CN117959428A - Light-responsive multifunctional nano-enzyme and preparation method and application thereof - Google Patents
Light-responsive multifunctional nano-enzyme and preparation method and application thereof Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims description 154
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 32
- 239000000725 suspension Substances 0.000 claims description 23
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 20
- 238000010438 heat treatment Methods 0.000 claims description 18
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 18
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 18
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 18
- BFPJYWDBBLZXOM-UHFFFAOYSA-L potassium tellurite Chemical compound [K+].[K+].[O-][Te]([O-])=O BFPJYWDBBLZXOM-UHFFFAOYSA-L 0.000 claims description 14
- 229960005070 ascorbic acid Drugs 0.000 claims description 13
- 239000002244 precipitate Substances 0.000 claims description 13
- 238000005406 washing Methods 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 11
- 239000002211 L-ascorbic acid Substances 0.000 claims description 10
- 235000000069 L-ascorbic acid Nutrition 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- NWAHZABTSDUXMJ-UHFFFAOYSA-N platinum(2+);dinitrate Chemical compound [Pt+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O NWAHZABTSDUXMJ-UHFFFAOYSA-N 0.000 claims description 9
- 238000009210 therapy by ultrasound Methods 0.000 claims description 9
- 238000001816 cooling Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 102000003992 Peroxidases Human genes 0.000 claims description 6
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 6
- 102000016938 Catalase Human genes 0.000 claims description 5
- 108030002440 Catalase peroxidases Proteins 0.000 claims description 5
- 102000006587 Glutathione peroxidase Human genes 0.000 claims description 5
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- 108090000854 Oxidoreductases Proteins 0.000 claims description 5
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- AGGKEGLBGGJEBZ-UHFFFAOYSA-N tetramethylenedisulfotetramine Chemical compound C1N(S2(=O)=O)CN3S(=O)(=O)N1CN2C3 AGGKEGLBGGJEBZ-UHFFFAOYSA-N 0.000 claims description 4
- 229940126585 therapeutic drug Drugs 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 abstract description 13
- 108090000790 Enzymes Proteins 0.000 abstract description 13
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- 238000011282 treatment Methods 0.000 abstract description 9
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- 229910052697 platinum Inorganic materials 0.000 abstract description 7
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- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 abstract description 4
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- 238000012360 testing method Methods 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
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- ZKSVYBRJSMBDMV-UHFFFAOYSA-N 1,3-diphenyl-2-benzofuran Chemical compound C1=CC=CC=C1C1=C2C=CC=CC2=C(C=2C=CC=CC=2)O1 ZKSVYBRJSMBDMV-UHFFFAOYSA-N 0.000 description 3
- 239000011668 ascorbic acid Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
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- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 2
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- PORWMNRCUJJQNO-UHFFFAOYSA-N tellurium atom Chemical compound [Te] PORWMNRCUJJQNO-UHFFFAOYSA-N 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
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- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
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- A61K33/00—Medicinal preparations containing inorganic active ingredients
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
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Abstract
The invention relates to a light-responsive multifunctional nano-enzyme, a preparation method and application thereof, and belongs to the technical field of nano-enzyme catalysis. The nano-enzyme can be prepared by using Pt and Te precursors through a hydrothermal synthesis method, and the synthesis method is simple, stable in condition and high in repeatability. The nano-enzyme has various enzyme activities such as oxidase-like, peroxidase-like, catalase-like, glutathione peroxidase-like and the like, and has the characteristics of good stability, good catalytic effect, high catalytic rate and the like. Meanwhile, the nanometer enzyme has good photo-thermal effect and photodynamic performance, and can realize the cooperative treatment of chemical power, photodynamic and photo-thermal, so that the nanometer enzyme becomes a potential material for effectively inhibiting the occurrence and development of tumors.
Description
Technical Field
The invention relates to the technical field of nano enzyme catalysis, in particular to a light-responsive multifunctional nano enzyme and a preparation method and application thereof.
Background
The nano enzyme, which is a nano inorganic catalytic material with good enzyme-like activity, can catalyze corresponding substrates to generate chemical reaction according to the principle of enzyme dynamics under physiological conditions. Compared with the traditional material, the nano-enzyme has the advantages of higher selectivity, physical and chemical stability, catalytic activity, low production cost, simple preparation and purification process and the like.
