Sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material and preparation method thereof

Abstract

On one hand, the invention relates to a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material which is prepared from an S, Se-CQDs solution and a Halicin solution in parts by weight as follows: 50-120 parts of the S, Se-CQDs solution, 0.1-0.3 parts of an EDC chemical reagent, 0.1-0.3 parts of an NHS chemical reagent, 125-300 parts of the Halicin solution and 12-35 parts of a NaOH solution, and on the other hand, the invention relates to a preparation method of the sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material. The characteristics that the sulfur-selenium-doped carbon dots are uniform in size and easy to enter cells are utilized, the sulfur-selenium-doped carbon dots can cooperate with the Halicin to enter the cells to destroy the flow of protons on cell membranes, so that the sterilization and antibacterial effects are achieved, the sulfur-selenium-doped carbon dots and the Halicin synergistic antibacterial material can form a hydration layer on the surface of a wound by utilizing the oxidation resistance of the sulfur-selenium-doped carbon dots, and the antibacterial effect of the wound is improved. The adhesion of bacteria is prevented, so that the antibacterial effect is enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field of medical antibacterial materials, and in particular to a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material and a preparation method thereof. Background Art

[0002] Since sulfur-selenium-doped carbon quantum dots have uniform size, strong fluorescence, simple preparation process, low cost, strong antioxidant ability, low toxicity and easy entry into cells, they have great application potential as biological antibacterial and antioxidant materials.

[0003] Halicin is a broad-spectrum antibiotic that can effectively inhibit a variety of pathogens, including those bacteria that are resistant to existing antibiotics. Halicin's mechanism of action is unique. It does not block conventional pathways such as cell wall biosynthesis, DNA repair or protein synthesis, but instead disrupts the flow of protons across the cell membrane. This disruption interferes with the bacteria's ability to maintain an electrochemical gradient, which is essential for the synthesis of ATP (the cell's energy currency). Once the electrochemical gradient is destroyed, the bacterial cell dies. Because of this unique mechanism of action, it may be difficult for bacteria to quickly develop resistance.

[0004] Sulfur-selenium doped carbon quantum dots and Halicin are both commonly used medical antibacterial materials, but the antibacterial effects of these two materials when used separately are not optimal. The present invention aims to provide a material with synergistic antibacterial properties of sulfur-selenium doped carbon dots and Halicin. Summary of the invention

[0005] In view of the shortcomings of the prior art, the present invention provides a sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material and a preparation method thereof. The specific technical scheme is as follows:

[0006] One of the purposes of the present invention is to provide a sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material, comprising an S, Se-CQDs solution and a Halicin solution, the weight fractions of which are: 50-120 parts of S, Se-CQDs solution, 0.1-0.3 parts of EDC chemical reagent, 0.1-0.3 parts of NHS chemical reagent, 125-300 parts of Halicin solution, and 12-35 parts of NaOH solution.

[0007] S, Se-CQDs stands for sulfur-selenium doped carbon quantum dots, or sulfur-selenium doped carbon dots for short. They have the characteristics of uniform size and strong fluorescence, strong antioxidant ability, low toxicity and are easy to enter cells.

[0008] Halicin stands for hailithromycin. As an antibiotic, it can effectively inhibit a variety of pathogens, including those bacteria that are resistant to existing antibiotics. It does this by disrupting the flow of protons across the cell membrane. This disruption interferes with the bacteria's ability to maintain an electrochemical gradient, which is essential for the synthesis of ATP (the energy currency of the cell). Once the electrochemical gradient is destroyed, the bacterial cells die. Because this mechanism of action is relatively unique, it may be difficult for bacteria to quickly develop drug resistance.

[0009] Therefore, the antibacterial material made by combining S, Se-CQDs and Halicin can not only allow S, Se-CQDs to cooperate with Halicin to enter the bacteria for effective sterilization, further improving the internal sterilization effect, but also utilize the antioxidant properties of S, Se-CQDs to enable the antibacterial material to form a hydration layer on the wound surface to prevent bacterial adhesion, thereby enhancing the external antibacterial effect.

[0010] As an improvement of the above technical solution, the composition weight fractions of S, Se-CQDs in the S, Se-CQDs solution are: 40-70 parts of pure water, 5-8 parts of citric acid, 0.8-1.2 parts of L-cysteine, and 0.06-0.1 parts of sodium selenite.

