Preparation method and application of metal ruthenium hydrogel drug delivery system

Abstract

The invention discloses a preparation method of a metal ruthenium hydrogel drug delivery system, which comprises the following steps: mixing a PDA-IR780 nanocapsule with a metal ruthenium hydrogel solution, quickly stirring, washing with distilled water, and freeze-drying to obtain the metal ruthenium hydrogel drug delivery system Ru (fmb) 3-HAADH (at) PDA-IR780. According to the invention, the drug targeting is improved, the pharmacokinetics of active ingredients is enhanced, a plurality of treatment modes are integrated, a photoresponse type metal ruthenium hydrogel drug loading system is constructed, and the killing effect on tumors is improved through photothermal and photodynamic combined treatment.

Description

Technical Field

[0001] The invention belongs to the technical field of drug delivery, and in particular relates to a preparation method and application of a metal ruthenium hydrogel drug-carrying system. Background Art

[0002] Hydrogels are a class of soft, biocompatible, three-dimensional (3D) porous materials that can effectively encapsulate various types of active drugs and are widely used in tumor therapy, drug controlled release, cell carriers, tissue engineering, biosensors, etc. By locally injecting hydrogel drug delivery systems into / around tumors, the biotoxicity of conventional chemotherapy and radiotherapy can be improved, drug targeting can be enhanced, and the pharmacokinetics of active ingredients can be enhanced. In addition, multiple treatment methods can be integrated simultaneously, thereby significantly improving the effect of cancer treatment.

[0003] Photoresponsive hydrogels have shown broad medical prospects. Through the hydrogel material itself or the internal encapsulation of photothermal agents and photosensitizers, PDT and PTT treatments for tumor cells are triggered under the action of light. This treatment has the advantages of few adverse reactions, high specificity, and good repeatability, and has obvious advantages over traditional tumor treatment methods. Among them, the photothermal agent polydopamine nanoparticles and the new photosensitizer IR780 are widely used in photothermal and photodynamic therapy of tumors, respectively, by generating reactive oxygen or converting light energy into heat energy under light triggering, exerting cytotoxic effects to kill tumor cells.

[0004] Ruthenium complexes have good biocompatibility and anti-tumor activity, and are recognized by the international medical community as anti-tumor drugs with great development potential. Summary of the invention

[0005] The purpose of the present invention is to provide a method for preparing a metal ruthenium hydrogel drug-carrying system.

[0006] Another object of the present invention is to provide a ruthenium hydrogel drug-carrying system prepared by the method for use in the preparation of anti-tumor drugs.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] The first aspect of the present invention provides a method for preparing a metal ruthenium hydrogel drug delivery system, comprising the following steps:

[0009] The first step is a method for preparing polydopamine (PDA) nanocapsules, comprising the following steps:

[0010] Dissolve ammonia water and dimethyldiethoxysilane (DMDES) in a volume ratio of 1:1 in ultrapure water, disperse by ultrasonic for 10 to 20 minutes (preferably 15 to 20 minutes), and stand for 2 to 8 hours (preferably 2 hours, 6 hours, or 8 hours) to obtain an emulsion mixture;

[0011] Add dopamine hydrochloride to a Tris buffer solution with a pH of 8.5 and a concentration of 1 to 20 mmol / mL (10 mmol / mL), wherein the molar ratio of dopamine hydrochloride to tris (hydroxymethylaminomethane) is 1:15, and uniformly disperse the dopamine hydrochloride by ultrasonication to obtain a mixed system. Add the mixed system to the emulsion mixture, wherein the volume ratio of the mixed system to the emulsion mixture is 1:1, and seal the mixture. Stir the mixture at a temperature of 20 to 30° C. (preferably 20 or 25° C.) for 1 to 24 h (preferably 6 h, 12 h, or 24 h);

[0012] An equal volume of anhydrous ethanol is added, ultrasonic dispersion is performed for 1 to 10 minutes (preferably 2, 5, or 10 minutes), the sample is allowed to stand at room temperature for 1 to 24 hours (preferably 8, 12, or 24 hours), centrifuged, the supernatant is removed, anhydrous ethanol is added to resuspend the sample and then centrifuged again, this operation is repeated for a total of at least three times, the sample is resuspended with ultrapure water and then centrifuged again, this operation is repeated for a total of at least three times, the sample is washed with ultrapure water at least twice and then dried to obtain the polydopamine (PDA) nanocapsules.

[0013] The second step is a method for preparing polydopamine (PDA) composite IR780 nanocapsules (abbreviated as PDA-IR780 nanocapsules), comprising the following steps:

[0014] The polydopamine (PDA) nanocapsule solution and the DMSO solution of IR780 are mixed, the mass ratio of polydopamine (PDA) nanocapsule to IR780 is 1-3:1 (preferably 1.7:1, 2:1, 3.3:1), and the mixture is vigorously stirred at room temperature for 1-24 hours (preferably 12, 24 hours), ultrasonicated, the supernatant solution is discarded, and the mixture is centrifuged. After removing the supernatant, anhydrous ethanol is added to resuspend the sample and then centrifuged again. This operation is repeated for at least two times, the anhydrous ethanol is removed, the sample is resuspended with ultrapure water and then centrifuged again, this operation is repeated for at least one time, and the mixture is dried to obtain PDA-IR780 nanocapsules.

[0015] The third step is a method for preparing the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780, comprising the following steps:

[0016] Dissolving the metal ruthenium hydrogel (Ru(fmb)3-HAADH) in distilled water to obtain a metal ruthenium hydrogel (Ru(fmb)3-HAADH) solution having a concentration of 5 to 50 mg / ml (preferably 15 or 25 mg / ml);

[0017] The PDA-IR780 nanocapsules are mixed with a metal ruthenium hydrogel (Ru(fmb)3-HAADH) solution with a concentration of 5 to 30 mg / ml (preferably 15 mg / ml) and rapidly stirred. The mass ratio of the PDA-IR780 nanocapsules to the metal ruthenium hydrogel (Ru(fmb)3-HAADH) is 1:2 to 20 (preferably 1:6, 1:10). The mixture is washed with distilled water and freeze-dried to obtain the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0018] The radius of the particle size of the PDA-IR780 nanocapsule is between 20 and 100 nm, and has a good particle size distribution.

[0019] In the second step, the polydopamine (PDA) nanocapsules prepared in the first step are resuspended in water to a concentration of 0.5 to 3 mg / mL (preferably 1 mg / mL) to obtain a polydopamine (PDA) nanocapsule solution.

