in particular relates to a metal iridium complex with a porphyrin part for multimodal thrombolysis, a preparation method, nanoparticles and applications. Background Art

[0002] Thrombotic cardiovascular and cerebrovascular diseases such as ischemic stroke, myocardial infarction and venous thromboembolism are among the most serious diseases with the highest morbidity and mortality worldwide. The development of new strategies for the treatment of thrombosis has always been the focus of clinical research. At present, the treatment strategies for thrombosis mainly include the use of thrombolytic drugs. Fibrinolytics, anticoagulants and antiplatelet drugs are commonly used antithrombotic drugs in clinical practice. These traditional thrombolytic drugs are not targeted and metabolized rapidly in the body. In order to achieve effective thrombolysis, large doses of continuous administration are required, which increases the risk of bleeding. In addition to drug treatment, local thermal therapy of thrombus can effectively loosen the thrombus by destroying the non-covalent interaction of fibrin. Under the excitation of external light sources, photothermal agents can induce photothermal thrombolysis, but single photothermal therapy is usually not enough to eradicate thrombus. In view of the problem that traditional antithrombotic drugs or photothermal thrombolysis are not ideal, combined therapy shows significant advantages in thrombosis treatment.

[0003] Another different treatment modality is ultrasound-triggered sonodynamic therapy (SDT). SDT therapy relies on oxygen in tissues to generate the ROS required for treatment and is a promising anti-tumor modality due to its non-invasive nature, deep tissue penetration, and high spatiotemporal accuracy. Since US radiation can penetrate deep tissues, ultrasound therapy can overcome the poor light penetration depth of traditional photodynamic therapy and photothermal therapy, and can treat deep thrombi. The synergistic approach of combining photothermal therapy PTT and SDT can overcome the inherent limitations of a single treatment modality while also improving treatment efficiency.

[0004] Organic sonosensitizers mainly include porphyrins and cyanines, but their ROS production capacity is limited, which greatly limits the effect of sonodynamic therapy. In recent years, metal complexes such as ruthenium and iridium have become a new generation of sonosensitizers because of their large Stokes shift, high acoustic stability, and good biosafety. In addition, flexible structural modifications can give metal complexes the desired physicochemical and biological properties. Therefore, building porphyrins into iridium complexes can not only achieve efficient sonodynamic therapy, but also utilize the absorption of porphyrins in the near-infrared region to enable photothermal therapy. By integrating PTT / SDT combined thrombolysis into a nanoplatform, it is expected to become a new strategy for the clinical treatment of thrombosis. Summary of the invention

[0005] The present invention provides a metal iridium complex with a porphyrin part for multimodal thrombolysis, a preparation method, nanoparticles and applications. Porphyrin derivatives and metal iridium can be used as sonosensitizers to achieve ultrasonic thrombolysis, and the porphyrin derivatives can be used as photothermal agents due to their absorption in the red light region. The metal iridium complex can simultaneously achieve PTT and SDT synergistic treatment of thrombus, further enhancing its thrombolytic effect.

[0006] The present invention first provides a metal iridium complex with a porphyrin moiety for multimodal thrombolysis, the structural formula of which is shown in Formula 1 or Formula 2:

[0007]

[0008] The present invention also provides a method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis, and the synthesis method is as follows:

[0009] Step 1: Under nitrogen protection, using tetrakistriphenylphosphine palladium as a catalyst, 4-boric acid triphenylamine and 1-chloroisoquinoline are heated under reflux to react to prepare a cyclometallated ligand TPA;

[0010] Step 2: Under nitrogen protection, IrCl3·3H2O and cyclometallated ligand TPA or 1-phenylisoquinoline ligand are heated under reflux to react to obtain quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2;

[0011] Step 3: dissolving 4-(10,15,20-triphenylporphyrin-5-yl)aniline and pyridine-2-carboxaldehyde in anhydrous ethanol and heating under reflux to obtain a porphyrin Schiff base ligand;

[0012] Step 4: Under nitrogen protection, the quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 obtained in step 2 and the porphyrin Schiff base ligand obtained in step 3 are dissolved in a mixed solvent of methanol and dichloromethane to reflux for reaction in the dark. After the reaction is completed, potassium hexafluorophosphate is added and stirred continuously to obtain the metal iridium complex Ir-TPA or Ir-PY.

