Photoresponsive multifunctional nanoscale enzyme and preparation method and application thereof

Abstract

The present application relates to a kind of light response multifunctional nano enzyme and its preparation method and application, belong to nano enzyme catalysis field.The nano enzyme described in the present application can utilize Pt, Te precursor, is prepared by hydrothermal synthesis method, synthesis method is simple, condition is stable, and repeatability is high.The nano enzyme exists multiple enzyme activity such as oxidase-like enzyme, peroxide enzyme-like, hydrogen peroxide enzyme-like, glutathione peroxidase-like, has good stability, catalytic effect, catalytic rate fast and the like characteristics.Meanwhile, by using the good photo-thermal effect and photodynamic performance of the nano enzyme, the synergy treatment of chemical dynamics, photodynamic and photo-thermal is realized, so that it becomes a potential material for effectively inhibiting the occurrence and development of tumor.

Description

Technical Field

[0001] This invention relates to the field of nanozyme catalysis technology, and in particular to a photoresponsive multifunctional nanozyme, its preparation method, and its applications. Background Technology

[0002] Nanozymes, as nanoscale inorganic catalytic materials with excellent enzyme-like activity, can catalyze chemical reactions of corresponding substrates under physiological conditions, following the principles of enzyme kinetics. Compared with traditional materials, nanozymes have advantages such as higher selectivity, physicochemical stability, and catalytic activity, as well as lower production costs and simpler preparation and purification processes.

[0003] In recent years, researchers have explored many nanozyme systems, which typically possess one or more enzyme-like activities. Typical nanozymes include CeO2, Fe3O4, V2O5, and MnO2. Furthermore, compared to traditional tumor treatments, their inherent physicochemical properties give them higher targeting capabilities. Currently, nanozymes have emerged as a potential tumor treatment strategy. For example, CeO2 possesses multiple catalytic activities, including those resembling oxidases, peroxidases, catalases, and superoxide dismutases. These activities can synergistically regulate reactive oxygen species levels, inducing oxidative stress in tumor cells while simultaneously reducing apoptosis in normal cells, thus providing effective protection for the body.

[0004] However, for nanozyme-based tumor therapy, it is essential to more systematically design the activity and structure of nanozymes according to the complexity of organisms in order to improve their efficiency and reduce biotoxicity. Summary of the Invention

[0005] Based on the current need to improve the working efficiency of nanozymes and reduce their biotoxicity in existing technologies, this invention provides a photoresponsive multifunctional nanozyme, its preparation method, and its applications.

[0006] This invention proposes a platinum-tellurium (Pt-Te) multifunctional nanozyme, its preparation method, and its applications. This invention utilizes its enzyme-like activity to specifically target the tumor microenvironment, improving treatment efficiency. Furthermore, it organically combines the catalytic therapeutic properties of the nanozyme with external stimuli to achieve a synergistic therapeutic effect.

[0007] The objective of this invention can be achieved through the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing a photoresponsive multifunctional nanozyme, comprising the following steps:

[0009] Step 1: Dissolve tetraammineplatinum nitrate in water to prepare solution A;

[0010] Step 2: Dissolve potassium tellurite in water to prepare solution B;

[0011] Step 3: Dissolve L-ascorbic acid in water to prepare solution C;

[0012] Step 4: Dissolve polyvinylpyrrolidone in water to prepare solution D;

[0013] Step 5: Mix solutions A, B, C and D thoroughly, then add ethylene glycol to prepare suspension E.

[0014] Step 6: Sonicate the suspension E until the solution system is evenly dispersed to obtain a clear solution F;

[0015] Step 7: Transfer the above solution F to a reaction vessel and heat it at high temperature, then cool it to room temperature to obtain solution G;

[0016] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate several times to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0017] In one embodiment of the present invention, in step 1, the concentration of tetraammineplatinum nitrate in solution A is 8-10 mg / mL.

[0018] In one embodiment of the present invention, in step 2, the concentration of potassium tellurite is 5-10 mg / mL.

