A ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation and its preparation method
Through the preparation of manganese -based multi -functional anti -tumor nano -preparations, combined with photos therapy of photos therapy and chemical dynamics, it solves the problem of combined therapy in existing technologies. Ability.
Patent Information
- Application Number
- CN202211448413.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-11-18
- Publication Date
- 2025-01-28
- Estimated Expiration
- 2042-11-18
AI Technical Summary
The prior art is difficult to achieve the combined effects of photothermal therapy, chemodynamic therapy and hunger therapy simultaneously in cancer treatment, and lacks effective tumor targeting ability and hypoxic microenvironment improvement ability.
The manganese dioxide -based multifunctional anti -tumor nano -preparation is adopted. By decentralized potassium permanganate and trichlot chloride as nanoparticles by dispersing yellow -brown suspension in the water, and the ultrasonic mixing method will be used. Its combination is then added with glucose oxidase and sodium hyaluronate, forming nanozons with optical and chemical dynamic properties.
A dual -oxygen effect is achieved, the tumor hypoxia is relieved, and the effect of light thermal effect and chemical dynamic therapy effect is possessed, and the anti -tumor efficacy is significantly improved through targeting and improving the micro -environment of oxygen.
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Figure CN115887650B_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of medical technology, and specifically relates to a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nano preparation and a preparation method thereof. Background Art
[0002] The incidence of cancer is increasing worldwide, posing a serious threat to human life and health. Due to the complexity of its pathogenesis and the heterogeneity of the tumor microenvironment, it is difficult to achieve satisfactory results using only a single therapy. The development of minimally invasive or non-invasive tumor treatment methods is a new trend in the current development of biomedicine and is generally considered to be a promising alternative to traditional treatments including surgery, chemotherapy and radiotherapy. In recent years, a series of new cancer treatment methods have emerged, including photothermal therapy, photodynamic therapy, sonodynamic therapy, chemodynamic therapy, gas therapy, immunotherapy, starvation therapy, etc. These are non-invasive or low-invasive treatment methods that improve the efficacy of tumor treatment, cause less damage to normal tissues, and improve patient compliance during treatment.
[0003] Ruthenium, as a precious metal, is a good photothermal agent that can absorb specific light irradiation and convert this energy into heat to initiate the photothermal effect to inhibit or even ablate tumors. Chemodynamic therapy is an emerging cancer treatment method that uses metal ions with Fenton or Fenton-like reactions to catalyze hydrogen peroxide to produce cytotoxic hydroxyl radicals to kill tumor cells. In an acidic and hydrogen peroxide environment, manganese dioxide has the ability to rapidly degrade and effectively self-generate oxygen. The Mn 2+ Manganese-based nanomaterials have excellent nuclear magnetic resonance imaging properties and are easily metabolized by the kidneys, so they can be developed into integrated drug delivery systems for diagnosis and treatment. In the tumor microenvironment, both manganese dioxide and ruthenium can catalyze hydrogen peroxide to produce oxygen and hydroxyl radicals through a series of reactions. The rapid proliferation of tumor cells is heavily dependent on aerobic glycolysis, which means that they are highly sensitive to changes in glucose levels. Therefore, inducing glucose decomposition can disrupt the glucose metabolism of tumor cells, thereby inducing starvation therapy. Glucose oxidase-catalyzed oxidation of glucose can cut off energy supply to inhibit tumor growth while producing glucuronic acid and hydrogen peroxide to promote Fenton-like reactions and enhance chemodynamic therapy. Hyaluronic acid has been shown to specifically bind to CD44 receptors overexpressed on the surface of cancer cells, and nanoformulations modified with hyaluronic acid are expected to improve tumor cell targeting capabilities.
