Antibacterial drug delivery system based on raw lacquer magnetofluid and preparation method thereof

Abstract

The invention provides a method for creating an antibacterial drug delivery system based on co-assembly of raw lacquer, glucan, superparamagnetic iron oxide nanoparticles (SPION) and other substances, the system takes raw lacquer magnetofluid as a core, and advanced material science and drug delivery technology are combined. The antibacterial drug delivery system comprises a raw lacquer magnetic fluid and an antibacterial drug active ingredient. The preparation method comprises the following steps: slowly adding raw lacquer and glucan into an SPION solution through a specific chemical self-assembly method, carrying out ultraviolet radiation self-assembly for a period of time to generate a product DU-Fe, and covalently modifying a surface hydroxyl group through glutaraldehyde to obtain the coating. And carrying out Schiff base reaction to graft D-alanyl-D-alanine (D-Ala-D-Ala) to prepare a carrier D-Ala-D-Ala (at) DU (at) Fe capable of immobilizing an antibacterial drug. And then, immobilizing an antibacterial drug (such as vancomycin) capable of being combined with D-Ala-D-Ala on the surface of the carrier onto the DU-Fe by utilizing an intermolecular acting force to obtain the antibacterial magnetic fluid V-DU-Fe. Finally, the stability and effectiveness of the antibacterial drug delivery system are ensured by evaluating the antibacterial performance after six times of recycling. The antibacterial drug delivery system has good biocompatibility and stability, the preparation method is simple and efficient, industrial production is easy to achieve, and a new choice is provided for killing bacteria in a water environment.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and specifically to an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof. The system uses raw lacquer magnetic fluid as a carrier and combines the active ingredients of antibacterial drugs to achieve accurate delivery and efficient treatment of antibacterial drugs. Background Art

[0002] Excessive bacteria in water bodies is a common problem, especially in the fields of drinking water treatment, food processing, and medical water. Excessive bacteria may cause serious health risks. Traditional methods of treating bacteria in water bodies can remove or kill bacteria in water to a certain extent, but there are problems such as low efficiency and easy secondary pollution. Traditional drug delivery methods have many limitations, such as uneven drug distribution, low bioavailability, and large side effects. These problems seriously affect the therapeutic effect of drugs and the quality of life of patients. Therefore, it is particularly important to develop a new, efficient, and low-side effect antibacterial drug delivery system.

[0003] At present, antibacterial drugs that can directly bind specifically to the bacterial cell wall precursor D-alanyl-D-alanine (D-Ala-D-Ala) and block the action of transglycosylase, transpeptidase and D,D-carboxypeptidase in peptidoglycan synthesis mainly include β-lactam antibiotics, such as penicillins and cephalosporins, and polypeptide antibiotics, such as vancomycin. These drugs inhibit the synthesis of bacterial cell wall peptidoglycan, resulting in bacterial cell wall defects, which in turn causes bacterial lysis and death.

[0004] As a new type of antimicrobial drug carrier, magnetic fluid has shown great application potential in the field of antimicrobial drug delivery due to its unique biocompatibility, magnetic response recovery performance and colloidal dispersibility. As a natural nonionic surfactant, raw lacquer is used as a hemostatic, anti-inflammatory, insecticidal and other medicinal materials. At the same time, raw lacquer also has good biocompatibility and degradability, and is one of the ideal raw materials for preparing magnetic fluid. Although some studies have reported drug delivery systems based on raw lacquer, most of these systems use synthetic polymer materials as carriers, which have problems such as poor biocompatibility and complex preparation process. In contrast, raw lacquer magnetic fluid has natural advantages, its preparation process is relatively simple, and its biocompatibility is better, making it more suitable for use in the biomedical field. Dextran, as a type of polysaccharide, is a polymer with good biocompatibility, non-toxicity and good water solubility. Dextran-coated SPIONs are negatively charged in physiological environments, have better biocompatibility and lower cytotoxicity. A large number of studies have shown that the mixed coating of dextran and polyphenols formed on the surface of nanoparticles can make the surface more hydrophilic and biocompatible. In addition, after coating, the van der Waals forces and steric repulsion between nanoparticles can reduce accumulation and thus be more conducive to antibacterial applications.

