Preparation method of novel nano-enzyme efficient antibacterial agent

Abstract

The invention provides a preparation method of a novel nano-enzyme efficient antibacterial agent, which comprises the following specific operation steps: 1, weighing CuAC2, and dissolving CuAC2 in N, N-dimethylformamide; 2, adding phenyl sodium dodecyl sulfate, stirring for one hour, and transferring into a reaction kettle; 3, adding thioacetamide, continuously stirring for 10 minutes, and sealing; and 4, performing constant-temperature heating in a 180-DEG C oven for 24 hours, taking out, cooling to room temperature, and performing centrifugal washing and airing to obtain powder, namely the novel nano-enzyme efficient antibacterial agent. The novel nano-enzyme prepared by the method disclosed by the invention is efficient and antibacterial, can generate sufficient ROS (hydroxyl free radicals) under a non-illumination condition, and is used for killing various typical drug-resistant bacteria.

Description

Technical Field

[0001] The present invention relates to the field of antibacterial agents, and in particular to a method for preparing a novel nanozyme high-efficiency antibacterial agent. Background Art

[0002] Pathogens pose a serious threat to global public health, often invading people with weakened immune systems and aggravating their condition. In modern medicine, the abuse of antibiotics is widespread, causing bacterial gene mutations, the formation of biofilm structures and drug resistance, which significantly reduces the efficacy of broad-spectrum antibiotics. Nanomaterials have attracted widespread attention as antibacterial systems due to their unique properties at the nanoscale. Some nanoparticles have shown good antibacterial activity against a variety of bacteria. Among them, the use of nanomaterials in photodynamic therapy is a very promising strategy, but improving the photoconversion efficiency of nanomaterials under specific light conditions to obtain excellent antibacterial activity remains a huge challenge. Cu2S is a common nanoenzyme with high efficiency and antibacterial effect, but the current Cu2S can only produce ROS under near-infrared light to achieve a sterilizing effect. The conditions for the production of ROS are relatively harsh and the antibacterial effect is poor.

[0003] This application adjusts the surface atomic arrangement and structure of nanomaterials, especially introduces defects for vacancy engineering, which can effectively change their electronic structure and regulate physical and chemical properties, which provides a new way to optimize the antibacterial properties of nanomaterials. Summary of the Invention

[0004] Based on the above technical problems, the present invention provides a method for preparing a novel nanozyme high-efficiency antibacterial agent, the specific steps of which are as follows: 1. Weigh CuAC2 and dissolve it in N,N-dimethylformamide; 2. Add sodium phenyl dodecyl sulfonate, stir for one hour and then transfer to the reactor; 3. Add thioacetamide, continue stirring for 10 minutes and then seal; 4. Heat in a constant temperature oven at 180 degrees for 24 hours, then take it out, cool it to room temperature, centrifuge it, wash it, and dry it. The powder obtained is the new nanozyme high-efficiency antibacterial product.

[0005] The CuAC 2,、 The mass ratio of N,N-dimethylformamide, sodium phenyl dodecyl sulfonate and thioacetamide is 35:3776:280:45.

[0006] In step 4, the material was heat-treated at 180°C in an oven for 24 hours to precisely manipulate the crystal structure of copper sulfide (Cu2S) and form sulfur vacancies. Cu2S and Cu2S-100 with different vacancy associations were successfully prepared.

[0007] N,N-dimethylformamide (DMF) is used as a solvent to dissolve the reactants and promote the reaction.

[0008] Sodium phenyl dodecyl sulfonate can act as a surfactant to help improve the dispersion of reactants and the reaction rate.

[0009] Thioacetamide provides the sulfur source.

[0010] The beneficial effects of the present invention are: The novel nanozyme prepared by the method of the present application is highly effective in antibacterial activities and can produce sufficient ROS (hydroxyl radicals) in the absence of light to kill a variety of typical drug-resistant bacteria.

[0011] Most of the synthesis methods on the market use thiourea as the sulfur source. However, this application uses thioacetamide in the synthesis. The advantages are as follows 1. Controllability of sulfide ion release Thioacetamide slowly releases sulfide ions (S 2- ), the reaction is mild and controllable. The release rate can be adjusted by pH and temperature. Thiourea will decompose under high temperature or strong acid conditions at 180℃ to release S 2- The rate is fast, which can easily lead to local supersaturation and may generate uneven precipitation or nanoparticles.

