CN113952984B - High-catalytic-activity molybdenum-based nanoenzyme and preparation method and application thereof - Google Patents
High-catalytic-activity molybdenum-based nanoenzyme and preparation method and application thereof Download PDFInfo
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- CN113952984B CN113952984B CN202010693159.1A CN202010693159A CN113952984B CN 113952984 B CN113952984 B CN 113952984B CN 202010693159 A CN202010693159 A CN 202010693159A CN 113952984 B CN113952984 B CN 113952984B
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- 229910052750 molybdenum Inorganic materials 0.000 title claims abstract description 37
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- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/16—Catalysts comprising hydrides, coordination complexes or organic compounds containing coordination complexes
- B01J31/22—Organic complexes
- B01J31/2204—Organic complexes the ligands containing oxygen or sulfur as complexing atoms
- B01J31/226—Sulfur, e.g. thiocarbamates
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- 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
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- B01J23/00—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00
- B01J23/16—Catalysts comprising metals or metal oxides or hydroxides, not provided for in group B01J21/00 of arsenic, antimony, bismuth, vanadium, niobium, tantalum, polonium, chromium, molybdenum, tungsten, manganese, technetium or rhenium
- B01J23/24—Chromium, molybdenum or tungsten
- B01J23/28—Molybdenum
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- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/003—Catalysts comprising hydrides, coordination complexes or organic compounds containing enzymes
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- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/02—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides
- B01J31/06—Catalysts comprising hydrides, coordination complexes or organic compounds containing organic compounds or metal hydrides containing polymers
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- B01J31/00—Catalysts comprising hydrides, coordination complexes or organic compounds
- B01J31/16—Catalysts comprising hydrides, coordination complexes or organic compounds containing coordination complexes
- B01J31/1616—Coordination complexes, e.g. organometallic complexes, immobilised on an inorganic support, e.g. ship-in-a-bottle type catalysts
- B01J31/1625—Coordination complexes, e.g. organometallic complexes, immobilised on an inorganic support, e.g. ship-in-a-bottle type catalysts immobilised by covalent linkages, i.e. pendant complexes with optional linking groups
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- B01J35/20—Catalysts, in general, characterised by their form or physical properties characterised by their non-solid state
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- B01J37/00—Processes, in general, for preparing catalysts; Processes, in general, for activation of catalysts
- B01J37/08—Heat treatment
- B01J37/10—Heat treatment in the presence of water, e.g. steam
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
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Abstract
The invention discloses a molybdenum-based nanoenzyme with high catalytic activity, a preparation method and application thereof, belongs to the technical field of nanoenzymes, solves the problem that in the prior art, the catalytic effect of single molybdenum oxide catalase-like enzyme is poor, and also solves the problems that the biotoxicity of copper ions is high, and the excessive dosage of copper ions can cause great side effect on human bodies. The molybdenum-based nano enzyme with high catalytic activity contains molybdenum oxide nano particles and copper ions, wherein the copper ions are anchored on the surfaces of the molybdenum oxide nano particles; the preparation raw materials of the molybdenum-based nanoenzyme with high catalytic activity comprise molybdenum oxide nanoparticles, cysteine, copper ions and a surfactant. The high catalytic activity molybdenum-based nanoenzyme has good catalytic activity and radiotherapy sensitization function, no obvious cytotoxicity and safe use.
Description
Technical Field
The invention relates to the technical field of nano-enzyme, in particular to molybdenum-based nano-enzyme with high catalytic activity and a preparation method and application thereof.
Background
Inorganic nanomaterials, especially transition metal oxides, have special physicochemical properties, and their preparation is simple and inexpensive, and they have adjustable local plasmon resonance phenomena similar to noble metals, leading to intensive research. Molybdenum oxide is a typical transition metal semiconductor inorganic nano material, and has wide application in the fields of color developers, photocatalysis, batteries, gas sensors and the like due to excellent electrochromic, photochromic, catalytic activity and electrode intercalation performance. In recent years, with the development of nanotechnology and nanomedicine, people find that the nano molybdenum oxide has the characteristics of low biotoxicity, easy surface modification, more oxygen holes, strong near-infrared photothermal conversion capability and the like, and the characteristics enable the nano molybdenum oxide to show wide application prospects in the biomedical field such as antibiosis, anti-tumor, biomolecule detection, biocatalysis and the like.
The tumor microenvironment-environmental (TME) -mediated nanocatalysis therapy mainly utilizes nontoxic or low-toxicity nanomaterials and selectively catalyzes and triggers specific chemical reactions in the TME to locally generate a considerable amount of specific reaction products such as Reactive Oxygen Species (ROS) tumor treatment strategies, so that a series of biological and pathological responses are realized, side effects on normal tissues are reduced, and the TME-mediated nanocatalysis therapy is a tumor-specific treatment mode. However, the efficiency of TME-mediated nanocatalysis therapy is generally limited by intratumoral H2O2Insufficient levels, acidity and oxygen deficiency. Therefore, whether the effect of tumor catalytic therapy can be enhanced by modulating TME has become one of the hot spots of the current research. The development of inorganic nano-enzyme (such as Peroxidase (POD), Oxidase (OXDD), Catalase (CAT) and the like) with potential high-efficiency enzyme-like catalytic activity provides a new opportunity for tumor catalytic treatment. For example: CAT-like nanoenzyme can catalyze and decompose highly expressed H in TME2O2(concentration range: 100. mu.M-1.0 mM) is oxygen, thereby alleviating hypoxia in the tumor. However, constructing highly catalytic inorganic nanoenzymes for precise TME response with little toxic side effects on normal tissues to achieve efficient tumor suppression remains a significant challenge.