[0020] In the second step, IR780 is dissolved in DMSO to a concentration of 0.5 to 3 mg / mL (preferably 1 or 1.2 mg / mL) to obtain a DMSO solution of IR780.

[0021] In the second step, the ultrasonic condition is: 300W ultrasonic for 5 minutes.

[0022] In the second step, the centrifugation condition is: centrifugation at 13000 rpm for 15 min or 10 min.

[0023] In the third step, the preparation method of the metal ruthenium hydrogel (Ru(fmb)3-HAADH) comprises the following steps:

[0024] The hydrazide hyaluronic acid solution and the polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution are mixed and rapidly stirred, the mass ratio of hydrazide hyaluronic acid to the polypyridine ruthenium metal complex Ru(fmb)3Cl2 is 1 to 30:1 (preferably 11.9:1, 14.8:1, 19.8:1), the stirring time is 1 to 3 minutes (preferably 2 minutes), dialyzed for purification, and freeze-dried to obtain the metal ruthenium hydrogel (Ru(fmb)3-HAADH).

[0025] The preparation method of the hydrazide hyaluronic acid solution comprises the following steps:

[0026] Hydrazide hyaluronic acid is dissolved in distilled water to obtain a hydrazide hyaluronic acid solution with a concentration of 10 to 50 mg / mL (preferably 40 mg / mL).

[0027] The preparation method of the polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution comprises the following steps:

[0028] The polypyridine ruthenium metal complex Ru(fmb)3Cl2 is dissolved in distilled water to obtain a polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution with a concentration of 1 to 5 mg / mL (preferably 2.025, 3.375, 2.7 mg / mL).

[0029] The preparation method of the polypyridine ruthenium metal complex Ru(fmb)3Cl2 comprises the following steps:

[0030] Under nitrogen protection, 4'-methyl-2,2'-bipyridine-4-carboxaldehyde is added to a Schlenk tube, ethanol and water are added in a volume ratio of 2 to 8:1 (preferably 6:1) to dissolve, and then RuCl3·3H2O is added, the molar ratio of 4'-methyl-2,2'-bipyridine-4-carboxaldehyde to RuCl3·3H2O is 1 to 6:1 (preferably 3:1, 3.5:1, 4.3:1), heated to reflux for 2 to 24 hours (preferably 12 hours, 24 hours), cooled to room temperature, filtered to obtain the solution, and spin-dried to obtain a crude product, which is dissolved in water, washed with excess dichloromethane at least 3 times, and purified by neutral alumina column chromatography to obtain the polypyridine ruthenium metal complex Ru(fmb)3Cl2.

[0031] The preparation method of the hydrazide hyaluronic acid (HAADH) comprises the following steps:

[0032] Dissolve low molecular weight sodium hyaluronate in ultrapure water and stir until completely dissolved, add adipic acid dihydrazide (ADH), the mass ratio of low molecular weight sodium hyaluronate to adipic acid dihydrazide (ADH) is 0.05-0.5:1 (preferably 0.12:1, 0.24:1), adjust the pH to 6.5-7 (preferably 6.8) with HCl or NaOH, then slowly add 1-hydroxybenzotriazole (HOBt), react for 15-60 minutes (preferably 30 minutes), then add 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDCi) and 1-hydroxybenzotriazole (HOBt) to the solution. The mass ratio of oxadiazole (HOBt) to adipic acid dihydrazide (ADH) is 0.01-1:1 (preferably 0.05:1, 0.1:1), the mass ratio of 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDCi) to adipic acid dihydrazide (ADH) is 0.01-1:1 (preferably 0.07:1, 0.14:1), and the pH is adjusted to 6.5-7 with HCl or NaOH every half an hour for 1-5 hours (preferably 4 hours), then stirred for 12-72 hours, dialyzed and purified for at least 3 times, and freeze-dried to obtain the hydrazide hyaluronic acid (HAADH).

[0033] The low molecular weight sodium hyaluronate was purchased from Bloomage Biotechnology Co., Ltd., HA-TLM 20-40, MW = 200 KDa-400 KDa.

[0034] Another object of the present invention is to provide a ruthenium hydrogel drug-carrying system prepared by the method for use in the preparation of anti-tumor drugs.

[0035] The tumor is selected from melanoma.

[0036] Due to the adoption of the above technical solution, the present invention has the following advantages and beneficial effects:

[0037] The present invention provides a method for preparing a metal ruthenium hydrogel drug delivery system. The metal ruthenium hydrogel drug delivery system prepared by the method uses polypyridyl ruthenium as a metal matrix, forms a metal ruthenium hydrogel with a three-dimensional porous structure by cross-linking with hyaluronic acid modified with hydrazide, and realizes photothermal and photodynamic combined treatment of tumors under light response by encapsulating polydopamine composite IR780 nanocapsules. The metal ruthenium hydrogel drug delivery system can respond to the weakly acidic environment of the tumor, slowly dissociate and release the polydopamine active nanoparticles encapsulated in the gel, and realize photothermal and photodynamic combined treatment of tumors under the stimulation of light signals.

[0038] The present invention provides a method for preparing a metal ruthenium hydrogel drug delivery system, wherein the metal ruthenium hydrogel drug delivery system prepared by the method has pH responsiveness. The gel skeleton thereof can intelligently respond to the weakly acidic environment inside the tumor, gradually dissociate and release the polydopamine active nanoparticles contained inside the gel, thereby realizing the precise delivery of combined photothermal and photodynamic therapy for the tumor.

[0039] The invention provides a method for preparing a metal ruthenium hydrogel drug delivery system. High-resolution scanning electron microscopy and rheology tests are performed on the metal ruthenium hydrogel drug delivery system prepared by the method. It is found that the metal ruthenium hydrogel drug delivery system prepared by the invention has a uniform three-dimensional pore distribution, confirms the effective loading of nanoparticles, and indicates that the metal ruthenium hydrogel drug delivery system has certain rheological properties, self-healing properties and injectability; dynamic light scattering and high-resolution transmission electron microscopy tests show that the particle sizes of the nanoparticles are all between 50 and 100 nm, with good particle size distribution; ultraviolet-visible absorption experiments confirm that the metal ruthenium hydrogel drug delivery system responds to environmental pH to achieve controllable release of photosensitizers; photothermal experiments show that the metal ruthenium hydrogel drug delivery system has good photothermal performance and repeatability; and cell ROS detection experiments show that the metal ruthenium hydrogel drug delivery system can effectively generate reactive oxygen species to kill cells.

