Magnesium oxide nanoparticle drug delivery system as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a magnesium oxide nanoparticle drug delivery system as well as a preparation method and application thereof. MgO is used as a drug-loading core, the outer layer of MgO is coated with the polydopamine organic coating and the modified hyaluronic acid supramolecular polymer, meanwhile, copper ions are introduced, and the obtained magnesium oxide drug-loading nanoparticle drug delivery system shows an excellent effect in an anti-atherosclerosis experiment. According to the invention, the magnesium oxide nano-particle drug delivery system is used for treating atherosclerosis for the first time, and a new thought and a new direction are provided for constructing a novel anti-atherosclerosis treatment drug.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to a magnesium oxide nanoparticle drug delivery system and a preparation method and application thereof. BACKGROUND

[0002] Nanoparticulate drug delivery systems (NDDS) are a frontier field of cross-fusion of nanobiotechnology and modern pharmacy, and the core thereof is to realize precise drug delivery, controllable release and improvement of bioavailability by designing nanocarriers (such as liposomes, nanoparticles, micelles, dendrimers and the like), with the rapid development of material science and nanotechnology, the NDDS shows great potential in the treatment of major diseases such as cancer and atherosclerosis.

[0003] At present, commonly used drug delivery carriers include organic materials such as lipids, peptides, glycosylated compounds and inorganic carriers such as iron oxide, zinc oxide, titanium dioxide, nanogold and the like, for example, patents CN117750943A, CN113827721A and CN105377244A disclose the same.

[0004] However, organic materials such as lipids and peptides have problems such as poor stability, limited drug loading capacity, easy phagocytosis by human mononuclear phagocyte system and immune response, which limits the drug delivery effect thereof. Although inorganic nanomaterials can avoid the above problems, however, some inorganic material carriers (such as iron oxide / nanogold) have poor biodegradability, long-term retention may have toxicity, and the inorganic material carriers themselves only have the effect of simply delivering drugs and have no regulating effect on the pathological process of diseases.

[0005] Based on the above, it is necessary to construct a nanodrug delivery system which is a carrier, an active ingredient and an environmental response device. SUMMARY

[0006] In order to solve the above technical problems, the application provides a magnesium oxide nanoparticle drug delivery system and a preparation method and application thereof.

[0007] Based on the good histocompatibility and biological safety of magnesium oxide nanoparticles, the application designs a drug delivery system taking magnesium oxide nanoparticles as the core of the drug delivery system, which can convert pro-inflammatory M1 macrophages into anti-inflammatory M2 macrophages, promote the release of nitric oxide of damaged endothelial cells, regulate the repair of damaged endothelial cells, and show good effects in anti-atherosclerosis or anti-atherosclerosis related diseases.

[0008] The first aspect of the present application provides a method for preparing a magnesium oxide nanoparticle drug delivery system, specifically comprising the following steps: S1, surface modification of MgO: first, the surface of the MgO nanoparticles is treated with a NaOH solution, then an organic coating solution is added for modification, and an organic coating is loaded on the surface to obtain organic coating modified magnesium oxide nanoparticles; S2, preparation of magnesium oxide / copper nanoparticles: mix the solution containing free copper ions with the organic coating solution described in S1, ultrasonically mix, then add ammonia water, and then add the organic coating modified magnesium oxide nanoparticles prepared in S1, ultrasonically mix, then react at room temperature, centrifuge, and obtain magnesium oxide / copper nanoparticles; S3, hyaluronic acid coating: add modified hyaluronic acid to the magnesium oxide / copper nanoparticles of S2 for reaction, then precipitate after the reaction is completed, and dry to obtain the magnesium oxide nanoparticle drug delivery system.

[0009] In the above preparation method provided by the present application, preferably, the mass-volume ratio of the MgO nanoparticles described in S1 to the NaOH solution is 1 g: 80-120 mL, and the concentration of the NaOH solution is 1-5 mol / L.

[0010] Further preferably, the concentration of the NaOH solution described in S1 is 1-3 mol / L.

[0011] Preferably, the mass-volume ratio of the MgO nanoparticles described in S1 to the organic coating solution is 1 g: 80-120 mL, and the concentration of the organic coating solution is 1-10 mg / mL.