Copper is a trace metal element in human body, and plays an important role in maintaining the health of cardiovascular system of human body. Copper ions have been shown to have excellent properties in stimulating angiogenesis, the collagen deposition process to promote wound healing; in addition, some new properties of copper compounds or ions are exploited and usedThe field of biomedicine. For example, Cu polyoxometalate catalysts (Cu-MOF) have good biocompatibility and excellent catalytic properties and are useful for drug delivery and tumor therapy (e.g., Cu2+Radiotherapy sensitization, photothermal therapy, etc.). However, due to the relatively high biological toxicity of copper ions, the use of excessive dosage can cause great side effects on human bodies.
Disclosure of Invention
In view of the above analysis, the present invention aims to provide a molybdenum-based nanoenzyme with high catalytic activity, a preparation method and applications thereof, which has low toxic and side effects and high catalytic activity, and can solve at least one of the following technical problems: (1) the catalase-like catalytic effect of the molybdenum oxide alone is poor; (2) the biological toxicity of copper ions is relatively high, and the use of excessive dosage in the anti-tumor process can produce great side effect on human body.
The invention is mainly realized by the following technical scheme:
in one aspect, the invention provides a molybdenum-based nanoenzyme with high catalytic activity, which comprises molybdenum oxide nanoparticles and copper ions, wherein the copper ions are anchored on the surfaces of the molybdenum oxide nanoparticles.
Further, the preparation raw materials of the molybdenum-based nano enzyme with high catalytic activity comprise molybdenum oxide nanoparticles, cysteine, copper ions and a surfactant.
Further, the surfactant is a polymer with good biocompatibility.
On the other hand, the invention provides a preparation method of molybdenum-based nanoenzyme with high catalytic activity, which comprises the following steps:
step S1, preparing molybdenum oxide nano particles;
step S2, adding cysteine and water-soluble copper salt into secondary water, ultrasonically stirring and chelating to synthesize white floccule L-Cys-Cu;
s3, putting the mixed solution of the molybdenum oxide nano-particles and L-Cys-Cu into a centrifugal tube, carrying out ultrasonic treatment in an ultrasonic pool, then putting the ultrasonic pool on a magnetic stirrer for stirring, and uniformly and slowly adding a surfactant in the stirring process;
and step S4, after the reaction is finished, centrifuging and taking out the precipitate, repeatedly centrifuging and washing the precipitate for 2-3 times by using secondary water, washing by using ethanol and centrifuging, and sucking away excessive ethanol to obtain the molybdenum-based nanoenzyme with high catalytic activity.
Further, in step S1, the step of preparing the molybdenum oxide nanoparticles includes:
s101, weighing ammonium molybdate powder according to a proportion, dissolving the ammonium molybdate powder in deionized water, and performing ultrasonic dissolution to obtain an ammonium molybdate aqueous solution;
s102, adding absolute ethyl alcohol into the ammonium molybdate aqueous solution obtained in the S101 by using a liquid transfer gun, performing ultrasonic dispersion, and stirring at room temperature to obtain a uniformly mixed solution;
s103, transferring the mixed solution into a hydrothermal reaction kettle, screwing the reaction kettle, placing the reaction kettle in an oven, setting the reaction temperature at 160-200 ℃ and the reaction time at 10-15 h;
and S104, after the reaction is finished, naturally cooling to room temperature, centrifuging, taking out the precipitate, repeatedly centrifuging and washing the precipitate for 2-5 times by using secondary water, and freeze-drying in a freeze dryer to obtain the molybdenum oxide nanoparticles.
Further, in S101 and S102, the ratio of ammonium molybdate: absolute ethanol: deionized water 0.7-1 mmoL: 10-15 mL: 20-25 mL.
Further, in step S2, the water-soluble copper salt is copper chloride, and the molar ratio of cysteine to copper chloride is 2: 1-1.5.
Further, in the step S3, the molar ratio of the molybdenum oxide nanoparticles to the L-Cys-Cu is 1: 1-3.
Further, in step S3, the surfactant is polyvinylpyrrolidone, and the mass ratio of the molybdenum oxide nanoparticles to the polyvinylpyrrolidone is 1: 1-2.
The invention also provides application of the high catalytic activity molybdenum-based nanoenzyme, wherein the high catalytic activity molybdenum-based nanoenzyme is used as an anti-tumor material.