[0012] 2. Homogeneous precipitation: TAA slowly releases S 2- The characteristics of sulfide nanoparticles are conducive to the formation of nanomaterials with uniform particle size and regular morphology (such as spherical and cubic sulfide particles).

[0013] 3. The by-product of TAA hydrolysis is acetamide (non-complexing), while thiourea decomposition may produce nitrogen-containing by-products (such as cyanamide), which may interfere with the coordination of metal ions.

[0014] In the conventional synthesis of Cu2S, the reaction reagent used previously was mostly N,N-dimethylacetamide (DMAc) rather than N,N-dimethylformamide (DMF). The use of N,N-dimethylformamide (DMF) in this application has three advantages: Since DMF is more volatile than DMAc, DMF is relatively easier to remove under subsequent high-temperature treatment, which is beneficial for the subsequent separation and purification steps to obtain purer Cu2S.

[0015] 2. Although DMF is less soluble than DMAc in general substances, it also avoids the intermediates and by-products that may appear in the reaction. In addition, through experimental data, we found that the solubility of sulfide in the two reagents and the reactivity of the ions will not affect the yield of Cu2S.

[0016] 3. DMF has low toxicity and low cost. For the future large-scale application, it requires lower production conditions, is cheaper, and has better economy and controllability. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the X-ray crystal diffraction analysis diagram of the present invention; Figure 2 It is the XRD analysis diagram of the synthetic product of the present invention; Figure 3 This is a performance test diagram of the peroxide oxidase product of the present invention; Figure 4 This is a performance test diagram of hydrogen peroxide oxidase, a product of the present invention; Figure 5 This is a paramagnetic detection diagram of the present invention under no light conditions; Figure 6 This is the inactivation rate result of the drug-resistant bacteria Pseudomonas aeruginosa of the present invention; Figure 7 This is the Klebsiella pneumoniae inactivation rate result of the present invention; Figure 8 This is the Klebsiella inactivation rate result of the present invention; Figure 9 This is the Staphylococcus aureus inactivation rate result of the present invention; Figure 10 This is the Staphylococcus aureus inactivation rate result of the present invention; Figure 11 This is the result of the inactivation rate of Pseudomonas aeruginosa of the present invention; Figure 12 This is the result of the inactivation rate of Pseudomonas aeruginosa of the present invention; Figure 13 This is the inactivation rate result of Acinetobacter baumannii of the present invention; Figure 14 This is the inactivation rate result of Acinetobacter baumannii of the present invention. DETAILED DESCRIPTION

[0018] Example 1

[0019] Prepare the raw materials: CuAC2 0.35g (anhydrous, 98%, CAS No.: 142-71-2), N,N-dimethylformamide 40ml (AR, CAS No.: 68-12-2), sodium phenyl dodecylsulfonate 2.8g (≥95.0%, mixture, CAS No.: 25155-30-0), thioacetamide 0.45g (≥98.0%, CAS No.: 62-55-5); Weigh a certain amount of 0.35g of CuAC 2,Dissolved in 40 ml of N,N-dimethylformamide, added a certain amount of 8 mmol (about 2.8 g) of sodium phenyldodecylsulfonate, stirred for one hour, transferred to a 50 ml reactor, added a certain amount of 0.45 g of thioacetamide, continued stirring for 10 minutes, and sealed. After constant temperature heating in an oven at 180 degrees for 24 hours, removed from the oven, cooled to room temperature, centrifuged, washed, and air-dried, Cu2S and Cu2S-100 with different vacancy associations were successfully prepared.

[0020] The above product was analyzed by instruments, and X-ray crystal diffraction analysis showed that the synthesized product was Cu2S (PDF#03-1071). Figure 1 Scanning electron microscopy analysis showed that the synthesized product was 70-80 nanometer particles (such as Figure 2 ); Analysis showed that the synthetic product showed peroxidase ( Figure 3 ) and hydrogen peroxide enzyme performance ( Figure 4 ), a multifunctional nanozyme. Paramagnetic detection under no light conditions showed a strong hydroxyl radical ( Figure 5 ).

