A phenylboronic acid functionalized magnetic nanoparticle, a preparation method and application thereof

Abstract

This application discloses phenylboronic acid-functionalized magnetic nanoparticles, their preparation method, and applications, relating to the field of biomimetic nanomaterials technology. The preparation method includes the following steps: dissolving carboxyl-containing magnetic nanoparticles with 3-aminophenylboronic acid and n-propylamine in water, stirring the reaction under catalytic conditions to obtain a reaction solution; purifying and freeze-drying the reaction solution to obtain the phenylboronic acid-functionalized magnetic nanoparticles. This application uses carboxyl-containing magnetic nanoparticles, 3-aminophenylboronic acid, and n-propylamine as raw materials, and obtains phenylboronic acid-functionalized magnetic nanoparticles (BMNPs) through an amide condensation reaction under the action of EDC-HCl and NHS catalysts. These functionalized magnetic nanoparticles can achieve transmembrane transport of glucose through the dynamic reversible interaction between phenylboronic acid and glucose. Furthermore, cell experiments show that the transport efficiency of BMNPs is significantly enhanced under the action of a magnetic field.

Description

Technical Field

[0001] This application relates to the field of biomimetic nanomaterials technology, and more specifically, to a phenylboronic acid functionalized magnetic nanoparticle, its preparation method, and its application. Background Technology

[0002] Cells are the basic building blocks of living organisms. They utilize channel proteins on their phospholipid membranes to exchange substances and information with the external environment and maintain the normal functioning of various life activities. For example, aquaporins promote the reabsorption and recycling of water by the kidneys, while the opening and closing of sodium and potassium ion channel proteins are closely related to physiological activities such as nerve signal transmission and muscle contraction. Glucose channel proteins facilitate the transport of glucose from the blood into cells, providing energy for the cells. However, abnormal function of channel proteins in the body can lead to certain channel-related diseases, such as arrhythmias, epilepsy, and cystic fibrosis.

[0003] Natural channel proteins are difficult to extract, expensive, and challenging to modify and functionalize. In recent years, chemists have developed a series of artificial transmembrane channel models using biomimetic approaches, such as artificial potassium and sodium ion channels and artificial water channels. Exploring the structure and properties of these artificial channels helps scientists gain a deeper understanding of the pathogenesis of certain diseases and the development of related channel-based drugs. It is well known that carbohydrates are the primary energy source for living organisms and participate extensively in various physiological activities. Insulin in the blood can activate glucose transporter 4 (GLUT4), promoting the passive transmembrane transport of glucose from the blood to the cytoplasm. This not only lowers blood sugar levels but also provides the energy needed for cellular life activities.

[0004] A literature review of existing technologies revealed various preparation schemes for different types of artificial ion channels and water channels. However, research on the preparation of biomimetic glucose channels is scarce. Only a few cases have achieved transmembrane transport of glucose molecules using large-diameter artificial nanopores. However, due to the large size of these artificial nanopores, the selectivity and efficiency for glucose molecule transport are lacking. Developing novel biomimetic glucose transmembrane transport systems is of great significance for understanding the functional mechanisms of natural glucose channel proteins and for drug development for related diseases. Summary of the Invention

[0005] The purpose of this application is to provide phenylboronic acid functionalized magnetic nanoparticles and their preparation method. By modifying magnetic nanoparticles with groups such as phenylboronic acid and alkylamine, the reversible covalent interaction between phenylboronic acid and glucose can be used to achieve controllable transmembrane transport of glucose molecules by the nanoparticles.

[0006] To solve the above-mentioned technical problems, the technical solution adopted in this application is as follows: On the one hand, this application provides a method for preparing phenylboronic acid functionalized magnetic nanoparticles, comprising the following steps: S1. Magnetic nanoparticles containing carboxyl groups are dissolved in water with 3-aminophenylboronic acid and n-propylamine, and the mixture is stirred under catalytic conditions to obtain a reaction solution. S2. After purifying and freeze-drying the reaction solution, the phenylboronic acid functionalized magnetic nanoparticles are obtained.

[0007] On the other hand, this application provides phenylboronic acid functionalized magnetic nanoparticles prepared by the above method.

[0008] On the other hand, this application provides the application of the above-mentioned phenylboronic acid functionalized magnetic nanoparticles as biomimetic glucose transporters in the field of glucose transmembrane transport.