[0013] Preferably, in the step 1, the molar ratio of 4-triphenylamine borate, 1-chloroisoquinoline and tetrakistriphenylphosphine palladium is 1.1:1:(0.05-0.06).

[0014] Preferably, in step 2, the molar ratio of IrCl3·3H2O to TPA ligand or 1-phenylisoquinoline ligand is 1:(2-2.5).

[0015] Preferably, in the step, the molar ratio of tripyridine-2-carboxaldehyde to 4-(10,15,20-triphenylporphyrin-5-yl)aniline is 2:1.

[0016] Preferably, the reaction temperature of step three is 75-80° C. and the reaction time is 8-12 h.

[0017] Preferably, in the step, the molar ratio of tetraquinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 to the porphyrin Schiff base ligand is 1:(2-2.2).

[0018] Preferably, the temperature of the reflux reaction in step 4 is 70-80° C., and the reaction time is 8-12 h.

[0019] The present invention also provides a nanoparticle prepared from a metal iridium complex with a porphyrin part shown in Formula 1 or Formula 2 for multimodal thrombolysis.

[0020] The present invention also provides the use of the nanoparticles in preparing drugs for treating thrombotic diseases.

[0021] Beneficial Effects of the Invention

[0022] The present invention provides a metal iridium complex with a porphyrin moiety for multimodal thrombolysis, a preparation method, nanoparticles and applications. The structural formula of the metal iridium complex is shown in Formula 1 or Formula 2. The present invention also provides a nanoparticle prepared from the metal iridium complex with a porphyrin moiety for multimodal thrombolysis shown in Formula 1 or Formula 2. The present invention also provides the use of the above-mentioned nanoparticles as a drug in the preparation and treatment of thrombotic diseases. The present invention synthesizes a metal iridium complex Ir-TPA and Ir-PY with a porphyrin moiety in a simple and convenient manner, introduces a porphyrin group into the metal iridium complex, improves its sonodynamic thrombolytic treatment effect, and utilizes the near-infrared absorption of the porphyrin derivative to achieve dual-modal photothermal / sound dynamic combined treatment, and the metal iridium complex Ir-TPA can be used as a chemiluminescent substrate to respond to ROS at the thrombus site to emit light, thereby achieving the purpose of accurate diagnosis and treatment. This type of strategy for the synergistic treatment of thrombosis-related diseases is of great significance in practical applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The NMR images of Ir-TPA and Ir-PY prepared in the present invention;

[0024] Figure 2 The particle size potential diagram of Ir-TPA NPs and Ir-PY prepared in the present invention;

[0025] Figure 3The ultraviolet absorption spectra and fluorescence emission spectra of Ir-TPA NPs and Ir-PY in solution prepared by the present invention are shown in FIG.

[0026] Figure 4 The photodynamic ROS generation diagram of Ir-TPA NPs and Ir-PY prepared in the present invention in solution;

[0027] Figure 5 The sonodynamic ROS generation diagram of Ir-TPA NPs and Ir-PY prepared in the present invention in solution;

[0028] Figure 6 This is the photothermal data diagram of Ir-TPA NPs prepared in the present invention in solution;

[0029] Figure 7 The photothermal images of Ir-TPA NPs and Ir-PY prepared in the present invention in the solution;

[0030] Figure 8 The acoustic thermal data diagram of Ir-TPA NPs prepared in the present invention in solution;

[0031] Fig. 9 The chemiluminescent properties of Ir-TPA NPs prepared in the present invention are shown in FIG.