[0040] The metal ruthenium hydrogel drug delivery system of the present invention can achieve combined photothermal and photodynamic therapy of tumors under light triggering, enhance the killing effect on tumor cells, and can respond to the weakly acidic environment of the tumor to achieve targeted drug release, reduce the biological toxicity of traditional tumor treatments, and provide new ideas for clinical tumor treatment.

[0041] This drug delivery system focuses on improving the responsiveness and targeting of the system.

[0042] The present invention aims to improve drug targeting, enhance the pharmacokinetics of active ingredients, and integrate multiple treatment methods to construct a light-responsive metal ruthenium hydrogel drug delivery system to improve the tumor-killing effect through combined photothermal and photodynamic therapy.

[0043] The metal ruthenium hydrogel drug delivery system of the present invention has injectability and photothermal and photodynamic properties, has light responsiveness, photothermal and photodynamic properties, and is injectable. The PDA-IR780 nanocapsules in the metal ruthenium hydrogel drug delivery system of the present invention can achieve photothermal and photodynamic combined treatment of tumors under light triggering.

[0044] In the present invention, it is the acylhydrazone functional group generated during the gel formation process that plays the pH response function and can slowly release the internal nanoparticles in the acidic environment of the tumor. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 Schematic diagram of the synthesis route of the metal ruthenium hydrogel drug delivery system.

[0046] Figure 2 This is a scanning electron microscope schematic diagram of the metal ruthenium hydrogel drug delivery system.

[0047] Figure 3 Schematic diagram of rheological test of the ruthenium metal hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0048] Figure 4 Schematic diagram of high-resolution transmission electron microscopy of PDA-IR780 nanocapsules.

[0049] Figure 5 Schematic diagram of the photothermal test curve results of the photoresponsive ruthenium metal hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0050] Figure 6 Schematic diagram of the reactive oxygen species detection results of the photoresponsive ruthenium metal hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0051] Figure 7Schematic diagram of the cytotoxicity test results of the ruthenium metal hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. DETAILED DESCRIPTION

[0052] In order to explain the present invention more clearly, the present invention is further described below in conjunction with preferred embodiments. It should be understood by those skilled in the art that the following specific description is illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0053] Example 1

[0054] A method for preparing a metal ruthenium hydrogel drug delivery system comprises the following steps:

[0055] Synthesis route such as Figure 1 As shown, Figure 1 Schematic diagram of the synthesis route of the metal ruthenium hydrogel drug delivery system.

[0056] The first step is a method for preparing a polypyridine ruthenium metal complex Ru(fmb)3Cl2, comprising the following steps:

[0057] Under nitrogen protection, add 4'-methyl-2,2'-bipyridine-4-carboxaldehyde (7.5mmol, 1.5g) to the Schlenk tube, add 60ml ethanol and 10ml water to dissolve, then add RuCl3·3H2O (1.75mmol, 456mg), heat and reflux for 24h, cool to room temperature, filter the solution, spin dry to obtain the crude product, brown-black solid. The crude product was dissolved in 50mL water, washed with excess dichloromethane 3 times, purified by neutral alumina column chromatography, and 614mg red-brown solid, i.e., polypyridyl ruthenium metal complex Ru(fmb)3Cl2, was obtained.

[0058] The second step is a method for preparing hydrazide hyaluronic acid (HAADH), comprising the following steps:

[0059] 1.2 g of low molecular weight sodium hyaluronate (Huaxi Biotechnology Co., Ltd., HA-TLM 20-40, MW = 200 KDa-400 KDa) was added to a round-bottom flask, 120 mL of ultrapure water was added and stirred until completely dissolved, adipic acid dihydrazide (ADH) (57.4 mmol, 10 g) was added, and the pH was adjusted to about 6.8 with 1 M NaOH aqueous solution. Then 1-hydroxybenzotriazole (HOBt) (3.6 mmol, 486.5 mg) was slowly added, reacted for half an hour, and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCi) (4.5 mmol, 698.6 mg) was added to the solution. The pH was adjusted to about 6.8 with 1 M NaOH for every half an hour of reaction, continued for 4 hours, and then stirred overnight. The next day, it was dialyzed and purified 3 times, and freeze-dried to obtain 1.4 g of hydrazide hyaluronic acid (HAADH).

[0060] The third step is a method for preparing a ruthenium metal hydrogel (Ru(fmb)3-HAADH), comprising the following steps:

[0061] Hydrazide hyaluronic acid (HAADH) (1.2 g) was dissolved in 30 ml of distilled water to obtain a 40 mg / mL hydrazide hyaluronic acid solution.

[0062] The polypyridine ruthenium metal complex Ru(fmb)3Cl2 (0.008 mmol, 5.4 mg) was dissolved in 2 ml of distilled water to obtain a polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution with a concentration of 2.7 mg / mL.

[0063] 2 mL of 40 mg / mL acylhydrazide hyaluronic acid solution was mixed with 2 mL of 2.7 mg / mL polypyridyl ruthenium metal complex Ru(fmb)3Cl2 solution and stirred rapidly, and a red gel was formed after 2 minutes. The red gel was dialyzed and purified three times, and freeze-dried to obtain 60 mg of metal ruthenium hydrogel (Ru(fmb)3-HAADH).

[0064] The fourth step is a method for preparing polydopamine (PDA) nanocapsules, comprising the following steps:

[0065] The concentration of ammonia water is 25% (m / m), and the concentration of dimethyldiethoxysilane (DMDES) is 99% (m / m). 300uL of ammonia water and 300uL of dimethyldiethoxysilane (DMDES) are added with ultrapure water to make the volume to 30mL, ultrasonically dispersed for 15 minutes, and allowed to stand for 6 hours to obtain an emulsion mixture.

[0066] Tris (hydroxymethylaminomethane) was added to ultrapure water to dissolve, and the volume was fixed to 30 mL with ultrapure water. The concentration of Tris (hydroxymethylaminomethane) was 10 mmol / mL, and a Tris buffer solution with a pH of 8.5 and a concentration of 10 mmol / mL was obtained.

[0067] Dopamine hydrochloride (20 mmol, 37 mg) was added to 30 mL of Tris buffer with a pH of 8.5 and a concentration of 10 mmol / mL, the molar ratio of dopamine hydrochloride to tris (hydroxymethylaminomethane) (Tris) being 1:15, and the mixture was uniformly dispersed by ultrasound to obtain a mixed system, and the mixed system was added to the above emulsion mixture, the volume ratio of the mixed system to the above emulsion mixture being 1:1, and the mixture was sealed and stirred at 25°C for 24 h.