[0019] In one embodiment of the present invention, in step 3, the concentration of L-ascorbic acid is 28-30 mg / mL.

[0020] In one embodiment of the present invention, in step 4, the molecular weight of polyvinylpyrrolidone is 58,000, and the concentration of polyvinylpyrrolidone is 60-100 mg / mL, for example, 80 mg / mL.

[0021] In one embodiment of the present invention, in step 5, solutions A, B, C and D are mixed in a volume ratio of 1:1:1:1, and the amount of ethylene glycol added is the total volume of solutions A, B, C and D.

[0022] In one embodiment of the present invention, in step 6, the suspension E is sonicated at room temperature for 1 to 2 hours.

[0023] In one embodiment of the present invention, in step 7, the heating temperature of solution F is 180-220°C, and the heating time is 4-6 hours.

[0024] In one embodiment of the present invention, in step 8, the precipitate is washed using an ethanol / acetone mixture, wherein the ethanol / acetone mixture is prepared in a volume ratio of 1:9.

[0025] Secondly, the present invention provides a photoresponsive multifunctional nanozyme obtained based on any of the above preparation methods.

[0026] Thirdly, the present invention provides photoresponsive multifunctional nanozymes obtained based on the above method for use as oxidases, peroxidases, catalases, and glutathione peroxidases.

[0027] Fourthly, the present invention provides the application of the above-obtained photoresponsive multifunctional nanozyme in the preparation of tumor therapeutic drugs. In this application, the photoresponsive multifunctional nanozyme is mainly used to generate reactive oxygen species for tumor treatment.

[0028] Compared with the prior art, the advantages and beneficial effects of the present invention are mainly reflected in the following aspects:

[0029] The nanozyme described in this invention can be prepared using Pt and Te raw materials via a hydrothermal synthesis method. The synthesis method is simple, the conditions are stable, and the reproducibility is high.

[0030] This nanozyme exhibits multiple enzyme activities, including oxidase-like, peroxidase-like, catalase-like, and glutathione peroxidase-like activities, and is characterized by good stability, good catalytic effect, and fast catalytic rate.

[0031] Meanwhile, the excellent photothermal and photodynamic properties of this nanozyme can be utilized to achieve synergistic therapy of chemodynamics, photodynamics, and photothermal effects, making it a potential material for effectively inhibiting tumor occurrence and development. Attached Figure Description

[0032] Figure 1 This is a transmission electron microscope image of the Pt-Te nanozyme prepared in Example 1.

[0033] Figure 2 This is an elemental distribution diagram of the Pt-Te nanozyme prepared in Example 1.

[0034] Figure 3 This is a comparison diagram of the peroxidase-like activities of the nanozymes prepared in Example 1 and Comparative Examples 1 and 2.

[0035] Figure 4 This is a peroxidase-like kinetic curve of the Pt-Te nanozyme prepared in Example 1.

[0036] Figure 5 This is a photodynamic performance diagram of the Pt-Te nanozyme prepared in Example 1.

[0037] Figure 6 This is a photothermal performance diagram of the Pt-Te nanozyme prepared in Example 1.

[0038] Figure 7This is a photothermal stability diagram of the Pt-Te nanozyme prepared in Example 1. Detailed Implementation

[0039] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0040] Example 1:

[0041] This embodiment provides a method for preparing a photoresponsive multifunctional nanozyme, the steps of which are as follows:

[0042] Step 1: Dissolve 30 mg of tetraammineplatinum nitrate in 3.75 mL of water to prepare solution A;

[0043] Step 2: Dissolve 38.4 mg of potassium tellurite in 3.75 mL of water to prepare solution B;

[0044] Step 3: Dissolve 106.8 mg L-ascorbic acid in 3.75 mL of water to prepare solution C;

[0045] Step 4: Dissolve 300 mg of polyvinylpyrrolidone in 3.75 mL of water to prepare solution D;

[0046] Step 5: Mix solutions A, B, C and D thoroughly, then add 15 mL of ethylene glycol to prepare suspension E.