[0012] Further preferably, the mass-volume ratio of the MgO nanoparticles described in S1 to the organic coating solution is 1 g: 90-110 mL, and the concentration of the organic coating solution is 1-5 mg / mL.

[0013] Preferably, the organic coating solution described in S1 is selected from any one of polyethylene glycol, polydopamine, polycaprolactone, chitosan, polylactic acid, and poly(lactic-co-glycolic acid) solution.

[0014] Preferably, when the organic coating solution described in S1 is a polydopamine solution, the polydopamine solution is obtained by dissolving dopamine in a mixed solvent, and the mixed solvent is a mixture of water, anhydrous ethanol, and ammonia water, wherein the volume ratio of water: anhydrous ethanol: ammonia water is 1-2: 6-9: 0.5-1.5.

[0015] Preferably, in the mixed solvent of the polydopamine solution, the volume ratio of water: anhydrous ethanol: ammonia water is 1:8:1.

[0016] Preferably, in S1, the temperature for treating the surface of the MgO nanoparticles with the NaOH solution is 25-28 DEG C, and the treatment time is 10-60 min.

[0017] Preferably, in S1, the temperature for the modification reaction of the organic coating solution is room temperature, and the modification reaction time is 1-10 h.

[0018] Further preferably, in S1, the modification reaction time of the organic coating solution is 3-8 h.

[0019] In the present application, the outer surface of the MgO nanoparticles is first treated with a NaOH solution to obtain nanoparticles with an inner core of MgO and an outer layer of Mg(OH)2. Then, dopamine is dissolved in a mixed solvent to obtain a polydopamine solution, and the MgO nanoparticles are modified with the polydopamine solution. In the present application, the use of a mixed solvent to dissolve dopamine before modification significantly improves the drug loading rate of the drug delivery system compared to the traditional use of Tris-HCl as a dopamine solvent. The drug loading rate is as high as 5.26±0.16%, while the drug loading rate of polydopamine modified by the traditional solvent is only 1.23±0.21%.

[0020] In addition, in the present application, the polydopamine organic coating on the outer layer of the MgO nanoparticles helps to slow down the release rate of magnesium ions, preventing premature release of magnesium oxide due to changes in the pH value in the body, thereby achieving better sustained release effect.

[0021] Preferably, in S2, the solution containing free copper ions is selected from at least one of copper nitrate solution, copper chloride solution, and copper sulfate solution, and the concentration of copper ions in the solution is 0.5-10 mM.

[0022] Further preferably, in S2, the solution containing free copper ions is copper nitrate solution, and the concentration of copper ions in the solution is 3-7 mM.

[0023] More preferably, in S2, the concentration of copper ions in the solution containing free copper ions is 5 mM.

[0024] Preferably, in S2, the volume ratio of the solution containing free copper ions to the organic coating solution is 0.1-10:1, and the volume ratio of the solution containing free copper ions to ammonia is 8-15:1.

[0025] In the present application, copper ions are further loaded on the outer layer of the polydopamine-modified MgO nanoparticles. The presence of copper ions can synergize with magnesium ions to regulate endothelial cells and immune cells.

[0026] As preferred, the modified hyaluronic acid in S3 is thiolated hyaluronic acid modified by thiolation, the mass-volume ratio of the magnesium / copper oxide nanoparticles to the modified hyaluronic acid is 1 g: 30-60 mL, and the concentration of the modified hyaluronic acid is 1-10 mg / mL.

[0027] As further preferred, in S3, the mass-volume ratio of the magnesium / copper oxide nanoparticles to the modified hyaluronic acid is 1 g: 50 mL, and the concentration of the modified hyaluronic acid is 1-5 mg / mL.

[0028] The diameter of the magnesium oxide nanoparticle delivery drug system obtained by the preparation method of the application is 50-150 nm.

[0029] As preferred, the diameter of the magnesium oxide nanoparticle delivery drug system is 90-120 nm.

