Preparation method of nanometer enzyme high-efficiency antibacterial agent

Abstract

The application provides a preparation method of a novel nano-enzyme high-efficiency antibacterial agent, and the specific operation steps are as follows: 1, CuAC2 is weighed and dissolved in N,N-dimethylformamide; 2, sodium phenyl dodecyl sulfonate is added, and after stirring for one hour, the mixture is moved into a reaction kettle; 3, thioacetamide is added, and after continuing to stir for 10 minutes, the mixture is sealed; 4, after constant temperature heating in a 180-degree oven for 24 hours, the mixture is taken out, cooled to room temperature, centrifuged, washed and dried, and the powder obtained is the novel nano-enzyme high-efficiency antibacterial agent. The novel nano-enzyme high-efficiency antibacterial agent prepared by the method can generate sufficient ROS (hydroxyl radicals) under the condition of no light, and is used for killing various typical drug-resistant bacteria.

Description

Technical Field

[0001] This invention relates to the field of antibacterial agents, and more particularly to a method for preparing a highly efficient antibacterial nanozyme agent. Background Technology

[0002] Pathogens pose a serious threat to global public health, often invading immunocompromised populations and exacerbating their conditions. In modern medicine, the widespread overuse of antibiotics leads to bacterial gene mutations, biofilm formation, and drug resistance, significantly reducing the efficacy of broad-spectrum antibiotics. Nanomaterials, due to their unique properties at the nanoscale, have attracted widespread attention as antibacterial systems. Some nanoparticles have shown good antibacterial activity against various bacteria. Among these, the use of nanomaterials in photodynamic therapy for antibacterial purposes is a promising strategy; however, improving the light conversion efficiency of nanomaterials under specific light conditions to achieve excellent antibacterial activity remains a significant challenge. Cu2S is a common and highly effective antibacterial nanoenzyme, but current Cu2S only generates ROS under near-infrared light irradiation to achieve sterilization, making the conditions for its generation relatively demanding and resulting in poor antibacterial efficacy.

[0003] This application modifies the surface atomic arrangement and structure of nanomaterials, particularly by introducing defects for vacancy engineering, which can effectively change their electronic structure and regulate their physicochemical properties. This provides a new approach to optimizing the antibacterial properties of nanomaterials. Summary of the Invention

[0004] To address the above technical problems, this invention provides a method for preparing a highly efficient antibacterial nanozyme agent, the specific steps of which are as follows:

[0005] 1. Weigh out CuAC2 and dissolve it in N,N-dimethylformamide;

[0006] 2. Add sodium phenyl dodecyl sulfonate, stir for one hour, and then transfer to a reaction vessel;

[0007] 3. After adding the thioacetamide, continue stirring for 10 minutes and then seal.

[0008] 4. After heating at a constant temperature of 180 degrees Celsius for 24 hours, remove the product, cool it to room temperature, centrifuge, wash, and dry it to obtain the powder, which is the new type of nanoenzyme high-efficiency antibacterial product.

[0009] The mass ratio of CuAC2, N,N-dimethylformamide, sodium phenyl dodecyl sulfonate, and thioacetamide is 35:3776:280:45.

[0010] In step 4, the material was heat-treated at 180°C for 24 hours in a constant temperature oven to precisely control the crystal structure of copper sulfide (Cu2S) and form sulfur vacancies, thus successfully preparing Cu2S and Cu2S-100 with different vacancy associations.

[0011] N,N-Dimethylformamide (DMF) can be used as a solvent to dissolve the reactants and promote the reaction.

[0012] Sodium phenyl dodecyl sulfonate can be used as a surfactant to help improve the dispersibility of reactants and the reaction rate.

[0013] Thioacetamide provides the sulfur source.

[0014] The beneficial effects of this invention are as follows:

[0015] The novel nanozyme prepared by the method described in this application has a high efficiency in antibacterial activity and can generate sufficient amounts of ROS (hydroxyl radicals) under light-free conditions, which can be used to kill a variety of typical drug-resistant bacteria.