[0032] Fig.10 This is an in vitro targeting test diagram of Ir-TPA NPs prepared by the present invention;

[0033] Fig.11 This is a hemolysis test diagram of Ir-TPA NPs and Ir-PY prepared in the present invention;

[0034] Fig.12 The thrombolysis data of Ir-TPA NPs and Ir-PY prepared in the present invention under different conditions;

[0035] Fig.13 This is the in vivo thrombolysis diagram of Ir-TPA NPs prepared by the present invention in mice;

[0036] Fig.14 This is the laser speckle pattern of Ir-TPA NPs prepared in the present invention in mice. DETAILED DESCRIPTION

[0037] The present invention first provides a metal iridium complex with a porphyrin moiety, Ir-TPA and Ir-PY, for multimodal thrombolysis. The molecular formula of Ir-TPA is C 104 H 72 Ir 10 , the relative molecular mass is 1654.01g / mol, and the molecular formula of Ir-PY is C 80 H54 IrN8, the relative molecular mass is 1319.5 g / mol and its structural formula is shown in Formula 1 or Formula 2:

[0038]

[0039] The present invention also provides a method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis, comprising:

[0040] Step 1: Under nitrogen protection, using tetrakistriphenylphosphine palladium as a catalyst, heating 4-boric acid triphenylamine and 1-chloroisoquinoline to reflux for reaction, the reaction temperature is preferably 110-120°C, the reaction time is preferably 36-48h, the solvent added during the reaction is preferably toluene, and sodium carbonate solution is preferably added during the reaction, and the concentration of the sodium carbonate solution is preferably 2mol / L. After the reaction is completed, post-treatment is performed, and the post-treatment is preferably, after cooling, extraction is performed with dichloromethane and water, the organic phase is dried with anhydrous magnesium sulfate, filtered, the organic phase is spin-dried, and the crude product is purified by silica gel column chromatography to obtain a cyclometal ligand TPA, and the molar ratio of the 4-boric acid triphenylamine, 1-chloroisoquinoline, and tetrakistriphenylphosphine palladium is preferably: 1.1:1:(0.05-0.06);

[0041] Step 2: Under nitrogen protection, IrCl3·3H2O and cyclometallated ligand TPA or 1-phenylisoquinoline ligand are heated to reflux for reaction, wherein the reaction temperature is preferably 120-130°C, the reaction time is preferably 24-30h, and the solvent added during the reaction is preferably a mixed solution of ethylene glycol ethyl ether and water, wherein the volume ratio of ethylene glycol ethyl ether to water is preferably 3:1; after stopping the reaction, standing and cooling to room temperature, then adding water, and continuing to stir, wherein the stirring time is preferably 30min, and after suction filtration and drying, quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 is obtained; the molar ratio of IrCl3·3H2O and TPA ligand or 1-phenylisoquinoline ligand is preferably 1:(2-2.5);

[0042] Step 3: dissolving 4-(10,15,20-triphenylporphyrin-5-yl)aniline and pyridine-2-carboxaldehyde in anhydrous ethanol and heating under reflux for reaction, wherein the reaction temperature is preferably 75-80°C, and the reaction time is preferably 8-12 hours. When the reaction is completed and cooled to room temperature, a large amount of ethanol is added to the reactor to precipitate the precipitate, which is filtered and washed with a large amount of ethanol solvent for multiple times to obtain a porphyrin Schiff base ligand; the molar ratio of pyridine-2-carboxaldehyde to 4-(10,15,20-triphenylporphyrin-5-yl)aniline is preferably 2:1;

[0043] Step 4: Under nitrogen protection, the quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 obtained in step 2 and the porphyrin Schiff base ligand obtained in step 3 are dissolved in a solvent in the dark for reflux reaction, wherein the solvent is preferably a mixed solvent of methanol and dichloromethane, wherein the volume ratio of methanol to dichloromethane is preferably 1:1, the temperature of the reflux reaction is preferably 70-80°C, the reaction time is preferably 8-12h, and after the reaction is completed, potassium hexafluorophosphate is added and stirred continuously, and the stirring time is preferably 30-40 minutes, after which the solvent in the system is removed by a rotary evaporator, and the excess potassium hexafluorophosphate solid is removed by extraction with dichloromethane and water, and the obtained substance is washed with petroleum ether and dried, and purified by column chromatography to obtain a metal iridium complex; the molar ratio of the quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 to the porphyrin Schiff base ligand is preferably 1:(2-2.2), and the molar ratio of the porphyrin Schiff base ligand to potassium hexafluorophosphate is preferably 1:(10-15).