Compared with the prior art, the invention can realize at least one of the following beneficial effects:
(1) the invention utilizes a simple one-step hydrothermal method to decompose ammonium molybdate and reform the ammonium molybdate into ammonium molybdate under the hydrothermal conditionMoO with porous nucleus growth2The nano-particles have high catalase-like catalytic activity, have degradation behavior in the catalytic process and can avoid the long-term toxicity of the nano-materials to organisms, and the nano-particles are safe and efficient H in TME2O2The responsive nano enzyme catalyst can improve hypoxic in tumors and enhance sensitivity for radiotherapy; in MoO2In the process of anchoring copper ions on the surfaces of the nano particles, only simple and safe ultrasonic stirring at room temperature is performed, and other high-temperature and high-pressure processes are not used for changing MoO2The shape and structure of the nano-particles are realized, and cysteine L-Cys is used as an intermediate linking agent for ion anchoring, so that a simple reference basis for a synthetic thought is provided for anchoring metal ions by other transition metal oxides, and the preparation method is simple and safe.
(2) The molybdenum-based nanoenzyme MoO with high catalytic activity of the invention2The catalase-like catalytic activity and the radiotherapy sensitization function of the-L-Cys-Cu-PVP (MCCP for short) are far higher than those of single molybdenum oxide and single L-Cys-Cu, and H2O2In TME, H in an amount of 100. mu.M-1.0 mM2O2In the high expression dose range of MCCP on the relatively low concentration of H2O2While the catalytic capability of catalase-like enzyme is improved, the dosage and H of copper ions with relatively high biological toxicity are reduced2O2The molybdenum-based nano enzyme with excellent catalase-like catalase-mimic catalytic activity is finally obtained. MCCP catalysis H2O2After oxygen is generated, the L-Cys-Cu is easily disassembled in TME under X-ray irradiation in the radiotherapy process, and the L-Cys-Cu is released from the composite material and starts to carry out Fenton-like reaction, so that more hydroxyl free radicals OH are generated, MCCP improves tumor hypoxia, simultaneously improves tumor sensitivity in radiotherapy, and synergistically enhances catalase-like catalytic decomposition H2O2The oxygen is generated to improve the effects of hypoxia in the tumor and sensitization radiotherapy, thereby having wide application prospect in the aspect of radiotherapy sensitization of the tumor.
(3) The high catalytic activity molybdenum-based nanoenzyme has no obvious cytotoxicity in the range of testing dosage, is safe to use, has obvious effect and is wide in application range.
In the invention, the above technical schemes can be combined with each other to realize more preferable combination schemes. Additional features and advantages of the invention will be set forth in the description which follows, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and drawings.
Drawings
The drawings are only for purposes of illustrating particular embodiments and are not to be construed as limiting the invention, wherein like reference numerals are used to indicate like parts throughout.
In FIG. 1, a is a scanning electron microscope image of the MCCP of the present invention; b is a transmission electron micrograph of an MCCP of the invention; c is an element energy spectrum of MCCP; d is the X-ray powder diffraction pattern of MCCP;
in fig. 2, a is the result of dissolved oxygen test of MP nanoparticles with different concentrations; b is a consumption curve of MCCP to hydrogen peroxide; c is hydrogen peroxide catalytic activity of three materials MP, MCCP, L-Cys-Cu to catalytically decompose H2O2Comparing the effects; d is the influence of temperature on the MCCP catalytic effect;
FIG. 3 is a photograph of a Confocal scanning microscope (CLSM) for detecting Reactive Oxygen Species (ROS) in MP, MCCP at the cell level;
FIG. 4 is a confocal scanning laser micrograph of an in vitro DNA double strand break test;
in fig. 5, a is a graph of GSH consumption of different concentrations of MCCP nanoparticles over the same time; b is control experiment of MP, PBS and MCCP; c is the consumption curve of GSH over time at the same MCCP nanoparticle concentration; d is copper ion and H under X-ray irradiation2O2The fluorescence emission spectrum of the product OH in the Fenton-like reaction of (1);
FIG. 6 is cytotoxicity data after 24-hour incubation of MCCP nanoparticles with human umbilical vein epithelial cells HUVEC;
FIG. 7 is MCCP catalysis H2O2Reaction machine for in vitro detection of generated hydroxyl free radicalsAnd (6) processing.
Detailed Description
The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, which form a part hereof, and which together with the embodiments of the invention serve to explain the principles of the invention and not to limit its scope.
The invention provides a molybdenum-based nanoenzyme with high catalytic activity, and the preparation raw materials of the molybdenum-based nanoenzyme with high catalytic activity comprise molybdenum oxide nanoparticles, cysteine and copper ions.
Specifically, the raw materials for preparing the molybdenum-based nanoenzyme with high catalytic activity also comprise a surfactant.
Specifically, the surfactant is a polymer having good biocompatibility, for example, polyvinylpyrrolidone (PVP).
