A drug delivery system, its preparation method and use
By loading budesonide onto black phosphorus quantum dots and encapsulating it with mesenchymal stem cell exosomes, combining photothermal effects and lung targeting, the problems of uncontrollable drug side effects and release rates in existing treatments for allergic asthma have been solved, achieving highly effective treatment for allergic asthma.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-11-21
- Publication Date
- 2026-03-17
AI Technical Summary
Existing medications for allergic asthma can only relieve symptoms and cannot address the underlying inflammation. They also suffer from side effects, drug resistance, and uncontrollable release rates. Local thermotherapy is inefficient and inconvenient to administer.
Budesonide was loaded onto black phosphorus quantum dots, and the drug-loaded black phosphorus quantum dots were encapsulated using mesenchymal stem cell exosomes. The photothermal effect was used to regulate lung temperature and control drug release rate. Combined with the lung-targeting ability of exosomes, the effective concentration of the drug in the target organ was achieved and the frequency of drug administration was reduced.
It achieves effective control of drug concentration in target organs, reduces the risk of side effects, improves treatment efficacy, reduces the frequency of medication, and relieves asthma symptoms through local heat therapy.
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Figure CN115708877B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical technology, specifically to a drug delivery system, its preparation method, and its application. Background Technology
[0002] Allergic asthma (AA) is the most common clinical phenotype of all types of asthma. It is a chronic inflammatory disease of the nasal mucosa mediated by serum immunoglobulin E, which seriously affects patients' daily lives and work. According to the World Health Organization, the economic cost of treating asthma globally each year far exceeds that of AIDS and tuberculosis. Currently, the most commonly used medications for treating asthma are anti-inflammatory drugs and bronchodilators. However, these drugs can only relieve symptoms and cannot address the root cause of inflammation in asthma patients. Therefore, research on allergic asthma is urgent and important. Decreased external temperature negatively impacts allergic diseases by affecting the body's immune response. Studies have found that in patients with allergic rhinitis, local hyperthermia (42°C) can reduce nasal congestion and vascular leakage caused by allergens, and localized hyperthermia can inhibit the production or release of allergic mediators. Local hyperthermia applied at temperatures above normal has shown beneficial effects on various respiratory diseases. Therefore, it is crucial to develop a heat-based delivery system that can synergize with existing drugs.
[0003] Black phosphorus quantum dots are novel nanomaterials with good drug loading capacity, strong photothermal effects, and excellent biocompatibility, making them ideal as drug carriers. However, black phosphorus quantum dots exhibit poor stability under physiological conditions, are prone to degradation and aggregation, and require additional coating for further application. Furthermore, black phosphorus quantum dots lack active targeting ability to target organs in vivo. Imparting active targeting ability through surface modification and other strategies can effectively improve their drug delivery performance. Exosomes are small vesicles secreted by cells, with diameters ranging from 30-150 nm, and possess a phospholipid bilayer. Due to their small size and unique biological homology, exosomes can be used to develop novel nanocarriers as advanced drug delivery systems. Mesenchymal stem cell exosomes inherit the immune-regulating and anti-inflammatory functions of their source cells and possess certain lung-targeting properties, thus showing potential applications in the clinical treatment and drug delivery of respiratory diseases.
[0004] Currently used anti-inflammatory and antihistamine drugs for treating allergic asthma only suppress clinical symptoms during asthma attacks. These drugs require long-term use and may cause side effects and drug resistance. Drug release rates are uncontrollable, requiring frequent use and carrying a high risk of side effects. Desensitization therapy is time-consuming, expensive, and has an effectiveness rate of only 40%. Although studies have shown that local hyperthermia can relieve allergy symptoms, most hyperthermia treatments use external heat conduction, which is inefficient, inconvenient to operate, and difficult to control precisely. Furthermore, there are currently no methods to utilize the thermal effect in conjunction with drug therapy. Summary of the Invention
[0005] To address the aforementioned technical problems, this invention develops a drug delivery system that loads budesonide onto black phosphorus quantum dots, and then encapsulates the drug-loaded black phosphorus quantum dots using mesenchymal stem cell exosomes. This drug delivery system can improve the effective drug concentration in the target organ and control the drug release rate, reducing the final drug concentration and dosing frequency, thereby reducing drug side effects. Simultaneously, it achieves a synergistic effect between mesenchymal stem cell exosomes, local hyperthermia, and the immunomodulatory effects of budesonide, resulting in better treatment outcomes for allergic asthma.
