CN113679838A - Vanadium nano enzyme and preparation method and application thereof - Google Patents
Vanadium nano enzyme and preparation method and application thereof Download PDFInfo
- Publication number
- CN113679838A CN113679838A CN202110942827.4A CN202110942827A CN113679838A CN 113679838 A CN113679838 A CN 113679838A CN 202110942827 A CN202110942827 A CN 202110942827A CN 113679838 A CN113679838 A CN 113679838A
- Authority
- CN
- China
- Prior art keywords
- vanadium
- nanoenzyme
- preparation
- ultrasonic treatment
- nano material
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nanotechnology (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Crystallography & Structural Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- General Physics & Mathematics (AREA)
- Manufacturing & Machinery (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a vanadium nano enzyme and a preparation method and application thereof, wherein the preparation method comprises the following steps: dispersing vanadium powder in an organic solvent, performing circulating ultrasonic treatment and centrifugation under an ice bath condition, and collecting precipitate to obtain a vanadium nano material; preparing a vanadium nano material into a dispersion liquid, adding amphiphilic molecules into the dispersion liquid under the ultrasonic condition, continuing ultrasonic treatment, stirring for reaction, centrifuging, and collecting precipitates to obtain the vanadium nano enzyme. The preparation method has the advantages that the source of the raw materials for preparing the vanadium nanoenzyme is wide, the preparation method is simple, and the size is controllable; the vanadium nano enzyme has the advantages of good biocompatibility and biodegradability, and simultaneously has better photothermal effect and catalytic activity, and experiments prove that the vanadium nano enzyme provided by the invention can obviously kill and eliminate tumor cells by cooperating with the photothermal effect and the catalytic activity, so that the effect of treating tumor diseases is achieved.
Description
Technical Field
The invention belongs to the technical field of biological medicines. More particularly, relates to a vanadium nano enzyme, a preparation method and an application thereof.
Background
Photothermal therapy is a therapeutic method in which a material having a high photothermal conversion efficiency is injected into the inside of a human body, is concentrated near tumor tissue by using a targeting recognition technology, and converts light energy into heat energy under the irradiation of an external light source (generally, near infrared light) to kill cancer cells. The commonly used photo-thermal materials comprise precious metals such as gold Au, silver Ag, platinum Pt and the like, and the photo-thermal conversion efficiency is higher. For example, chinese patent application CN101711872A discloses a nano gold nano material, which selects gold as a research content, has a quasi-spherical shell shape, has a hollow structure inside, can not only increase the temperature in a local range after near-infrared laser irradiation, but also be used together with an antitumor drug to kill tumor cells, and can also be applied to an infrared tomography technology as a developer. However, the main material adopted by the material is gold, and the raw material is expensive and has high cost; on the other hand, the anti-tumor effect achieved by using the photo-thermal material gold alone is limited, the tumor cells cannot be completely eliminated, and the tumor cells can be well killed by combining an anti-tumor drug. Therefore, it is urgently needed to provide a material having significant killing and removing effects on tumor cells.
Disclosure of Invention
The invention aims to solve the technical problems of high cost and limited tumor removal effect of the existing anti-tumor photothermal material, and provides the vanadium nanoenzyme which has low cost, catalytic action and can significantly remove tumor cells.
The invention aims to provide a preparation method of vanadium nanoenzyme.
The invention also aims to provide the application of the vanadium nanoenzyme in preparing the medicines for preventing and treating tumors.
The invention also aims to provide the application of the vanadium nanoenzyme as a catalyst in catalytic oxidation-reduction reaction.
The above purpose of the invention is realized by the following technical scheme:
the invention provides a preparation method of vanadium nanoenzyme, which comprises the following steps:
s1, dispersing vanadium powder in an organic solvent, performing circulating ultrasonic treatment and centrifugation under an ice bath condition, and collecting precipitates to obtain a vanadium nano material;
s2, preparing the vanadium nano material into dispersion, adding amphiphilic molecules into the dispersion under the ultrasonic condition, continuing ultrasonic treatment, stirring for reaction, centrifuging, and collecting precipitates to obtain the vanadium nano enzyme.
Further, in step S2, the amphiphilic molecule is phospholipid-polyethylene glycol, polyvinylpyrrolidone or hydroxyethyl starch.
Further, in step S1, the organic solvent is N-methylpyrrolidone, dimethylformamide or dimethylsulfoxide.