[0044] The specific reaction process is as follows:

[0045]

[0046] The present invention also provides a nanoparticle prepared from a metal iridium complex with a porphyrin part shown in Formula 1 or Formula 2 for multimodal thrombolysis.

[0047] The present invention also provides a method for preparing nanoparticles, comprising:

[0048] The metal iridium complex with a porphyrin moiety for multimodal thrombolysis shown in Formula 1 or Formula 2 and distearoyl phosphatidylethanolamine-polyethylene glycol are dissolved in a solvent, preferably tetrahydrofuran, and ultrapure water is added dropwise with stirring, preferably stirred overnight at room temperature, and after the solvent is evaporated, the prepared nanoparticles are dialyzed to obtain nanoparticles with uniform particle size. The mass ratio of the metal iridium complex with a porphyrin moiety for multimodal thrombolysis and distearoyl phosphatidylethanolamine-polyethylene glycol is 1:2.

[0049] The present invention also provides the use of the nanoparticles in preparing drugs for treating thrombotic diseases.

[0050] The present invention is further described by way of the following examples, which do not limit the present invention in any way. Without departing from the technical solution of the present invention, any modification or change made to the present invention that is easily implemented by a person of ordinary skill in the art will fall within the scope of the claims of the present invention.

[0051] Example 1

[0052] Step 1: Dissolve 4-boric acid triphenylamine (0.972 g, 3.36 mmol) and 1-chloroisoquinoline (0.496 g, 3.06 mmol) in 30 mL of toluene, and add tetrakistriphenylphosphine palladium (0.188 g, 0.16 mmol) as a catalyst. Add 20 mL of 2 mol / L sodium carbonate solution, protect with nitrogen atmosphere, and reflux at 110 ° C for 48 h. After the reaction is completed, cool and extract with dichloromethane and water. Dry the organic phase with anhydrous magnesium sulfate, filter, spin dry the organic phase, and purify the crude product by silica gel column chromatography to obtain a light yellow solid, which is TPA (1).

[0053] Step 2: Dissolve TPA (0.930 g, 2.5 mmol) (1) and IrCl3·H2O (0.317 g, 1 mmol) (2) in a mixed solution of 30 mL of ethylene glycol ether and 10 mL of water, react under nitrogen protection, and reflux at 120°C for 24 h. Stop the reaction, let it stand and cool to room temperature, add 15 mL of water to a round-bottom flask, continue stirring for 30 min, filter with a Buchner funnel, put the obtained solid in an oven for 24 h, and dry the obtained dark red solid, which is TPA dichloro bridge [Ir(TPA)2Cl2]2 (2).

[0054] Step 3: Pyridine-2-carboxaldehyde (0.2 mmol, 0.0214 g) and 4-(10,15,20-triphenylporphyrin-5-yl)aniline (0.1 mmol, 0.0629 g) were added to a 100 mL single-mouth bottle, respectively, and 30 mL of ethanol was used as the solvent of the reaction system, and the reaction was carried out at 80°C for 12 h. After the reaction was completed and cooled to room temperature, a large amount of ethanol was added to the solution in the bottle to precipitate the precipitate, which was filtered and washed with a large amount of ethanol solvent for multiple times to obtain a violet porphyrin Schiff base ligand (3).

[0055] Step 4: TPA dichloro bridge (0.097 g, 0.05 mmol) [Ir(TPA)2Cl2]2 (2) and porphyrin Schiff base ligand (0.1 mmol, 0.0718 g) (3) were added to a 100 mL single-mouth bottle, and 60 mL of dichloromethane and methanol (v:v=1:1) were used as solvents for the reaction system. Under the protection of inert gas N2, the reaction was placed in a dark place and refluxed for 8 hours. When the reaction was completed and cooled to room temperature, potassium hexafluorophosphate solid (1 mmol, 0.184 g) was added to the solution in the bottle, and stirring was continued at room temperature for 30 minutes. The solvent in the system was removed by rotary evaporator, and the excess potassium hexafluorophosphate solid was extracted with dichloromethane and water, and the obtained substance was washed with petroleum ether and dried, and purified by column chromatography to obtain a purple-red solid, which is Ir-TPA. C 104 H 72 Ir10 The relative molecular mass is 1654.01 g / mol. The H NMR mass spectrum of the metal iridium complex Ir-TPA with a porphyrin moiety prepared in Example 1 is as follows: Figure 1 As shown in A.