The invention also provides a preparation method of the molybdenum-based nanoenzyme with high catalytic activity, which comprises the following steps:
step S1, preparing molybdenum oxide nano particles;
step S2, adding cysteine (L-Cys) and water-soluble copper salt into 20mL of secondary water solution, and carrying out ultrasonic stirring and chelation to obtain white floccule L-Cys-Cu; wherein the water-soluble copper salt is cupric chloride (CuCl)2) (ii) a Cysteine (L-Cys) in an amount of 2mmoL, copper chloride (CuCl)2) In an amount of 1 mmoL;
step S3, mixing the molybdenum oxide nano-particles and L-Cys-Cu, placing the mixture into a centrifuge tube, placing the centrifuge tube into an ultrasonic pool, carrying out ultrasonic treatment for 3-8min, then placing the centrifuge tube on a magnetic stirrer for stirring, uniformly and slowly adding a surfactant PVP (polyvinyl pyrrolidone) in the stirring process, and carrying out self-assembly;
step S4, centrifuging to take out the precipitate after the reaction is finished, repeatedly centrifuging and washing the precipitate for 2-3 times by using secondary water, centrifuging for 1 time by using ethanol, and absorbing excessive ethanol to obtain the molybdenum-based nanoenzyme (MoO) with high catalytic activity2-L-Cys-Cu-PVP)。
Specifically, in step S1, the step of preparing the molybdenum oxide nanoparticles includes:
s101, weighing ammonium molybdate powder according to a proportion, dissolving the ammonium molybdate powder in deionized water, and performing ultrasonic dissolution to obtain an ammonium molybdate aqueous solution;
s102, slowly adding absolute ethyl alcohol into the ammonium molybdate aqueous solution obtained in the S101 by using a liquid transfer gun, performing ultrasonic dispersion, and stirring at room temperature for 30-40min to obtain a uniformly mixed solution;
s103, transferring the mixed solution into a hydrothermal reaction kettle with a polytetrafluoroethylene inner container, screwing the reaction kettle, and placing the reaction kettle in an oven, wherein the reaction temperature is set to 160-200 ℃, and the reaction time is 10-15 h;
s104, after the reaction is finished, naturally cooling to room temperature, centrifuging, taking out the precipitate, repeatedly centrifuging and washing the precipitate for 2-5 times by using secondary water, and freeze-drying the precipitate in a freeze dryer for 45-50h to obtain molybdenum oxide (MoO)2) And (3) nanoparticles.
In S101 and S102, ammonium molybdate: absolute ethanol: deionized water 0.7-1 mmoL: 10-15 mL: 20-25 mL.
Specifically, in S104, molybdenum oxide (MoO)2) The surface of the nano-particles is of a rough porous structure.
Specifically, in step S2, the molar ratio of cysteine to copper chloride is controlled to be 2: 1-1.5. Preferably, the molar ratio of cysteine to copper chloride is 2: 1.
specifically, in step S3, the molar ratio of the molybdenum oxide nanoparticles to L-Cys-Cu is controlled to be 1: 1-3.
Specifically, in step S3, the mass ratio of the molybdenum oxide nanoparticles to the PVP is controlled to be 1: 1-2.
Specifically, in step S4, the structure of the molybdenum-based nanoenzyme with high catalytic activity is that copper ions are dispersed and adsorbed on the surface of the molybdenum oxide nanoparticle, that is, the porous molybdenum oxide nanoparticle and cysteine (L-Cys) -copper ions are effectively anchored.
On the other hand, the invention also provides application of the molybdenum-based nanoenzyme with high catalytic activity as an anti-tumor material.
Compared with the prior art, the method utilizes a simple one-step hydrothermal method to decompose ammonium molybdate and re-nucleate and grow porous MoO under the hydrothermal condition2Nanoparticles having a high catalase-like catalytic activity in a catalytic processHas degradation action and can avoid the long-term toxicity of the nano material to the organism, which is safe and efficient H in TME2O2The responsive nano enzyme catalyst can improve hypoxic in tumors and enhance sensitivity for radiotherapy; in MoO2In the process of anchoring copper ions on the surfaces of the nano particles, only simple and safe ultrasonic stirring is carried out at room temperature, and other high-temperature and high-pressure processes are not used for changing MoO2The shape and structure of the nano-particles are anchored by cysteine L-Cys serving as an intermediate linking agent, so that a simple reference basis for a synthetic thought is provided for anchoring metal ions by other transition metal oxides.
The molybdenum-based nanoenzyme (MCCP) with high catalytic activity has the synergistic enhancement of catalase-like catalytic decomposition H2O2The oxygen is generated to improve the effects of hypoxia in the tumor and sensitization radiotherapy. In the same MoO2In two aqueous solutions (MP, MCCP) of the mass content, MCCP catalytically decomposes H in the same time2O2The content is about twice that of a single MP experimental group without L-Cys-Cu; at the same H2O2The aqueous solution of L-Cys-Cu alone at the experimental concentration did not have obvious catalase-like catalytic effect, probably because the L-Cys-Cu is adsorbed on the MoO with large surface area due to larger particles and fewer exposed active sites2After the surface of the nano-particles, the high dispersibility of L-Cys-Cu is realized, and more H is exposed2O2The contacted copper active sites realize the effective catalase-like catalytic activity of MCCP under low concentration, generate a large amount of oxygen, improve hypoxia in tumors and realize radiotherapy sensitization.
The catalase catalytic activity and the radiotherapy sensitization function of the molybdenum-based nanoenzyme with high catalytic activity provided by the invention are far higher than those of single molybdenum oxide and single L-Cys-Cu, and H2O2In TME, H is used in a dose of 100. mu.M to 1.0mM2O2In a high expression dose range of increasing MCCP to a relatively low concentration of H2O2While the catalytic ability of the catalase-like enzyme is improved, the dosage and H of copper ions with relatively high biological toxicity are reduced2O2The molybdenum-based nanoenzyme with excellent catalytic activity of the catalase pseudoenzyme is finally obtained. MCCP catalysis H2O2After oxygen is generated, the L-Cys-Cu is easily disassembled in TME under X-ray irradiation in the radiotherapy process, and the L-Cys-Cu is released from the composite material and starts to carry out Fenton-like reaction, so that more hydroxyl free radicals OH are generated, and MCCP improves tumor oxygen lack and tumor sensitivity in radiotherapy, thereby having wide application prospect in the aspect of radiotherapy sensitization of tumors.