[0016] Most commercially available synthetic methods use thiourea as the sulfur source. However, this application uses thioacetamide in its synthesis. The advantages are as follows:

[0017] 1. Controllability of sulfide ion release

[0018] Thioacetamide slowly releases sulfide ions (S²⁻) via hydrolysis in acidic or neutral aqueous solutions; the reaction is mild and controllable. The release rate can be adjusted by pH and temperature. Thiourea decomposes under high temperature or strongly acidic conditions. At 180°C, the reaction releases S²⁻ at a faster rate, which can easily lead to local supersaturation and may generate uneven precipitates or nanoparticles.

[0019] 2. Homogeneous precipitation: The slow release of S²⁻ by TAA is beneficial for generating nanomaterials with uniform particle size and regular morphology (such as spherical and cubic sulfide particles).

[0020] 3. The hydrolysis byproduct of TAA is acetamide (which has no complexing properties), while the decomposition of thiourea may generate nitrogen-containing byproducts (such as cyanamide), which may interfere with the coordination of metal ions.

[0021] In the conventional synthesis of Cu₂S, the reagents previously used were mostly N,N-dimethylacetamide (DMAc) rather than N,N-dimethylformamide (DMF). This application uses N,N-dimethylformamide (DMF) for three advantages:

[0022] 1. Since DMF is more volatile than DMAc, it is easier to remove under subsequent high-temperature treatment, which is beneficial for obtaining purer Cu2S in subsequent separation and purification steps.

[0023] 2. Although DMF is less soluble than DMAc for most substances, this avoids the intermediate and byproducts that may occur in the reaction. Furthermore, experimental data showed that the solubility of sulfides and the reactivity of ions in both reagents did not affect the yield of Cu2S.

[0024] 3. DMF has low toxicity and low cost. For future large-scale applications, it has lower production requirements, lower manufacturing costs, and better economic efficiency and controllability. Attached Figure Description

[0025] Figure 1 This is an X-ray crystal diffraction analysis diagram of the present invention;

[0026] Figure 2 This is the XRD pattern of the synthesized product of this invention;

[0027] Figure 3 This is a graph showing the performance test of the peroxide oxidase product of this invention;

[0028] Figure 4 This is a graph showing the performance test of the peroxidase product of this invention;

[0029] Figure 5 This is a paramagnetic detection diagram under no-light conditions according to the present invention;

[0030] Figure 6 The results show the inactivation rate of the drug-resistant Pseudomonas aeruginosa in this invention.

[0031] Figure 7 This is the result of the Klebsiella pneumoniae inactivation rate of the present invention;

[0032] Figure 8 This is the result of the inactivation rate of Klebsiella pneumoniae in this invention;

[0033] Figure 9 This is the result of the Staphylococcus aureus inactivation rate in this invention;

[0034] Figure 10 This is the result of the Staphylococcus aureus inactivation rate in this invention;

[0035] Figure 11 This is the result of the Pseudomonas aeruginosa inactivation rate of the present invention;

[0036] Figure 12 This is the result of the Pseudomonas aeruginosa inactivation rate of the present invention;

[0037] Figure 13 This is the result of the Acinetobacter baumannii inactivation rate in this invention;

[0038] Figure 14 The results show the inactivation rate of Acinetobacter baumannii in this invention. Detailed Implementation Example

[0039] Prepare the following raw materials: CuAC2 0.35g (anhydrous, 98%, CAS No.: 142-71-2), N,N-dimethylformamide 40ml (AR, CAS No.: 68-12-2), sodium phenyl dodecyl sulfonate 2.8g (≥95.0%, mixture, CAS No.: 25155-30-0), and thioacetamide 0.45g (≥98.0%, CAS No.: 62-55-5).