[0056] Example 2

[0057] Step 1: Add IrCl3·3H2O (0.1mmol, 0.0352g) and 1-phenylisoquinoline ligand (0.25mmol, 0.05125g) to a round-bottom flask containing 30mL ethylene glycol ether and 10mL water, and heat under reflux for 24h under N2 protection. Stop the reaction, let it stand and cool to room temperature, add 15mL water to the round-bottom flask, continue stirring for 30min, use a Buchner funnel for suction filtration, put the obtained solid in an oven for 24h, and dry the obtained dark red solid, which is 1-phenylisoquinoline dichlorobridge [Ir(pqy)2Cl2]2(4).

[0058] Step 2: Pyridine-2-carboxaldehyde (0.2 mmol, 0.0214 g) and 4-(10,15,20-triphenylporphyrin-5-yl)aniline (0.1 mmol, 0.0629 g) were added to a 100 mL single-mouth bottle, and 30 mL of ethanol was used as the solvent of the reaction system. The reaction was carried out at 80°C for 12 h. After the reaction was completed and cooled to room temperature, a large amount of ethanol was added to the solution in the bottle to precipitate the precipitate, which was filtered and washed with a large amount of ethanol solvent for multiple times to obtain a violet porphyrin Schiff base ligand (3).

[0059] Step 3: Add 1-phenylisoquinoline dichloro bridge (0.05mmol g, 0.0635g) (4) and porphyrin Schiff base ligand (0.1mmol, 0.0718g) (3) to a 100mL single-mouth bottle, and use 60mL of dichloromethane and methanol (v:v=1:1) as the solvent of the reaction system. Under the protection of inert gas N2, place the reaction in a dark place and reflux for 8h. When the reaction is completed and cooled to room temperature, add potassium hexafluorophosphate solid (1mmol, 0.184g) to the solution in the bottle, continue to stir at room temperature for 30min, use a rotary evaporator to remove the solvent in the system, extract with dichloromethane and water to remove excess potassium hexafluorophosphate solid, and wash the obtained substance with petroleum ether and dry it, purify it by column chromatography, and obtain a purple-red solid, which is Ir-PY. The molecular formula is C 80 H 54 IrN8, relative molecular mass is 1319.5g / mol. NMR hydrogen spectrum mass spectrum is as follows Figure 1 As shown in B.

[0060] Example 3

[0061] The preparation process and conditions are the same as those in Example 1, except that the reaction time in step 1 is 40 h.

[0062] Example 4

[0063] The preparation process and conditions are the same as those in Example 1, except that the reaction time in step 2 is 28 h and the reaction temperature is 125° C.

[0064] Example 5

[0065] The preparation process and conditions are the same as those in Example 1, except that in step 2, TPA is (0.744 g, 2 mmol) and IrCl3·H2O is (0.317 g, 1 mmol).

[0066] Example 6

[0067] The preparation process and conditions are the same as those of Example 1, except that the reaction time in step 4 is 12 h. The metal iridium complexes prepared in Example 1 and Example 2 were characterized for performance as follows:

[0068] 1. Preparation and properties of Ir-TPA NPs and Ir-PY NPs:

[0069] The preparation method of Ir-TPA NPs and Ir-PY NPs in the present invention is to prepare nanoparticles in a simple and convenient manner. The specific operation is as follows:

[0070] Ir-TPA (0.0010 g) or Ir-PY (0.0010 g) and distearoyl phosphatidylethanolamine-polyethylene glycol (0.0020 g) were added to a 20 mL glass bottle and dissolved in 1 ml of tetrahydrofuran, and then added dropwise into 10 mL of ultrapure water. The mixture was stirred overnight at room temperature. After the tetrahydrofuran was evaporated, the prepared nanoparticles were dialyzed to obtain Ir-TPANPs and Ir-PY NPs with uniform particle size and stability.