Further, in step S1, the cyclic ultrasonic treatment is performed at a power of 100-1000W with an on/off cycle: 2-8 s/2-8 s are circulated, and the ultrasonic treatment is carried out for 12-72 h. Preferably, the conditions of the cyclic sonication are, at a power of 700W, in on/off cycles: 5s/5s is circulation, and ultrasonic treatment is carried out for 48 hours; the centrifugation is performed for the first time at 3000-8000 rpm/min, supernatant is collected, and then the second centrifugation is performed at 10000-50000 rpm/min; preferably, the first centrifugation time is 5-20 min, and the second centrifugation time is 15-30 min.
Further, in step S1, the mass-to-volume ratio of the vanadium powder to the organic solvent is (0.1 to 10) g: (50-200) mL. Preferably, the mass volume ratio of the vanadium powder to the organic solvent is (0.1-5) g: (50-100) mL; more preferably, the mass volume ratio of the vanadium powder to the organic solvent is 1 g: 100 mL.
Further, in step S2, the mass ratio of the vanadium nanomaterial to the amphiphilic molecule is 1: (0.5 to 4). Preferably, the mass ratio of the vanadium nanomaterial to the amphiphilic molecule is 1: (0.5 to 2); more preferably, the mass ratio of the vanadium nanomaterial to the amphiphilic molecule is 1: 2; the centrifugal rotating speed is 10000-50000 rpm/min.
Preferably, in step S2, the concentration of the vanadium nanomaterial in the vanadium nanomaterial dispersion liquid is 0.5-4 mg/mL, and the solvent is ethanol; the step of adding the amphiphilic molecules into the dispersion liquid is to add an amphiphilic molecule solution into the dispersion liquid, wherein the concentration of the amphiphilic molecules in the amphiphilic molecule solution is 0.5-4 mg/mL, and the solvent is an acetone-ethanol mixed solution (acetone: ethanol is 2: 3).
Preferably, in step S2, the stirring reaction is performed by stirring with a magnetic stirrer at a rotation speed of 600 to 1500rpm overnight.
Preferably, step S2 further includes washing the collected precipitate, wherein the solvent for washing is ethanol.
Further, in step S2, the step of adding the amphipathic molecule to the dispersion under the ultrasonic condition and continuing the ultrasonic treatment includes adding the amphipathic molecule to the dispersion under the ice bath and the power of 100-500W and continuing the ultrasonic treatment for a period of time.
Furthermore, the invention also provides the vanadium nanoenzyme prepared by the preparation method. Vanadium in the vanadium nano enzyme is one or more of pentavalent, tetravalent and trivalent; the average particle size of the obtained vanadium nanoenzyme is 50-800 nm.
In addition, the invention also provides application of the vanadium nanoenzyme in preparing a medicament for preventing and treating tumors.
Preferably, the medicine can be combined with a photothermal therapy method to treat tumors, and can achieve better tumor removal and treatment effects.
In addition, the invention also provides application of the vanadium nanoenzyme as a catalyst in catalytic oxidation-reduction reaction. Research proves that the vanadium nanoenzyme has the activity of one or more of peroxidase-like enzyme, horseradish-like peroxidase, oxidase-like enzyme, superoxide dismutase-like enzyme, glutathione oxidase-like enzyme and catalase-like enzyme as a catalyst, and is an excellent natural enzyme substitute.
The vanadium nano enzyme has peroxidase-like activity and canCatalyzing hydrogen peroxide (H) in the tumor microenvironment2O2) Generating a large amount of hydroxyl radicals (. OH) to kill tumor cells; has glutathione oxidase-like activity, and can catalyze reduced Glutathione (GSH) to change into oxidized glutathione; has catalase-like activity, and can catalyze H in tumor microenvironment2O2Generating oxygen; the vanadium nanoenzyme with various enzyme activities can form a nano platform for self-cascade regulation of tumor microenvironment, generate a large amount of OH and exhaust GSH in cells, thereby achieving the effect of promoting tumor cell apoptosis. Meanwhile, the vanadium nanoenzyme has a photothermal effect, can further cauterize tumors by combining photothermal therapy, enhances the activity of the nanoenzyme, and realizes an excellent tumor treatment effect under the synergistic effect.