[0068] An equal volume of anhydrous ethanol was added, ultrasonic dispersion was performed for 5 minutes, the mixture was allowed to stand at room temperature for 12 hours, and the supernatant was removed. Anhydrous ethanol was added to resuspend the sample and centrifuged again. This operation was repeated for a total of three times. The sample was resuspended in ultrapure water and centrifuged again. This operation was repeated for a total of three times. The mixture was washed twice with ultrapure water and dried to obtain 20 mg of polydopamine (PDA) nanocapsules.

[0069] The fifth step is a method for preparing polydopamine (PDA) composite IR780 nanocapsules (abbreviated as PDA-IR780 nanocapsules), comprising the following steps:

[0070] The polydopamine (PDA) nanocapsules were resuspended in water to a concentration of 1 mg / mL to obtain a polydopamine (PDA) nanocapsule solution.

[0071] Indogreen fluorescent probe IR780 (0.03 mmol, 20 mg) was dissolved in 20 mL of DMSO to a concentration of 1 mg / mL to obtain a DMSO solution of IR780.

[0072] 5 mL of 1 mg / mL polydopamine (PDA) nanocapsule solution and 2.5 mL of 1 mg / mL IR780 DMSO solution were mixed and stirred vigorously at room temperature for 24 h. Ultrasonication was performed at 300 W for 5 min, the supernatant solution was discarded, the mixed solution was transferred to a high-speed centrifuge tube, and centrifuged at 13000 rpm for 15 min. After removing the supernatant, 20 mL of anhydrous ethanol was added to resuspend the sample and then centrifuged at 13000 rpm for 10 min. This resuspension and centrifugation step was repeated twice. The anhydrous ethanol was removed, and the sample was resuspended and washed with 20 mL of ultrapure water and centrifuged again. This resuspension and centrifugation step was repeated once and the sample was dried to obtain 6.2 mg PDA-IR780 nanocapsules.

[0073] The sixth step is a method for preparing the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780, comprising the following steps:

[0074] 60 mg of the metal ruthenium hydrogel (Ru(fmb)3-HAADH) obtained in the third step was dissolved in 2 mL of distilled water to obtain a metal ruthenium hydrogel (Ru(fmb)3-HAADH) solution.

[0075] 5 mg of the PDA-IR780 nanocapsules obtained in the fifth step were mixed with the above-mentioned ruthenium hydrogel (Ru(fmb)3-HAADH) solution and quickly stirred, washed three times with distilled water, and freeze-dried to obtain about 65 mg of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0076] Example 2

[0077] A method for preparing a metal ruthenium hydrogel drug delivery system comprises the following steps:

[0078] The first step is a method for preparing a polypyridine ruthenium metal complex Ru(fmb)3Cl2, comprising the following steps:

[0079] Under nitrogen protection, add 4'-methyl-2,2'-bipyridine-4-carboxaldehyde (5.25mmol, 1.05g) to the Schlenk tube, add 60ml ethanol and 10ml water to dissolve, then add RuCl3·3H2O (1.75mmol, 456mg), heat and reflux for 12h, cool to room temperature, filter the solution, spin dry to obtain the crude product, brown-black solid. The crude product was dissolved in 50mL water, washed with excess dichloromethane 3 times, purified by neutral alumina column chromatography, and 509mg of red-brown solid, i.e., polypyridyl ruthenium metal complex Ru(fmb)3Cl2, was obtained.

[0080] The second step is a method for preparing hydrazide hyaluronic acid (HAADH), comprising the following steps:

[0081] 1.2g of low molecular weight sodium hyaluronate is added to a round-bottom flask, 120mL ultrapure water is added and stirred until all dissolved, adipic acid dihydrazide (ADH) (28.7mmol, 5g) is added, and pH is adjusted to about 6.8 with 1M NaOH aqueous solution. Then 1-hydroxybenzotriazole (HOBt) (3.6mmol, 486.5mg) is slowly added, reacted for half an hour, and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCi) (4.5mmol, 698.6mg) is added to the solution. pH is adjusted to about 6.8 with 1M NaOH for half an hour of each reaction, continued for 4h, and then stirred overnight. Dialysis purification 3 times the next day, freeze-dried, obtained 1.3g hydrazide hyaluronic acid (HAADH).

[0082] The third step is a method for preparing a ruthenium metal hydrogel (Ru(fmb)3-HAADH), comprising the following steps:

[0083] Hydrazide hyaluronic acid (HAADH) (1.2 g) was dissolved in 30 ml of distilled water to obtain a 40 mg / mL hydrazide hyaluronic acid solution.

[0084] The polypyridine ruthenium metal complex Ru(fmb)3Cl2 (0.006 mmol, 4.05 mg) was dissolved in 2 ml of distilled water to obtain a polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution with a concentration of 2.025 mg / mL.

[0085] 2 mL of 40 mg / mL acylhydrazide hyaluronic acid solution was mixed with 2 mL of 2.025 mg / mL polypyridyl ruthenium metal complex Ru(fmb)3Cl2 solution and stirred rapidly, and a red gel was formed after 2 minutes. The red gel was dialyzed and purified 3 times, and freeze-dried to obtain 55 mg of metal ruthenium hydrogel (Ru(fmb)3-HAADH).

[0086] The fourth step is a method for preparing polydopamine (PDA) nanocapsules, comprising the following steps:

[0087] The concentration of ammonia water is 25% (m / m), and the concentration of dimethyldiethoxysilane (DMDES) is 99% (m / m). 300uL of ammonia water and 300uL of dimethyldiethoxysilane (DMDES) are added with ultrapure water to make the volume to 30mL, ultrasonically dispersed for 10 minutes, and allowed to stand for 2 hours to obtain an emulsion mixture.

[0088] Tris (hydroxymethylaminomethane) was added to ultrapure water to dissolve, and the volume was fixed to 30 mL with ultrapure water. The concentration of Tris (hydroxymethylaminomethane) was 10 mmol / mL, and a Tris buffer solution with a pH of 8.5 and a concentration of 10 mmol / mL was obtained.

[0089] Dopamine hydrochloride (20 mmol, 37 mg) was added to 30 mL of Tris buffer with a pH of 8.5 and a concentration of 10 mmol / mL, the molar ratio of dopamine hydrochloride to tris (hydroxymethylaminomethane) (Tris) being 1:15, and the mixture was uniformly dispersed by ultrasound to obtain a mixed system, and the mixed system was added to the above emulsion mixture, the volume ratio of the mixed system to the above emulsion mixture being 1:1, and the mixture was sealed and stirred at 20°C for 12 h.