[0071] The particle size and potential of Ir-TPA NPs and Ir-PY NPs in the present invention, Figure 2 is the particle size of Ir-TPA NPs and Ir-PY NPs of the present invention ( Figure 2 A, 2B) and potential diagram ( Figure 2C), as can be seen from the figure, the particle size of Ir-TPA NPs in aqueous solution is 79.75nm, and the particle size of Ir-PY NPs is 82.504nm. At the same time, the potential of Ir-TPA NPs is -5.05mV, and the potential of Ir-PYNPs is -2.72mV. The particle size of Ir-TPA NPs and Ir-PY NPs is <100nm, and the uniform size indicates that they have good biocompatibility, the potential is negative, and they can have a good circulation effect in the body.

[0072] 2. Photophysical properties of metal iridium complexes:

[0073] Figure 3 is the ultraviolet absorption and fluorescence emission of the Ir-TPA NPs and Ir-PY NPs of the present invention in DMSO solution, Figure 3 A is its ultraviolet absorption spectrum, Figure 3 B is the fluorescence emission spectrum;

[0074] 3. Singlet oxygen generation ability of Ir-TPA NPs:

[0075] Figure 4 The photodynamic performance test of Ir-TPA NPs and Ir-PY NPs prepared by the present invention is as follows: Figure 4 A is the use of 1,3-diphenylisobenzofuran (DPBF) as an indicator to evaluate the ROS production ability of Ir-TPA NPs and Ir-PY NPs. The absorbance of DPBF degrades after illumination in the presence of Ir-TPANPs, Ir-PY NPs, and DPBF, indicating the generation of ROS. Figure 4 B is the time-dependent generation 1 In the O2 generation kinetics curve, the larger the slope, the stronger the ROS generation ability. The ROS generation ability of Ir-TPANPs and Ir-PY NPs was significantly higher than that of the control group, indicating their excellent photodynamic properties, which can be used for subsequent thrombolysis tests.

[0076] Figure 5 The acoustic dynamic performance test diagram of Ir-TPA NPs and Ir-PY NPs. Figure 5 A is the in vitro singlet oxygen generation experiment of Ir-TPA NPs and Ir-PYNPs under ultrasound and DPBF conditions. Figure 5 B related kinetic curves.

[0077] 4. Photothermal properties of Ir-TPA NPs and Ir-PY NPs:

[0078] Figure 6The photothermal properties of Ir-TPANPs and Ir-PY NPs were investigated using PBS as a control. Figure 6 AAt the same laser power and irradiation time, the temperature of the control group hardly changed, while the temperature of the Ir-TPANPs group increased rapidly. Compared with the control group, Ir-TPA NPs had better photothermal generation ability. Then, the temperature changes of Ir-TPA NPs solutions with different concentrations under 635nm laser irradiation were measured. Figure 6 B shows that as the concentration of Ir-TPANPs increases, the temperature of the system also increases significantly under the same laser power. In order to reduce the damage to the organism caused by excessive laser power, Figure 6 C investigated the effect of laser power on the system temperature. The higher the laser power, the higher the system temperature. When the laser power was 0.8W / cm 2 The solution temperature can rise to about 60°C after 5 minutes of irradiation. Good photothermal stability is crucial for subsequent in vivo treatment. Figure 6 D shows that the maximum temperature of Ir-TPA NPs remained basically unchanged during 5 laser on-off cycles, and even increased slightly, indicating that Ir-TPA NPs have good photothermal stability and can be used for photothermal therapy.

[0079] Figure 7 The thermal images of Ir-TPA NPs and Ir-PY NPs corresponding to the photothermal data show that the change trend is similar to Figure 6 The temperature change curve in A is consistent.