[0030] The magnesium oxide nanoparticle delivery drug system provided by the application has a whole layer shell structure, with MgO as the core structure of the carrier, the next outer layer being wrapped with polydopamine and loaded with copper ions, and the outermost layer being wrapped with thiol-modified hyaluronic acid. The above hierarchical structure makes the obtained delivery drug system not only have the effects of reducing inflammatory response, reducing ROS production, and preventing damage caused by oxidative stress, but also has excellent biocompatibility. In addition, the introduction of copper ions can synergize with magnesium ions to play a regulatory role on endothelial cells and immune cells, and synergize with therapeutic drugs to achieve better therapeutic effect.

[0031] Naturally, the magnesium oxide nanoparticle delivery drug system obtained by the above preparation method is also the focus of protection of the application.

[0032] The second aspect of the application is to provide the use of the magnesium oxide nanoparticle delivery drug system, and more specifically, to provide the use of the magnesium oxide nanoparticle delivery drug system in the preparation of drugs for resisting atherosclerosis or atherosclerosis-related diseases.

[0033] The atherosclerosis-related diseases are selected from one or more of the following: coronary atherosclerotic heart disease, cerebral atherosclerosis, and peripheral vascular atherosclerosis.

[0034] The third aspect of the present application provides an anti-atherosclerosis drug, which comprises the magnesium oxide nanoparticle delivery drug system prepared by the above method, a therapeutic preparation, and a pharmaceutically acceptable adjuvant; wherein the therapeutic preparation is a preparation for treating atherosclerosis or a disease related to atherosclerosis, and the therapeutic preparation is selected from any one of statin preparations, fibrate preparations, anti-platelet preparations, anticoagulant preparations, angiotensin converting enzyme inhibitors, calcium ion antagonists, beta receptor blockers, glucocorticoids and pharmaceutically acceptable salts thereof, active oxygen scavengers, and traditional Chinese medicine preparations.

[0035] Preferably, the traditional Chinese medicine preparation comprises any one of salvianolic acid and puerarin.

[0036] Preferably, in the drug, the mass-volume ratio of the magnesium oxide nanoparticle delivery drug system to the therapeutic preparation is 1 g: 30-60 mL.

[0037] Further preferably, in the drug, the mass-volume ratio of the magnesium oxide nanoparticle delivery drug system to the therapeutic preparation is 1 g: 50 mL.

[0038] Preferably, the applicable concentration of the drug in the treatment of atherosclerosis is 1-500 μg / mL.

[0039] In addition, the preparation method of the anti-atherosclerosis drug comprises the following steps: (1) Surface modification of MgO: first, the surface of MgO nanoparticles is treated with a NaOH solution, then polydopamine solution is added for modification to obtain polydopamine-modified magnesium oxide nanoparticles, wherein the polydopamine solution is obtained by dissolving dopamine in a mixed solvent, and the volume ratio of water: anhydrous ethanol: ammonia in the mixed solvent is 1-2: 6-9: 0.5-1.5; (2) Preparation of magnesium oxide / copper nanoparticles: mix the copper nitrate solution with the dopamine solution in (1), ultrasonically mix, then add ammonia water, and then add the polydopamine-modified magnesium oxide nanoparticles prepared in (1), ultrasonically mix, react at room temperature, centrifuge, and obtain magnesium oxide / copper nanoparticles; (3) Hyaluronic acid coating: add thiol-modified hyaluronic acid to the magnesium oxide / copper nanoparticles prepared in (2) to react, precipitate after the reaction is completed, and dry to obtain the magnesium oxide nanoparticle delivery drug system; (4) Loading of the therapeutic preparation: in the present application, the loading of the therapeutic preparation can be selected from any one of the following two methods: Method one: directly mix the therapeutic preparation with the magnesium oxide nanoparticle delivery drug system; Method two: the therapeutic preparation is directly mixed with the modified hyaluronic acid and magnesium oxide / copper nanoparticles in step (3), and is dried after reaction.

[0040] Among them, method two is preferred, which is more conducive to loading the drug component in the delivery system, thereby improving the drug preparation loading rate.

