CN114917354B - Preparation and application of Cu monoatomic nano-enzyme - Google Patents
Preparation and application of Cu monoatomic nano-enzyme Download PDFInfo
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- XCJYREBRNVKWGJ-UHFFFAOYSA-N copper(II) phthalocyanine Chemical compound [Cu+2].C12=CC=CC=C2C(N=C2[N-]C(C3=CC=CC=C32)=N2)=NC1=NC([C]1C=CC=CC1=1)=NC=1N=C1[C]3C=CC=CC3=C2[N-]1 XCJYREBRNVKWGJ-UHFFFAOYSA-N 0.000 claims abstract description 14
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- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 1
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- MDMUYJRRYYXDLZ-UHFFFAOYSA-N tert-butyl 2-methyl-1-oxido-3,4-dihydropyrrol-1-ium-2-carboxylate Chemical compound CC(C)(C)OC(=O)C1(C)CCC=[N+]1[O-] MDMUYJRRYYXDLZ-UHFFFAOYSA-N 0.000 description 1
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- MFLKDEMTKSVIBK-UHFFFAOYSA-N zinc;2-methylimidazol-3-ide Chemical class [Zn+2].CC1=NC=C[N-]1.CC1=NC=C[N-]1 MFLKDEMTKSVIBK-UHFFFAOYSA-N 0.000 description 1
Classifications
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- 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
- A61K47/52—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 inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/409—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil having four such rings, e.g. porphine derivatives, bilirubin, biliverdine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/34—Copper; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/36—Hydrogen production from non-carbon containing sources, e.g. by water electrolysis
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Abstract
The invention discloses a Cu monoatomic nano-enzyme which is prepared from a compound Cu-N 4 ClG; the preparation method comprises the following steps: s21, adding acidified graphene and copper phthalocyanine CuPc into triple distilled water for centrifugation; s22, carrying out ultrasonic treatment on the product obtained in the step S21; s23, washing and drying the product obtained in the step S22 to obtain a CuPc functionalized G composite material; s24, carrying out Ar atmosphere heat treatment on the CuPc functionalized G composite material; s25, cooling the product obtained in the step S24, and then placing the cooled product in an HCl solution for stirring treatment; step S26, washing and drying to obtain the monoatomic nano-enzyme Cu-N 4 ClG. The Cu-N4ClG monoatomic nano-enzyme designed by the application has excellent CAT and SOD-like activity, and provides an effective strategy and a new idea for treating osteoarthritis and chronic diseases related to oxidative stress in the future.
Description
Technical Field
The invention belongs to the field of nano-enzyme preparation, and particularly relates to preparation and application of Cu single-atom nano-enzyme.
Background
Osteoarthritis is known as a chronic joint disease, and currently, for the treatment of osteoarthritis, oral non-steroidal anti-inflammatory analgesics, local microcirculation improving drugs are mainly used, and intra-articular injection of hyaluronic acid or corticosteroids may be used. However, these methods only alleviate the symptoms of the patient to a different extent, and it is difficult to prevent the progression of the disease, which results in the osteoarthritis patient developing to a later stage, requiring a lot of time and money for treatment and care, and even performing expensive joint replacement surgery. Therefore, there is an urgent need in the field of clinical treatment of orthopedics to develop new safe and effective osteoarthritis treatment strategies.
Studies have shown that excessive accumulation of free radicals such as reactive oxygen species in the joints is an important cause of osteoarthritis, can cause oxidative stress damage to biological membranes, DNA, lipids and proteins, and lead to over-expression of many inflammatory factors, leading to cartilage degradation and osteoarthritis pathogenesis. Strategies based on active oxygen radical scavengers, which can scavenge excess active oxygen radicals and inhibit oxidative stress, have proven to be promising tools for the treatment of osteoarthritis. The nano enzyme is used as a potential active oxygen scavenger, has high catalase-like and superoxide dismutase-like enzyme activities, can effectively remove active oxygen free radicals, and is of great interest in the treatment of osteoarthritis. For example, mnO reported in the related studies 2 The nanometer particles, the hollow Prussian blue nanometer enzyme, the modified ZIF-8 nanometer particles and the dopamine melanin nanometer particles all show that the nanometer particles have good therapeutic effect on osteoarthritis. However, the existing nano-enzyme has a series of problems of strong cytotoxicity, low selectivity, low catalytic activity, low metal atom utilization rate and the like, and the performance and further application of the nano-enzyme are severely limited.