[0011] As an improvement of the above technical solution, the concentration range of the NaOH solution is between 0.01 mol / L and 0.5 mol / L.

[0012] Another object of the present invention is to provide a method for preparing a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material, comprising the following steps:

[0013] S1: Prepare component A: Add EDC chemical reagent and NHS chemical reagent into S, Se-CQDs solution and stir thoroughly to dissolve;

[0014] S1: preparing component B: adding a NaOH solution with a concentration ranging from 0.01 mol / L to 0.5 mol / L to component A, adjusting the initial pH of the resulting solution to 7.0, and then shaking the solution at a constant temperature of 25°C for 0.5 hours;

[0015] S3: Preparation of component C: Slowly add the Halicin solution dropwise to component B under stirring conditions in the dark and stir thoroughly for 4 h;

[0016] S4: Collection: After taking out the prepared C component, centrifuge at 10000 rpm for 3 minutes to collect the S, Se-CQDs-Hal nanoparticles by centrifugation, wash them with deionized water several times to remove unreacted EDC, NHS and S, Se-CQDs, and then dry them in a vacuum drying oven.

[0017] As an improvement of the above technical solution, the preparation of S, Se-CQDs includes the following steps:

[0018] (1) Dissolve citric acid, L-cysteine ​​and sodium selenite in pure water, then transfer the solution into a polytetrafluoroethylene liner, insert the liner into a stainless steel, and finally place the stainless steel reactor into an oven and react at 150°C for 150 min;

[0019] (2) The reaction solution obtained in step (1) was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200 Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S and Se-CQDs.

[0020] Beneficial effects of the present invention:

[0021] By utilizing the characteristics of sulfur-selenium-doped carbon dots due to their uniform size and easy entry into cells, they can cooperate with Halicin to enter cells and disrupt the flow of protons on the cell membrane, thereby achieving a sterilization and antibacterial effect. In addition, by utilizing the antioxidant properties of sulfur-selenium-doped carbon dots, the sulfur-selenium-doped carbon dots and Halicin antibacterial materials can cooperate to form a hydration layer on the wound surface to prevent bacterial adhesion, thereby enhancing the antibacterial effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The following are the antibacterial effect diagrams of different antibacterial materials of the present invention under a microscope. DETAILED DESCRIPTION

[0023] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0024] Embodiment 1:

[0025] A sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material comprises an S, Se-CQDs solution and a Halicin solution, wherein the weight fractions of the composition are: 50-120 parts of the S, Se-CQDs solution, 0.1-0.3 parts of an EDC chemical reagent, 0.1-0.3 parts of an NHS chemical reagent, 125-300 parts of a Halicin solution, and 12-35 parts of a NaOH solution.

[0026] S, Se-CQDs stands for sulfur-selenium doped carbon quantum dots, or sulfur-selenium doped carbon dots for short. They have the characteristics of uniform size and strong fluorescence, strong antioxidant ability, low toxicity and are easy to enter cells.

[0027] Halicin stands for hailithromycin. As an antibiotic, it can effectively inhibit a variety of pathogens, including those bacteria that are resistant to existing antibiotics. It does this by disrupting the flow of protons across the cell membrane. This disruption interferes with the bacteria's ability to maintain an electrochemical gradient, which is essential for the synthesis of ATP (the energy currency of the cell). Once the electrochemical gradient is destroyed, the bacterial cells die. Because this mechanism of action is relatively unique, it may be difficult for bacteria to quickly develop drug resistance.

[0028] Therefore, the antibacterial material made by combining S, Se-CQDs and Halicin is S, Se-CQDs-Hal, which can not only enable S, Se-CQDs to cooperate with Halicin to enter the bacteria for effective sterilization, further improving the internal sterilization effect, but also utilize the antioxidant properties of S, Se-CQDs to enable the antibacterial material to form a hydration layer on the wound surface to prevent bacterial adhesion, thereby enhancing the external antibacterial effect.

[0029] Among them, the EDC chemical reagent, namely 1-ethyl-(3-dimethylaminopropyl)carbodiimide, is an organic compound with a chemical formula of C8H17N3. It can be used as a carboxyl activator and can also be used to activate phosphate groups.