[0021] Example 2

[0022] Prepare the raw materials: CuAC2 0.7g (anhydrous, 98%, CAS No.: 142-71-2), N,N-dimethylformamide 80ml (AR, CAS No.: 68-12-2), sodium phenyl dodecylsulfonate 5.6g (≥95.0%, mixture, CAS No.: 25155-30-0), thioacetamide 0.9g (≥98.0%, CAS No.: 62-55-5); Weigh a certain amount of 0.7g of CuAC 2, Dissolved in 80 ml of N,N-dimethylformamide, added a certain amount of 16 mmol (about 5.6 g) of sodium phenyldodecylsulfonate, stirred for one hour, transferred to a 100 ml reactor, added a certain amount of 0.9 g of thioacetamide, continued stirring for 10 minutes, and then sealed. The reaction was heated in a constant temperature oven at 180 degrees for 24 hours, removed, cooled to room temperature, centrifuged, washed, and air-dried. Cu2S and Cu2S-100 with different vacancy associations were successfully prepared.

[0023] Example 3

[0024] Inactivation of drug-resistant bacteria: Under the condition of 50ppm and no light, the inactivation rate of drug-resistant bacteria Pseudomonas aeruginosa is greater than 99.99%. Figure 6 .

[0025] Example 4

[0026] Inactivation of drug-resistant bacteria: Under the condition of 50ppm and no light, the inactivation rate of Klebsiella pneumoniae is greater than 99.99%. Figure 7 、 8 .

[0027] Example 5

[0028] Inactivation of drug-resistant bacteria: Under the condition of 50ppm and no light, the inactivation rate of Staphylococcus aureus is greater than 99.99%. Figure 9 、 10 .

[0029] Example 6

[0030] Inactivation of drug-resistant bacteria: Under the condition of 50ppm and no light, the inactivation rate of Pseudomonas aeruginosa is greater than 99.99%. Figure 11 、 12 .

[0031] Example 7

[0032] Inactivation of drug-resistant bacteria: Under the condition of 50ppm and no light, the inactivation rate of Acinetobacter baumannii is greater than 99.99%. Figure 13 、 14 .

[0033] The above shows and describes the basic principles, main features, and advantages of the present invention. The various components mentioned in the present invention are conventional technologies in the prior art. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and improvements may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and improvements are intended to fall within the scope of the invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a novel nanozyme high-efficiency antibacterial agent, the specific steps of which are: (1) Weigh CuAC2 and dissolve it in N,N-dimethylformamide; (2) Add sodium phenyl dodecyl sulfonate, stir for one hour and then transfer to the reactor; (3) Add thioacetamide, continue stirring for 10 minutes and then seal; (4) After heating in a constant temperature oven at 180 degrees for 24 hours, take it out, cool it to room temperature, centrifuge it, wash it, and dry it. The powder obtained is the new nanozyme high-efficiency antibacterial product.

2. A method for preparing a novel nanozyme high-efficiency antibacterial agent according to claim 1, characterized in that The CuAC 2,、 The mass ratio of N,N-dimethylformamide, sodium phenyl dodecyl sulfonate and thioacetamide is 35:3776:280:

45.

3. A method for preparing a novel nanozyme high-efficiency antibacterial agent according to claim 1, characterized in that In step 4, the material was heat-treated at 180°C in an oven for 24 hours to precisely control the crystal structure of copper sulfide (Cu2S) and form sulfur vacancies. Cu2S and Cu2S-100 with different vacancy associations were successfully prepared.

4. A method for preparing a novel nanozyme high-efficiency antibacterial agent according to claim 1, characterized in that N,N-dimethylformamide (DMF) is used as a solvent to dissolve the reactants and promote the reaction.

5. A method for preparing a novel nanozyme high-efficiency antibacterial agent according to claim 1, characterized in that Sodium phenyl dodecyl sulfonate acts as a surfactant to help improve the dispersion of reactants and the reaction rate.

6. A method for preparing a novel nanozyme high-efficiency antibacterial agent according to claim 1, characterized in that Thioacetamide provides the sulfur source.

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