[0090] An equal volume of anhydrous ethanol was added, ultrasonic dispersion was performed for 2 minutes, the mixture was allowed to stand at room temperature for 8 hours, and the supernatant was removed. Anhydrous ethanol was added to resuspend the sample and centrifuged again. This operation was repeated for a total of three times. The sample was resuspended in ultrapure water and centrifuged again. This operation was repeated for a total of three times. The mixture was washed twice with ultrapure water and dried to obtain 15 mg of polydopamine (PDA) nanocapsules.

[0091] The fifth step is a method for preparing polydopamine (PDA) composite IR780 nanocapsules (abbreviated as PDA-IR780 nanocapsules), comprising the following steps:

[0092] The polydopamine (PDA) nanocapsules were resuspended in water to a concentration of 1 mg / mL to obtain a polydopamine (PDA) nanocapsule solution.

[0093] Indogreen fluorescent probe IR780 (0.03 mmol, 20 mg) was dissolved in 20 mL of DMSO to a concentration of 1 mg / mL to obtain a DMSO solution of IR780.

[0094] 5 mL of 1 mg / mL polydopamine (PDA) nanocapsule solution and 1.5 mL of 1 mg / mL IR780 DMSO solution were mixed and stirred vigorously at room temperature for 12 h. Ultrasonication was performed at 300 W for 5 min, the supernatant solution was discarded, the mixed solution was transferred to a high-speed centrifuge tube, and centrifuged at 13000 rpm for 15 min. After removing the supernatant, 20 mL of anhydrous ethanol was added to resuspend the sample and then centrifuged at 13000 rpm for 10 min. This resuspension and centrifugation step was repeated twice. The anhydrous ethanol was removed, and the sample was resuspended and washed with 20 mL of ultrapure water and then centrifuged again. This resuspension and centrifugation step was repeated once and the sample was dried to obtain 5.6 mg PDA-IR780 nanocapsules.

[0095] The sixth step is a method for preparing the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780, comprising the following steps:

[0096] 30 mg of the metal ruthenium hydrogel (Ru(fmb)3-HAADH) obtained in the third step was dissolved in 2 mL of distilled water to obtain a metal ruthenium hydrogel (Ru(fmb)3-HAADH) solution.

[0097] 5 mg of the PDA-IR780 nanocapsules obtained in the fifth step were mixed with the above-mentioned ruthenium hydrogel (Ru(fmb)3-HAADH) solution and quickly stirred, washed three times with distilled water, and freeze-dried to obtain about 33 mg of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0098] Example 3

[0099] A method for preparing a metal ruthenium hydrogel drug delivery system comprises the following steps:

[0100] The first step is a method for preparing a polypyridine ruthenium metal complex Ru(fmb)3Cl2, comprising the following steps:

[0101] Under nitrogen protection, add 4'-methyl-2,2'-bipyridine-4-carboxaldehyde (6.2mmol, 1.2g) to the Schlenk tube, add 60ml ethanol and 10ml water to dissolve, then add RuCl3·3H2O (1.75mmol, 456mg), heat and reflux for 12h, cool to room temperature, filter the solution, spin dry to obtain the crude product, brown-black solid. The crude product was dissolved in 50mL water, washed with excess dichloromethane 3 times, purified by neutral alumina column chromatography, and 572mg of red-brown solid, i.e., polypyridyl ruthenium metal complex Ru(fmb)3Cl2, was obtained.

[0102] The second step is a method for preparing hydrazide hyaluronic acid (HAADH), comprising the following steps:

[0103] 1.2g of low molecular weight sodium hyaluronate was added to a round-bottomed flask, 120mL of ultrapure water was added and stirred until all dissolved, adipic acid dihydrazide (ADH) (57.4mmol, 10g) was added, and the pH was adjusted to about 6.8 with a 1M NaOH aqueous solution. Then 1-hydroxybenzotriazole (HOBt) (3.6mmol, 486.5mg) was slowly added, reacted for half an hour, and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCi) (4.5mmol, 698.6mg) was added to the solution. The pH was adjusted to about 6.8 with 1M NaOH for half an hour of each reaction, and continued for 4h. After continuing to stir for 48h, it was purified by dialysis 3 times and freeze-dried to obtain 1.5g hydrazide hyaluronic acid (HAADH).

[0104] The third step is a method for preparing a ruthenium metal hydrogel (Ru(fmb)3-HAADH), comprising the following steps:

[0105] Hydrazide hyaluronic acid (HAADH) (1.2 g) was dissolved in 30 ml of distilled water to obtain a 40 mg / mL hydrazide hyaluronic acid solution.

[0106] The polypyridine ruthenium metal complex Ru(fmb)3Cl2 (0.01 mmol, 6.75 mg) was dissolved in 2 ml of distilled water to obtain a polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution with a concentration of 3.375 mg / mL.

[0107] 2 mL of 40 mg / mL acylhydrazide hyaluronic acid solution was mixed with 2 mL of 3.375 mg / mL polypyridyl ruthenium metal complex Ru(fmb)3Cl2 solution and stirred rapidly, and a red gel was formed after 2 minutes. The red gel was dialyzed and purified three times, and freeze-dried to obtain 63 mg of metal ruthenium hydrogel (Ru(fmb)3-HAADH).

[0108] The fourth step is a method for preparing polydopamine (PDA) nanocapsules, comprising the following steps:

[0109] The concentration of ammonia water is 25% (m / m), and the concentration of dimethyldiethoxysilane (DMDES) is 99% (m / m). 300uL of ammonia water and 300uL of dimethyldiethoxysilane (DMDES) are added with ultrapure water to make the volume to 30mL, ultrasonically dispersed for 20 minutes, and allowed to stand for 8 hours to obtain an emulsion mixture.

[0110] Tris (hydroxymethylaminomethane) was added to ultrapure water to dissolve, and the volume was fixed to 30 mL with ultrapure water. The concentration of Tris (hydroxymethylaminomethane) was 10 mmol / mL, and a Tris buffer solution with a pH of 8.5 and a concentration of 10 mmol / mL was obtained.

[0111] Dopamine hydrochloride (20 mmol, 37 mg) was added to 30 mL of Tris buffer with a pH of 8.5 and a concentration of 10 mmol / mL, the molar ratio of dopamine hydrochloride to tris (hydroxymethylaminomethane) (Tris) being 1:15, and the mixture was uniformly dispersed by ultrasound to obtain a mixed system, and the mixed system was added to the above emulsion mixture, the volume ratio of the mixed system to the above emulsion mixture being 1:1, and the mixture was sealed and stirred at 25°C for 6 h.