Chinese patent CN113457659a discloses a transition metal monoatomic nano-enzyme, and a preparation method and use thereof, wherein the transition metal monoatomic nano-enzyme is obtained by heat treatment of a transition metal doped metal organic framework material, and the transition metal doped metal organic framework material is prepared by using water as a solvent. The prepared transition metal monoatomic nano enzyme has uniform morphology, specific surface area and aperture, has atomically dispersed active sites, and provides a platform for further research of metal active centers and catalytic mechanisms of monoatomic catalysts. Experimental results show that the prepared transition metal monoatomic nano-enzyme has good oxidase-like activity, peroxidase-like activity and halogen-like peroxidase-like activity, can be used for preparing a high-catalytic-activity enzyme-like preparation, and has wide application prospects in the fields of antibiosis, anti-tumor, wastewater treatment, immunoblotting analysis and the like.
In the research, the applicant found that the single-atom nano-enzyme has very excellent metal atom utilization rate, and the utilization rate can reach up to 100%. Meanwhile, the catalyst also has strong metal-carrier interaction, low coordination environment, clear electronic structure and geometric structure, high selectivity and metal atom distribution stability, and the excellent performances enable the catalyst to simulate a natural highly evolved enzyme catalysis center, and take single atoms as an enzyme design concept, so that the catalyst becomes a good substitute for traditional enzymes, and in a plurality of single-atom enzyme design systems, the Cu single-atom nano-enzyme is considered as one of the single-atom nano-enzymes with the most potential for treating oxidative stress diseases by utilizing CAT and SOD-like enzyme activities, and the Cu single-atom catalyst has high photocatalytic hydrogen production activity, so that the photocatalytic activity can be improved. However, how to use Cu to prepare monoatomic nanoenzyme for treating osteoarthritis is a difficulty of current research.
Disclosure of Invention
In order to solve the problems, the technical effect of treating osteoarthritis is achieved by preparing the Cu monoatomic nano-enzyme and further effectively removing active oxygen.
In order to achieve the effects, the invention designs a preparation method and application of Cu monoatomic nano-enzyme.
The method comprises the following steps ofCu monoatomic nanoenzyme composed of compound Cu-N 4 ClG.
Preferably, the preparation method of the Cu monoatomic nano-enzyme comprises the following steps of:
step S21, adding graphene and copper phthalocyanine CuPc into triple distilled water for centrifugation;
s22, carrying out ultrasonic treatment on the product obtained in the step S21;
s23, washing and drying the product obtained in the step S22 to obtain a CuPc functionalized G composite material;
s24, carrying out Ar atmosphere heat treatment on the CuPc functionalized G composite material;
s25, cooling the product obtained in the step S24, and then placing the cooled product in an HCl solution for stirring treatment;
step S26, washing and drying to obtain the monoatomic nano-enzyme Cu-N 4 ClG。
Preferably, in the step S21, the mass ratio of graphene to copper phthalocyanine is 1-4: 2 to 10.
Preferably, in the step S22, the ultrasonic treatment time is 1 to 6 hours.
Preferably, in the step S23, the washing method is as follows: repeatedly washing with ethanol;
the drying treatment method comprises the following steps: vacuum drying at 40℃for 12h.
Preferably, in the step S24, the method of Ar atmosphere heat treatment is as follows: and (3) heating the product obtained in the step S23 to 600-900 ℃ in Ar atmosphere, and performing heat treatment for 1-5h.
Preferably, in the step S25, the concentration of the HCl solution is 3mol/L HCl solution.
Preferably, in the step S25, the stirring method is stirring for 6-18 hours at 40-80 ℃.
Preferably, in the step S26, the method of washing and drying is as follows: the solution was washed to neutrality and dried under vacuum at 60 ℃ for 24h.
A medicine prepared from Cu monoatomic nano-enzyme can be used for treating osteoarthritis.
The advantages and effects of the application are as follows:
1. the application uses graphene as a carrier, and uses copper phthalocyanine with a specific Cu coordination environment as Cu and N sources to construct the Cl N-doped graphene-loaded Cu monoatomic nano enzyme Cu-N 4 ClG, which has excellent CAT and SOD-like activity, can be of great significance for the treatment of osteoarthritis.
2. The application proves that the Cu-N is proved by experimental results 4 ClG can effectively remove O 2 ·-、·OH、H 2 O 2 And the equivalent free radicals have excellent CAT and SOD enzyme activities. At the same time, cu-N 4 ClG has good biological safety and can effectively remove active oxygen in chondrocytes in osteoarthritis.