[0005] Therefore, the development of an antimicrobial drug delivery system based on raw lacquer and dextran composite magnetic fluid is not only theoretically feasible, but also has practical application value. This system can utilize the magnetic responsiveness of raw lacquer magnetic fluid to achieve precise positioning and controlled release of antimicrobial drugs through an external magnetic field, thereby improving the therapeutic effect and bioavailability of antimicrobial drugs while reducing side effects. Summary of the invention 1. Summary of the Invention The present invention proposes an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof. The system uses raw lacquer magnetic fluid as a carrier and combines the active ingredients of antibacterial drugs to achieve precise delivery and efficient treatment of antibacterial drugs. Raw lacquer magnetic fluid has unique physical and chemical properties, such as good biocompatibility, magnetic responsiveness and colloidal dispersibility, making it an ideal choice for antibacterial drug delivery. 2. Summary of the invention The present invention provides an antimicrobial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, which solves the industrial application problems such as high-efficiency antibacterial and precise recovery of antimicrobial drugs, reduces production and application costs, and improves production economic benefits.

[0008] The technical solution adopted by the present invention is as follows: an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, the steps are: 1. The present invention discloses an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, wherein the specific steps are as follows: at room temperature, raw lacquer and dextran are slowly added to a SPION solution, and the product DU@Fe is generated after ultraviolet irradiation self-assembly for a period of time. Then, glutaraldehyde is slowly added dropwise, and high-speed stirring is continued for a period of time to covalently modify the surface hydroxyl groups. Next, based on the Schiff base reaction, D-alanyl-D-alanine (D-Ala-D-Ala) groups are grafted to prepare a carrier D-Ala-D-Ala@DU@Fe that can immobilize antibacterial drugs. Subsequently, the antibacterial drug (taking vancomycin as an example) that can bind to D-Ala-D-Ala on the carrier surface is immobilized on DU@Fe by using intermolecular forces to obtain the antibacterial magnetic fluid V@DU@Fe. Finally, the antibacterial performance is evaluated after 6 cycles of use to ensure the stability and effectiveness of the antibacterial drug delivery system.

[0009] 2. In a preferred embodiment of the present invention, the raw lacquer magnetic fluid carrier is composed of a magnetic fluid obtained by specifically treating raw lacquer phenol and dextran (10,000 Da to 100,000 Da), and has good colloidal dispersibility and biocompatibility.

[0010] 3. In a preferred embodiment of the present invention, the raw lacquer magnetic fluid carrier has adjustable magnetic responsiveness (-90 ~ +90 emu / g) and can be controlled to be recycled and reused by an external magnetic field.

[0011] 4. In a preferred embodiment of the present invention, the active ingredients of the antibacterial drugs include but are not limited to β-lactam antibiotics that can specifically bind to D-alanyl-D-alanine (D-Ala-D-Ala), such as penicillins and cephalosporins, and polypeptide antibiotics, such as vancomycin, etc., which are selected according to treatment needs.

[0012] 5. An antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, characterized in that the method comprises the following steps: Step A: Raw lacquer and dextran are placed in a buffer solution (pH 3-6) and subjected to ultraviolet irradiation (100-500W) to synthesize raw lacquer magnetic fluid, and covalently bind glutaraldehyde and D-Ala-D-Ala on the surface to facilitate binding with active molecules of antibacterial drugs; Step B: using intermolecular forces to specifically combine the antibacterial drug and D-Ala-D-Ala, so as to achieve the purpose of grafting onto the raw lacquer magnetic fluid carrier; Step C: After the prepared antimicrobial drug delivery system is used cyclically (1-10 times), quality control and performance evaluation are performed to ensure its colloidal dispersion stability and antimicrobial effectiveness.

[0013] 6. Further preferably, the surface modification preparation conditions in step A include but are not limited to reaction speed (100-800 rpm / min), reaction temperature (0-80°C), reaction time (60-600 min), etc., so as to improve the surface properties and biocompatibility of the raw lacquer magnetic fluid.

[0014] VII. In a preferred embodiment of the present invention, the method for loading the antimicrobial drug in step B includes but is not limited to van der Waals forces, dipole-dipole interactions, hydrogen bonds, etc., and is selected based on the properties and needs of the active ingredients of the antimicrobial drug.

[0015] 8. Further preferably, the colloidal nanoparticle size in step C is 30-300 nm to ensure the stable combination of the antibacterial drug molecules and the raw lacquer magnetic fluid carrier, and the antibacterial activity is greater than 90%.

[0016] The beneficial effects of the present invention are: the position and movement of the raw lacquer magnetic fluid in the water body are controlled by an external magnetic field, so as to achieve accurate recovery and improve multiple reuse. The present invention provides an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, which has the advantages of accurate delivery, efficient treatment, reduced side effects and good biocompatibility, and provides a new and efficient drug delivery method for the biomedical field. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Attached Figure 1 : Schematic diagram of the synthesis of raw lacquer magnetic fluid.