[0041] The beneficial effects of the present application are: Firstly, in the present application, MgO nanoparticles are used as the carrier core for drug loading, and the surface thereof is modified to sequentially coat a polydopamine layer and a hyaluronic acid layer on the outer layer of the MgO nanoparticles, and copper ions are introduced and coated between the outermost layer and the second outer layer. The obtained magnesium oxide nanoparticle drug delivery system with a multilayer structure has a particle size of about 110 nm, and a drug loading amount of 5.26±0.16%; Secondly, the magnesium oxide nanoparticle drug delivery system constructed in the present application not only can regulate the number of M1 / M2 type macrophages, but also can promote the release of NO by endothelial cells, inhibit the uptake of lipids by macrophages to form foam cells, and inhibit the formation of plaques, thereby showing a significant anti-atherosclerotic effect. The experimental results show that, compared with the control group added with phosphate buffer solution (PBS), the concentration of NO released by endothelial cells is increased by 70% after treatment with the drug system of the present application, the oil red O staining area of macrophages in vitro is reduced by 50%, and the plaque area of the mouse aorta is reduced by 50%. BRIEF DESCRIPTION OF DRAWINGS

[0042] Figure 1 Structure characterization of the magnesium oxide nanoparticle drug delivery system prepared in Example 1 of the present application, wherein a is a transmission electron microscope image, and b is an X-ray diffraction element peak fitting diagram; Figure 2 Ability of the anti-atherosclerotic drug containing the magnesium oxide nanoparticle drug delivery system in Example 2 of the present application to polarize macrophages; Figure 3 Ability of the anti-atherosclerotic drug containing the magnesium oxide nanoparticle drug delivery system in Example 3 of the present application to promote the release of nitric oxide; Figure 4 Effect of the anti-atherosclerotic drug containing the magnesium oxide nanoparticle drug delivery system in Example 4 of the present application on anti-atherosclerosis; Figure 5 Effect of the copper ion concentration on the magnesium oxide nanoparticle drug delivery system in Test Example 1 of the present application, wherein a is the effect of the magnesium oxide nanoparticle drug delivery system on the survival of endothelial cells, and b is the effect of the magnesium oxide nanoparticle drug delivery system on the release of NO; Figure 6 The effects of different drug systems on the inhibition of foam cell formation in vitro were compared in Test Example 2 of the present application.

[0043] Figure 7 The X-ray diffraction pattern element peak fitting diagram of the magnesium oxide nanoparticle drug delivery system prepared when the organic coating of Comparative Example 1 of the present application was replaced with polycaprolactone. DETAILED DESCRIPTION

[0044] In order to enable those skilled in the art to better understand the present application, the present application will be further described in conjunction with specific embodiments.

[0045] It should be noted that the following examples in the present application are only used to illustrate the technical solutions of the present application and do not limit the present application, although the present application has been described in detail through the following preferred examples, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined in the claims of the present application.

[0046] Example 1 The anti-atherosclerotic drug containing the magnesium oxide nanoparticle drug delivery system was prepared as follows: (1) Surface modification of MgO: 0.10 g of MgO nanoparticles was added to 10 mL of 1 M NaOH solution, and after treatment at room temperature for 10 min, centrifugal treatment was performed at a speed of 10000 rpm for 5 min, and the temperature of centrifugation was 10°C. The precipitate obtained was washed twice with anhydrous ethanol, and then 10 mL of 5 mg / mL polydopamine solution (solvent: 10% water, 80% anhydrous ethanol, 10% ammonia water) was added, and the reaction was carried out at room temperature for 4 h. The obtained product was centrifuged at a speed of 10000 rpm for 5 min, and the temperature of centrifugation was 10°C. After the precipitate was separated and washed twice with anhydrous ethanol, polydopamine-modified magnesium oxide nanoparticles were obtained; (2) Preparation of magnesium oxide / copper nanoparticles: 5 mL of 2 mM copper nitrate solution (solvent: anhydrous ethanol) and 5 mL of 5 mg / mL polydopamine solution (solvent: same as step (1)) were mixed and ultrasonically mixed for 5 min, and finally 500 μL of ammonia water was added to the polydopamine-modified magnesium oxide nanoparticles prepared in step (1), and after ultrasonic mixing, the reaction was carried out at room temperature for 2 h. The same centrifugation and washing conditions as in step (1) were used for treatment, and then the precipitate was placed in a freeze-drying machine and dried for 6 h to obtain magnesium oxide / copper nanoparticles; (3) Hyaluronic acid coating and loading of therapeutic formulation: 5 mL of 4 mg / mL thiolated hyaluronic acid (the thiolated hyaluronic acid is a commercially available product) was mixed with 5 mL of traditional Chinese medicine Danshensu B solution, and added to 0.1 g of magnesium oxide / copper nanoparticles prepared in step (2). The mixture was stirred at room temperature for 2 h. After separation by centrifugation (10000 rpm, 5 min, 10℃), the product was washed twice with phosphate buffer. Finally, the precipitate was dried in a freeze dryer for 6 h to obtain the magnesium oxide nanoparticle drug delivery system.