[0006] To achieve the above objectives, the technical solution adopted by this invention is as follows: a drug delivery system comprising exosomes, black phosphorus quantum dots, and budesonide. Through extensive experimentation, the inventors of this application have discovered that using exosomes to prepare a drug delivery system enables targeted lung delivery; the phospholipid bilayer of exosomes can slow down the degradation and metabolism of black phosphorus and budesonide in vivo, achieving a sustained-release effect; using black phosphorus quantum dots to prepare a drug delivery system allows for precise control of lung temperature rise in vitro via near-infrared light irradiation, alleviating asthma symptoms through local hyperthermia and regulating the release rate of budesonide in the lungs. Ultimately, the prepared drug delivery system increases the effective concentration of the drug in the target organ and controls the drug release rate, reducing drug side effects.
[0007] In a preferred embodiment of the drug delivery system described in this invention, the exosomes are mesenchymal stem cell exosomes. Through extensive experiments, the inventors of this application have discovered that the drug delivery system prepared using mesenchymal stem cell exosomes has lung-targeting properties, can improve the lung delivery effect of black phosphorus and budesonide, and that mesenchymal stem cell exosomes can synergistically treat allergic asthma with budesonide.
[0008] In a preferred embodiment of the drug delivery system of the present invention, the mass ratio of black phosphorus quantum dots to mesenchymal stem cell exosomes is 1:2.
[0009] The present invention also provides a method for preparing the drug delivery system, comprising the following steps:
[0010] (1) Preparation of black phosphorus quantum dots;
[0011] (2) Preparation of mesenchymal stem cell exosomes;
[0012] (3) Budesonide was loaded onto the black phosphorus quantum dots from step (1) to obtain drug-loaded black phosphorus quantum dots.
[0013] (4) The drug delivery system is obtained by encapsulating drug-loaded black phosphorus quantum dots with mesenchymal stem cell exosomes.
[0014] As a preferred embodiment of the preparation method of the drug delivery system of the present invention, the specific preparation method of the black phosphorus quantum dots in step (1) is as follows: black phosphorus block crystals are suspended in isopropanol, ground into black phosphorus powder and transferred to a brown bottle, and peeled off by ultrasound; the obtained black phosphorus solution is centrifuged at 12000 rpm for 20 minutes and then centrifuged at 18000 rpm for 20 minutes to obtain the black phosphorus quantum dots.
[0015] As a preferred embodiment of the preparation method of the drug delivery system of the present invention, the preparation method of the drug-loaded black phosphorus quantum dots in step (3) is as follows: centrifuge the supernatant containing black phosphorus quantum dots and discard the supernatant to enrich the black phosphorus quantum dots; then mix the black phosphorus quantum dots and polyethylene glycol evenly, add budesonide and perform ice bath sonication to obtain the drug-loaded black phosphorus quantum dots.
[0016] In a preferred embodiment of the preparation method of the drug delivery system of the present invention, the mass ratio of black phosphorus quantum dots to polyethylene glycol is 1:10.
[0017] In a preferred embodiment of the preparation method of the drug delivery system of the present invention, the mass ratio of black phosphorus quantum dots to budesonide is 1:1 to 6.
[0018] As a preferred embodiment of the preparation method of the drug delivery system of the present invention, the method of encapsulating drug-loaded black phosphorus quantum dots in mesenchymal stem cell exosomes in step (4) is as follows: drug-loaded black phosphorus quantum dots and mesenchymal stem cell exosomes are mixed evenly and subjected to ice bath sonication. After sonication, PBS is added and centrifuged.
[0019] The present invention also provides the use of the drug delivery system described herein in the preparation of a medicament for treating allergic asthma.
[0020] The beneficial effects of the present invention are as follows: The present invention provides a drug delivery system in which budesonide is loaded onto black phosphorus quantum dots, and then the drug-loaded black phosphorus quantum dots are encapsulated by mesenchymal stem cell exosomes. (1) The drug delivery system of the present invention is the first to use the photothermal effect of black phosphorus quantum dots in asthma to achieve precise control of lung temperature and realize local thermotherapy for allergic asthma; (2) The drug delivery system of the present invention uses the photothermal effect of black phosphorus quantum dots in the carrier to control the release rate of budesonide, which is conducive to maintaining the optimal drug concentration in the body, and is expected to reduce the dosage and frequency of medication and reduce the risk of side effects; (3) The drug delivery system of the present invention contains mesenchymal stem cell exosomes, which have lung targeting and can synergistically exert an anti-inflammatory effect with budesonide. Attached Figure Description
[0021] Figure 1 A shows the high-performance liquid chromatography results (with peak area representing drug concentration) after loading 20 μg of black phosphorus quantum dots with different concentrations of budesonide; B shows the loading efficiency of budesonide at different concentrations; C shows the transmission electron microscopy image of the drug delivery system prepared in Example 1.