[0047] Step 6: Sonicate the above suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed to obtain a clear solution F;

[0048] Step 7: Transfer the above solution F to a reaction vessel and heat at 200°C for 5 hours. Cool to room temperature to obtain solution G.

[0049] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate three times with a mixture of ethanol / acetone in a volume ratio of 1:9 to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0050] like Figure 1 As shown: The transmission electron microscopy image of the Pt-Te nanozyme prepared in Example 1 shows that the size of the nanozyme is approximately 100–150 nm. Figure 2 As shown: The elemental analysis diagram of the Pt-Te nanozyme prepared in Example 1 shows that Pt and Te are uniformly distributed in the material.

[0051] Example 2

[0052] This embodiment provides a method for preparing a photoresponsive multifunctional nanozyme, which includes the following steps:

[0053] Step 1: Dissolve tetraammineplatinum nitrate in water to prepare solution A;

[0054] Step 2: Dissolve potassium tellurite in water to prepare solution B;

[0055] Step 3: Dissolve L-ascorbic acid in water to prepare solution C;

[0056] Step 4: Dissolve polyvinylpyrrolidone in water to prepare solution D;

[0057] Step 5: Mix solutions A, B, C and D thoroughly, then add ethylene glycol to prepare suspension E.

[0058] Step 6: Sonicate the suspension E until the solution system is evenly dispersed to obtain a clear solution F;

[0059] Step 7: Transfer the above solution F to a reaction vessel and heat it at high temperature, then cool it to room temperature to obtain solution G;

[0060] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate several times to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0061] In step 1, the concentration of tetraammineplatinum nitrate in solution A is 8 mg / mL. In step 2, the concentration of potassium tellurite is 5 mg / mL. In step 3, the concentration of L-ascorbic acid is 28 mg / mL. In step 4, the molecular weight of polyvinylpyrrolidone is 58,000, and the concentration of polyvinylpyrrolidone is 60 mg / mL. In step 5, solutions A, B, C, and D are mixed in a volume ratio of 1:1:1:1, and the amount of ethylene glycol added is the total volume of solutions A, B, C, and D. In step 6, suspension E is sonicated at room temperature for 1 hour. In step 7, solution F is heated to 220°C for 4 hours. In step 8, the precipitate is washed using an ethanol / acetone mixture, prepared in a volume ratio of 1:9.

[0062] The photoresponsive multifunctional nanozyme obtained in this embodiment is used as an oxidase, peroxidase, catalase, and glutathione peroxidase.

[0063] This embodiment also provides the application of photoresponsive multifunctional nanozymes in the preparation of tumor therapeutic drugs. In this application, the photoresponsive multifunctional nanozymes are mainly used to generate reactive oxygen species for tumor treatment.

[0064] Example 3

[0065] This embodiment provides a method for preparing a photoresponsive multifunctional nanozyme, which includes the following steps:

[0066] Step 1: Dissolve tetraammineplatinum nitrate in water to prepare solution A;

[0067] Step 2: Dissolve potassium tellurite in water to prepare solution B;

[0068] Step 3: Dissolve L-ascorbic acid in water to prepare solution C;

[0069] Step 4: Dissolve polyvinylpyrrolidone in water to prepare solution D;

[0070] Step 5: Mix solutions A, B, C and D thoroughly, then add ethylene glycol to prepare suspension E.

[0071] Step 6: Sonicate the suspension E until the solution system is evenly dispersed to obtain a clear solution F;

[0072] Step 7: Transfer the above solution F to a reaction vessel and heat it at high temperature, then cool it to room temperature to obtain solution G;

[0073] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate several times to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0074] In step 1, the concentration of tetraammineplatinum nitrate in solution A is 10 mg / mL. In step 2, the concentration of potassium tellurite is 10 mg / mL. In step 3, the concentration of L-ascorbic acid is 30 mg / mL. In step 4, the molecular weight of polyvinylpyrrolidone is 58,000, and the concentration of polyvinylpyrrolidone is 100 mg / mL. In step 5, solutions A, B, C, and D are mixed in a volume ratio of 1:1:1:1, and the amount of ethylene glycol added is the total volume of solutions A, B, C, and D. In step 6, suspension E is sonicated at room temperature for 2 hours. In step 7, solution F is heated to 180°C for 4 hours. In step 8, the precipitate is washed using an ethanol / acetone mixture, prepared in a volume ratio of 1:9.