[0112] An equal volume of anhydrous ethanol was added, ultrasonic dispersion was performed for 10 minutes, the mixture was allowed to stand at room temperature for 24 hours, and the supernatant was removed. Anhydrous ethanol was added to resuspend the sample and centrifuged again. This operation was repeated for a total of three times. The sample was resuspended in ultrapure water and centrifuged again. This operation was repeated for a total of three times. The mixture was washed twice with ultrapure water and then dried to obtain 13 mg of polydopamine (PDA) nanocapsules.

[0113] The fifth step is a method for preparing polydopamine (PDA) composite IR780 nanocapsules (abbreviated as PDA-IR780 nanocapsules), comprising the following steps:

[0114] The polydopamine (PDA) nanocapsules were resuspended in water to a concentration of 1 mg / mL to obtain a polydopamine (PDA) nanocapsule solution.

[0115] Indo green fluorescent probe IR780 (0.036 mmol, 24 mg) was dissolved in 20 mL of DMSO to a concentration of 1.2 mg / mL to obtain a DMSO solution of IR780.

[0116] 5 mL of 1 mg / mL polydopamine (PDA) nanocapsule solution and 2.5 mL of 1.2 mg / mL IR780 DMSO solution were mixed and stirred vigorously at room temperature for 24 h. Ultrasonication was performed at 300 W for 5 min, the supernatant solution was discarded, the mixed solution was transferred to a high-speed centrifuge tube, and centrifuged at 8000 rpm for 15 min. After removing the supernatant, 20 mL of anhydrous ethanol was added to resuspend the sample and then centrifuged at 8000 rpm for 10 min. This resuspension and centrifugation step was repeated twice. The anhydrous ethanol was removed, and the sample was resuspended and washed with 20 mL of ultrapure water and then centrifuged again. This resuspension and centrifugation step was repeated once and the sample was dried to obtain 5.2 mg PDA-IR780 nanocapsules.

[0117] The sixth step is a method for preparing the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780, comprising the following steps:

[0118] 50 mg of the metal ruthenium hydrogel (Ru(fmb)3-HAADH) obtained in the third step was dissolved in 2 mL of distilled water to obtain a metal ruthenium hydrogel (Ru(fmb)3-HAADH) solution.

[0119] 5 mg of the PDA-IR780 nanocapsules obtained in the fifth step were mixed with the above-mentioned ruthenium hydrogel (Ru(fmb)3-HAADH) solution and quickly stirred, washed three times with distilled water, and freeze-dried to obtain about 55 mg of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

[0120] Example 4

[0121] SEM and rheological characterization

[0122] The ruthenium metal hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in Example 1 was observed by high-resolution scanning electron microscopy (SEM). Figure 2 As shown, Figure 2 The following are SEM images of the ruthenium hydrogel drug delivery system. (i) is a schematic diagram of the SEM morphology of Ru(fmb)3-HAADH at a scale of 500um, (ii) is a schematic diagram of the SEM morphology of Ru(fmb)3-HAADH at a scale of 50um, (iii) is a schematic diagram of the SEM morphology of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 at a scale of 500um, and (iv) is a schematic diagram of the SEM morphology of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 at a scale of 50um. It can be clearly seen that the metal ruthenium hydrogel (Ru(fmb)3-HAADH) has a three-dimensional network structure, and after loading PDA@IR780 nanocapsules, the pores of the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 have obvious morphological changes and the texture becomes rough.

[0123] Rheological test results are as follows Figure 3 As shown, Figure 3 Schematic diagram of rheological test of Ru(fmb)3-HAADH@PDA-IR780, a is a schematic diagram of rheological curve of Ru(fmb)3-HAADH@PDA-IR780, b is a schematic diagram of strain scanning curve of Ru(fmb)3-HAADH@PDA-IR780, c is a schematic diagram of continuous step strain curve of Ru(fmb)3-HAADH@PDA-IR780, and d is a schematic diagram of thixotropy curve of Ru(fmb)3-HAADH@PDA-IR780. As can be seen from the figure, the strain scanning curve shows that the modulus of the Ru(fmb)3-HAADH@PDA-IR780 metal ruthenium hydrogel drug delivery system changes under a large stress, indicating that the Ru(fmb)3-HAADH@PDA-IR780 metal ruthenium hydrogel drug delivery system has a high mechanical strength. The continuous step strain curve and the thixotropy curve both reflect that the viscosity of the Ru(fmb)3-HAADH@PDA-IR780 metal ruthenium hydrogel drug delivery system decreases and the fluidity increases after the shear stress is applied. However, after the shear stress is removed, the material will gradually return to its original high viscosity state, indicating that the Ru(fmb)3-HAADH@PDA-IR780 metal ruthenium hydrogel drug delivery system has good self-healing properties. The Ru(fmb)3-HAADH@PDA-IR780 metal ruthenium hydrogel has good rheological characteristics and self-healing properties.

[0124] The PDA-IR780 nanocapsules prepared in Example 1 were observed by high-resolution scanning electron microscopy (SEM). Figure 4As shown, Figure 4 The high-resolution transmission electron microscopy diagram of PDA-IR780 nanocapsules shows that the diameter of PDA-IR780 nanocapsules is 41.77 nm, and the particle radius is between 20 and 100 nm.

[0125] Example 5

[0126] Photothermal performance experiment

[0127] The ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in Example 1 was diluted into 200ug / mL, 500ug / mL, 750ug / mL, 1000ug / mL, 1500ug / mL, and 2000ug / mL (PDA nanocapsule concentration) PBS solutions, and the PBS solution was used as a blank control. The wavelength was λ=808nm, the power density was P=1.5W / cm 2 The solution was irradiated with laser for 540 seconds, and its temperature rise process was monitored and a temperature rise curve was drawn. The results are shown in Figure 5 As shown in (i), Figure 5 Figure 3 is a schematic diagram of the photothermal test curve results of the photoresponsive ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. From the analysis in (i), it can be seen that at a fixed light power, as the concentration of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 increases, the concentration of PDA nanocapsules in the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 increases, and the solution's heating ability becomes stronger. When the concentration of PDA nanocapsules in the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 reaches 1500μg / mL, the heating ability is basically saturated, and the heating ability no longer increases with the increase of PDA nanocapsule concentration.