3. The method creatively adjusts the single-atom copper activity center through the accurate coordination of chlorine and nitrogen, and establishes the atomic engineering Cu-N 4 ClG monoatomic nano-enzyme has excellent CAT and SOD-like activity and is first applied to the treatment of osteoarthritis; however, studies on a preparation method of graphene-supported Cu monoatomic nanoenzyme precisely coordinated with chlorine and nitrogen and application thereof to OA treatment have not been reported in literature and patent; the application not only provides scientific basis for further application and expansion of monoatomic nano-enzyme, but also provides effective strategies and new ideas for treating osteoarthritis and chronic diseases related to oxidative stress in the future.
The foregoing description is only a summary of the technical solutions of the present application, so that the technical means of the present application may be implemented according to the content of the specification, and so that the foregoing and other objects, features and advantages of the present application may be more clearly understood, the following detailed description of the preferred embodiments of the present application is given in conjunction with the accompanying drawings.
The above and other objects, advantages and features of the present application will become more apparent to those skilled in the art from the following detailed description of the specific embodiments thereof, taken in conjunction with the accompanying drawings.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, and it is obvious that the drawings in the following description are some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. Like elements or portions are generally identified by like reference numerals throughout the several figures. In the drawings, elements or portions thereof are not necessarily drawn to scale.
FIG. 1 shows a Cu-N structure according to the present invention 4 ClG SAzyme high resolution TEM images;
FIG. 2 shows a Cu-N structure according to the present invention 4 ClG and G, evaluating their ability to scavenge ROS;
FIG. 3 shows a Cu-N structure according to the present invention 4 ClG is subjected to H2O2 decomposition comparison with G, and the activity power diagram of CAT enzyme is evaluated;
FIG. 4 shows a Cu-N structure according to the present invention 4 ClG and G are subjected to CCK-8 experiments, and the biocompatibility diagram is evaluated;
FIG. 5 shows a Cu-N structure according to the present invention 4 ClG and G were subjected to DCFH-DA probe detection to evaluate the fluorescence profile of their ability to scavenge ROS under different control materials and treatment conditions.
Detailed Description
For the purposes of making the objects, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions of the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is apparent that the described embodiments are some embodiments of the present application, but not all embodiments. In the following description, specific details such as specific configurations and components are provided merely to facilitate a thorough understanding of embodiments of the present application. It will therefore be apparent to those skilled in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the application. In addition, descriptions of well-known functions and constructions are omitted in the embodiments for clarity and conciseness.
It should be appreciated that reference throughout this specification to "one embodiment" or "the present embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present application. Thus, the appearances of the "one embodiment" or "this embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
Furthermore, the present application may repeat reference numerals and/or letters in the various examples. This repetition is for the purpose of simplicity and clarity and does not in itself dictate a relationship between the various embodiments and/or configurations discussed.
The term "and/or" is herein merely an association relationship describing an associated object, meaning that there may be three relationships, e.g., a and/or B, may represent: the terms "/and" herein describe another associative object relationship, indicating that there may be two relationships, e.g., a/and B, may indicate that: the character "/" herein generally indicates that the associated object is an "or" relationship.
The term "at least one" is herein merely an association relation describing an associated object, meaning that there may be three kinds of relations, e.g., at least one of a and B may represent: a exists alone, A and B exist together, and B exists alone.
It is further noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprise," "include," or any other variation thereof, are intended to cover a non-exclusive inclusion.
Example 1
This example mainly describes a Cu monoatomic nanoenzyme and a method for preparing the same.
A Cu monoatomic nano-enzyme is prepared from Cu-N compound 4 ClG.
A preparation method of Cu monoatomic nano-enzyme, which comprises the following steps:
step S21, adding graphene and copper phthalocyanine CuPc into triple distilled water for centrifugation;
s22, carrying out ultrasonic treatment on the product obtained in the step S21;
s23, washing and drying the product obtained in the step S22 to obtain a CuPc functionalized G composite material;
s24, carrying out Ar atmosphere heat treatment on the CuPc functionalized G composite material;
s25, cooling the product obtained in the step S24, and then placing the cooled product in an HCl solution for stirring treatment;
step S26, washing and drying to obtain the monoatomic nano-enzyme Cu-N 4 ClG。
Further, in the step S21, the mass ratio of graphene to copper phthalocyanine is 1-4: 2 to 10.