[0075] The photoresponsive multifunctional nanozyme obtained in this embodiment is used as an oxidase, peroxidase, catalase, and glutathione peroxidase.

[0076] This embodiment also provides the application of photoresponsive multifunctional nanozymes in the preparation of tumor therapeutic drugs. In this application, the photoresponsive multifunctional nanozymes are mainly used to generate reactive oxygen species for tumor treatment.

[0077] Example 4

[0078] This embodiment provides a method for preparing a photoresponsive multifunctional nanozyme, which includes the following steps:

[0079] Step 1: Dissolve tetraammineplatinum nitrate in water to prepare solution A;

[0080] Step 2: Dissolve potassium tellurite in water to prepare solution B;

[0081] Step 3: Dissolve L-ascorbic acid in water to prepare solution C;

[0082] Step 4: Dissolve polyvinylpyrrolidone in water to prepare solution D;

[0083] Step 5: Mix solutions A, B, C and D thoroughly, then add ethylene glycol to prepare suspension E.

[0084] Step 6: Sonicate the suspension E until the solution system is evenly dispersed to obtain a clear solution F;

[0085] Step 7: Transfer the above solution F to a reaction vessel and heat it at high temperature, then cool it to room temperature to obtain solution G;

[0086] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate several times to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0087] In step 1, the concentration of tetraammineplatinum nitrate in solution A is 9 mg / mL. In step 2, the concentration of potassium tellurite is 8 mg / mL. In step 3, the concentration of L-ascorbic acid is 29 mg / mL. In step 4, the molecular weight of polyvinylpyrrolidone is 58,000, and the concentration of polyvinylpyrrolidone is 80 mg / mL. In step 5, solutions A, B, C, and D are mixed in a volume ratio of 1:1:1:1, and the amount of ethylene glycol added is the total volume of solutions A, B, C, and D. In step 6, suspension E is sonicated at room temperature for 1.5 hours. In step 7, solution F is heated to 200°C for 5 hours. In step 8, the precipitate is washed using an ethanol / acetone mixture, prepared in a volume ratio of 1:9.

[0088] The photoresponsive multifunctional nanozyme obtained in this embodiment is used as an oxidase, peroxidase, catalase, and glutathione peroxidase.

[0089] This embodiment also provides the application of photoresponsive multifunctional nanozymes in the preparation of tumor therapeutic drugs. In this application, the photoresponsive multifunctional nanozymes are mainly used to generate reactive oxygen species for tumor treatment.

[0090] Comparative Example 1:

[0091] This comparative example provides a method for preparing nanozymes, the steps of which are as follows:

[0092] Step 1: Dissolve 30 mg of tetraammineplatinum nitrate in 3.75 mL of water to prepare solution A;

[0093] Step 2: Dissolve 19.2 mg of potassium tellurite in 3.75 mL of water to prepare solution B;

[0094] Step 3: Dissolve 106.8 mg L-ascorbic acid in 3.75 mL of water to prepare solution C;

[0095] Step 4: Dissolve 300 mg of polyvinylpyrrolidone in 3.75 mL of water to prepare solution D;

[0096] Step 5: Mix solutions A, B, C and D thoroughly, then add 15 mL of ethylene glycol to prepare suspension E.

[0097] Step 6: Sonicate the above suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed to obtain a clear solution F;

[0098] Step 7: Transfer the above solution F to a reaction vessel and heat at 200°C for 5 hours. Cool to room temperature to obtain solution G.