[0047] The results showed that the drug loading rate of the obtained magnesium oxide nanoparticle drug delivery system was 5.26 ± 0.16%.

[0048] The magnesium oxide nanoparticle drug delivery system prepared according to the method of this embodiment is shown in the transmission electron microscope image below. Figure 1 As shown in Figure a, the diameter of this magnesium oxide nanoparticle drug delivery system is approximately 110 nm.

[0049] Elemental peak fitting analysis of X-ray diffraction pattern is shown in […]. Figure 1 As shown in Figure b, the magnesium oxide nanoparticle drug delivery system contains elements such as Cu, O, and N.

[0050] Example 2 The following steps were taken to evaluate the effect of the magnesium oxide nanoparticle drug delivery system prepared in Example 1 on the drug polarization of macrophages against atherosclerosis: 5×10 6 RAW 264.7 macrophages were seeded in 6-well plates for 24 h. Then, 100 ng / mL lipopolysaccharide (LPS) or 20 ng / mL interleukin-4 (IL-4) and the magnesium oxide nanoparticle drug delivery system obtained in Example 1 (50 μg / mL) or an equal amount of salvianolic acid B loaded in the 50 μg / mL magnesium oxide nanoparticle drug delivery system were added and incubated for 24 h. Macrophages were then harvested and incubated with anti-CD86-APC and anti-CD163-PE antibodies. Finally, the polarization effect of RAW 264.7 cells was detected by flow cytometry.

[0051] The magnesium oxide nanoparticle drug delivery system prepared in this invention, after loading a therapeutic formulation, exhibits the ability to polarize macrophages with anti-atherosclerotic drugs, as shown in the following example. Figure 2 As shown.

[0052] from Figure 2As can be seen, the proportion of M1 macrophages (CD86+CD163-) in the blank control group was 42.5%, while the proportion of M1 macrophages with only salvianolic acid B was 39.1%. However, after adding the magnesium oxide nanoparticle drug delivery system, the proportion decreased to 20.4%, which was significantly lower than the blank control group and the group with only salvianolic acid B. The proportion of M2 macrophages (CD86-CD163+) in the blank control group was 12.9%, while the proportion of M2 macrophages with only salvianolic acid B was 13.9%. However, after adding the magnesium oxide nanoparticle drug delivery system, the proportion decreased to 16.9%, which was significantly higher than the blank control group and the group with only salvianolic acid B. The above results are sufficient to prove that the magnesium oxide nanoparticle drug delivery system prepared in this invention has excellent ability to polarize macrophages and can polarize pro-inflammatory M1 macrophages into anti-inflammatory M2 macrophages.

[0053] Example 3 The following steps were taken to evaluate the effect of the magnesium oxide nanoparticle drug delivery system prepared in Example 1 on promoting the release of NO from endothelial cells after loading a therapeutic formulation: 5×10 5 After 24 h of seeding personal umbilical vein endothelial cells (HUVECs) in 6-well plates, 500 ng / mL lipopolysaccharide, 50 μg / mL oxidized low-density lipoprotein, and the magnesium oxide nanoparticle drug delivery system obtained in Example 1 (50 μg / mL) or an equal amount of salvianolic acid B loaded with the 50 μg / mL magnesium oxide nanoparticle drug delivery system were added and incubated for 24 h. Then, the culture supernatant was harvested, and the release of NO from the endothelial cells was detected using a NO detection kit.

[0054] The magnesium oxide nanoparticle drug delivery system prepared in this invention, after loading a therapeutic formulation, exhibits the following ability to promote NO release in order to combat atherosclerosis: Figure 3 As shown.