[0018] Attached Figure 2 : Surface electron microscopy morphology of delivery system nanoparticles V@DU@Fe (A), transmission electron microscopy morphology of V@DU@Fe (B), magnified transmission electron microscopy morphology of V@DU@Fe (C) and dynamic light scattering (DLS) particle size distribution diagram of V@DU@Fe (D).

[0019] Attached Figure 3 : Particle size, polymer dispersibility index (PDI) and zeta potential (Zeta potential) of vancomycin-loaded delivery system nanoparticles prepared by different processes.

[0020] Attached Figure 4 : FT-IR spectrum (A), hysteresis loop (B), XRD spectrum (C) and full scan XPS spectrum (D).

[0021] Attached Figure 5 : Cell morphological changes (A) and cell survival rate (B) of normal hepatocytes after V@U@Fe acted on LO2. DETAILED DESCRIPTION The technical solution of the present invention is further illustrated and described below by specific implementation modes in combination with the accompanying drawings. The present invention is further described in detail by the following examples, but the present invention is not limited thereto.

[0023] Embodiment 1: The synthesis method of a raw lacquer magnetic fluid of this embodiment is carried out according to the following steps: Step 1: Weigh 2.4g Fe3O4 and 2.6g FeCl3 and dissolve them in 500ml aqueous solution under magnetic stirring, slowly add 10% sodium hydroxide, and then adjust the pH value to 10 with 2mol / L NaOH solution. Under ultrasonic stirring, slowly add 15% dextran solution, mix well, and stir and react at 80℃ in N2 environment for 2h to obtain a black-brown suspension. After the solution is naturally cooled, centrifuge at 5000 r / min for 10 min, discard the precipitate, dialyze the supernatant with a 3.5K MWCO molecular weight dialysis bag, and freeze-dry the precipitate.

[0024] Step 2: Dissolve 5.0g of concentrated urushiol in 100mL of 50% ethanol solution and stir at 150rpm for 30min to obtain a brown mixed solution. Then, under mechanical stirring at 100rpm, slowly drip the above solution into 500ml of a buffer solution (10mM Tris-HCl, pH 8.5) containing 5mM Fe3O4 nanoparticles prepared in step 1. After ultrasonic treatment for 20min, the obtained solution was irradiated with 50W ultraviolet light for polymerization for 2 hours. After the reaction, the product was separated and collected by a magnet and rinsed alternately with ethanol and ultrapure water for 3 times. Finally, the raw lacquer magnetic fluid product was dried in a vacuum oven at 40℃ overnight to obtain dextran / urushiol-superparamagnetic magnet (DU@Fe).

[0025] Example 2: Functionalized surface modification of the raw lacquer magnetic fluid prepared in Example 1: 0.2 mg of DU@Fe was added to 80 ml of 5% glutaraldehyde solution, and a few drops of glacial acetic acid were added to adjust the pH of the solution to 5.0. The mixture was then stirred at 45 °C for 15 hours. After the reaction, the product was collected with a magnet and rinsed with excess ultrapure water to remove unbound glutaraldehyde. Then 0.5 g of the prepared glutaraldehyde-modified raw lacquer magnetic fluid was dispersed in 50 ml of PBS buffer solution (40 mM, pH8.0), and then 50 ml of 5% Lys-D-Ala-D-Ala solution was added thereto and oscillated at 35 °C for 8 hours. After the reaction, the functionalized lacquer phenol magnetic fluid was magnetically separated and washed with excess PBS buffer. After vacuum freeze-drying, the ligand-functionalized DU@Fe (Lys-D-Ala-D-Ala@DU@Fe) was obtained.

[0026] Example 3: Preparation of drug-loaded lacquer antibacterial delivery nanoparticles: 50 mg of Lys-D-Ala-D-Ala@DU@Fe was added to 50 ml of 500 mg / L vancomycin buffer solution (10 mM PBS, pH 6.5), and the mixed solution was shaken for 10 h at room temperature. After the reaction, the product was separated by magnetic absorption, washed with PBS, and vacuum dried to obtain antibacterial delivery nanoparticles (V@DU@Fe), which were sealed and stored for later use.