In recent years, researchers have explored many nanoenzyme systems, and they typically have one or more enzyme-like activities. Among them, typical nanoezymes include CeO 2、Fe3O4、V2O5、MnO2 and the like. In addition, compared with the traditional tumor treatment mode, the inherent physical and chemical properties of the nano-enzyme enable the nano-enzyme to have higher targeting property, so that the nano-enzyme becomes a potential tumor treatment strategy at present, for example, ceO 2 has multiple catalytic activities of oxidase-like, peroxidase-like, catalase-like and superoxide dismutase-like, and can synergistically regulate and control the content of active oxygen to enable tumor cells to generate oxidative stress, reduce apoptosis of normal cells and effectively protect organisms.
However, for nanoenzyme-based tumor therapy, it is necessary to more systematically design the activity and structure of nanoenzymes according to the complexity of organisms to increase their working efficiency and reduce biotoxicity.
Disclosure of Invention
Based on the current situation that the working efficiency of the nano-enzyme needs to be improved and the biotoxicity needs to be reduced in the prior art, the invention provides a light-responsive multifunctional nano-enzyme, and a preparation method and application thereof.
The invention provides a platinum tellurium (Pt-Te) multifunctional nano-enzyme, a preparation method and application thereof. The invention can utilize the activity of the enzyme, specifically acts on the tumor microenvironment, improves the treatment efficiency, organically combines the catalytic treatment performance of the nano-enzyme with external stimulus, and realizes a treatment mode of mutual synergistic promotion.
The aim of the invention can be achieved by the following technical scheme:
In a first aspect, the present invention provides a method for preparing a light-responsive multifunctional nano-enzyme, comprising the steps of:
step 1, dissolving tetrammine platinum nitrate in water to prepare a solution A;
Step2, dissolving potassium tellurite in water to prepare a solution B;
step 3, dissolving L-ascorbic acid in water to prepare a solution C;
Step 4, dissolving polyvinylpyrrolidone in water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E until the solution system is uniformly dispersed to obtain a clear solution F;
Step 7, transferring the solution F into a reaction kettle, heating at high temperature, and cooling to room temperature to obtain a solution G;
And 8, centrifugally collecting the solution G, washing the precipitate for several times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
In one embodiment of the present invention, in the step 1, the concentration of platinum tetrammine nitrate in the solution A is 8 to 10mg/mL.
In one embodiment of the present invention, in the step 2, the concentration of potassium tellurite is 5 to 10mg/mL.
In one embodiment of the present invention, the concentration of L-ascorbic acid in the step 3 is 28 to 30mg/mL.
In one embodiment of the present invention, in the step 4, the molecular weight of polyvinylpyrrolidone is 58000 and the concentration of polyvinylpyrrolidone is 60 to 100mg/mL, for example, 80mg/mL.
In one embodiment of the present invention, in the step 5, the solution a, the solution B, the solution C and the solution D are mixed according to a volume ratio of 1:1:1:1, and the addition amount of the ethylene glycol is the total volume of the solution a, the solution B, the solution C and the solution D.
In one embodiment of the invention, in step 6, suspension E is sonicated for a period of 1 to 2 hours at room temperature.
In one embodiment of the present invention, in the step 7, the heating temperature of the solution F is 180 to 220℃and the heating time is 4 to 6 hours.
In one embodiment of the present invention, in the step 8, the washing precipitate uses an ethanol/acetone mixture, and the ethanol/acetone mixture is prepared in a manner that the volume ratio of ethanol to acetone is 1:9.
In a second aspect, the invention provides a light-responsive multifunctional nano-enzyme obtained based on any one of the preparation methods.
In a third aspect, the present invention provides the use of the light-responsive multifunctional nanoenzyme obtained based on the above method as an oxidase, peroxidase, catalase and glutathione peroxidase.
In a fourth aspect, the invention provides an application of the obtained light-responsive multifunctional nano-enzyme in preparing a tumor therapeutic drug. In the application, active oxygen is mainly generated by utilizing the light-responsive multifunctional nano-enzyme to treat tumors.
Compared with the prior art, the invention has the advantages and beneficial effects that:
The nano enzyme can be prepared from Pt raw material and Te raw material by a hydrothermal synthesis method, and the synthesis method is simple, stable in condition and high in repeatability.
The nano-enzyme has various enzyme activities such as oxidase-like, peroxidase-like, catalase-like, glutathione peroxidase-like and the like, and has the characteristics of good stability, good catalytic effect, high catalytic rate and the like.