[0022] Figure 2 The expression levels of eosinophil chemokine-3 mRNA in BAES-2B airway epithelial cells after different treatments.
[0023] Figure 3 Fluorescence signal detection in different organs of mice after tail vein injection of exosomes. Detailed Implementation
[0024] To more concisely and clearly demonstrate the technical solution, purpose, and advantages of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.
[0025] Example 1
[0026] One embodiment of the drug delivery system includes black phosphorus quantum dots, exosomes, and budesonide; wherein the exosomes are mesenchymal stem cell exosomes.
[0027] The preparation method of the drug delivery system in this embodiment includes the following steps:
[0028] (1) Preparation of black phosphorus quantum dots: 100 mg of black phosphorus block crystals were suspended in isopropanol and then ground in an agate mortar for 40 minutes; then the black phosphorus quantum dot powder was transferred to a 100 mL brown bottle and peeled off by an ultrasonic crusher at a power of 650 W for 12 hours; the obtained black phosphorus solution was centrifuged at a speed of 12000 rpm for 20 minutes; finally, the black phosphorus quantum dot suspension was further centrifuged at a speed of 18000 rpm for 20 minutes, and the obtained black phosphorus quantum dots were resuspended in isopropanol for later use.
[0029] (2) Preparation of mesenchymal stem cell exosomes: Collect mesenchymal stem cell culture supernatant from 10 T175 culture flasks, centrifuge at 3000 rpm for 20 min at 4℃ to remove dead cells and precipitate, centrifuge at 10000g to remove large vesicles, centrifuge at 110000g, discard the supernatant, resuspend in 200ul sterile PBS to obtain exosomes;
[0030] (3) Black phosphorus quantum dots loaded with budesonide: The supernatant containing black phosphorus quantum dots was centrifuged at 18,000 rpm for 1 h at 4 °C and the supernatant was discarded to enrich the black phosphorus quantum dots; then the mixture was mixed at a mass ratio of black phosphorus quantum dots to polyethylene glycol of 1:10, and finally the drug was added at a mass ratio of black phosphorus quantum dots to budesonide of 1:5. The mixture was sonicated in an ice bath for 30 min and shaken at 4 °C for 3 h.
[0031] (4) Exosome-encapsulated drug-loaded black phosphorus quantum dots: The mixture from step 3 was mixed at a mass ratio of black phosphorus quantum dots to exosomes of 1:2 and sonicated in an ice bath for 10 min. PBS was added to the mixture, and the mixture was centrifuged at 110,000 g at 4 °C for 1 h. The supernatant was discarded, and the mixture was resuspended in 500 μL of PBS to obtain the drug delivery system.
[0032] Example 2
[0033] The only difference between the drug delivery system of this embodiment and that of embodiment 1 is the mass ratio of black phosphorus quantum dots to budesonide in step (3). The mass ratio of black phosphorus quantum dots to budesonide in the drug delivery system of this embodiment is 1:1.
[0034] Example 3
[0035] The only difference between the drug delivery system in this embodiment and that in embodiment 1 is the mass ratio of black phosphorus quantum dots to budesonide in step (3). The mass ratio of black phosphorus quantum dots to budesonide in the drug delivery system of this embodiment is black phosphorus quantum dots: budesonide = 1:2.
[0036] Example 4
[0037] The only difference between the drug delivery system in this embodiment and that in embodiment 1 is the mass ratio of black phosphorus quantum dots to budesonide in step (3). The mass ratio of black phosphorus quantum dots to budesonide in the drug delivery system of this embodiment is black phosphorus quantum dots: budesonide = 1:3.
[0038] Example 5
[0039] The only difference between the drug delivery system in this embodiment and that in embodiment 1 is the mass ratio of black phosphorus quantum dots to budesonide in step (3). The mass ratio of black phosphorus quantum dots to budesonide in the drug delivery system of this embodiment is 1:4.
[0040] Example 6
[0041] The only difference between the drug delivery system in this embodiment and that in embodiment 1 is the mass ratio of black phosphorus quantum dots to budesonide in step (3). The mass ratio of black phosphorus quantum dots to budesonide in the drug delivery system of this embodiment is black phosphorus quantum dots: budesonide = 1:6.
[0042] Experimental Example 1
[0043] In this experimental example, the loading rate of budesonide in the drug delivery systems prepared in Examples 1-6 was determined using high-performance liquid chromatography. The results are as follows: Figure 1 A and Figure 1 As shown in B. Figure 1 It can be seen that the higher the amount of budesonide added, the higher the loading efficiency of the final drug delivery system.