The invention has the following beneficial effects:
the vanadium nanoenzyme provided by the invention has the advantages that the source of the required raw materials is wide, the preparation method is simple, and the size is controllable; the vanadium nano enzyme has the advantages of good biocompatibility and biodegradability, and simultaneously has better photothermal effect and catalytic activity, and experiments prove that the vanadium nano enzyme provided by the invention can obviously kill and eliminate tumor cells by cooperating with the photothermal effect and the catalytic activity, so that the effect of treating tumor diseases is achieved.
Drawings
FIG. 1 is an electron microscope scanning image of the vanadium nanoenzyme obtained in example 1 of the present invention.
FIG. 2 is an element distribution analysis mapping map of the vanadium nanoenzyme obtained in example 1 of the present invention.
FIG. 3 is the X-ray photoelectron spectrum of the vanadium nanoenzyme and vanadium powder obtained in example 1 of the present invention.
FIG. 4 is an X-ray diffraction pattern of vanadium nanoenzyme and vanadium powder obtained in example 1 of the present invention.
FIG. 5 is a UV-Vis spectrum of the vanadium nanoenzyme obtained in example 1 of the present invention.
FIG. 6 is a statistical chart of the photo-thermal performance study of the vanadium nanoenzyme obtained in example 1 of the present invention.
FIG. 7 is a statistical chart of the enzyme activity study of vanadium nanoenzymes obtained in example 1 of the present invention; wherein, A-type catalase, B-type peroxidase and C-type glutathione oxidase.
FIG. 8 is a statistical chart of the tumor cytotoxicity test study of vanadium nanoenzymes obtained in example 1 of the present invention; a-single enzyme catalysis therapy and B-enzyme catalysis-photothermal synergistic therapy.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1
The embodiment is a preparation method of vanadium nanoenzyme, which comprises the following steps:
s1, weighing 1g of vanadium powder, dispersing the vanadium powder in 100mL of N-methylpyrrolidone (NMP), and carrying out on-off cycling on the mixture under the ice bath condition of 700W: performing probe ultrasonic treatment for 48h in a circulation mode of 5s/5s, centrifuging for 10min at 5000rpm/min by using a low-temperature ultracentrifuge, removing non-stripped vanadium crystals, collecting supernatant, centrifuging for 20min at 20000rpm/min, and collecting precipitate to obtain a vanadium nano material;
s2, preparing the vanadium nano material obtained in the step S1 and ethanol into 1mg/mL vanadium nano material dispersion liquid, under the condition of ice water bath 300W ultrasound, dropwise adding 1mL distearoyl phosphatidyl ethanolamine polyethylene glycol (DSPE-PEG) solution (2mg/mL, the solvent is acetone: ethanol ═ 2:3) into 1mL vanadium nano material dispersion liquid, continuing ultrasound for 30min, magnetically stirring at 600-1500 rpm for overnight reaction, centrifuging at 20000rpm for 20min after the reaction is finished, and washing with ethanol twice to obtain the vanadium nano material dispersion liquid.
Example 2
The embodiment is a preparation method of vanadium nanoenzyme, which comprises the following steps:
s1, weighing 1g of vanadium powder, dispersing the vanadium powder in 100mL of dimethylformamide, and carrying out on-off cycle under ice bath conditions of 1000W: performing probe ultrasonic treatment for 12h in 8s/8s cycle, centrifuging for 5min at 8000rpm/min of a low-temperature ultracentrifuge, removing non-stripped vanadium crystals, collecting supernatant, centrifuging for 15min at 50000rpm/min, and collecting precipitate to obtain vanadium nano material;
s2, preparing the vanadium nano material obtained in the step S1 and ethanol into 1mg/mL vanadium nano material dispersion liquid, under the condition of ice water bath 300W ultrasound, dropwise adding 1mL distearoyl phosphatidyl ethanolamine polyethylene glycol (DSPE-PEG) solution (2mg/mL, the solvent is acetone: ethanol ═ 2:3) into 1mL vanadium nano material dispersion liquid, continuing ultrasound for 30min, magnetically stirring at 600-1500 rpm for overnight reaction, centrifuging at 20000rpm for 20min after the reaction is finished, and washing with ethanol twice to obtain the vanadium nano material dispersion liquid.