[0055] from Figure 3 As can be seen, compared with the blank control group, the addition of only salvianolic acid B did not significantly improve the concentration of NO released by endothelial cells, while the concentration of NO released by endothelial cells increased by 70% after treatment with the magnesium oxide nanoparticle drug delivery system, proving that the magnesium oxide nanoparticle drug delivery system has the ability to promote the repair of damaged endothelial cells and improve the integrity of vascular endothelium.

[0056] Example 4 The following steps were taken to evaluate the effect of the magnesium oxide nanoparticle drug delivery system prepared in Example 1 on inhibiting the formation of atherosclerotic plaques: In vitro inhibition of foam cell formation was achieved by using RAW 264.7 macrophages at a concentration of 5 × 10⁻⁶ cells.6 After 24 h of inoculation in 6-well plates, 500 ng / mL lipopolysaccharide, 50 μg / mL oxidized low-density lipoprotein, and the magnesium oxide nanoparticle drug delivery system (50 μg / mL) obtained in Example 1 or an equivalent amount of salvianolic acid B loaded in the magnesium oxide nanoparticle drug delivery system at 50 μg / mL were added, and incubation was performed for 24 h. Then, the cells were subjected to oil red O staining, and the formation of foam cells was observed by taking pictures under an optical microscope.

[0057] The in vivo inhibition of atherosclerotic plaque formation was performed as follows: First, a mouse atherosclerosis model was constructed, and 6-week-old APOE- / - mice were fed with high-fat feed for 12 weeks. The aortas were subjected to gross oil red O staining, and the plaque area was semi-quantitatively observed by taking pictures. After 1 week of adaptation to high-fat feed, the mice were injected with physiological saline (control group), an equivalent amount of salvianolic acid B carried by the drug delivery system, and the magnesium oxide nanoparticle drug delivery system (20 mg / kg, solvent: physiological saline) via the tail vein. The drug administration frequency was once every 3 days, and the mice were fed with high-fat feed for 12 weeks. The aortas of the mice were then taken for evaluation.

[0058] After the magnesium oxide nanoparticle drug delivery system obtained in the present application was loaded with a therapeutic preparation, the anti-atherosclerotic effect of the obtained anti-atherosclerotic drug was as shown in Figure 4 .

[0059] The in vitro inhibition of foam cell formation was as shown in Figure 4 a and b, and it can be seen from the figures that, compared with the blank control group, the addition of salvianolic acid B alone can reduce the oil red O staining area of macrophages, and the oil red O staining area of macrophages is significantly reduced after treatment with the magnesium oxide nanoparticle drug delivery system, about 50% of that of the normal control group, proving that the magnesium oxide nanoparticle drug delivery system has the effect of inhibiting the formation of foam cells.

[0060] The in vivo inhibition of atherosclerotic plaque formation was as shown in Figure 4 c and d, and it can be seen from the figures that, compared with the control group, the administration of salvianolic acid B alone can reduce the plaque area of the aortas of the mice (without statistical difference), and the plaque area of the aortas of the mice is reduced by 50% after treatment with the magnesium oxide nanoparticle drug delivery system, proving that the magnesium oxide nanoparticle drug delivery system has the effect of inhibiting the formation of atherosclerotic plaques.

[0061] In summary, the magnesium oxide nanoparticle drug delivery system prepared in the present application can achieve precise drug delivery, prevent premature degradation of the drug, adjust immunity to reverse pro-inflammatory cells into anti-inflammatory cells, promote the release of endothelial cell nitric oxide and the repair of damaged endothelium, and achieve more effective inhibition of the progression of atherosclerosis.

[0062] Test Example 1 In this test example, the copper ion concentration in the preparation of the magnesium oxide nanoparticle drug delivery system was screened. Unlike Example 1, a magnesium oxide nanoparticle drug delivery system was prepared without loading salvianolic acid B, and the copper ion concentration was set to 1 mM, 3 mM, 5 mM, 7 mM, and 10 mM, respectively, to evaluate the effect of the obtained magnesium oxide nanoparticle drug delivery system on endothelial cell survival rate and NO release.