[0080] 5. Acoustic and thermal properties of Ir-TPA NPs;

[0081] Figure 8 The acoustic-thermal properties of Ir-TPA NPs show that the Ir-TPANPs group has better acoustic-thermal generation capability than the control group;

[0082] 6. Chemiluminescence experiment of Ir-TPANPs:

[0083] Fig. 9 For the relevant test of Ir-TPA NPs chemiluminescence of the present invention, Fig. 9 A is to add a variety of reactive oxygen species into Ir-TPA NPs to explore the selectivity of Ir-TPA NPs chemiluminescence to several oxidants. It can be seen that compared with other types of reactive oxygen species, ONOO - It is a more effective chemiluminescent initiator. Next, the effect of ONOO- concentration on the chemiluminescent signal of Ir-TPA NPs was investigated. Fig. 9 As shown in B, the chemiluminescence signal of Ir-TPA NPs is related to ONOO- The concentration showed a very good linear correlation. Then the chemiluminescence intensity of Ir-TPA NPs under ultrasound stimulation was verified. As shown in 9C, the chemiluminescence intensity of Ir-TPA NPs was the strongest when ultrasound was applied for 4 minutes. Fig. 9 D verifies the chemiluminescence intensity of Ir-TPA NPs under different illumination times. It can be seen from the figure that the luminescence intensity is highest when the illumination time is 120s. Fig. 9 E explored and then verified the chemiluminescence signal attenuation of Ir-TPA NPs, such as Fig. 9 F shows the addition of ONOO - The luminescence signal of the group was significantly stronger than that of the laser irradiation group, and the continuous luminescence signal of Ir-TPANPs showed a decay half-life of about 3 minutes, which was conducive to subsequent in vivo imaging. - The penetration depth and sensitivity of the chemiluminescence were then compared with the fluorescence signal. Fig. 9 G shows that the chemiluminescence and fluorescence signals of Ir-TPA NPs decrease with the increase of chicken breast tissue thickness, and there is still a chemiluminescence signal when the chicken breast thickness is 9 mm. Chemiluminescence can eliminate the autofluorescence of tissues and avoid the photon scattering of excitation light, which is beneficial to improve the imaging depth.

[0084] Fig.10 This is an in vitro targeting test of the Ir-TPA NPs of the present invention. The Ir-TPA@RGD NPs obtained by encapsulating the cRGD targeting peptide with Ir-TPA were co-incubated with thrombi, and the cRGD-encapsulated group showed a more obvious chemiluminescent signal. And as time goes by, the chemiluminescent intensity of the thrombus increases, which is because the encapsulated cRGD peptide can bind to the protein on the surface of platelets and specifically enrich on the surface of platelet-rich thrombi.

[0085] 7. Hemolysis experiment of Ir-TPA NPs and Ir-PY NPs:

[0086] Fig.11 For the hemolysis test of Ir-TPA NPs and Ir-PY NPs of the present invention, 100 μL of red blood cells were incubated with different concentrations of Ir-TPA NPs or Ir-PY NPs in a constant temperature water box at 37°C for 2 h. The PBS group was the negative control group, and the water group was the positive control group. The measured hemolysis rate of the drug was about 5% at 100 μg / mL, indicating that Ir-TPA NPs and Ir-PY NPs have good biosafety.

[0087] 8. In vitro thrombolytic experiments of Ir-TPA NPs and Ir-PY NPs:

[0088] Fig.12 This is an in vitro thrombolytic test of the Ir-TPA NPs of the present invention (wherein P+u+L represents the Ir-PY NPs group, and T+u+L represents the Ir-TPA NPs group). Under the simultaneous action of 635nm laser and ultrasound, the Ir-TPA NPs group started the SDT / PTT dual-mode thrombolytic therapy, which produced a significant thrombolytic effect. Fig.12 A is the photo of hemolytic clot and solution after different treatments. Fig.12 B is the thrombolysis rate of blood clots. Compared with the single treatment group, the thrombolysis rate of Ir-TPA NPs under both laser and ultrasound was higher, reaching 73%. Fig.12 D is the measurement of hemoglobin and fibrin content in the supernatant after thrombolysis. The above experimental results show that Ir-TPA NPs have good in vitro thrombolytic ability, and the combined treatment effect of SDT / PTT is better than that of the single treatment group.