It is worth noting that the principle of the application of the molybdenum-based nanoenzyme with high catalytic activity as the anti-tumor material provided by the invention is as follows: using intratumoral high expression of H2O2Or H supplied from an external source2O2(for example, X-ray irradiation during radiotherapy can increase H in tumor2O2Expression of) and binding Cu2+And Cu1+The valence conversion (namely, Fenton-like reaction) of the two can circularly catalyze H2O2The decomposition reaction of (2) generates highly toxic hydroxyl radical (. OH), the. OH has strong oxidizing property and can kill cancer cells more effectively, and the reaction process can generate O2。
Fenton-like reaction of copper ions:
Cu2++H2O2＝Cu1++O2+H2O
Cu2++H2O2＝Cu1++·OH
example 1
The embodiment provides a molybdenum oxide nanoparticle, and the preparation method comprises the following steps:
(1) weighing 0.7mmol (0.0865g) of ammonium molybdate powder, dissolving in 20mL of deionized water, and performing ultrasonic dissolution to obtain an ammonium molybdate aqueous solution;
(2) slowly adding 10mL of absolute ethyl alcohol into the ammonium molybdate aqueous solution obtained in the step (1) by using a 5mL liquid-transferring gun, performing ultrasonic dispersion, and stirring at room temperature for 30 minutes to obtain a completely and uniformly mixed solution; wherein the volume ratio of the absolute ethyl alcohol to the water is 1:2, and the total volume of the mixed solution is 30 mL;
(3) transferring 30mL of the mixed solution into a 50mL hydrothermal reaction kettle with a polytetrafluoroethylene inner container, screwing the reaction kettle, placing the reaction kettle in a drying oven, setting the reaction temperature to be 160 ℃, and setting the reaction time to be 12 hours;
(4) after the reaction is finished, naturally cooling to room temperature, centrifuging, taking out the precipitate, repeatedly centrifuging and washing for three times by using secondary water, and freeze-drying for 48 hours in a freeze-drying machine to obtain the molybdenum oxide nanoparticles (MoO)2)。
Example 2
The embodiment provides a molybdenum-based nanoenzyme with high catalytic activity, which is prepared by the following steps:
(1) MoO synthesized in example 12And L-Cys-Cu in a molar ratio of 1: 1 in 20mL of aqueous solution, performing ultrasonic treatment in a centrifuge tube for 3 minutes, and then stirring on a magnetic stirrer;
(2) during the stirring process, the surface active agent PVP (MoO) is uniformly and slowly added2The mass ratio of the PVP to the PVP is 1: 1) (ii) a
(3) After the reaction is finished, centrifuging, taking out the precipitate, repeatedly centrifuging and washing for 2-3 times by using secondary water, washing by using ethanol and centrifuging, and absorbing excess ethanol to obtain the molybdenum-based nanoenzyme (MoO) with high catalytic activity2-L-Cys-Cu-PVP, abbreviated MCCP).
The term (3) includes MoO at the bottom of the obtained centrifugal tube2Storing in a refrigerator at 4 deg.C for use.
Comparative example 1
This comparative example provides a MoO2-PVP, prepared as follows:
100-300mg of MoO synthesized in example 1 was added at room temperature2Adding the powder into 20mL of secondary aqueous solution, and uniformly and slowly adding a surface active agent PVP in the stirring process to obtain MoO2PVP (abbreviation: MP). Wherein, MoO2The mass ratio of the PVP to the PVP is 1: 1.
test example 1
Scanning electron microscope and transmission electron microscope.
As shown in fig. 1: a and b are respectively the scanning electron microscope and transmission electron microscope images of the MCCP nano-particles of the inventionThe size of the nano particles is about 150nm, each nano particle is composed of particles with smaller sizes, and a certain gap is formed between the small particles, so that an obvious porous structure is displayed; c is an element energy spectrum (EDX) of the MCCP, and indicates that the main elements of the MCCP comprise Mo and Cu; d is X-ray powder diffraction (XRD) pattern of MCCP with peak position corresponding to MoO2Corresponds to the standard card (JCPDS: 50-0739).
Test example 2
MoO2Catalase-like catalytic activity evaluation of L-Cys-Cu and MCCP.