[0009] Compared with the prior art, the embodiments of this application have at least the following advantages or beneficial effects: This application uses carboxyl-containing magnetic nanoparticles, 3-aminophenylboronic acid (APBA), and n-propylamine as raw materials, and obtains phenylboronic acid-functionalized magnetic nanoparticles (BMNPs) through an amide condensation reaction under the action of catalysts such as EDC-HCl and NHS. These functionalized magnetic nanoparticles can achieve transmembrane transport of glucose through the dynamic and reversible interaction between phenylboronic acid and glucose. Furthermore, cell experiments show that the transport efficiency of BMNPs is significantly enhanced under the action of a magnetic field. Attached Figure Description

[0010] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0011] Figure 1 The chemical reaction equations for the preparation methods in the embodiments of this application are as follows; Figure 2 This is a comparison of the infrared spectra of the raw materials and products in Example 1 of the experimental examples of this application; Figure 3 Zeta potential analysis of raw materials and products (including different feed ratios) in the experimental examples of this application; Figure 4 This is a schematic diagram of the U-tube experiment in the experimental examples of this application and a fluorescence test of the glucose transport properties of BMNPs; Figure 5 This study presents cellular experiments to test the transmembrane transport properties of glucose by BMNPs under both magnetic field and non-magnetic field conditions in the experimental examples of this application. Detailed Implementation

[0012] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0013] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present application will now be described in detail with reference to specific embodiments.

[0014] A method for preparing phenylboronic acid functionalized magnetic nanoparticles includes the following steps: S1. Magnetic nanoparticles containing carboxyl groups are dissolved in water with 3-aminophenylboronic acid and n-propylamine, and the mixture is stirred under catalytic conditions to obtain a reaction solution. S2. After purifying and freeze-drying the reaction solution, the phenylboronic acid functionalized magnetic nanoparticles are obtained.

[0015] In some embodiments of this application, the magnetic nanoparticles containing carboxyl groups in step S1 above include one or more of Fe3O4 magnetic nanoparticles, γ-Fe2O3 magnetic nanoparticles, magnetic iron nanoparticles, nickel-zinc ferrite magnetic nanoparticles, manganese-zinc ferrite magnetic nanoparticles, and nickel-copper-zinc ferrite magnetic nanoparticles, wherein the particle size of the magnetic nanoparticles is 100-200 nm.

[0016] In some embodiments of this application, the equivalent ratio of the above-mentioned 3-aminophenylboronic acid, n-propylamine, and carboxyl-containing magnetic nanoparticles is 1:1:(10) -2 ~10 -4 )eq.

[0017] In some embodiments of this application, the catalyst is N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide; the equivalence ratio of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide, and the total amino group in the raw materials is 2:2:1. The raw materials refer to the reaction materials in step S1, including magnetic nanoparticles containing carboxyl groups, 3-aminophenylboronic acid, and n-propylamine.

[0018] In some embodiments of this application, the stirring reaction temperature in step S1 is 10°C to 20°C, and the reaction time is 2 to 3 days.

[0019] In some embodiments of this application, the purification in step S2 is specifically as follows: the reaction solution is placed in a 2000-3000 Da dialysis bag and dialyzed in deionized water for 2-3 days, with the dialysis solution being changed every 4-6 hours during dialysis.

[0020] In some embodiments of this application, the freeze-drying temperature in step S2 above is -100 to -120°C, and the time is 1 to 2 days.

[0021] A phenylboronic acid-functionalized magnetic nanoparticle was prepared using the method described above.

[0022] Application of phenylboronic acid-functionalized magnetic nanoparticles as biomimetic glucose transporters in the field of glucose transmembrane transport.

[0023] In some embodiments of this application, the transmembrane transport environment of the above-mentioned phenylboronic acid functionalized magnetic nanoparticles is: magnetic field strength of 80-100 mT.