[0128] The wavelength λ is 808nm, the power density P is 1.5W / cm 2 , 2.0W / cm 2 , 2.5W / cm 2 、3.0W / cm 2 The laser irradiated 1000ug / mL ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 for 540s, monitored its heating process, and plotted the heating curve. The results are as follows: Figure 5As shown in (ii), from the analysis in (ii), it can be seen that the laser light power can greatly affect the photothermal heating of the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. The heating ability increases with the increase of power. However, in actual use, it is not advisable to use lasers with excessive power to avoid burns to the skin and other tissues.

[0129] Wavelength λ = 808nm, power density P = 1.5W / cm 2 The laser irradiated the 1000ug / mL system for 240s, then allowed it to cool naturally at room temperature, and then repeated the irradiation-cooling process, monitored the temperature change, and drew a heating curve to explore the photothermal elasticity of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. The results are shown in Figure 5 As shown in (iii), from the analysis in (iii), it can be seen that this gel system has photothermal elasticity and can still maintain excellent photothermal performance after repeated photothermal heating-cooling. It can be repeatedly illuminated during the use cycle without repeated injection, and biosafety is improved.

[0130] Figure 5 In (iv) and (v), the results of in vitro photothermal imaging were shown. 2 mL of PBS, polydopamine (PDA) nanocapsule solution (PDA is shown in the figure) (1000 μg / mL), IR780 DMSO solution (IR780 is shown in the figure) (500 μg / mL), ruthenium hydrogel (Ru(fmb)3-HAADH) solution (Ru(fmb)3-HAADH is shown in the figure) (2000 μg / mL), PDA-IR780 nanocapsule solution (PDA@IR780 is shown in the figure) (1000 μg / mL, 500 μg / mL) and ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 (Ru(fmb)3-HAADH@PDA-IR780 is shown in the figure) (2000 μg / mL, 1000 μg / mL, 500 μg / mL) were taken in quartz cuvettes respectively. At the same concentration of PDA-IR780 nanocapsule solution, the photothermal performance of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 is the strongest. The system temperature rises by 24.5°C within 2 minutes after illumination, and the PDA-IR780 system temperature rises by 22.9°C in 2 minutes. The results show that the prepared ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 has a good photothermal effect and excellent photothermal elasticity, and can be repeatedly heated and cooled.

[0131] Example 6

[0132] Detection of intracellular reactive oxygen species (ROS) in the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in Example 1

[0133] Mouse melanoma cells were selected, and logarithmic phase B16F10 cells were taken. 5 / mL was inoculated in a confocal culture dish for 24h, and the culture medium was discarded. Free IR780, PDA-IR780 nanocapsules and the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in Example 1 were added respectively, and a group of blank cells was set up, for a total of four groups. After continuing to culture in the dark for 24h, the culture medium was discarded, and the cells were washed three times with PBS to remove the unabsorbed drugs. 2mL2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) reactive oxygen species detection probe (DCFH-DA: culture medium 3:1000) was added to each well, and the wavelength λ=808nm and the power density P=1.5W / cm 2 The cells were irradiated with laser for 120 seconds. After culturing in the incubator for 20 minutes, the excess staining probe was washed away with PBS and the staining was observed with a confocal microscope.

[0134] Test results such as Figure 6 As shown, Figure 6 The figure is a schematic diagram of the reactive oxygen detection results of the photoresponsive ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. It can be seen from the figure that the reactive oxygen content in the cells of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 illumination group is significantly higher than the reactive oxygen produced by the nanocapsules in the free IR780 illumination group and the PDA-IR780 illumination group. In addition, in the environment of pH = 5.5, the intracellular ROS content of Ru(fmb)3-HAADH@PDA-IR780 after illumination is higher than that in the environment of pH = 7.4, which proves that the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 has a strong oxygen enhancement ability and releases the encapsulated drug faster in response to pH in a weakly acidic environment.

[0135] Example 7

[0136] Cytotoxicity experiment of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in Example 1

[0137] Mouse melanoma cells were selected, and logarithmic phase B16F10 cells were taken. The cell concentration was 1×10 4Each well was inoculated in four 96-well plates and cultured in groups for 24 hours. The culture medium was discarded and five parallel groups were set up, namely, Ru(fmb)3-HAADH@PDA-IR780 illumination group, Ru(fmb)3-HAADH@PDA-IR780 light-proof group, PDA-IR780 nanocapsule illumination group, PDA-IR780 nanocapsule light-proof group, and free IR780 illumination group. Different concentration gradients were added, which were equivalent to the concentrations of photosensitizers of 50μg / mL, 25μg / mL, 12.5μg / mL, 6.25μg / mL, 3.13μg / mL, 1.56μg / mL, 0.78μg / mL and 0μg / mL. Six replicate wells were set for each gradient and cultured in the dark for 24 hours. The culture medium was discarded, washed with PBS three times, and 100μL complete culture medium was added. The illumination group was at a wavelength of λ=808nm and a power density of P=1.5W / cm 2 After 120s of laser irradiation, the cells were cultured in the dark for 24h. The cells in the light-protected group were washed with PBS three times, and 100μL of complete culture medium was added to continue the culture for 24h. 100μL of serum-free culture medium containing 10% CCK-8 was added to both groups, and the cells were cultured in the dark for 2h. The absorbance of each well at 450nm was measured using an enzyme-labeled instrument.

[0138] The results of cytotoxicity experiments were Figure 7 As shown, Figure 7 The schematic diagram of the cytotoxicity test results of the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780. As can be seen from the figure, the median lethal concentration (IC 50 ) value is significantly higher than the IC value of Ru(fmb)3-HAADH@PDA-IR780 in the light-shielded group. 50 The value is lower, indicating that the gel system in the Ru(fmb)3-HAADH@PDA-IR780 illumination group has a stronger killing effect on tumors; at the same time, the Ru(fmb)3-HAADH@PDA-IR780 illumination group is more lethal than the PDA@IR780 illumination group, indicating that the Ru(II) ligand released by the Ru(fmb)3-HAADH@PDA-IR780 gel has a killing effect on tumors.

[0139] The ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in the embodiment of the present invention has a significantly better killing effect on tumor cells than the free IR780, and is also significantly better than the killing effect of PDA-IR780 nanocapsules, indicating that the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780 prepared in the embodiment of the present invention is beneficial to enhancing the endocytosis of IR780, and exhibits a stronger tumor cell killing ability enhanced after illumination.