[0004] Compared with single photothermal therapy or chemodynamic therapy, the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation constructed this time is expected to achieve good anti-tumor effects by combining photothermal therapy, chemodynamic therapy and starvation therapy, and endowing certain tumor targeting capabilities and tumor hypoxic microenvironment improvement capabilities, which has important social significance. Summary of the invention
[0005] The purpose of the present invention is to provide a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation and a preparation method thereof. The prepared ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation has both photothermal and chemical kinetic properties, simple preparation process, good biocompatibility, uniform particle size distribution, high photothermal conversion efficiency, and has a broad application prospect in the field of tumor treatment. To achieve the above purpose, the technical solution adopted by the present invention is:
[0006] A ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation and a preparation method thereof, characterized in that the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation is a yellow-brown suspension with good dispersibility in water, and the preparation method comprises the following steps:
[0007] (1) Weigh a certain amount of potassium permanganate, dissolve it in ultrapure water, add formamide, heat to 37-45° C., react for 5-10 hours, and prepare manganese dioxide nanoparticles;
[0008] (2) Weigh a certain amount of ruthenium trichloride, dissolve it in ultrapure water, add sodium borohydride and polyvinyl pyrrolidone, and react for 0.5 to 2 hours to prepare ruthenium nanoparticles;
[0009] (3) dispersing the products obtained in step (1) and step (2) in a certain amount of ultrapure water respectively, adding the product of step (2) to the dispersion of the product of step (1) under ultrasonic conditions, and continuously stirring for 10 to 24 hours to obtain ruthenium-loaded manganese dioxide nanoparticles;
[0010] (4) Weigh a certain amount of glucose oxidase and dissolve it in water, mix it with the aqueous dispersion of the product obtained in step (3), add it to a sodium hyaluronate solution of a certain concentration, and react for 10 to 24 hours to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation.
[0011] A further improvement of the technical solution of the present invention is that in the step (1), the ratio of potassium permanganate, ultrapure water and formamide is 30-300 mg: 16-160 mL: 0.5-5.0 mL.
[0012] A further improvement of the technical solution of the present invention is that in the step (2), the mass ratio of ruthenium trichloride, polyvinyl pyrrolidone and sodium borohydride is 12-96:25-200:4-32.
[0013] The further improvement of the technical solution of the present invention lies in that: in the step (3), the volume ratio of the manganese dioxide nanoparticle dispersion liquid to the ruthenium nanoparticle dispersion liquid is 5-20:2.5-10; the mass ratio of the manganese dioxide nanoparticles to the ruthenium nanoparticles is 5-24:2.5-12.
[0014] A further improvement of the technical solution of the present invention is that: the concentration of sodium hyaluronate in step (4) is 1-2 mg / mL; the mass ratio of ruthenium-loaded manganese dioxide nanoparticles, glucose oxidase and sodium hyaluronate is 5-20:2.5-10:10-40; the volume ratio of ruthenium-loaded manganese dioxide nanoparticles, glucose oxidase and sodium hyaluronate dispersion is 0.5-2:0.5-2:5-20.
[0015] A further improvement of the technical solution of the present invention is that the particle size of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nano-preparation is 80 to 300 nm.
[0016] Due to the adoption of the above technical solution, the technical progress achieved by the present invention is:
[0017] The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation provided by the present invention can produce a dual oxygenation effect and alleviate the hypoxia of tumor tissue; the loaded ruthenium nanoparticles are a good photothermal agent and can produce a good photothermal effect under infrared laser irradiation; the modified hyaluronic acid can target and deliver the nanoparticles to the tumor site, and in the tumor microenvironment, both the manganese dioxide nanoparticles and the ruthenium nanoparticles can undergo a series of reactions to generate oxygen and hydroxyl free radicals; the loaded glucose oxidase can utilize the increased oxygen to consume glucose in the tumor site, reduce energy supply, and generate hydrogen peroxide at the same time, promoting the Fenton-like reaction to generate hydroxyl free radicals, thereby achieving a good anti-tumor effect.
[0018] The invention has the advantages of good biocompatibility, small particle size, good dispersibility, good stability and high photothermal conversion efficiency.