Further, in the step S22, the ultrasonic treatment time is 1 to 6 hours.
Further, in the step S23, the washing method is as follows: repeatedly washing with ethanol;
the drying treatment method comprises the following steps: vacuum drying at 40℃for 12h.
Further, in the step S24, the method of heat treatment in Ar atmosphere is as follows: and (3) heating the product obtained in the step S23 to 600-900 ℃ in Ar atmosphere, and performing heat treatment for 1-5h.
Further, in the step S25, the concentration of the HCl solution is 3mol/LHCl solution.
Further, in the step S25, the stirring method is stirring for 6-18h at 40-80 ℃.
Further, in the step S26, the method of washing and drying is as follows: the solution was washed to neutrality and dried under vacuum at 60 ℃ for 24h.
A medicine prepared by Cu monoatomic nano-enzyme, which is used for preparing osteoarthritis.
Example 2
This example mainly describes the preparation of a Cu monoatomic nanoenzyme.
The method comprises the steps of firstly adding 5-20mg of graphene and 10-50mg of copper phthalocyanine (CuPc) into water containing 8mL of three timesUltrasonic treatment is carried out for 1-6h, then repeated washing is carried out by ethanol, and vacuum drying is carried out for 12h at 40 ℃ to obtain the CuPc functionalized G composite material. Then placing the mixture in Ar atmosphere, heating to 600-900 ℃ (5 ℃/min), and performing heat treatment for 1-5h. After cooling to room temperature, placing the sample in 3mol/LHCl solution, stirring at 40-80deg.C for 6-18h, washing until the solution is neutral, and vacuum drying at 60deg.C for 24h to obtain N-doped graphene loaded N and Cl coordinated Cu monoatomic nanoenzyme (Cu-N) 4 ClG SAzyme)。
Example 3
Based on the above example 1, this example mainly describes the effect verification of Cu monoatomic nanoenzyme in osteoarthritis treatment.
FIG. 1 shows Cu-N 4 High resolution TEM image of ClG nanoenzyme. TEM images clearly show that Cu monoatoms are uniformly distributed on the graphene surface, illustrating the formation of Cu monoatoms. Meanwhile, cu-N in FIG. 1 4 ClG also demonstrates that only a broad peak located around 25℃is detected, due to the characteristic diffraction of graphitic carbon, no diffraction peak of Cu nanoparticles is observed, indicating Cu-N 4 The Cu atoms in ClG did not significantly aggregate.
FIG. 2 shows the Cu-N of the prepared material 4 ClG As shown in FIG. 2a, cu-N was studied by Electron Spin Resonance (ESR) using 5-tert-butylcarbonyl-5-methyl-1-pyrroline oxide (BMPO) as a trapping agent and H2O2 as a generator of O2. Cndot. -radicals 4 ClG scavenging of O2-free radicals. When Cu-N is not added 4 ClG, the ESR signal of O2-is strong, indicating that an excessive amount of O2-is present, and Cu-N is added 4 After ClG, the ESR signal of O2. Cndot. -was gradually decreased with increasing concentration, and almost all of the ESR signal was completely attenuated when the concentration was increased to 150ug/mL, indicating excellent scavenging of OH radicals. But with Cu-N 4 ClG still has a pronounced OHER signal peak (FIG. 2 b), indicating that the addition of Cu monoatoms plays a decisive role in increasing the scavenging capacity of the O2. Cndot. -radicals. Also, we have examined Cu-N 4 ClG scavenging action on OH. Also, we have examined Cu-N 4 ClG scavenging of O2-. As shown in FIG. 2c, cu-N 4 ClG also scavenges OH radicals effectively in a concentration dependent manner. When 100ug/mLCu-N are added respectively 4 After ClG and G (FIG. 2 d), the ESR signal of G remains strong, in contrast, at Cu-N 4 ClG shows that the ESR signal of OH is almost lost, and that the excellent SOD-like activity is confirmed.