Meanwhile, the nanometer enzyme has good photo-thermal effect and photodynamic performance, and can realize the cooperative treatment of chemical power, photodynamic and photo-thermal, so that the nanometer enzyme becomes a potential material for effectively inhibiting the occurrence and development of tumors.
Drawings
FIG. 1 is a transmission electron microscope image of the Pt-Te nano-enzyme prepared in example 1.
FIG. 2 is an elemental distribution diagram of the Pt-Te nano-enzyme prepared in example 1.
FIG. 3 is a graph showing the comparison of the peroxidase-like activities of the nanoelectrozymes obtained in example 1 and comparative examples 1 and 2.
FIG. 4 is a graph showing the peroxidase-like kinetics of the Pt-Te nano-enzyme prepared in example 1.
FIG. 5 is a graph showing photodynamic properties of the Pt-Te nano-enzyme prepared in example 1.
FIG. 6 is a photo-thermal property diagram of the Pt-Te nano-enzyme prepared in example 1.
FIG. 7 is a photo-thermal stability graph of the Pt-Te nano-enzyme prepared in example 1.
Detailed Description
The invention will now be described in detail with reference to the drawings and specific examples.
Example 1:
the embodiment provides a preparation method of light-responsive multifunctional nano-enzyme, which comprises the following steps:
step 1, dissolving 30mg of platinum tetrammine nitrate in 3.75mL of water to prepare a solution A;
step2, dissolving 38.4mg of potassium tellurite in 3.75mL of water to prepare a solution B;
step 3, dissolving 106.8 mgL-ascorbic acid in 3.75mL of water to prepare a solution C;
step 4, 300mg of polyvinylpyrrolidone is dissolved in 3.75mL of water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding 15mL of ethylene glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed, so as to obtain a clear solution F;
and 7, transferring the solution F into a reaction kettle, heating for 5 hours at 200 ℃, and cooling to room temperature to obtain a solution G.
And 8, centrifugally collecting the solution G, washing the precipitate with mixed solution with the volume ratio of ethanol to acetone of 1:9 for three times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
As shown in fig. 1: the transmission electron microscope image of the Pt-Te nanoenzyme prepared in example 1 shows that the nanoenzyme is about 100 to 150nm in size. As shown in fig. 2: the elemental analysis map of the Pt-Te nanoenzyme prepared in example 1 shows that Pt, te is uniformly distributed in the material.
Example 2
The embodiment provides a preparation method of a light-responsive multifunctional nano-enzyme, which comprises the following steps:
step 1, dissolving tetrammine platinum nitrate in water to prepare a solution A;
Step2, dissolving potassium tellurite in water to prepare a solution B;
step 3, dissolving L-ascorbic acid in water to prepare a solution C;
Step 4, dissolving polyvinylpyrrolidone in water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E until the solution system is uniformly dispersed to obtain a clear solution F;
Step 7, transferring the solution F into a reaction kettle, heating at high temperature, and cooling to room temperature to obtain a solution G;
And 8, centrifugally collecting the solution G, washing the precipitate for several times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
Wherein in the step 1, the concentration of the tetramine platinum nitrate in the solution A is 8mg/mL. In the step 2, the concentration of potassium tellurite is 5mg/mL. In the step 3, the concentration of the L-ascorbic acid is 28mg/mL. In the step 4, the molecular weight of polyvinylpyrrolidone is 58000, the concentration of polyvinylpyrrolidone is 60mg/mL, and in the step 5, the solution A, the solution B, the solution C and the solution D are mixed according to the volume ratio of 1:1:1:1, and the addition amount of ethylene glycol is the total volume of the solution A, the solution B, the solution C and the solution D. In step 6, suspension E was sonicated for 1 hour at room temperature. In the step 7, the heating temperature of the solution F is 220 ℃ and the heating time is 4 hours. In the step 8, the washing precipitate is prepared by using an ethanol/acetone mixed solution, wherein the preparation mode of the ethanol/acetone mixed solution is in accordance with the volume ratio of ethanol/acetone being 1:9.
The light-responsive multifunctional nano-enzyme obtained in this example was used as oxidase, peroxidase, catalase and glutathione peroxidase.
The embodiment also provides application of the light-responsive multifunctional nano-enzyme in preparing tumor treatment medicines. In the application, active oxygen is mainly generated by utilizing the light-responsive multifunctional nano-enzyme to treat tumors.