[0019] The manganese dioxide nanoparticles and ruthenium nanoparticles in the present invention are obtained by a simple one-step reduction method, and the remaining steps are prepared by a simple physical stirring method. The preparation method is simple and easy, and can achieve large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 is a particle size distribution diagram of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation obtained in Example 8 of the present invention;
[0021] Figure 2 The photothermal performance of the ruthenium-loaded manganese dioxide-based multifunctional antitumor nanoformulation obtained in Example 8 of the present invention;
[0022] Figure 3 It is the hydroxyl radical detection of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation obtained in Example 8 of the present invention;
[0023] Figure 4 It is the dissolved oxygen detection of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation obtained in Example 8 of the present invention;
[0024] Figure 5 This is a graph showing the pH change after the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation obtained in Example 8 of the present invention is mixed with a glucose solution;
[0025] Figure 6 This is an investigation of the cytotoxicity of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation obtained in Example 8 of the present invention combined with laser irradiation. DETAILED DESCRIPTION
[0026] The present invention is further described in detail below by examples:
[0027] Example 1
[0028] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0029] Weigh 30 mg of potassium permanganate, dissolve it in 16 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 0.5 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 12 mg of ruthenium trichloride, dissolve it in 2 mL of water, add 50 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (4 mg / mL), continue to react for 30 minutes, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 5 mg of manganese dioxide nanoparticles and disperse them in 5 mL of water, take 2.5 mg of ruthenium nanoparticles and disperse them in 1 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 10 mg of sodium hyaluronate, stir and dissolve in 5 mL of water, disperse 5 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 12 hours, then collect by centrifugation and wash once with water to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0030] Example 2
[0031] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0032] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water under stirring at room temperature, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 12 mg of ruthenium trichloride, dissolve it in 2 mL of water, add 50 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (4 mg / mL), continue to react for 30 minutes, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 5 mg of manganese dioxide nanoparticles and disperse them in 5 mL of water, take 2.5 mg of ruthenium nanoparticles and disperse them in 1 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 10 mg of sodium hyaluronate, stir and dissolve in 5 mL of water, disperse 5 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 12 hours, then collect by centrifugation and wash once with water to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0033] Example 3
[0034] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0035] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 24 mg of ruthenium trichloride, dissolve it in 4 mL of water, add 50 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 2 mL of sodium borohydride solution (4 mg / mL), continue to react for 50 min, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 5 mg of manganese dioxide nanoparticles and disperse them in 5 mL of water, take 2.5 mg of ruthenium nanoparticles and disperse them in 1 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 10 mg of sodium hyaluronate, stir and dissolve in 5 mL of water, disperse 5 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 12 hours, then collect by centrifugation and wash once with water to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0036] Example 4
[0037] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0038] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 24 mg of ruthenium trichloride, dissolve it in 4 mL of water, add 50 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 2 mL of sodium borohydride solution (4 mg / mL), continue to react for 50 min, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 2 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 40 mg of sodium hyaluronate, stir and dissolve in 20 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 20 hours, then collect by centrifugation and wash once to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0039] Example 5
[0040] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0041] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 2 mL of sodium borohydride solution (8 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 2 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 40 mg of sodium hyaluronate, stir and dissolve in 20 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 20 hours, then collect by centrifugation and wash once to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0042] Example 6
[0043] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0044] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 1 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 40 mg of sodium hyaluronate, stir and dissolve in 20 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 2 mL of water, add them to the sodium hyaluronate solution, continue stirring for 20 hours, then collect by centrifugation and wash once to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0045] Example 7
[0046] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0047] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 40 mg of sodium hyaluronate, stir and dissolve in 20 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 20 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0048] Example 8
[0049] This embodiment is a method for preparing a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, comprising the following steps:
[0050] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0051] Example 9
[0052] This example is an investigation of the photothermal performance of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulations:
[0053] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0054] The power density is 1.0 W / cm 2 The 808nm laser was used to irradiate the aqueous dispersion of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation and record the temperature changes, such as Figure 2 As shown, the temperature gradually increased with the increase of illumination time, indicating that the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation has photothermal properties.
[0055] Example 10
[0056] This example is an investigation of the generation of hydroxyl radicals by a Fenton-like reaction of a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation:
[0057] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0058] The 3,3',5,5'-tetramethylbenzidine (TMB) method was used to detect the generation of hydroxyl radicals in ruthenium-loaded manganese dioxide-based multifunctional antitumor nanoformulations under hydrogen peroxide and slightly acidic (pH 6.5) conditions. Figure 3 As shown, compared with the control group, the preparation group has an obvious characteristic absorption peak at around 560nm, indicating that the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation can generate hydroxyl radicals.
[0059] Embodiment 11
[0060] This example is an investigation of the ability of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation to produce oxygen:
[0061] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0062] A portable dissolved oxygen meter was used to measure the ability of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation to react with hydrogen peroxide to generate oxygen in a slightly acidic environment (pH 6.5). Figure 4 As shown, compared with the control group, the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation can react with hydrogen peroxide to produce oxygen.
[0063] Example 12
[0064] This example is a verification that the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation can catalyze the oxidation of glucose to generate gluconic acid and hydrogen peroxide, resulting in a decrease in the pH of the mixed solution:
[0065] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0066] The glucose solution was mixed with the dispersion of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, and samples were centrifuged at different time points. The pH value was measured using a pH meter, such as Figure 5 As shown, compared with the control group (glucose solution only), the pH of the mixed solution of glucose and ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation was significantly reduced, indicating that the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation has glucose oxidase activity.