[0089] 9. In vivo experiments on mice using Ir-TPA NPs:

[0090] Fig.13 This is an in vivo thrombolytic test diagram of Ir-TPANPs of the present invention. The present invention adopts a rat carotid artery thrombosis model induced by FeCl3. After the arterial thrombosis model is established, laser irradiation is given and SDT / PTT dual-mode thrombosis treatment is started. After the treatment of the experimental group, the thrombus is significantly dissolved.

[0091] Fig.14 The blood flow of Ir-TPANPs in the present invention is recorded during the laser speckle monitoring of thrombolysis. After inducing thrombosis, the blood flow changes are monitored after administration through the tail vein. Fig.14 A) In the PBS treatment group, there was no significant change in blood flow after treatment with 635nm laser. Fig.14 B) is the UK group. After UK treatment, blood flow gradually recovered, but blood flow decreased again 30 minutes later. This may be because UK is easily cleared from the blood. Fig.14 C) Ir-TPANPs alone were irradiated with laser. Fig.14 D) Ir-TPANPs were stimulated with ultrasound alone. Fig.14 E) is the Ir-TPANPs PTT / SDT dual-mode treatment group, Fig.14 F) is a graph of blood perfusion under different conditions. The blood perfusion of the experimental group gradually recovered to more than 70% within 2 hours, and no re-embolism occurred. It had a better therapeutic effect than the single treatment group, indicating the effectiveness of dual-mode treatment of thrombosis.

Claims

1. A metal iridium complex with a porphyrin moiety for multimodal thrombolysis, characterized in that: Its structural formula is shown in Formula 1 or Formula 2:

2. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: include: Step 1: Under nitrogen protection, using tetrakistriphenylphosphine palladium as a catalyst, 4-boric acid triphenylamine and 1-chloroisoquinoline are heated under reflux to react to prepare a cyclometallated ligand TPA; Step 2: Under nitrogen protection, IrCl3·3H2O and cyclometallated ligand TPA or 1-phenylisoquinoline ligand are heated under reflux to react to obtain quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2; Step 3: dissolving 4-(10,15,20-triphenylporphyrin-5-yl)aniline and pyridine-2-carboxaldehyde in anhydrous ethanol and heating under reflux to obtain a porphyrin Schiff base ligand; Step 4: Under nitrogen protection, the quinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 obtained in step 2 and the porphyrin Schiff base ligand obtained in step 3 are dissolved in a mixed solvent of methanol and dichloromethane to reflux for reaction in the dark. After the reaction is completed, potassium hexafluorophosphate is added and stirred continuously to obtain the metal iridium complex Ir-TPA or Ir-PY.

3. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: In the step 1, the molar ratio of 4-triphenylamine borate, 1-chloroisoquinoline and tetrakistriphenylphosphine palladium is 1.1:1:(0.05-0.06).

4. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: In the step 2, the molar ratio of IrCl3·3H2O to TPA ligand or 1-phenylisoquinoline ligand is 1:(2-2.5).

5. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: In the step, the molar ratio of tripyridine-2-carboxaldehyde to 4-(10,15,20-triphenylporphyrin-5-yl)aniline is 2:

1.

6. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: The reaction temperature of step three is 75-80°C and the reaction time is 8-12h.

7. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: In the step, the molar ratio of tetraquinoline triphenylamine iridium dichloro bridge [Ir(TPA)2Cl2]2 or 1-phenylisoquinoline iridium dichloro bridge [Ir(pqy)2Cl2]2 to the porphyrin Schiff base ligand is 1:(2-2.2).

8. The method for preparing a metal iridium complex with a porphyrin moiety for multimodal thrombolysis according to claim 1, characterized in that: The temperature of the reflux reaction in step 4 is 70-80° C., and the reaction time is 8-12 hours.

9. A nanoparticle, characterized in that: The invention is prepared from the metal iridium complex with a porphyrin part for multimodal thrombolysis represented by formula 1 or formula 2 as described in claim 1.

10. Use of the nanoparticles according to claim 9 in the preparation of drugs for treating thrombotic diseases.

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