[0044] Experiment Example 2
[0045] In this experimental example, the drug delivery system prepared in Example 1 was characterized using transmission electron microscopy (TEM), and the results are as follows: Figure 1 As shown in Figure C, electron microscopy results show clear black bright spots inside the exosomes, indicating that black phosphorus was successfully encapsulated by the exosomes.
[0046] Experimental Example 3
[0047] Eosinophil chemokine 3 (Eotaxin-3) is a typical inflammatory factor highly expressed by epithelial cells during airway inflammation. Stimulation of BEAS-2B airway epithelial cells with IL-4 significantly increased Eotaxin-3 expression, exhibiting an inflammatory phenotype. BEAS-2B airway epithelial cells stimulated with IL-4 were divided into three groups and treated with 10 μg / ml budesonide, mesenchymal stem cell exosomes, and the drug delivery system of Example 1 with a final budesonide concentration of 6.60 μg / ml, respectively. The expression of Eotaxin-3 in BEAS-2B airway epithelial cells of each group was then detected.
[0048] The measurement results are as follows Figure 2 As shown. By Figure 2It was found that budesonide (10 ug / ml) treatment decreased Eotaxin-3 expression (Group 3), while exosome-only treatment showed no change (Group 4). However, the drug delivery system of Example 1, with a final budesonide concentration of 6.60 ug / ml, significantly downregulated Eotaxin-3 expression (Group 5), and the effect was superior to Group 3. This indicates that black phosphorus-budesonide and mesenchymal stem cell exosomes have a synergistic anti-inflammatory effect. Furthermore, the final budesonide concentration in Group 5 was 6.60 ug / ml, lower than that in Group 3 (10 ug / ml), preliminarily confirming that exosomes loaded with black phosphorus-budesonide can reduce the required budesonide concentration.
[0049] Experiment Example 4
[0050] In this experiment, PKH26 dye was used to label the mesenchymal stem cell exosomes from Example 1, and these exosomes were injected via the tail vein into mice with an allergic asthma model. Eight hours later, the mice's organs were removed and photographed. The results are as follows: Figure 3 As shown. By Figure 3 It is known that exosomes can specifically accumulate in the lungs. Figure 3 A), heart, intestines, spleen, kidneys ( Figure 3 No signal was detected in the BE (Below-the-Lung) signal, indicating that the drug delivery system of the present invention has lung targeting.
[0051] Finally, it should be noted that the above-described embodiments are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be pointed out that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.
Claims
1. A drug delivery system for the treatment of allergic asthma, characterized in that, The drug delivery system is composed of black phosphorus quantum dots, exosomes and budesonide; the black phosphorus quantum dots load budesonide to obtain drug-loaded black phosphorus quantum dots; the mass ratio of the black phosphorus quantum dots to budesonide is 1:1-6; the exosomes are mesenchymal stem cell exosomes; the mass ratio of the black phosphorus quantum dots to the mesenchymal stem cell exosomes is 1:2; the mesenchymal stem cell exosomes encapsulate the drug-loaded black phosphorus quantum dots to obtain the drug delivery system.
2. The method of claim 1, wherein the drug delivery system is prepared by, The method comprises the following steps: (1) preparing black phosphorus quantum dots; (2) preparing mesenchymal stem cell exosomes; (3) the preparation method of the drug-loaded black phosphorus quantum dots is as follows: centrifuging the supernatant of the supernatant containing black phosphorus quantum dots to discard the supernatant and enrich the black phosphorus quantum dots; then uniformly mixing the black phosphorus quantum dots and polyethylene glycol, adding budesonide for ice bath ultrasonication to obtain the drug-loaded black phosphorus quantum dots; (4) the preparation method of the mesenchymal stem cell exosomes encapsulating the drug-loaded black phosphorus quantum dots is as follows: uniformly mixing the drug-loaded black phosphorus quantum dots and the mesenchymal stem cell exosomes and performing ice bath ultrasonication, and then adding PBS for centrifugation after the ultrasonication is completed.
3. The method of claim 2, wherein the drug delivery system is prepared by, The preparation method of the black phosphorus quantum dots in step (1) is as follows: suspending black phosphorus bulk crystals in isopropyl alcohol, grinding into black phosphorus powder and transferring to a brown bottle, and performing peeling through ultrasonication; centrifuging the obtained black phosphorus solution at 12000 rpm for 20 minutes and then at 18000 rpm for 20 minutes to obtain the black phosphorus quantum dots.
4. The method of claim 2, wherein the drug delivery system is prepared by, The mass ratio of the black phosphorus quantum dots to polyethylene glycol in step (3) is 1:
10.
5. The use of the drug delivery system according to claim 1 in the preparation of a drug for treating allergic asthma.
Citation Information
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