[0063] The specific operation steps of this test example are as follows: Determination of endothelial cell survival rate: 1 x 10 4 After 24 h of inoculation of 1 x 10

[0064] The determination experiment of NO release was the same as Example 3, except that the concentration of the magnesium oxide nanoparticle drug delivery system was 100 μg / mL, and the copper ion concentration was 1-10 mM.

[0065] In this test example, the effects of the magnesium oxide nanoparticle drug delivery system loaded with different concentrations of copper ions on endothelial cell survival and NO release capacity are shown in a and b of Figure 5

[0066] Figure 5 As shown in a of FIG. 1,

[0067] Figure 5 As shown in b of FIG. 1, the magnesium oxide nanoparticle drug delivery system promoted the release of NO from endothelial cells most strongly at a copper ion concentration of 1-5 mM, and the ability to promote the release of NO from endothelial cells gradually decreased as the concentration increased at a copper ion concentration of > 5 mM.

[0068] Test Example 2 The effect of the magnesium oxide nanoparticle drug delivery system loaded with different types of therapeutic agents on the inhibition of atherosclerotic plaque formation was evaluated as follows: Inhibition of foam cell formation in vitro, macrophage RAW 264.7 was cultured at a concentration of 5 x 10 6 ​After 24 h of inoculation in 6-well plates, 500 ng / mL lipopolysaccharide, 50 μg / mL oxidized low-density lipoprotein, and 100 μg / mL of the magnesium oxide nanoparticle drug delivery system loaded with different drugs were added, wherein the drug components were: statin atorvastatin, active oxygen scavenger Tempol, and traditional Chinese medicine Danshensuan B, and the cells were incubated for 24 h. Then, the cells were stained with oil red O, and the percentage of the area of oil red O-stained cells to the total area of the cells was calculated.

[0069] As can be seen from Figure 6 Compared with the control group added with phosphate buffer solution (PBS), the oil red O staining area of macrophages was reduced to different degrees after treatment with single drugs and magnesium oxide nanoparticle drug delivery systems loaded with different therapeutic agents, and the reduction of the oil red O staining area of macrophages was more obvious after treatment with magnesium oxide nanoparticle drug delivery systems loaded with different drugs than after treatment with single drugs.

[0070] Comparative Example 1 In this test example, polycaprolactone was selected as the organic coating layer during the preparation of the magnesium oxide nanoparticle drug delivery system, and the rest of the preparation method was the same as that in Example 1. The preparation was evaluated by X-ray diffraction pattern element peak fitting analysis.

[0071] The specific operation steps of this test example are as follows: The polycaprolactone (PCL) (Mn = 60000) with a mass fraction of 6-10% was dissolved in ethyl acetate under the conditions of magnetic stirring at 40±1℃ and 1300 rpm, and then the sample was soaked in the PCL solution for 100 s, and the rest of the treatment conditions were the same as those for the preparation of the polydopamine coating.

[0072] As can be seen from Figure 7 It can be seen that when the polycaprolactone organic coating layer with a concentration of 8% is selected, the X-ray diffraction pattern element peak fitting analysis of the prepared magnesium oxide nanoparticle drug delivery system contains C-O, C=O, and C-H structures, proving that the preparation is successful.

[0073] The drug loading rate of the obtained nanoparticle delivery system was only 2.84±0.19%, while the drug loading rate of the polydopamine coating was 5.26±0.16%. The effect of the polycaprolactone coating was worse than that of the polydopamine coating, and was only 1 / 2 of that of the polydopamine coating.

[0074] Comparative Example 2 Different from the embodiment 1, in (1), the traditional Tris-HCl aqueous solution is used as the solvent of dopamine to modify the surface of the magnesium oxide nanoparticles with polydopamine, and the specific operation is as follows: a 10 mM Tris-HCl aqueous solution is prepared, hydrochloric acid is used to adjust the pH value to 8.5, dopamine is dissolved in the 10 mM Tris-HCl (pH=8.5) solution to obtain a polydopamine solution, and the rest of the reaction conditions and reaction steps are the same as those in the embodiment 1.