[0030] NHS chemical reagent, namely N-hydroxysulfosuccinimide, is a polypeptide condensation agent that can form a stable active ester intermediate and is used to prepare hydrophilic active esters, such as a protein cross-linking agent.

[0031] The composition weight fractions of S, Se-CQDs in the S, Se-CQDs solution are: 40-70 parts of pure water, 5-8 parts of citric acid, 0.8-1.2 parts of L-cysteine, and 0.06-0.1 parts of sodium selenite.

[0032] Among them, citric acid is an important organic acid, also known as citric acid, colorless crystal, often containing one molecule of crystal water, odorless, with a strong sour taste, easily soluble in water, and its calcium salt is more soluble in cold water than in hot water. This property is often used to identify and separate citric acid. Anhydrous citric acid can be obtained by controlling the appropriate temperature during crystallization. It has a wide range of uses in industry, food industry, cosmetics industry, etc.

[0033] L-cysteine ​​is a common amino acid in organisms. It exists in many proteins and glutathione. It can form insoluble thiolates with metal ions such as Ag+, Hg+, and Cu+.

[0034] Sodium selenite is white crystal or crystalline powder, soluble in water but insoluble in ethanol. It will precipitate selenium when it meets reducing agent.

[0035] A dilute NaOH solution is used, and the concentration range of the NaOH solution is between 0.01 mol / L and 0.5 mol / L.

[0036] A method for preparing a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material comprises the following steps:

[0037] S1: Prepare component A: Add EDC chemical reagent and NHS chemical reagent into S, Se-CQDs solution and stir thoroughly to dissolve;

[0038] S1: preparing component B: adding a NaOH solution with a concentration ranging from 0.01 mol / L to 0.5 mol / L to component A, adjusting the initial pH of the resulting solution to 7.0, and then shaking the solution at a constant temperature of 25°C for 0.5 hours;

[0039] S3: Preparation of component C: Slowly add the Halicin solution dropwise to component B under stirring conditions in the dark and stir thoroughly for 4 h;

[0040] S4: Collection: After taking out the prepared C component, centrifuge at 10000 rpm for 3 minutes to collect the S, Se-CQDs-Hal nanoparticles by centrifugation, wash them with deionized water several times to remove unreacted EDC, NHS and S, Se-CQDs, and then dry them in a vacuum drying oven.

[0041] The preparation of S, Se-CQDs includes the following steps:

[0042] (1) Dissolve citric acid, L-cysteine ​​and sodium selenite in pure water, then transfer the solution into a polytetrafluoroethylene liner, insert the liner into a stainless steel, and finally place the stainless steel reactor into an oven and react at 150°C for 150 min;

[0043] (2) The reaction solution obtained in step (1) was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200 Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S and Se-CQDs.

[0044] Embodiment 2:

[0045] A sulfur-selenium-doped carbon dot and Halicin synergistic antibacterial material, which is composed of 50 parts of S, Se-CQDs solution, 0.1 part of EDC chemical reagent, 0.1 part of NHS chemical reagent, 125 parts of Halicin solution, and 12 parts of NaOH solution in weight fractions.

[0046] The composition weight fractions of S, Se-CQDs in the S, Se-CQDs solution are: 40 parts of pure water, 5 parts of citric acid, 0.8 parts of L-cysteine, and 0.06 parts of sodium selenite.

[0047] A method for preparing a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material comprises the following steps:

[0048] S1: Prepare component A: Add EDC chemical reagent and NHS chemical reagent into S, Se-CQDs solution and stir thoroughly to dissolve;

[0049] S1: preparing component B: adding a NaOH solution with a concentration ranging from 0.01 mol / L to 0.5 mol / L to component A, adjusting the initial pH of the resulting solution to 7.0, and then shaking the solution at a constant temperature of 25°C for 0.5 hours;

[0050] S3: Preparation of component C: Slowly add the Halicin solution dropwise to component B under stirring conditions in the dark and stir thoroughly for 4 h;

[0051] S4: Collection: After taking out the prepared C component, centrifuge at 10000 rpm for 3 minutes to collect the S, Se-CQDs-Hal nanoparticles by centrifugation, wash them with deionized water several times to remove unreacted EDC, NHS and S, Se-CQDs, and then dry them in a vacuum drying oven.