FIG. 3 shows a Cu-N structure according to the present invention 4 ClG and G are subjected to H2O2 decomposition and comparison effect graph, which gives Cu-N of the prepared material 4 ClG can effectively bind H in a concentration-dependent manner (50, 100, 150, 200 ug/mL) 2 O 2 Decompose into O2 (FIG. 3 a), and decompose O 2 The yield of (C) is far higher than that of G and pure H 2 O 2 Group (fig. 3 b), demonstrating excellent CAT-like activity. Supplement of specific concentration scheme with the accompanying drawings
FIG. 4 is a diagram showing the synthesis of Cu-N 4 ClG the biological activity and the biological safety of the material to the cells are the key of primary concern. Cu-N was first tested using the CellCounting kit-8 (CCK-8) assay System 4 ClG toxicity to chondrocytes. As shown in FIG. 4, cu-N 4 ClG and G are in the concentration range of 200ug/mL (50, 100, 150, 200 ug/mL), the cell activity reaches more than 95%, and no obvious cytotoxicity exists. Supplement of specific concentration scheme with the accompanying drawings
FIG. 5 is a schematic diagram of H 2 O 2 Induction of chondrocytes construction of an OA in vitro cell model followed by further investigation of Cu-N using the active oxygen fluorescent probe 2',7' -dichlorodiacetate (DCFH-DA) 4 ClG ability to clear ROS in chondrocytes. The DCFH-DA chemical probe can be oxidized by the ROS in the cells, so that green fluorescence is generated, and the fluorescence intensity can reflect the ROS content in the cells. As shown in the figure, H 2 O 2 Stimulation can significantly increase green fluorescence intensity, indicating that excessive ROS is generated. And G+H 2 O 2 The group (100 ug/mL) still maintains a strong green fluorescence intensity, consistent with H 2 O 2 The green fluorescence content of the group was close. But when Cu-N is added 4 After ClG (100 ug/mL), the green fluorescence was significantly reduced by more than 70%. Thus, it was revealed that Cu-N having excellent CAT and SOD-like enzymatic activities 4 ClG is capable of substantially scavenging ROS in OA chondrocytes.
According to the preparation method, graphene is used as a carrier, copper phthalocyanine with a specific Cu coordination environment is used as a Cu source and an N source, the Cl N-doped graphene loaded Cu monoatomic nano enzyme is constructed, and the catalyst has excellent CAT and SOD-like activity and is first applied to treatment of OA. Experimental results prove that Cu-N 4 ClG can effectively remove O 2 ·-、·OH、H 2 O 2 And the equivalent free radicals have excellent CAT and SOD enzyme activities.
The above description is only of the preferred embodiments of the present invention and it is not intended to limit the scope of the present invention, but various modifications and variations can be made by those skilled in the art. Variations, modifications, substitutions, integration and parameter changes may be made to these embodiments by conventional means or may be made to achieve the same functionality within the spirit and principles of the present invention without departing from such principles and spirit of the invention.
Claims (6)
1. A Cu monoatomic nano-enzyme is characterized by comprising a compound Cu-N 4 ClG;
the preparation method of the Cu monoatomic nano-enzyme comprises the following steps:
step S21, adding graphene and copper phthalocyanine CuPc into triple distilled water for centrifugation, wherein the mass ratio of the graphene to the copper phthalocyanine is 1-4: 2-10;
s22, carrying out ultrasonic treatment on the product obtained in the step S21;
s23, washing and drying the product obtained in the step S22 to obtain a CuPc functionalized G composite material;
s24, carrying out Ar atmosphere heat treatment on the CuPc functionalized G composite material, heating the product obtained in the S23 to 600-900 ℃ in Ar atmosphere, and carrying out heat treatment for 1-5 h;
s25, cooling the product obtained in the step S24, and then placing the cooled product in an HCl solution for stirring treatment, wherein the concentration of the HCl solution is 3 mol/L;
step S26, washing and drying to obtain the monoatomic nano-enzyme Cu-N 4 ClG。
2. The Cu monoatomic nano-enzyme according to claim 1, wherein in step S22, the ultrasonic treatment time is 1 to 6 hours.
3. The Cu monoatomic nanoenzyme according to claim 1, wherein in step S23, the washing method is: repeatedly washing with ethanol;
the drying treatment method comprises the following steps: dried under vacuum at 40 ℃ for 12h.
4. The Cu monoatomic nanoenzyme of claim 1, wherein in step S25, the stirring is performed at 40-80 ℃ for 6-18h.
5. The Cu monoatomic nano-enzyme according to claim 1, wherein in step S26, the method of washing and drying is: the solution was washed to neutrality and dried under vacuum at 60 ℃ for 24h.
6. The use of a Cu monoatomic nanoenzyme according to claim 1 for the manufacture of a medicament for osteoarthritis.
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