[0067] Embodiment 13
[0068] This example is a study on the cytotoxicity of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulations:
[0069] Weigh 60 mg of potassium permanganate, dissolve it in 32 mL of ultrapure water at room temperature with stirring, continue stirring for 30 min, then transfer to a 40 ° C water bath, slowly add 1 mL of formamide under stirring, continue to react for 8 hours, centrifuge and collect, wash 3 times with water to obtain manganese dioxide nanoparticles. Weigh 48 mg of ruthenium trichloride, dissolve it in 10 mL of water, add 100 mg of polyvinyl pyrrolidone under stirring, after polyvinyl pyrrolidone is completely dissolved, add 1 mL of sodium borohydride solution (16 mg / mL), continue to react for 1 hour, centrifuge and collect, wash 3 times with water to obtain ruthenium nanoparticles. Take 10 mg of manganese dioxide nanoparticles and disperse them in 10 mL of water, take 5 mg of ruthenium nanoparticles and disperse them in 5 mL of water, add the ruthenium nanoparticle dispersion to the manganese dioxide nanoparticle dispersion under ultrasound, continue to stir for 12 hours, centrifuge and collect, wash 3 times with water to obtain ruthenium-loaded manganese dioxide nanoparticles. Weigh 20 mg of sodium hyaluronate, stir and dissolve in 10 mL of water, disperse 10 mg of ruthenium-loaded manganese dioxide nanoparticles in 1 mL of water, mix evenly with 1 mL of glucose oxidase solution (5 mg / mL), and then add to the sodium hyaluronate solution. After continuous stirring for 12 hours, collect by centrifugation, wash once with water, and obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanopreparation.
[0070] Mouse breast cancer cells (4T1 cells) were used to determine the cell-killing effect of different concentrations of ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticles combined with laser irradiation. The laser power density was 1.0 W / cm 2 The 808nm laser was used for 5min. Figure 6 As shown, when the concentration reaches 60 μg / mL, the cells are basically dead, indicating that the ruthenium manganese dioxide-based multifunctional anti-tumor nanoformulation combined with laser irradiation has a strong killing effect on 4T1 cells.
Claims
1. A ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoparticle preparation, the preparation method of which comprises the following steps: (1) Weigh a certain amount of potassium permanganate, dissolve it in ultrapure water, add formamide, heat to 37-45° C., react for 5-10 hours, and prepare manganese dioxide nanoparticles; (2) Weigh a certain amount of ruthenium trichloride, dissolve it in ultrapure water, add sodium borohydride and polyvinyl pyrrolidone, and react for 0.5 to 2 hours to prepare ruthenium nanoparticles; (3) dispersing the products obtained in step (1) and step (2) in a certain amount of ultrapure water respectively, adding the product of step (2) to the dispersion of the product of step (1) under ultrasonic conditions, and continuously stirring for 10 to 24 hours to obtain ruthenium-loaded manganese dioxide nanoparticles; (4) Weigh a certain amount of glucose oxidase and dissolve it in water, mix it with the aqueous dispersion of the product obtained in step (3), add it to a sodium hyaluronate solution of a certain concentration, and react for 10 to 24 hours to obtain a ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation.
2. The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation according to claim 1, characterized in that: In the preparation step (1), the ratio of potassium permanganate, ultrapure water and formamide is 30-300 mg: 16-160 mL: 0.5-5.0 mL.
3. The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation according to claim 1, characterized in that: In the preparation step (2), the mass ratio of ruthenium trichloride, polyvinyl pyrrolidone and sodium borohydride is 12-96:25-200:4-32.
4. The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation according to claim 1, characterized in that: In the preparation step (3), the volume ratio of the manganese dioxide nanoparticle dispersion liquid to the ruthenium nanoparticle dispersion liquid is 5-20:2.5-10; the mass ratio of the manganese dioxide nanoparticles to the ruthenium nanoparticles is 5-24:2.5-12.
5. The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation according to claim 1, characterized in that: The concentration of sodium hyaluronate in the preparation step (4) is 1-2 mg / mL; the mass ratio of ruthenium-loaded manganese dioxide nanoparticles, glucose oxidase and sodium hyaluronate is 5-20:2.5-10:10-40; the volume ratio of ruthenium-loaded manganese dioxide nanoparticle dispersion, glucose oxidase dispersion and sodium hyaluronate dispersion is 0.5-2:0.5-2:5-20.
6. The ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nanoformulation according to claim 1, characterized in that: The particle size of the ruthenium-loaded manganese dioxide-based multifunctional anti-tumor nano preparation is 80-300 nm.