[0024] The features and performance of this application will be further described in detail below with reference to the embodiments. Example 1

[0025] This embodiment provides phenylboronic acid-functionalized magnetic nanoparticles, the synthesis equation of which is as follows: Figure 1 As shown, the specific preparation method is as follows: Fe3O4 magnetic nanoparticles (Fe3O4-(COOH)) n The particle size of Fe3O4-(COOH) is 100-200 nm, and the concentration is 5 mg / mL. n 5.1 mg (approximately 1 mL, 22 μM) of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC-HCl) (1.69 g, 8.8 mM) and N-hydroxysuccinimide (NHS) (1.01 g, 8.8 mM) were stirred overnight in water. Then, 3-aminophenylboronic acid (300 mg, 2.2 mM) and n-propylamine (130 mg, 2.2 mM) were added, and the mixture was stirred at 10°C for 2 days to obtain the reaction solution. The reaction solution was then dialyzed in deionized water for 3 days using a dialysis bag (molecular weight cutoff 2000 Da), with the dialysate changed every 4 hours. Finally, the dialyzed liquid was lyophilized and thoroughly ground to obtain a gray powder product, phenylboronic acid functionalized magnetic nanoparticles, denoted as BMNPs-1. Example 2

[0026] This embodiment provides phenylboronic acid-functionalized magnetic nanoparticles, the preparation method of which is as follows: Fe3O4-(COOH) nThe particle size is 100-200 nm, and the concentration is 5 mg / mL. Take this Fe3O4-(COOH)... n 1 mg of solution (approximately 0.2 mL, 4.4 μM), EDC-HCl (1.69 g, 8.8 mM), and NHS (1.01 g, 8.8 mM) were stirred overnight in water. Then, 3-aminophenylboronic acid (300 mg, 2.2 mM) and n-propylamine (130 mg, 2.2 mM) were added, and the mixture was stirred at 10°C for 2 days. The solution was then dialyzed in deionized water for 3 days using a dialysis bag (molecular weight cutoff 2000 Da), with the dialysate changed every 4 hours. Finally, the dialyzed liquid was lyophilized and thoroughly ground to obtain a gray powder product, BMNPs-2. Example 3

[0027] This embodiment provides phenylboronic acid-functionalized magnetic nanoparticles, the preparation method of which is as follows: Fe3O4-(COOH) n The particle size is 100-200 nm, and the concentration is 5 mg / mL. Take this Fe3O4-(COOH)... n 0.51 mg (approximately 0.1 mL, 2.2 μM) of solution, EDC-HCl (1.69 g, 8.8 mM), and NHS (1.01 g, 8.8 mM) were stirred overnight in water. Then, 3-aminophenylboronic acid (300 mg, 2.2 mM) and n-propylamine (130 mg, 2.2 mM) were added, and the mixture was stirred at 10°C for 2 days. The solution was then dialyzed in deionized water for 3 days using a dialysis bag (molecular weight cutoff 2000 Da), with the dialysate changed every 4 hours. Finally, the dialyzed liquid was lyophilized and thoroughly ground to obtain a gray powder product, BMNPs-3.

[0028] Experimental Example

[0029] 1. The infrared absorption spectra of the prepared samples were measured using a Thermo Nicolet IS5 Fourier Transform Infrared Spectrometer (FTIR). The infrared absorption spectra of the product sample (BMNPs) prepared in Example 1 and the raw materials Fe3O4 magnetic nanoparticles (Fe3O4) and 3-aminophenylboronic acid (APBA) are shown below. Figure 2 As shown, the results of the prepared sample were found at 3299 cm⁻¹. -1 1643cm -1 A strong secondary amide absorption peak was observed at the [location]. This indicates that 3-aminophenylboronic acid successfully reacted with iron oxide nanoparticles with carboxyl groups on their surface to obtain phenylboronic acid-functionalized magnetic nanoparticles (BMNPs).

[0030] 2. The particle size distribution of 3-aminophenylboronic acid and Fe3O4-(COOH) was determined using a Zetasizer Nano ZSE nanoparticle size analyzer. n The surface zeta potentials of nanoparticles and BMNPs prepared in Examples 1-3, such as Figure 3 As shown, the zeta potential of BMNPs increased significantly compared with that of the raw materials, indicating that the carboxyl groups with negative potential on the surface of iron nanoparticles combined with amino groups, and the target product was successfully obtained through chemical reaction between the raw materials.

[0031] 3. The glucose-carrying capacity and transmembrane capacity of the samples prepared in this invention were tested using a U-tube experiment to simulate a cellular environment. The left side of the U-tube contained an aqueous solution of 0.01 mg / mL 2-NBDG (a fluorescent glucose analog), the right side contained a pure aqueous solution, and the middle contained a dichloromethane solution containing BMNPs. This simulated the hydrophilic nature of the cell membrane, with hydrophobic sides and a hydrophobic center. The glucose concentration in the pure aqueous solution on the right side was measured after 2 hours to determine whether BMNPs could transport glucose across the membrane. The results are as follows: Figure 4 We found that the dichloromethane solution with added BMNPs significantly increased the glucose transport efficiency compared to the pure dichloromethane solution. This indicates that the prepared BMNPs can achieve the goal of transmembrane glucose transport.