Example 3
The embodiment is a preparation method of vanadium nanoenzyme, which comprises the following steps:
s1, weighing 1g of vanadium powder, dispersing the vanadium powder in 100mL of dimethyl sulfoxide, and carrying out on-off cycle under ice bath conditions of 300W: performing probe ultrasonic treatment for 72h in a circulation mode of 2s/2s, centrifuging for 20min by using a low-temperature ultracentrifuge at 3000rpm/min, removing vanadium crystals which are not stripped, collecting supernate, centrifuging for 20min at 25000rpm/min, and collecting precipitates to obtain a vanadium nano material;
s2, preparing the vanadium nano material obtained in the step S1 and ethanol into 1mg/mL vanadium nano material dispersion liquid, under the condition of ice water bath 300W ultrasound, dropwise adding 1mL distearoyl phosphatidyl ethanolamine polyethylene glycol (DSPE-PEG) solution (2mg/mL, the solvent is acetone: ethanol ═ 2:3) into 1mL vanadium nano material dispersion liquid, continuing ultrasound for 30min, magnetically stirring at 600-1500 rpm for overnight reaction, centrifuging at 20000rpm for 20min after the reaction is finished, and washing with ethanol twice to obtain the vanadium nano material dispersion liquid.
The vanadium nanoenzyme prepared in example 1 is used as an example for structural characteristic and performance test, and other examples can achieve similar effects.
Test example 1 determination of physicochemical Properties of vanadium Nanolase
The vanadium nanoenzyme obtained in example 1 was analyzed by scanning electron microscopy, and the results are shown in FIG. 1. As can be seen from the figure, the vanadium nanoenzyme modified by the polyethylene glycol obtained in the example 1 has good dispersibility, and the particle size is 180-220 nm.
The vanadium nanoenzyme obtained in example 1 was subjected to element distribution analysis and mapping, and the result is shown in FIG. 2. As can be seen from the figure, V, C, N, O elements exist in the polyethylene glycol modified vanadium nanoenzyme, which indicates that the vanadium nanoenzyme of the invention is successfully prepared.
The vanadium nanoenzyme and vanadium powder obtained in example 1 were subjected to X-ray photoelectron spectroscopy and X-ray diffraction spectroscopy, respectively, and the results are shown in fig. 3 to 4. As can be seen from the figure, V, C, N, O elements exist in both the polyethylene glycol modified vanadium nanoenzyme and the vanadium powder, which indicates that the vanadium nanoenzyme is successfully prepared and does not dope byproducts; the characteristic absorption peak of the vanadium nanoenzyme is completely consistent with the characteristic peak of the vanadium powder, which indicates that the vanadium nanoenzyme is successfully prepared and the crystal structure is not changed.
Experimental example 2 determination of optical Properties of vanadium nanoenzyme
The vanadium nanoenzyme obtained in example 1 was evaluated for optical absorption and the results are shown in FIG. 5. As can be seen from the figure, the polyethylene glycol modified vanadium nanoenzyme has strong optical absorption in ultraviolet, visible light and near infrared regions, and shows concentration dependence.
The photo-thermal performance of the vanadium nanoenzyme modified by polyethylene glycol was evaluated under the condition of 808nm wavelength laser, and the results are shown in fig. 6.
As can be seen from FIG. 6A, the vanadium nanoenzyme modified by polyethylene glycol has higher temperature rise under the excitation of 808nm laser, and has excellent photo-thermal performance; as can be seen from FIG. 6B, the vanadium nanoenzyme modified by polyethylene glycol has good photo-thermal stability. Therefore, the vanadium nanoenzyme modified by the polyethylene glycol has the temperature required by photo-thermal treatment of cauterization tumors.
Experimental example 3 measurement of catalytic Properties of vanadium nanoenzyme
The catalase-like activity of the vanadium nanoenzyme obtained in example 1 was tested by the following method: vanadium nanoenzymes (0, 12.5, 25, 50. mu.g/mL) were dispersed in phosphate buffer (pH7.4) at various concentrations and H was added once with a syringe2O2(10mM), the oxygen concentration of the solution was measured at various time points using a JPSJ-605F portable dissolved oxygen meter, and the results are shown in FIG. 7A.
The peroxidase-like activity of the vanadium nanoenzyme obtained in example 1 was tested by the following method: vanadium nanoenzyme (0, 12.5, 25) with different concentrations50, 100. mu.g/mL) in phosphate buffer (pH7.4), followed by the addition of 10. mu.g/mL methylene blue indicator and finally H2O2(10mM), and the UV absorption of the solution was measured by UV-visible spectrometer at different time points, and the results are shown in FIG. 7B.