[0075] The results show that when the polydopamine solution prepared by dissolving dopamine in the traditional solvent is used to modify the magnesium oxide nanoparticles, the drug loading rate of the obtained nanoparticle delivery system is only 1.23±0.21%, while after the solvent of the polydopamine solution is adjusted and improved in the embodiment 1 of the present application, the drug loading rate of the obtained nanoparticle delivery system is as high as 5.26±0.16%, which is 5 times of that of the traditional method.

Claims

1. A method for preparing a magnesium oxide nanoparticle delivery drug system, characterized by, The method comprises the following steps: S1, surface modification of MgO: first, the surface of MgO nanoparticles is treated with NaOH solution, then an organic coating solution is added for modification, and an organic coating is loaded on the surface to obtain organic coating modified MgO nanoparticles; S2, preparation of MgO / copper nanoparticles: a solution containing free copper ions is mixed with the organic coating solution described in S1, and after ultrasonic mixing, ammonia water is added, then the organic coating modified MgO nanoparticles prepared in S1 are added, and after ultrasonic mixing, reaction is carried out at room temperature, and centrifugation is carried out to obtain MgO / copper nanoparticles; S3, hyaluronic acid coating: modified hyaluronic acid is added to the MgO / copper nanoparticles of S2 for reaction, and after reaction, precipitation is carried out, and drying is carried out to obtain the MgO nanoparticle drug delivery system.

2. The production method according to claim 1, wherein The mass-volume ratio of the MgO nanoparticles described in S1 to the NaOH solution is 1 g:80-120 mL, and the concentration of the NaOH solution is 1-5 mol / L; the mass-volume ratio of the MgO nanoparticles to the organic coating solution is 1 g:80-120 mL, and the concentration of the organic coating solution is 1-10 mg / mL.

3. The production method according to claim 1, wherein The organic coating solution described in S1 is selected from any one of polyethylene glycol, polydopamine, polycaprolactone, chitosan, polylactic acid, and poly(lactic-co-glycolic acid) solution.

4. The production method according to claim 3, wherein In S1, when the organic coating solution is a polydopamine solution, the polydopamine solution is obtained by dissolving dopamine in a mixed solvent, and the mixed solvent is a mixture of water, anhydrous ethanol, and ammonia water, wherein the volume ratio of water:anhydrous ethanol:ammonia water is 1-2:6-9:0.5-1.

5.

5. The production method according to claim 1, wherein The solution containing free copper ions described in S2 is selected from at least one of copper nitrate solution, copper chloride solution, and copper sulfate solution, and the concentration of copper ions in the solution is 0.5-10 mM; the volume ratio of the solution containing free copper ions to the organic coating solution is 0.1-10:1, and the volume ratio of the solution containing free copper ions to ammonia water is 8-15:

1.

6. The production method according to claim 1, wherein The modified hyaluronic acid described in S3 is thiolated hyaluronic acid, and the mass-volume ratio of the MgO / copper nanoparticles to the modified hyaluronic acid is 1 g:30-60 mL; and the concentration of the modified hyaluronic acid is 1-10 mg / mL.

7. Use of a nanoparticulate delivery drug system prepared by a process according to any one of claims 1 to 6 for the preparation of a medicament against atherosclerosis or against atherosclerosis-related diseases, characterized in that, The atherosclerosis-related disease is selected from one or more of the following: coronary atherosclerotic heart disease, cerebral atherosclerosis, and peripheral vascular atherosclerosis.

8. An anti-atherosclerotic medicament, characterized by, The method comprises the following steps: The therapeutic preparation is a preparation for treating atherosclerosis or a disease related to atherosclerosis, and the therapeutic preparation is selected from any one of statin preparations, fibrate preparations, anti-platelet preparations, anticoagulant preparations, angiotensin converting enzyme inhibitors, calcium ion antagonists, beta receptor blockers, glucocorticoids, pharmaceutically acceptable salts thereof, active oxygen scavengers, and traditional Chinese medicine preparations.

9. An anti-atherosclerotic medicament according to claim 8, wherein the compound is ###00003### The mass / volume ratio of the magnesium oxide nanoparticle delivery drug system to the therapeutic preparation is 1 g:30-60 mL.

10. The anti-atherosclerotic medicament according to claim 8, wherein the compound is ###00003### The applicable concentration of the drug is 1-500 μg / mL.

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