Firstly, MP with different masses (the final concentration is 2.5, 5, 10, 20 mu g/mL) is ultrasonically dissolved in deionized water, a reset dissolved oxygen tester probe is immersed in an MP water solution, after the reading of the instrument is stable, the reading of the instrument at the moment is recorded, namely the initial oxygen content in the water solution, namely the initial zero-time point in the catalytic oxygen production process, hydrogen peroxide with the final concentration of 1.0mM is added, and after the catalytic process is started, data recording is carried out at intervals of 15 seconds. As shown in a in fig. 2, the results of the dissolved oxygen test of the MP nanoparticles with different concentrations show that the catalase-like catalytic activity of the MP nanoparticles is positively correlated with the concentration of MP; in order to prove that MCCP obtained after loading L-Cys-Cu has enhanced catalase-like catalytic effect compared with MP, the following comparative experiment is carried out: three experimental groups, namely MP, MCCP and L-Cys-Cu are set, and the specific process is as follows: the same volume of 50. mu.g/mL MoO2The aqueous solution and the 400 mu g/mL L-Cys-Cu aqueous solution with equal volume are respectively divided into two parts and placed in a centrifuge tube, and then one part of each of the two solutions is stirred and synthesized into MCCP nanoparticles (MoO in the MCCP nanoparticle composition)2The concentration became 25. mu.g/mL, L-Cys-Cu became 200. mu.g/mL), and then one part of MoO remained2Synthesizing MP nano particles by aqueous solution, and adding deionized water to MoO2The concentration of the L-Cys-Cu solution is 25 mu g/mL, the rest 400 mu g/mL only needs to be added with the same volume of deionized water to reduce the concentration to 200 mu g/mL, and the three groups of solutions are MCCP (MoO)225. mu.g/mL and L-Cys-Cu 200. mu.g/mL), MP (MoO)225 μ g/mL), L-Cys-Cu aqueous solution (200 μ g/mL), and then, following the previous stepsAnd (5) measuring the oxygen production by a dissolved oxygen tester. As shown in fig. 2, b is the consumption curve of MCCP to hydrogen peroxide using titanium sulfate as indicator, wherein the inset is the consumption curve of MCCP to H under the indication of titanium sulfate2O2Colorimetric optical photograph of the color change of the indicator during consumption (titanium sulfate solution itself is colorless, H)2O2It is yellow (407 nm absorption peak visible in ultraviolet) after reacting with titanium sulfate, and is accompanied by H2O2Titanium sulfate remained colorless upon consumption of MCCP), the inset color changed from yellow to colorless over time, indicating that H changed over time2O2Is continuously consumed; in FIG. 2, c is the catalase-like catalytic activity of three materials MP, MCCP, L-Cys-Cu to catalytically decompose H2O2Comparing the effects; the results show that L-Cys-Cu and MoO2The MCCP obtained by compounding can obviously enhance catalase-like activity of the two, i.e. catalyzing and decomposing H2O2(ii) ability of; in fig. 2, d is the effect of temperature on the catalytic effect of MCCP; indicating that the reaction is an endothermic reaction, with the rate of reaction increasing with increasing temperature.
Test example 3
And evaluating the content of Reactive Oxygen Species (ROS) in the cells.
The catalytic activity of MCCP nanoparticles under X-ray irradiation was assessed using ROS probe 2 ', 7' -dichlordihydrofluorescin diacetate (H2DCF-DA,10 μ M, Beyotime Co, Sigma-Aldrich, USA), ROS was able to deacetylate H2DCF-DA and oxidize it to 2 ', 7' -Dichlorfluorofluorescin (DCF) with high fluorescence properties. First, adenocarcinoma human alveolar basal epithelial cells (a549 cells) were cultured at 2 × 10 using complete medium5Cell concentration per dish was cultured for 24 hours adherent culture, and then MCCP was added to two groups of cells: 50. mu.g/mL, MP: 50 μ g/mL, another group of cells without material, with the same volume of complete medium and as a control, after 5 hours, washed twice with PBS, 200 μ L of probe solution (1 μ L of RPMI cell culture medium, 1 μ L of LDCFH-DA +999 μ L) per dish incubated in the incubator for 20 minutes, then washed with PBS, while the experimental group was irradiated with X-ray (50kv, 49 μ A) for 15 minutes, after which cells were incubated with Hoechst33342(Beyotime)Nuclear staining was carried out for 15 minutes. Finally, the cells were washed twice with PBS and observed under a laser confocal microscope (A1/LSM-Kit, Nikon/Pico Quant GmbH, Japan/Germany).
FIG. 3 is a photograph of a Confocal scanning microscope (CLSM) of MP, MCCP at the cell level for Reactive Oxygen Species (ROS) detection, showing that MCCP has the strongest free radical generating ability under X-ray excitation, probably due to the concerted catalysis of hydrogen peroxide by copper ions and molybdenum oxide to generate O by oxygen peroxide2The increased level improves tumor hypoxia, which in turn promotes the production of ROS, and the other reason is that the copper ions on L-Cys-Cu are released from the gradually degraded molybdenum oxide surface and then undergo a light-excited Fenton-like reaction under the activation of X-ray to produce more OH, thereby sensitizing the radiotherapy to cancer cells.
Test example 4
Evaluation of killing of cancer cells by in vitro DNA double strand break assay.