[0052] The preparation of S, Se-CQDs includes the following steps:

[0053] (1) Dissolve citric acid, L-cysteine ​​and sodium selenite in pure water, then transfer the solution into a polytetrafluoroethylene liner, insert the liner into a stainless steel, and finally place the stainless steel reactor into an oven and react at 150°C for 150 min;

[0054] (2) The reaction solution obtained in step (1) was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200 Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S and Se-CQDs.

[0055] The sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material prepared by the above method, Halicin alone, the prepared S, Se-CQDs and PBS were added to the culture dishes of the same pathogen at the same concentration, and the inactivation rate of the pathogens in the culture dishes was detected after 30 minutes.

[0056] PBS is phosphate buffer saline, which is generally used as a solvent to dissolve and protect reagents. It can provide a stable ionic environment and is essential for maintaining the structure and function of biologically active substances. It is the most widely used buffer in biochemical research.

[0057]

[0058]

[0059] Embodiment three:

[0060] The difference from Example 2 is that the amounts of the components are different, as follows:

[0061] A sulfur-selenium-doped carbon dot and Halicin synergistic antibacterial material, which is composed of 120 parts of S, Se-CQDs solution, 0.3 parts of EDC chemical reagent, 0.3 parts of NHS chemical reagent, 300 parts of Halicin solution, and 35 parts of NaOH solution in weight fractions.

[0062] The composition weight fractions of S, Se-CQDs in the S, Se-CQDs solution are: 70 parts of pure water, 8 parts of citric acid, 1.2 parts of L-cysteine, and 0.1 parts of sodium selenite.

[0063] A method for preparing a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material comprises the following steps:

[0064] S1: Prepare component A: Add EDC chemical reagent and NHS chemical reagent into S, Se-CQDs solution and stir thoroughly to dissolve;

[0065] S1: preparing component B: adding a NaOH solution with a concentration ranging from 0.01 mol / L to 0.5 mol / L to component A, adjusting the initial pH of the resulting solution to 7.0, and then shaking the solution at a constant temperature of 25°C for 0.5 hours;

[0066] S3: Preparation of component C: Slowly add the Halicin solution dropwise to component B under stirring conditions in the dark and stir thoroughly for 4 h;

[0067] S4: Collection: After taking out the prepared C component, centrifuge at 10000 rpm for 3 minutes to collect the S, Se-CQDs-Hal nanoparticles by centrifugation, wash them with deionized water several times to remove unreacted EDC, NHS and S, Se-CQDs, and then dry them in a vacuum drying oven.

[0068] The preparation of S, Se-CQDs includes the following steps:

[0069] (1) Dissolve citric acid, L-cysteine ​​and sodium selenite in pure water, then transfer the solution into a polytetrafluoroethylene liner, insert the liner into a stainless steel, and finally place the stainless steel reactor into an oven and react at 150°C for 150 min;

[0070] (2) The reaction solution obtained in step (1) was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200 Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S and Se-CQDs.

[0071] The sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material prepared by the above method, Halicin alone, the prepared S, Se-CQDs and PBS were added to the culture dishes of the same pathogen at the same concentration, and the inactivation rate of the pathogens in the culture dishes was detected after 30 minutes.

[0072] PBS is phosphate buffer saline, which is generally used as a solvent to dissolve and protect reagents. It can provide a stable ionic environment and is essential for maintaining the structure and function of biologically active substances. It is the most widely used buffer in biochemical research.

[0073]

[0074] Embodiment 4:

[0075] like Figure 1 As shown, Figure 1 The following are the antibacterial effect diagrams of different antibacterial materials of the present invention under a microscope.

[0076] The materials were prepared in precise amounts according to the ratios in Example 1, as follows:

[0077] Citric acid (5 g), L-cysteine ​​(0.8 g) and sodium selenite (0.06 g) were dissolved in 40 mL of pure water, and then the solution was transferred into a 50 mL polytetrafluoroethylene liner, and the liner was inserted into stainless steel. Finally, the stainless steel reactor was placed in an oven and reacted at 150 ° C for 150 min. After taking out, the reaction solution was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S, Se-CQDs.