[0140] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. A method for preparing a metal ruthenium hydrogel drug delivery system, characterized in that: The following steps are involved: The first step is a method for preparing polydopamine nanocapsules, comprising the following steps: Dissolve ammonia water and dimethyldiethoxysilane in a volume ratio of 1:1 in ultrapure water, disperse by ultrasonic for 10 to 20 minutes, and let stand for 2 to 8 hours to obtain an emulsion mixture; Add dopamine hydrochloride to a Tris buffer solution with a pH of 8.5 and a concentration of 1 to 20 mmol / mL, wherein the molar ratio of dopamine hydrochloride to tris(hydroxymethyl)aminomethane is 1:15, and uniformly disperse the dopamine hydrochloride by ultrasonication to obtain a mixed system. Add the mixed system to the emulsion mixture, wherein the volume ratio of the mixed system to the emulsion mixture is 1:1, seal the mixture, and stir the mixture at a temperature of 20 to 30° C. for 1 to 24 hours; Add an equal volume of anhydrous ethanol, ultrasonically disperse for 1 to 10 minutes, stand at room temperature for 1 to 24 hours, centrifuge, remove the supernatant, add anhydrous ethanol to resuspend the sample and then centrifuge again, repeat this operation of resuspending and centrifuging at least three times, resuspend the sample with ultrapure water and then centrifuge again, repeat this operation of resuspending and centrifuging at least three times, wash with ultrapure water at least twice and then dry to obtain the polydopamine nanocapsules; The second step is a method for preparing polydopamine composite IR780 nanocapsules, comprising the following steps: The polydopamine nanocapsule solution and the DMSO solution of IR780 are mixed, the mass ratio of the polydopamine nanocapsule to IR780 is 1 to 3:1, and the mixture is vigorously stirred at room temperature for 1 to 24 hours, ultrasonicated, and the supernatant solution is discarded and centrifuged. After the supernatant is removed, anhydrous ethanol is added to resuspend the sample and then centrifuged again. This operation is repeated for at least two times, the anhydrous ethanol is removed, the sample is resuspended with ultrapure water and then centrifuged again, and this operation is repeated for at least one time, and the sample is dried to obtain PDA-IR780 nanocapsules; The third step is a method for preparing the metal ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780, comprising the following steps: Dissolving the metal ruthenium hydrogel Ru(fmb)3-HAADH in distilled water to obtain a metal ruthenium hydrogel Ru(fmb)3-HAADH solution with a concentration of 5 to 50 mg / ml; The PDA-IR780 nanocapsules were mixed with a 5-30 mg / ml ruthenium hydrogel Ru(fmb)3-HAADH solution and stirred rapidly, the mass ratio of the PDA-IR780 nanocapsules to the ruthenium hydrogel Ru(fmb)3-HAADH was 1:2-20, washed with distilled water, and freeze-dried to obtain the ruthenium hydrogel drug delivery system Ru(fmb)3-HAADH@PDA-IR780.

2. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: In the second step, the polydopamine nanocapsules prepared in the first step are resuspended in water to a concentration of 0.5 to 3 mg / mL to obtain a polydopamine nanocapsule solution.

3. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: In the second step, IR780 is dissolved in DMSO to a concentration of 0.5 to 3 mg / mL to obtain a DMSO solution of IR780.

4. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: In the second step, the ultrasonic conditions are: 300W ultrasonic for 5 min; In the second step, the centrifugation condition is: centrifugation at 13000 rpm for 15 min or 10 min.

5. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: In the third step, the preparation method of the metal ruthenium hydrogel includes the following steps: mixing the hydrazide hyaluronic acid solution and the polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution and rapidly stirring, the mass ratio of the hydrazide hyaluronic acid to the polypyridine ruthenium metal complex Ru(fmb)3Cl2 is 1 to 30:1, the stirring time is 1 to 3 minutes, dialysis purification, freeze drying, and obtaining the metal ruthenium hydrogel Ru(fmb)3-HAADH.

6. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: The preparation method of the hydrazide hyaluronic acid solution comprises the following steps: dissolving the hydrazide hyaluronic acid in distilled water to obtain the hydrazide hyaluronic acid solution with a concentration of 10 to 50 mg / mL.

7. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: The preparation method of the polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution comprises the following steps: dissolving the polypyridine ruthenium metal complex Ru(fmb)3Cl2 in distilled water to obtain a polypyridine ruthenium metal complex Ru(fmb)3Cl2 solution with a concentration of 1 to 5 mg / mL.

8. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: The preparation method of the polypyridine ruthenium metal complex Ru(fmb)3Cl2 comprises the following steps: under nitrogen protection, adding 4'-methyl-2,2'-bipyridine-4-carboxaldehyde into a Schlenk tube, adding ethanol and water in a volume ratio of 2 to 8:1 to dissolve, then adding RuCl3·3H2O, the molar ratio of 4'-methyl-2,2'-bipyridine-4-carboxaldehyde to RuCl3·3H2O being 1 to 6:1, heating and refluxing for 2 to 24 hours, cooling to room temperature, filtering to obtain a solution, spin-drying to obtain a crude product, dissolving the crude product in water, washing with excess dichloromethane for at least 3 times, and purifying by neutral alumina column chromatography to obtain the polypyridine ruthenium metal complex Ru(fmb)3Cl2.

9. The method for preparing the metal ruthenium hydrogel drug delivery system according to claim 1, characterized in that: The preparation method of hydrazide hyaluronic acid comprises the following steps: Dissolve low molecular weight sodium hyaluronate in ultrapure water and stir until completely dissolved, add adipic acid dihydrazide, the mass ratio of low molecular weight sodium hyaluronate to adipic acid dihydrazide is 0.05-0.5:1, adjust the pH to 6.5-7 with HCl or NaOH, then slowly add 1-hydroxybenzotriazole, react for 15-60 minutes, then add 1-ethyl-(3-dimethylaminopropyl)carbodiimide to the solution, the mass ratio of 1-hydroxybenzotriazole to adipic acid dihydrazide is 0.01-1:1, the mass ratio of 1-ethyl-(3-dimethylaminopropyl)carbodiimide to adipic acid dihydrazide is 0.01-1:1, adjust the pH to 6.5-7 with HCl or NaOH every half an hour, continue for 1-5 hours, then stir for 12-72 hours, dialysis purification for at least 3 times, and freeze-dry to obtain the hydrazide hyaluronic acid.

10. Use of a ruthenium hydrogel drug delivery system prepared by the method according to any one of claims 1 to 9 in the preparation of anti-tumor drugs.

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