Example 3
The embodiment provides a preparation method of a light-responsive multifunctional nano-enzyme, which comprises the following steps:
step 1, dissolving tetrammine platinum nitrate in water to prepare a solution A;
Step2, dissolving potassium tellurite in water to prepare a solution B;
step 3, dissolving L-ascorbic acid in water to prepare a solution C;
Step 4, dissolving polyvinylpyrrolidone in water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E until the solution system is uniformly dispersed to obtain a clear solution F;
Step 7, transferring the solution F into a reaction kettle, heating at high temperature, and cooling to room temperature to obtain a solution G;
And 8, centrifugally collecting the solution G, washing the precipitate for several times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
Wherein in the step 1, the concentration of the tetramine platinum nitrate in the solution A is 10mg/mL. In the step 2, the concentration of potassium tellurite is 10mg/mL. In the step 3, the concentration of the L-ascorbic acid is 30mg/mL. In the step 4, the molecular weight of polyvinylpyrrolidone is 58000, the concentration of polyvinylpyrrolidone is 100mg/mL, and in the step 5, the solution A, the solution B, the solution C and the solution D are mixed according to the volume ratio of 1:1:1:1, and the addition amount of ethylene glycol is the total volume of the solution A, the solution B, the solution C and the solution D. In step 6, suspension E was sonicated for 2 hours at room temperature. In the step 7, the heating temperature of the solution F is 180 ℃ and the heating time is 4 hours. In the step 8, the washing precipitate is prepared by using an ethanol/acetone mixed solution, wherein the preparation mode of the ethanol/acetone mixed solution is in accordance with the volume ratio of ethanol/acetone being 1:9.
The light-responsive multifunctional nano-enzyme obtained in this example was used as oxidase, peroxidase, catalase and glutathione peroxidase.
The embodiment also provides application of the light-responsive multifunctional nano-enzyme in preparing tumor treatment medicines. In the application, active oxygen is mainly generated by utilizing the light-responsive multifunctional nano-enzyme to treat tumors.
Example 4
The embodiment provides a preparation method of a light-responsive multifunctional nano-enzyme, which comprises the following steps:
step 1, dissolving tetrammine platinum nitrate in water to prepare a solution A;
Step2, dissolving potassium tellurite in water to prepare a solution B;
step 3, dissolving L-ascorbic acid in water to prepare a solution C;
Step 4, dissolving polyvinylpyrrolidone in water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E until the solution system is uniformly dispersed to obtain a clear solution F;
Step 7, transferring the solution F into a reaction kettle, heating at high temperature, and cooling to room temperature to obtain a solution G;
And 8, centrifugally collecting the solution G, washing the precipitate for several times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
Wherein in the step 1, the concentration of the tetramine platinum nitrate in the solution A is 9mg/mL. In the step 2, the concentration of potassium tellurite is 8mg/mL. In the step 3, the concentration of the L-ascorbic acid is 29mg/mL. In the step 4, the molecular weight of polyvinylpyrrolidone is 58000, and the concentration of polyvinylpyrrolidone is 80mg/mL. In the step 5, the solution A, the solution B, the solution C and the solution D are mixed according to the volume ratio of 1:1:1:1, and the addition amount of the ethylene glycol is the total volume of the solution A, the solution B, the solution C and the solution D. In step 6, suspension E was sonicated for 1.5 hours at room temperature. In the step 7, the heating temperature of the solution F is 200 ℃ and the heating time is 5 hours. In the step 8, the washing precipitate is prepared by using an ethanol/acetone mixed solution, wherein the preparation mode of the ethanol/acetone mixed solution is in accordance with the volume ratio of ethanol/acetone being 1:9.
The light-responsive multifunctional nano-enzyme obtained in this example was used as oxidase, peroxidase, catalase and glutathione peroxidase.
The embodiment also provides application of the light-responsive multifunctional nano-enzyme in preparing tumor treatment medicines. In the application, active oxygen is mainly generated by utilizing the light-responsive multifunctional nano-enzyme to treat tumors.
Comparative example 1:
the comparative example provides a preparation method of nano-enzyme, which comprises the following steps:
step 1, dissolving 30mg of platinum tetrammine nitrate in 3.75mL of water to prepare a solution A;
step2, dissolving 19.2mg of potassium tellurite in 3.75mL of water to prepare a solution B;
step 3, dissolving 106.8 mgL-ascorbic acid in 3.75mL of water to prepare a solution C;
step 4, 300mg of polyvinylpyrrolidone is dissolved in 3.75mL of water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding 15mL of ethylene glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed, so as to obtain a clear solution F;
and 7, transferring the solution F into a reaction kettle, heating for 5 hours at 200 ℃, and cooling to room temperature to obtain a solution G.