First, a549 cells were seeded in a 24-well plate at a cell concentration of 3 × 104Wells, after 24 hours incubation for cell attachment, were divided into three groups including: wherein MCCP is added into two groups of cells respectively: 50. mu.g/mL, MP: 50 μ g/mL, and another group of cells, to which no material was added, and to which only the same volume of complete medium was added as a control group, after 5 hours, X-ray (50kv, 49 μ A) irradiation was performed for 15 minutes. Cells were washed twice with PBS and fixed with 4% paraformaldehyde, then washed three times with PBS, each time with shaking for 3 minutes on a shaker, pretreated with 0.2% Triton-X-100(300 μ L,10 minutes) to increase membrane permeability, and primary antibody (β -Actin, a scaffold protein antibody) diluted with blocking solution (5% FBS, 1% Triton-X-100) at a dilution ratio of 1: 100, prepare primary anti-dilution, discard the block from wells after dilution, add 200. mu.L of primary anti-dilution per well, and finally wrap the 24-well plate in PBS soaked gauze overnight in a refrigerator at 4 ℃. The 24-well plate was removed from the refrigerator and washed three times with PBS in a shaker for 3 minutes each. Subsequently, a secondary antibody dilution (anti-rabbitAlexaFluor-488conjugate igg: PBS ═ 1: 500, 200 μ L per well, temperature 37 ℃, duration 1 hour) was added, followed by three washes with PBSAnd then, performing Hoechst staining on cell nuclei, finally washing the cell nuclei by PBS for three times, sealing the cell nuclei by using a cover slip, observing the cell nuclei under a laser confocal microscope, imaging and recording data.
Fig. 4 is a laser confocal scanning micrograph of an in vitro DNA double strand break test, and the result shows that, consistent with ROS analysis in vitro, MCCP nanoparticles have stronger DNA damage capability, i.e., stronger anti-tumor capability.
Test example 5
MCCP catalysis H2O2In vitro detection of hydroxyl radicals produced.
Design considerations: taking into account that not only H is present in the tumour2O2High expression, and GSH is used as an antioxidant and is also expressed in tumor, therefore, the GSH highly expressed in the tumor microenvironment is simulated in the experiment, and can act as a reducing agent to reduce Cu in MCCP2+Reduction to Cu1+And is of Cu1+Has stronger Fenton-like reaction capability under the action of X-ray, thereby being capable of leading H to react2O2Decomposed into hydroxyl radicals. First, a carbonate buffer solution (BBS) with a concentration of 0.1mol/L is prepared, then a DTNB (a probe for detection-SH) solution with a concentration of 100mM, a Glutathione (GSH) solution with a concentration of 1.0mM, and MP and MCCP solutions with different concentrations are prepared by using a BBS solution, a Tris HCL solution with a concentration of 100mM is prepared by using secondary water, after all the solutions are prepared, Ellman's analysis is carried out, firstly 225 μ L of the GSH solution is mixed with 225 μ L of the MCCP/MP solution, shaking for 3 hours in the dark, then 785 μ L of Tris diluent and 15 μ L of the DTNB solution are added, each group has a volume of 1250 μ L, and the absorbance is detected at 412 nm. The reaction mechanism is shown in FIG. 7.
The OH produced by MCCP under the action of X-ray was detected by OH probe (terephthalic acid, TA,5mM, AlfaAesar), and TA reacted with OH to produce 2-hydroxyterephthalic acid (TAOH, maximum fluorescence peak 435 nm). Firstly, the prepared TA solution is added with the divided H or not2O2The concentration of hydrogen peroxide is 1.0mM, wherein the MCCP solution reacts with excessive GSH to ensure that Cu is contained2+All conversion to Cu1+Then placed on a shaker at 37 ℃ overnight and then onThe change in fluorescence emission front was recorded at 435 nm.
As shown in fig. 5: a is the consumption curve of the MCCP nanoparticles with different concentrations in the same time for GSH, which shows that the consumption speed of the GSH is positively correlated with the concentration of the MCCP nanoparticles; b is a control experiment of MP, PBS and MCCP, and shows that only MCCP experimental group can consume GSH, namely Cu2+With GSH to undergo a redox reaction; c is a GSH consumption curve along with time under the same MCCP nanoparticle concentration, which shows that the GSH consumption is positively correlated with the time; d is Cu under X-ray irradiation2+And H2O2The fluorescence emission spectrum of the product OH in the Fenton-like reaction of (1), TA + H2O2+ MCCP group ratio TA + H2O2+ MP clearly has higher fluorescence intensity, indicating that the Fenton-like reaction of copper ions in MCCP under X-ray irradiation produces OH, but in contrast to TA + H2O2The reason for this low may be part H2O2Is catalyzed to generate O2. Resulting in a fixed total amount of H2O2The content of (b) is reduced, and relatively less OH is generated.
As shown in fig. 6, fig. 6 is the cytotoxicity data of MCCP nanoparticles incubated with human umbilical vein epithelial cells HUVEC for 24 hours, and the test results show that MCCP has no significant cytotoxicity within the tested dose range.
While the invention has been described with reference to specific preferred embodiments, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the following claims.
Claims (8)
1. The molybdenum-based nanoenzyme with high catalytic activity is characterized by comprising molybdenum oxide nanoparticles and copper ions, wherein the copper ions are anchored on the surfaces of the molybdenum oxide nanoparticles;
the preparation method of the molybdenum-based nanoenzyme with high catalytic activity comprises the following steps:
step S1, preparing molybdenum oxide nano particles;
step S2, adding cysteine and water-soluble copper salt into secondary water, ultrasonically stirring and chelating to synthesize white floccule L-Cys-Cu;
s3, placing the molybdenum oxide nano-particles and the L-Cys-Cu mixed solution into a centrifuge tube, carrying out ultrasonic treatment in an ultrasonic pool, then placing the centrifuge tube on a magnetic stirrer for stirring, and uniformly and slowly adding a surfactant in the stirring process;
and step S4, after the reaction is finished, centrifuging to take out the precipitate, repeatedly centrifuging and washing the precipitate for 2-3 times by using secondary water, washing by using ethanol and centrifuging, and sucking off redundant ethanol to obtain the molybdenum-based nanoenzyme with high catalytic activity.