[0027] Example 4: Quality control and performance evaluation of drug-loaded raw lacquer magnetic fluid: Figure 1 Schematic diagram of the synthesis of raw lacquer magnetic fluid. TEM, SEM and particle size distribution characterization of V@DU@Fe nanoparticles Figure 2 As shown in the figure, as well as the Fourier transform infrared spectra, XRD graphs, XPS graphs, and hysteresis loop VSM graphs of Fe3O4 nanoparticles, DU@Fe, and Lys-D-Ala-D-Ala@DU@Fe in the embodiments of the present invention. Figure 3 The above results show that the synthesis of V@DU@Fe was successful, and the saturation magnetization (Ms) was 53 emu / g. Therefore, the magnetic response performance of the prepared drug-loaded raw lacquer magnetic fluid was good, which met the requirements of the magnetic targeting performance of the drug carrier. Figure 4 The biocompatibility evaluation showed that V@DU@Fe had good blood compatibility. The antibacterial activity of V@DU@Fe was confirmed by cell viability detection and hemolysis test, which also showed that it has broad application prospects in the field of biomedicine.

Claims

1. The present invention discloses an antibacterial drug delivery system based on raw lacquer magnetic fluid and a preparation method thereof, and the specific steps are as follows: at room temperature, raw lacquer and dextran are slowly added to the SPION solution, and the product DU@Fe can be generated after ultraviolet irradiation self-assembly for a period of time. Then, glutaraldehyde is slowly added dropwise, and high-speed stirring is continued for a period of time to covalently modify the surface hydroxyl groups. Next, based on the Schiff base reaction, D-alanyl-D-alanine (D-Ala-D-Ala) groups are grafted to prepare a carrier D-Ala-D-Ala@DU@Fe that can immobilize antibacterial drugs. Subsequently, the antibacterial drug (taking vancomycin as an example) that can bind to the D-Ala-D-Ala on the carrier surface is immobilized on DU@Fe by using intermolecular forces to obtain the antibacterial magnetic fluid V@DU@Fe. Finally, the antibacterial performance is evaluated after 6 cycles of use to ensure the stability and effectiveness of the antibacterial drug delivery system.

2. The antimicrobial drug delivery system according to claim 1, characterized in that: The system includes: a raw lacquer magnetic fluid carrier, which is composed of a magnetic fluid obtained by specifically treating raw lacquer phenol and dextran (10,000 Da to 100,000 Da) and has good colloidal dispersibility and biocompatibility.

3. The antimicrobial drug delivery system according to claim 1, characterized in that: The raw lacquer magnetic fluid carrier has adjustable magnetic responsiveness (-90 ~ +90 emu / g) and can be controlled to be recycled and reused through an external magnetic field.

4. The antimicrobial drug delivery system according to claim 1, characterized in that: The active ingredients of the antibacterial drugs include, but are not limited to, β-lactam antibiotics that can specifically bind to D-alanyl-D-alanine (D-Ala-D-Ala), such as penicillins and cephalosporins, and polypeptide antibiotics, such as vancomycin, etc., which are selected according to treatment needs.

5. An antibacterial drug delivery system based on raw lacquer magnetic fluid and its preparation method, characterized in that: The method comprises the following steps: Step A: Raw lacquer and dextran are placed in a buffer solution (pH 3-6) and subjected to ultraviolet irradiation (100-500W) to synthesize raw lacquer magnetic fluid, and covalently bind glutaraldehyde and D-Ala-D-Ala on the surface to facilitate binding with active molecules of antibacterial drugs; Step B: using intermolecular forces to specifically combine the antibacterial drug and D-Ala-D-Ala, so as to achieve the purpose of grafting onto the raw lacquer magnetic fluid carrier; Step C: After the prepared antimicrobial drug delivery system is used cyclically (1-10 times), quality control and performance evaluation are performed to ensure its colloidal dispersion stability and antimicrobial effectiveness.

6. The preparation method according to claim 5, characterized in that: The surface modification preparation conditions in step A include but are not limited to reaction speed (100-800 rpm / min), reaction temperature (0-80°C), reaction time (60-600 min), etc., to improve the surface properties and biocompatibility of the raw lacquer magnetic fluid.

7. The preparation method according to claim 5, characterized in that: The antimicrobial drug loading method in step B includes, but is not limited to, van der Waals force, dipole-dipole interaction force, and hydrogen bond, and is selected based on the properties and needs of the active ingredients of the antimicrobial drug.

8. The preparation method according to claim 5, characterized in that: The colloidal nanoparticle size in step C is 30-300 nm to ensure the stable combination of the antibacterial drug molecules and the raw lacquer magnetic fluid carrier, and the antibacterial activity is greater than 90%.

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