[0032] 4. The glucose transport capacity of the samples prepared in this invention in real cells was tested using glial cells. Specific steps: Glial cells were cultured adherently for 48 h, then co-cultured with BMNPs and 2-NBDG prepared in Example 1 for 20 min. After incubation, the cells were washed twice with PBS, then PBS was added again, and the cells were observed under a confocal microscope (Zeiss LSM 900 (AiryScan 2)) for imaging. The results are as follows: Figure 5 As shown, compared with the blank control, the intracellular fluorescence of the BMNPs-added group was enhanced, indicating that BMNPs facilitated the transport of glucose from the extracellular space to the intracellular space. Furthermore, after applying a magnetic field to the test system (the magnetic field was provided by a circular strong magnet with a diameter of 48 mm and a thickness of 9 mm, placed directly below the eight-well plate, with a magnetic field strength of 90 mT), a significant increase in intracellular fluorescence was observed under a microscope, demonstrating the magnetic field-sensitive nature of BMNPs for glucose transmembrane transport.

[0033] In summary, the phenylboronic acid-functionalized magnetic nanoparticles, their preparation method, and their applications described in this application have the following advantages: This application uses carboxyl-containing magnetic nanoparticles, 3-aminophenylboronic acid (APBA), and n-propylamine as raw materials. Under the action of catalysts such as EDC-HCl and NHS, amide condensation reaction is used to ultimately obtain phenylboronic acid-functionalized magnetic nanoparticles (BMNPs). These functionalized magnetic nanoparticles can achieve transmembrane transport of glucose through the dynamic and reversible interaction between phenylboronic acid and glucose. Furthermore, cell experiments have shown that the transport efficiency of BMNPs is significantly enhanced under the action of a magnetic field.

[0034] The embodiments described above are some, but not all, of the embodiments of this application. The detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

Claims

1. A method for preparing phenylboronic acid functionalized magnetic nanoparticles, characterized in that, Includes the following steps: S1. Magnetic nanoparticles containing carboxyl groups are dissolved in water with 3-aminophenylboronic acid and n-propylamine, and the mixture is stirred under catalytic conditions to obtain a reaction solution. S2. After purifying and freeze-drying the reaction solution, the phenylboronic acid functionalized magnetic nanoparticles are obtained.

2. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, The magnetic nanoparticles containing carboxyl groups in step S1 include one or more of Fe3O4 magnetic nanoparticles, γ-Fe2O3 magnetic nanoparticles, magnetic iron nanoparticles, nickel-zinc ferrite magnetic nanoparticles, manganese-zinc ferrite magnetic nanoparticles, and nickel-copper-zinc ferrite magnetic nanoparticles, wherein the particle size of the magnetic nanoparticles is 100-200 nm.

3. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, The equivalent ratio of the 3-aminophenylboronic acid, n-propylamine, and carboxyl-containing magnetic nanoparticles is 1:1:(10). -2 ~10 -4 )eq.

4. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, The catalyst is N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide; the equivalent ratio of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and the total amino group in the raw material is 2:2:

1.

5. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, The stirring reaction temperature in step S1 is 10℃~20℃, and the time is 2~3 days.

6. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, The purification process in step S2 involves placing the reaction solution into a 2000–3000 Da dialysis bag and dialyzing it in deionized water for 2–3 days, changing the dialysis solution every 4–6 hours during dialysis.

7. The method for preparing phenylboronic acid functionalized magnetic nanoparticles according to claim 1, characterized in that, In step S2, the freeze-drying temperature is -100 to -120°C, and the time is 1 to 2 days.

8. A phenylboronic acid-functionalized magnetic nanoparticle, characterized in that, It is prepared by any one of the methods of claims 1 to 7.

9. The application of phenylboronic acid functionalized magnetic nanoparticles as described in claim 8 as a biomimetic glucose transporter in the field of glucose transmembrane transport.

10. The application according to claim 9, characterized in that, The transmembrane transport environment of the phenylboronic acid functionalized magnetic nanoparticles is a magnetic field strength of 80–100 mT.

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