And 8, centrifugally collecting the solution G, washing the precipitate with mixed solution with the volume ratio of ethanol to acetone of 1:9 for three times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
Comparative example 2:
the comparative example provides a preparation method of nano-enzyme, which comprises the following steps:
step 1, dissolving 30mg of platinum tetrammine nitrate in 3.75mL of water to prepare a solution A;
step2, 28.8mg of potassium tellurite is dissolved in 3.75mL of water to prepare a solution B;
step 3, dissolving 106.8 mgL-ascorbic acid in 3.75mL of water to prepare a solution C;
step 4, 300mg of polyvinylpyrrolidone is dissolved in 3.75mL of water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding 15mL of ethylene glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E at room temperature for 1.5 hours until the solution system is uniformly dispersed, so as to obtain a clear solution F;
and 7, transferring the solution F into a reaction kettle, heating for 5 hours at 200 ℃, and cooling to room temperature to obtain a solution G.
And 8, centrifugally collecting the solution G, washing the precipitate with mixed solution with the volume ratio of ethanol to acetone of 1:9 for three times to obtain the light-responsive multifunctional nano-enzyme, namely the Pt-Te nano-enzyme.
The nano-enzymes prepared in example 1 and comparative examples 1 and 2 were subjected to peroxidase activity detection, and the specific steps are as follows: the nanoezymes prepared in example 1 and comparative examples 1 and 2 were first formulated into a suspension of 80. Mu.g/mL. Then, 80. Mu.L of each of the above-mentioned suspensions of the mimic enzymes was added to a 96-well plate, and 40. Mu.L of 10mM H 2O2 and 40. Mu.L of 4mM 3,3', 5' -Tetramethylbenzidine (TMB) were added thereto, and the optical density value (OD 652) at 652nm was measured. As can be seen from FIG. 3, the OD 652 of the nanoenzyme obtained in example 1 was significantly increased, while the OD 652 of the comparative examples 1 and 2 was smaller, which indicates that the amount of potassium tellurite added as a raw material would affect the peroxidase-like activity of the Pt-Te nanoenzyme.
In order to verify the beneficial effects of the invention, the following test experiments are specially made.
Test experiment 1: peroxidase-like kinetic calculation of Pt-Te nanoenzyme
To calculate the peroxidase-like kinetic related parameters of the Pt-Te nanoenzyme prepared in example 1, the specific steps were as follows: the nanoenzyme prepared in example 1 was first prepared as a dispersion of 80. Mu.g/mL. Then, 80. Mu.L of the above nano-enzyme dispersion was added to a 96-well plate, and 40. Mu.L of 4mM TMB was added thereto, followed by adding 40. Mu.L of 6mM, 8mM, 10mM, 20mM, 40mM, 60mM, 80mM H 2O2, respectively, and after reacting at room temperature for 5 minutes, the optical density value (OD 652) at 652nm was measured. As a result, as shown in FIG. 4, the rate of the enzymatic reaction gradually increased with an increase in the concentration of H 2O2, and the Michaelis constant K m of the nano-enzyme was 6.319mM and the maximum reaction rate was 6.1965X 10 -7 M/s as calculated by curve fitting.
Test experiment 2: photodynamic performance test of Pt-Te nanoenzyme
To test the photodynamic properties of the Pt-Te nanoenzymes prepared in example 1, the specific procedure was as follows: the Pt-Te nanoenzyme prepared in example 1 was first formulated into a dispersion of 40. Mu.g/mL. Then taking 2mL of a sample to be detected, adding 60 mu L of 1mg/mL of 1, 3-diphenyl isobenzofuran (DPBF) into the sample, respectively irradiating a 1064nm laser with the power density of 1.0W/cm 2 for 0min, 1min, 2min, 3min, 4min and 5min, and measuring the ultraviolet absorption spectrum of the reaction system under different laser irradiation time. As shown in FIG. 5, the characteristic absorption value of DPBF significantly decreases with the increase of the laser irradiation time, so that the crystal form Pt-Te nano-enzyme has a certain photodynamic property.