[0099] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate three times with a mixture of ethanol / acetone in a volume ratio of 1:9 to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0100] Comparative Example 2:

[0101] This comparative example provides a method for preparing nanozymes, the steps of which are as follows:

[0102] Step 1: Dissolve 30 mg of tetraammineplatinum nitrate in 3.75 mL of water to prepare solution A;

[0103] Step 2: Dissolve 28.8 mg of potassium tellurite in 3.75 mL of water to prepare solution B;

[0104] Step 3: Dissolve 106.8 mg L-ascorbic acid in 3.75 mL of water to prepare solution C;

[0105] Step 4: Dissolve 300 mg of polyvinylpyrrolidone in 3.75 mL of water to prepare solution D;

[0106] Step 5: Mix solutions A, B, C and D thoroughly, then add 15 mL of ethylene glycol to prepare suspension E.

[0107] Step 6: Sonicate the above suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed to obtain a clear solution F;

[0108] Step 7: Transfer the above solution F to a reaction vessel and heat at 200°C for 5 hours. Cool to room temperature to obtain solution G.

[0109] Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate three times with a mixture of ethanol / acetone in a volume ratio of 1:9 to obtain the photoresponsive multifunctional nanozyme, namely Pt-Te nanozyme.

[0110] The nanozymes prepared in Example 1 and Comparative Examples 1 and 2 were subjected to peroxidase activity assays. The specific steps are as follows: First, the nanozymes prepared in Example 1 and Comparative Examples 1 and 2 were prepared into suspensions of 80 μg / mL. Then, 80 μL of each of the above-mentioned enzyme suspensions was added to a 96-well plate, and 40 μL of 10 mM H2O2 and 40 μL of 4 mM 3,3',5,5'-tetramethylbenzidine (TMB) were added. The optical density (OD) at 652 nm was measured. 652 ).from Figure 3 It can be seen that the nanozyme OD obtained in Example 1 652 The value increased significantly, while in comparative examples 1 and 2, the OD value increased significantly. 652 The value is relatively small, indicating that the amount of potassium tellurite added will affect the peroxidase-like activity of Pt-Te nanozymes.

[0111] To verify the beneficial effects of the present invention, the following test experiments were conducted.

[0112] Test Experiment 1: Peroxidase-like Kinetic Calculation of Pt-Te Nanozymes

[0113] To calculate the peroxidase-like kinetic parameters of the Pt-Te nanozyme prepared in Example 1, the specific steps are as follows: First, the nanozyme obtained in Example 1 was prepared into an 80 μg / mL dispersion. Then, 80 μL of the above nanozyme dispersion was added to a 96-well plate, followed by 40 μL of 4 mM TMB, and then 40 μL of 6 mM, 8 mM, 10 mM, 20 mM, 40 mM, 60 mM, and 80 mM H2O2, respectively. After reacting at room temperature for 5 min, the optical density (OD) value at 652 nm was measured. 652 The result is as follows: Figure 4 As shown, the rate of the enzymatic reaction gradually increases with the increase of H2O2 concentration, and the Michaelis constant K of the nanozyme can be calculated by curve fitting. m The concentration was 6.319 mM, and the maximum reaction rate was 6.1965 × 10⁻⁶. -7 M / s.

[0114] Test Experiment 2: Photodynamic Performance Test of Pt-Te Nanozymes

[0115] To test the photodynamic properties of the Pt-Te nanozyme prepared in Example 1, the specific steps were as follows: First, the Pt-Te nanozyme prepared in Example 1 was prepared into a 40 μg / mL dispersion. Then, 2 mL of sample was taken for testing, and 60 μL of 1 mg / mL 1,3-diphenylisobenzofuran (DPBF) was added to it. Then, a power density of 1.0 W / cm² was used. 2 The reaction system was irradiated with a 1064 nm laser for 0 min, 1 min, 2 min, 3 min, 4 min, and 5 min, and the ultraviolet absorption spectra of the system at different irradiation times were measured. The results are as follows: Figure 5 As shown, the characteristic absorption value of DPBF decreases significantly with increasing laser irradiation time, indicating that this Pt-Te nanozyme has certain photodynamic properties.