The activity of glutathione-like oxidase of the vanadium nanoenzyme obtained in example 1 is tested by the following method: vanadium nanoenzymes (0, 12.5, 25, 50, 100. mu.g/mL) were dispersed in phosphate buffer (pH7.4) at different concentrations, then 0.1mM GSH was added, and finally the consumption of GSH was detected using 5,5' -dithiobis (2-nitrobenzoic acid) probe, as shown in FIG. 7C.
As can be seen from FIG. 7A, the polyethylene glycol-modified vanadium nanoenzyme has strong oxygen generation capacity in a hydrogen peroxide environment, has concentration dependence and shows strong catalase-like activity; as can be seen from FIG. 7B, the polyethylene glycol-modified vanadium nanoenzyme has significant OH generating ability, has concentration dependence and shows stronger peroxidase-like activity; as can be seen from FIG. 7C, the polyethylene glycol-modified vanadium nanoenzyme has an obvious GSH oxidation ability, can reduce the tumor oxidation resistance, and shows a strong glutathione-like oxidase activity. In conclusion, the polyethylene glycol modified vanadium nanoenzyme has the capabilities of catalase-like enzyme, peroxidase-like enzyme and glutathione-like oxidase, and has the capabilities of regulating and controlling a tumor microenvironment to kill tumors and prevent tumor recurrence and metastasis.
Experimental example 4 Activity measurement of vanadium nanoenzyme on tumor cells
The toxicity of the vanadium nanoenzyme obtained in the example 1, which is caused by enzyme catalysis on human breast cancer cells MCF-7, is tested by the following test method: MCF-7 cells were plated at 8X 10 per well3The density of individual cells was seeded in 96-well plates and cultured for 12 hours. After washing once with PBS, cells were incubated with a gradient concentration (0, 3.13, 6.25, 12.5, 25, 50. mu.g/mL) of vanadium nanoenzyme for 12 or 24 hours, respectively. After washing with PBS, Cell Counting Kit-8(CCK-8) was incubated for 2 hours, absorbance was measured at 450nm wavelength, and standard Cell viability assays were performed to determine relative Cell viability, the results of which are shown in FIG. 8A.
Example 1 resultsThe vanadium nanoenzyme is used for testing toxicity induced by enzyme catalysis-photothermal synergy of human breast cancer cells MCF-7, and the testing method comprises the following steps: MCF-7 cells were plated at 8X 10 per well3The density of individual cells was seeded in 96-well plates and cultured for 12 hours. After washing once with PBS, the cells were incubated with a gradient concentration (0, 3.13, 6.25, 12.5, 25, 50. mu.g/mL) of vanadium nanoenzyme for 6 hours, followed by 808nm laser irradiation (1W/cm)25 min). The incubation was continued for 18 hours, washed with PBS, and then incubated for 2 hours using Cell Counting Kit-8(CCK-8), absorbance was measured at a wavelength of 450nm, and a standard Cell viability assay was performed to determine relative Cell viability, the results of which are shown in FIG. 8B.
As can be seen from fig. 8A, under the single enzyme catalysis treatment, the vanadium nanoenzyme modified by polyethylene glycol shows a certain toxicity to tumor cells and shows concentration dependence; as can be seen from FIG. 8B, under the photo-thermal and enzyme catalysis synergistic treatment, the toxicity of the polyethylene glycol-modified vanadium nanoenzyme to tumor cells is greatly enhanced, and a significant tumor killing effect is achieved.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. The preparation method of the vanadium nanoenzyme is characterized by comprising the following steps of:
s1, dispersing vanadium powder in an organic solvent, performing circulating ultrasonic treatment and centrifugation under an ice bath condition, and collecting precipitates to obtain a vanadium nano material;
s2, preparing the vanadium nano material into dispersion, adding amphiphilic molecules into the dispersion under the ultrasonic condition, continuing ultrasonic treatment, stirring for reaction, centrifuging, and collecting precipitates to obtain the vanadium nano enzyme.
2. The method of claim 1, wherein in step S2, the amphiphilic molecule is phospholipid-polyethylene glycol, polyvinylpyrrolidone or hydroxyethyl starch.
3. The method according to claim 1, wherein in step S1, the organic solvent is N-methylpyrrolidone, dimethylformamide or dimethylsulfoxide.