[0040] Weigh out 0.35 g of CuAC2 and dissolve it in 40 mL of N,N-dimethylformamide. Add 8 mmol (about 2.8 g) of sodium phenyl dodecyl sulfonate and stir for one hour. Transfer the mixture to a 50 mL reactor and add 0.45 g of thioacetamide. Continue stirring for 10 minutes and then seal the reactor. Heat the reactor at 180 degrees Celsius for 24 hours. Remove the reactor, cool it to room temperature, centrifuge, wash and dry it. Cu2S and Cu2S-100 with different vacancy associations were successfully prepared.

[0041] The above products were analyzed using instruments. X-ray crystallography analysis showed that the synthesized product was Cu₂S (PDF#03-1071), and XRD (as shown) Figure 1 Scanning electron microscopy analysis showed that the synthesized product consisted of 70-80 nanometer particles (e.g., ...). Figure 2 Analysis showed that the synthesized product contained peroxidase (…). Figure 3 ) and the performance of peroxidase ( Figure 4 This is a multifunctional nanozyme. Paramagnetic detection under no-light conditions reveals a strong hydroxyl radical (H2O) reaction. Figure 5 ). Example

[0042] Prepare the following raw materials: CuAC2 0.7g (anhydrous, 98%, CAS No.: 142-71-2), N,N-dimethylformamide 80ml (AR, CAS No.: 68-12-2), sodium phenyl dodecyl sulfonate 5.6g (≥95.0%, mixture, CAS No.: 25155-30-0), and thioacetamide 0.9g (≥98.0%, CAS No.: 62-55-5).

[0043] Weigh out 0.7 g of CuAC2 and dissolve it in 80 mL of N,N-dimethylformamide. Add 16 mmol (approximately 5.6 g) of sodium phenyl dodecyl sulfonate and stir for one hour. Transfer the mixture to a 100 mL reactor and add 0.9 g of thioacetamide. Continue stirring for 10 minutes and then seal the reactor. Heat the reactor at 180°C for 24 hours. Remove the reactor, cool it to room temperature, centrifuge, wash, and air dry. Cu2S and Cu2S-100 with different vacancy associations were successfully prepared. Example

[0044] Under conditions of 50 ppm inactivation and no light, the inactivation rate of drug-resistant Pseudomonas aeruginosa was greater than 99.99%, as shown in the results. Figure 6 . Example

[0045] Under conditions of 50 ppm inactivation of drug-resistant bacteria and no light, the inactivation rate of Klebsiella pneumoniae was greater than 99.99%, as shown in the results. Figure 7 , 8 . Example

[0046] Under conditions of 50 ppm inactivation of drug-resistant bacteria and no light, the inactivation rate of Staphylococcus aureus was greater than 99.99%, as shown in the results. Figure 9 , 10 . Example

[0047] Under conditions of 50 ppm inactivation of drug-resistant bacteria and no light, the inactivation rate of *Pseudomonas aeruginosa* was greater than 99.99%, as shown in the results. Figure 11 , 12 . Example

[0048] Under conditions of 50 ppm inactivation of drug-resistant bacteria and no light, the inactivation rate of Acinetobacter baumannii was greater than 99.99%, as shown in the results. Figure 13 , 14 .

[0049] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. The various components mentioned in this invention are common technologies in the existing field. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a highly efficient antibacterial nanozyme, the specific steps of which are as follows: (1) Weigh out CuAC2 and dissolve it in N,N-dimethylformamide; (2) Add sodium phenyl dodecyl sulfonate, stir for one hour, and then transfer it into the reaction vessel; (3) After adding thioacetamide, continue stirring for 10 minutes and then seal. (4) After heating in a 180-degree oven for 24 hours, remove the product, cool it to room temperature, centrifuge, wash and dry it to obtain the powder, which is the nano-enzyme high-efficiency antibacterial product. The CuAC 2、 The mass ratio of N,N-dimethylformamide, sodium phenyl dodecyl sulfonate, and thioacetamide is 35:3776:280:45; In step 4, the material was heat-treated at 180°C for 24 hours in a constant temperature oven to precisely control the crystal structure of copper sulfide Cu2S, forming sulfur vacancies, and successfully preparing Cu2S and Cu2S-100 with different vacancy associations.

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