[0078] EDC (1 mmol) and NHS (1 mmol) were added to 5 mL of S, Se-CQDs solution (1 mg / mL), stirred thoroughly to dissolve, and the initial pH of the resulting solution was adjusted to 7.0 using a dilute NaOH solution with a concentration range of 0.01 mol / L-0.5 mol / L. The solution was then shaken at a constant temperature of 25°C for 0.5 h. Under dark stirring conditions, 12.5 ml of Halicin (1 mg / ml) solution was slowly added dropwise and stirred thoroughly for 4 h. After taking it out, it was centrifuged at a speed of 10000 rpm for 3 minutes. S, Se-CQDs-Hal nanoparticles were collected by centrifugation, washed several times with deionized water to remove unreacted EDC, NHS and S, Se-CQDs, and then dried in a vacuum drying oven.

[0079] The sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material prepared by the above method, Halicin alone, the prepared S, Se-CQDs and PBS were added to the culture dishes of the same pathogen at the same concentration, and the inactivation rate of the pathogens in the culture dishes was detected after 30 minutes.

[0080] PBS is phosphate buffer saline, which is generally used as a solvent to dissolve and protect reagents. It can provide a stable ionic environment and is essential for maintaining the structure and function of biologically active substances. It is the most widely used buffer in biochemical research.

[0081]

[0082] in Figure 1 The Coli in "Escherichia coli" refers to Escherichia coli, a common Gram-negative bacillus that lives in the intestines of humans and animals. Most E. coli are harmless, but some serotypes of E. coli can cause diarrhea and other diseases in humans.

[0083] Figure 1 The aureus in the word refers to Staphylococcus aureus, which is a common pathogen that can be found in humans and animals. It is a Gram-positive bacterium that is usually spherical and can form yellow colonies on laboratory culture media, hence the name "golden" Staphylococcus. This bacterium can cause a variety of infections, including skin infections, food poisoning, pneumonia, etc.

[0084] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material, characterized in that: It comprises S, Se-CQDs solution and Halicin solution, and its weight fractions are: 50-120 parts of S, Se-CQDs solution, 0.1-0.3 parts of EDC chemical reagent, 0.1-0.3 parts of NHS chemical reagent, 125-300 parts of Halicin solution, and 12-35 parts of NaOH solution.

2. The sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material according to claim 1, characterized in that: The composition weight fractions of S, Se-CQDs in the S, Se-CQDs solution are: 40-70 parts of pure water, 5-8 parts of citric acid, 0.8-1.2 parts of L-cysteine, and 0.06-0.1 parts of sodium selenite.

3. The sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material according to claim 1, characterized in that: The concentration of the NaOH solution is in the range of 0.01 mol / L to 0.5 mol / L.

4. A method for preparing a sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material, based on the sulfur-selenium doped carbon dot and Halicin synergistic antibacterial material according to any one of claims 1 to 3, characterized in that: The following steps are involved: S1: Prepare component A: Add EDC chemical reagent and NHS chemical reagent into S, Se-CQDs solution and stir thoroughly to dissolve; S1: preparing component B: adding a NaOH solution with a concentration ranging from 0.01 mol / L to 0.5 mol / L to component A, adjusting the initial pH of the resulting solution to 7.0, and then shaking the solution at a constant temperature of 25°C for 0.5 hours; S3: Preparation of component C: Slowly add the Halicin solution dropwise to component B under stirring conditions in the dark and stir thoroughly for 4 h; S4: Collection: After taking out the prepared C component, centrifuge at 10000 rpm for 3 minutes to collect the S, Se-CQDs-Hal nanoparticles by centrifugation, wash them with deionized water several times to remove unreacted EDC, NHS and S, Se-CQDs, and then dry them in a vacuum drying oven.

5. The method for preparing a sulfur-selenium doped carbon dots and Halicin synergistic antibacterial material according to claim 4, characterized in that: The preparation of S, Se-CQDs includes the following steps: (1) Dissolve citric acid, L-cysteine ​​and sodium selenite in pure water, then transfer the solution into a polytetrafluoroethylene liner, insert the liner into a stainless steel, and finally place the stainless steel reactor into an oven and react at 150°C for 150 min; (2) The reaction solution obtained in step (1) was centrifuged at 8000 r / min for 20 min, the supernatant was separated, and dialyzed with a 200 Da dialysis bag for 2 days, changing it every 5 hours, and freeze-dried to obtain S and Se-CQDs.

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