Test experiment 3: photothermal performance test of Pt-Te nanoenzyme
To test the photo-thermal properties of the Pt-Te nanoenzymes prepared in example 1, the specific procedure was as follows: the Pt-Te nanoenzymes prepared in example 1 were first formulated into dispersions of 25. Mu.g/mL, 50. Mu.g/mL, 100. Mu.g/mL, and 200. Mu.g/mL, respectively. Then sequentially added to a 96-well plate, each was irradiated with a 1064nm laser having a power density of 1.0W/cm 2 for 10 minutes while recording the corresponding temperature change with a FOTRIC photothermal camera. The results are shown in FIG. 6, in which the temperature change of the Pt-Te nano-enzyme is enlarged with the increase of the concentration thereof by using the aqueous solution as a control, which indicates that the photo-thermal performance thereof is gradually enhanced with the increase of the concentration thereof.
Test experiment 4: photo-thermal stability test of Pt-Te nanoenzyme
To test the photo-thermal stability of the Pt-Te nanoenzyme prepared in example 1, the specific procedure was as follows: the Pt-Te nanoenzymes prepared in example 1 were first formulated into a dispersion of 200. Mu.g/mL, irradiated with a 1064nm laser having a power density of 1.0W/cm 2 for 12.5 minutes, while recording the corresponding temperature changes with a FOTRIC photothermal camera. The laser was then turned off and again turned on for 12.5 minutes after the temperature had fallen to the initial temperature, and the cycle was repeated 4 times. As shown in FIG. 7, the temperature of the Pt-Te nano-enzyme was increased almost equally in 4 cycles of the switching laser, which indicates that the Pt-Te nano-enzyme has good photo-thermal stability.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments, and those skilled in the art, based on the present disclosure, should make improvements and modifications without departing from the scope of the present invention.
Claims (10)
1. The preparation method of the light-responsive multifunctional nano-enzyme is characterized by comprising the following steps of:
step 1, dissolving tetrammine platinum nitrate in water to prepare a solution A;
Step2, dissolving potassium tellurite in water to prepare a solution B;
step 3, dissolving L-ascorbic acid in water to prepare a solution C;
Step 4, dissolving polyvinylpyrrolidone in water to prepare a solution D;
step 5, uniformly mixing the solution A, the solution B, the solution C and the solution D, and adding glycol into the mixture to prepare a suspension E;
Step 6, carrying out ultrasonic treatment on the suspension E until the solution system is uniformly dispersed to obtain a clear solution F;
Step 7, transferring the solution F into a reaction kettle, heating at high temperature, and cooling to room temperature to obtain a solution G;
And 8, centrifugally collecting the solution G, and washing the precipitate for several times to obtain the light-responsive multifunctional nano-enzyme.
2. The method for preparing the light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 1, the concentration of the tetramine platinum nitrate in the solution A is 8-10 mg/mL.
3. The method for preparing the light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 2, the concentration of potassium tellurite is 5-10 mg/mL.
4. The method for preparing a light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 3, the concentration of L-ascorbic acid is 28-30 mg/mL.
5. The method for preparing a light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 4, the molecular weight of polyvinylpyrrolidone is 58000, and the concentration of polyvinylpyrrolidone is 60-100 mg/mL, for example, 80mg/mL.
6. The method for preparing the light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 5, the solution A, the solution B, the solution C and the solution D are mixed according to a volume ratio of 1:1:1:1, and the addition amount of the ethylene glycol is the total volume of the solution A, the solution B, the solution C and the solution D.
7. The method for preparing the light-responsive multifunctional nano-enzyme according to claim 1, wherein in the step 6, the suspension E is subjected to ultrasonic treatment at room temperature for 1-2 hours;
in the step 7, the heating temperature of the solution F is 180-220 ℃ and the heating time is 4-6 hours;
In the step 8, the washing precipitate is prepared by using an ethanol/acetone mixed solution, wherein the preparation mode of the ethanol/acetone mixed solution is in accordance with the volume ratio of ethanol/acetone being 1:9.
8. A light-responsive multifunctional nanoenzyme obtained based on the preparation method of any one of claims 1 to 7.
9. The use of the light-responsive multifunctional nanoenzyme of claim 8 as an oxidase, peroxidase, catalase and glutathione peroxidase.
10. The use of the light-responsive multifunctional nano-enzyme according to claim 8 for preparing a tumor therapeutic drug.
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