2. The molybdenum-based nanoenzyme with high catalytic activity as claimed in claim 1, wherein the surfactant is a polymer with good biocompatibility.
3. The method for preparing molybdenum-based nanoenzyme with high catalytic activity as claimed in claim 1, comprising the steps of:
step S1, preparing molybdenum oxide nano particles;
step S2, adding cysteine and water-soluble copper salt into secondary water, ultrasonically stirring and chelating to synthesize white floccule L-Cys-Cu;
s3, putting the mixed solution of the molybdenum oxide nano-particles and L-Cys-Cu into a centrifugal tube, carrying out ultrasonic treatment in an ultrasonic pool, then putting the ultrasonic pool on a magnetic stirrer for stirring, and uniformly and slowly adding a surfactant in the stirring process;
and step S4, after the reaction is finished, centrifuging and taking out the precipitate, repeatedly centrifuging and washing the precipitate for 2-3 times by using secondary water, washing by using ethanol and centrifuging, and sucking away excessive ethanol to obtain the molybdenum-based nanoenzyme with high catalytic activity.
4. The method according to claim 3, wherein the step S1 of preparing the molybdenum oxide nanoparticles comprises:
s101, weighing ammonium molybdate powder according to a proportion, dissolving the ammonium molybdate powder in deionized water, and performing ultrasonic dissolution to obtain an ammonium molybdate aqueous solution;
s102, adding absolute ethyl alcohol into the ammonium molybdate aqueous solution obtained in the S101 by using a liquid-transferring gun, performing ultrasonic dispersion, and stirring at room temperature to obtain a uniformly mixed solution;
s103, transferring the mixed solution into a hydrothermal reaction kettle, screwing the reaction kettle, placing the reaction kettle in an oven, setting the reaction temperature to 160-200 ℃, and the reaction time to 10-15 h;
and S104, after the reaction is finished, naturally cooling to room temperature, centrifuging, taking out the precipitate, repeatedly centrifuging and washing the precipitate for 2-5 times by using secondary water, and freeze-drying in a freeze dryer to obtain the molybdenum oxide nanoparticles.
5. The method according to claim 4, wherein in S101 and S102, the ratio of ammonium molybdate: anhydrous ethanol: deionized water =0.7-1 mmoL: 10-15 mL: 20-25 mL.
6. The method according to claim 3, wherein in step S2, the water-soluble copper salt is cupric chloride, and the molar ratio of cysteine to cupric chloride is 2: 1-1.5.
7. The method according to claim 3, wherein in the step S3, the molar ratio of the molybdenum oxide nanoparticles to L-Cys-Cu is 1: 1-3.
8. The method according to any one of claims 3 to 7, wherein in the step S3, the surfactant is polyvinylpyrrolidone, and the mass ratio of the molybdenum oxide nanoparticles to the polyvinylpyrrolidone is 1: 1-2.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106841188A (en) * | 2017-03-01 | 2017-06-13 | 福建医科大学 | A kind of paper micro-fluidic chip that intracellular hydrogen peroxide is detected for nanoscale molybdenum oxide analogue enztme |
| CN109928427A (en) * | 2019-03-21 | 2019-06-25 | 暨南大学 | A kind of double ion intercalation molybdenum oxide nanometer sheet, hetero-junctions and preparation method thereof |
| CN110819339A (en) * | 2019-10-14 | 2020-02-21 | 浙江师范大学 | Cu-amino acid composite up-conversion nano material and preparation method thereof |
| CN110872131A (en) * | 2019-12-06 | 2020-03-10 | 中国科学院长春应用化学研究所 | Cu2MoS4Nano enzyme, preparation method thereof, nano medicine and application |
| CN111017996A (en) * | 2019-09-25 | 2020-04-17 | 青岛大学 | Synthesis of MoO with double simulated enzyme activity3-XMethod for producing antimicrobial material |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106841188A (en) * | 2017-03-01 | 2017-06-13 | 福建医科大学 | A kind of paper micro-fluidic chip that intracellular hydrogen peroxide is detected for nanoscale molybdenum oxide analogue enztme |
| CN109928427A (en) * | 2019-03-21 | 2019-06-25 | 暨南大学 | A kind of double ion intercalation molybdenum oxide nanometer sheet, hetero-junctions and preparation method thereof |
| CN111017996A (en) * | 2019-09-25 | 2020-04-17 | 青岛大学 | Synthesis of MoO with double simulated enzyme activity3-XMethod for producing antimicrobial material |
| CN110819339A (en) * | 2019-10-14 | 2020-02-21 | 浙江师范大学 | Cu-amino acid composite up-conversion nano material and preparation method thereof |
| CN110872131A (en) * | 2019-12-06 | 2020-03-10 | 中国科学院长春应用化学研究所 | Cu2MoS4Nano enzyme, preparation method thereof, nano medicine and application |
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