[0116] Test Experiment 3: Photothermal Performance Test of Pt-Te Nanozymes

[0117] To test the photothermal properties of the Pt-Te nanozyme prepared in Example 1, the specific steps are as follows: First, the Pt-Te nanozyme prepared in Example 1 was prepared into dispersions of 25 μg / mL, 50 μg / mL, 100 μg / mL, and 200 μg / mL, respectively. Then, these dispersions were sequentially added to 96-well plates, and the plates were heated with a power density of 1.0 W / cm². 2 A 1064nm laser was used to irradiate the sample for 10 minutes, and the corresponding temperature changes were recorded using a FOTRIC photothermal camera. The results are as follows: Figure 6 As shown, with aqueous solution as a control, the temperature change of Pt-Te nanozyme increases with increasing concentration, indicating that its photothermal properties gradually increase with increasing concentration.

[0118] Test Experiment 4: Photothermal Stability Test of Pt-Te Nanozymes

[0119] To test the photothermal stability of the Pt-Te nanozyme prepared in Example 1, the specific steps are as follows: First, the Pt-Te nanozyme prepared in Example 1 was prepared into a dispersion of 200 μg / mL, and then... 2 A 1064nm laser was used to irradiate the area for 12.5 minutes, and the corresponding temperature changes were recorded using a FOTRIC photothermal camera. The laser was then turned off, and after the temperature dropped to its initial value, it was turned back on for another 12.5 minutes. This cycle was repeated four times. The results are as follows: Figure 7 As shown, the temperature rise of the Pt-Te nanozyme was almost equal during the four cycles of switching the laser, indicating that the Pt-Te nanozyme has good photothermal stability.

[0120] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. The application of a photoresponsive multifunctional nanozyme in the preparation of tumor therapeutic drugs, characterized in that, The preparation method of the photoresponsive multifunctional nanozyme includes the following steps: Step 1: Dissolve tetraammineplatinum nitrate in water to prepare solution A; Step 2: Dissolve potassium tellurite in water to prepare solution B; Step 3: Dissolve L-ascorbic acid in water to prepare solution C; Step 4: Dissolve polyvinylpyrrolidone in water to prepare solution D; Step 5: Mix solutions A, B, C and D thoroughly, then add ethylene glycol to prepare suspension E. Step 6: Sonicate the suspension E until the solution system is evenly dispersed to obtain a clear solution F; Step 7: Transfer the above solution F to a reaction vessel and heat it at high temperature, then cool it to room temperature to obtain solution G; Step 8: Centrifuge the above solution G to collect the precipitate, wash the precipitate several times to obtain the photoresponsive multifunctional nanozyme.

2. The application according to claim 1, characterized in that, In step 1, the concentration of tetraammineplatinum nitrate in solution A is 8-10 mg / mL.

3. The application according to claim 1, characterized in that, In step 2, the concentration of potassium tellurite is 5~10 mg / mL.

4. The application according to claim 1, characterized in that, In step 3, the concentration of L-ascorbic acid is 28~30 mg / mL.

5. The application according to claim 1, characterized in that, In step 4, the molecular weight of polyvinylpyrrolidone is 58,000, and the concentration of polyvinylpyrrolidone is 60~100 mg / mL.

6. The application according to claim 1, characterized in that, In step 5, solutions A, B, C, and D are mixed in a volume ratio of 1:1:1:1, and the amount of ethylene glycol added is the total volume of solutions A, B, C, and D.

7. The application according to claim 1, characterized in that, In step 6, suspension E is sonicated at room temperature for 1-2 hours; In step 7, the heating temperature of solution F is 180~220 ℃, and the heating time is 4~6 hours; In step 8, the precipitate is washed using an ethanol / acetone mixture, which is prepared in a volume ratio of 1:9.

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