4. The method according to claim 1, wherein the cyclic ultrasonic treatment is performed at a power of 100-1000W in an on/off cycle of step S1: 2-8 s/2-8 s are circulation, and ultrasonic treatment is carried out for 12-72 h; the centrifugation is performed by performing first centrifugation at 3000-8000 rpm/min, collecting supernatant, and performing second centrifugation at 10000-50000 rpm/min.
5. The preparation method according to claim 1, wherein in the step S1, the mass-to-volume ratio of the vanadium powder to the organic solvent is (0.1-10) g: (50-200) mL.
6. The preparation method according to claim 1, wherein in the step S2, the mass ratio of the vanadium nano material to the amphiphilic molecule is 1: (0.5 to 4); the centrifugal rotating speed is 10000-50000 rpm/min.
7. A vanadium nanoenzyme prepared by the preparation method of any one of claims 1 to 6.
8. The use of the vanadium nanoenzyme of claim 7 in the preparation of a medicament for the prevention or treatment of tumors.
9. Use of the vanadium nanoenzyme of claim 7 as a catalyst in catalytic oxidation-reduction reactions.
10. The use of claim 9, wherein the vanadium nanoenzyme has as a catalyst one or more of peroxidase-like, horseradish-like peroxidase, oxidase-like, superoxide dismutase-like, glutathione oxidase-like, catalase-like activities.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110942827.4A CN113679838B (en) | 2021-08-17 | 2021-08-17 | Vanadium nano-enzyme and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110942827.4A CN113679838B (en) | 2021-08-17 | 2021-08-17 | Vanadium nano-enzyme and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113679838A true CN113679838A (en) | 2021-11-23 |
| CN113679838B CN113679838B (en) | 2023-09-22 |
Family
ID=78580324
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110942827.4A Active CN113679838B (en) | 2021-08-17 | 2021-08-17 | Vanadium nano-enzyme and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113679838B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114177288A (en) * | 2021-12-14 | 2022-03-15 | 合肥工业大学 | Modified vanadium carbide nanodot for chemical power and photo-thermal synergistic antibacterial and preparation method and application thereof |
| CN114470310A (en) * | 2021-12-20 | 2022-05-13 | 山西医科大学 | Self-adhesive hydrogel based on tetraase activity nanoenzyme, and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001053494A2 (en) * | 2000-01-24 | 2001-07-26 | The Regents Of The University Of California | Cloning and overexpression vanadium bromoperoxidase from marine algae |
| US20070292532A1 (en) * | 2003-12-04 | 2007-12-20 | Rudiger Woscholski | Vanadium Compounds as Inhibitors of Phosphatases |
| CN103692763A (en) * | 2013-12-06 | 2014-04-02 | 深圳先进技术研究院 | Peeling method of two-dimensional layered nano material |
| CN108926712A (en) * | 2017-05-27 | 2018-12-04 | 首都师范大学 | Vfanadium compound is as the application in optical-thermal conversion material |
| CN110479241A (en) * | 2019-08-02 | 2019-11-22 | 电子科技大学 | Improve the active processing method of peroxidase and product of nano enzyme |
| US20210000955A1 (en) * | 2017-12-08 | 2021-01-07 | Nanyang Technological University | Photodynamically active organosilica nanoparticles and medical uses thereof |
-
2021
- 2021-08-17 CN CN202110942827.4A patent/CN113679838B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001053494A2 (en) * | 2000-01-24 | 2001-07-26 | The Regents Of The University Of California | Cloning and overexpression vanadium bromoperoxidase from marine algae |
| US20070292532A1 (en) * | 2003-12-04 | 2007-12-20 | Rudiger Woscholski | Vanadium Compounds as Inhibitors of Phosphatases |
| CN103692763A (en) * | 2013-12-06 | 2014-04-02 | 深圳先进技术研究院 | Peeling method of two-dimensional layered nano material |
| CN108926712A (en) * | 2017-05-27 | 2018-12-04 | 首都师范大学 | Vfanadium compound is as the application in optical-thermal conversion material |
| US20210000955A1 (en) * | 2017-12-08 | 2021-01-07 | Nanyang Technological University | Photodynamically active organosilica nanoparticles and medical uses thereof |
| CN110479241A (en) * | 2019-08-02 | 2019-11-22 | 电子科技大学 | Improve the active processing method of peroxidase and product of nano enzyme |
Non-Patent Citations (5)
| Title |
|---|
| JIANG OUYANG等: "Cryogenic Exfoliation of 2D Stanene Nanosheets for Cancer Theranostics", 《NANO-MICRO LETT.》 * |
| LIANGMIAO ZHANG等: "Synthesis and formation mechanism of VO2(A) nanoplates with intrinsic peroxidase-like activity", 《RSC ADVANCES》 * |
| RAJARSHI SANKAR RAY等: "Vanadium，a versatile biochemical effector in chemical rat mammary carcinogenesis", 《NUTR. CANCER》 * |
| YICHU NIE等: "Novel biodegradable two-dimensional vanadene augmented photoelectro-fenton process for cancer catalytic therapy", 《BIOMATERIALS》 * |
| 董海姣等: "纳米酶及其细胞活性氧调控", 生物化学与生物物理进展 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114177288A (en) * | 2021-12-14 | 2022-03-15 | 合肥工业大学 | Modified vanadium carbide nanodot for chemical power and photo-thermal synergistic antibacterial and preparation method and application thereof |
| CN114470310A (en) * | 2021-12-20 | 2022-05-13 | 山西医科大学 | Self-adhesive hydrogel based on tetraase activity nanoenzyme, and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113679838B (en) | 2023-09-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dan et al. | Ultrasmall theranostic nanozymes to modulate tumor hypoxia for augmenting photodynamic therapy and radiotherapy | |
| CN113679838A (en) | Vanadium nano enzyme and preparation method and application thereof | |
| CN109364245A (en) | A kind of poly-dopamine nanometer diagnosis and treatment agent and preparation method thereof | |
| Lu et al. | Photosynthetic Oxygenation‐Augmented Sonodynamic Nanotherapy of Hypoxic Tumors | |
| CN108836949B (en) | Preparation method of Prussian blue nano-particles wrapped by Ce6 embedded red cell membranes | |
| WO2019184201A1 (en) | Titanium quantum dot-based nano titanium photo-thermal preparation and preparation method therefor | |
| Cao et al. | A multimodal imaging-guided nanoreactor for cooperative combination of tumor starvation and multiple mechanism-enhanced mild temperature phototherapy | |
| CN112807430A (en) | Application of nano enzyme-based material | |
| CN111632040A (en) | Manganese dioxide-coated drug-loaded mesoporous titanium dioxide nanoparticle and preparation method and application thereof | |
| CN111558032A (en) | Protein nano-drug and preparation method and application thereof | |
| CN113694197A (en) | Photothermal/photodynamic synergistic tumor phototherapy reagent and preparation method and application thereof | |
| CN115317607A (en) | Monoatomic iron-doped graphite-phase carbon nitride nanocomposite and preparation method and application thereof | |
| Gong et al. | Phthalocyanine iron nanodots for combined chemodynamic-sonodynamic cancer therapy | |
| Kang et al. | Photothermally Enhanced Dual Enzyme‐mimic Activity of Gold‐Palladium Hybrid Nanozyme for Cancer Therapy | |
| Zhang et al. | DNA‐Templated Ag@ Pd Nanoclusters for NIR‐II Photoacoustic Imaging‐Guided Photothermal‐Augmented Nanocatalytic Therapy | |
| CN113117077A (en) | Platinum-based monatomic nanoenzyme for tumor combined treatment and preparation method thereof | |
| CN117482231A (en) | Porous copper-manganese bimetallic nano material applied to tumor immunotherapy and preparation method thereof | |
| CN109010829B (en) | Self-assembled porphyrin nano material with photothermal and chemotherapy dual functions as well as preparation method and application thereof | |
| CN108771760B (en) | Platinum sulfide protein nanoparticle with near infrared photothermal effect and multi-mode imaging function, and preparation method and application thereof | |
| CN114668841B (en) | Activatable nano photosensitizer and preparation method and application thereof | |
| CN114601925B (en) | Hyaluronic acid and RSL3 co-modified photosensitive nanomaterial, preparation method and application thereof | |
| Loke et al. | ROS-generating alginate-coated gold nanorods as biocompatible nanosonosensitisers for effective sonodynamic therapy of cancer | |
| CN108069458A (en) | A kind of ultra micro nano-level sphere bismuth tungstate crystal grain and preparation method and application | |
| CN113786485A (en) | Bi-based nano composite material and preparation method and application thereof | |
| Wu et al. | Second near-infrared photoactivatable nanomedicines for enhanced photothermal–chemodynamic therapy of cancer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |