Tripterine-loaded nanoparticles as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to tripterine-loaded nano-particles as well as a preparation method and application thereof, and the tripterine-loaded nano-particles (Cel-GDNPs) comprise ginger exosome (GDNPs) and tripterine (Cel) encapsulated in the ginger exosome. According to the invention, the Cel is used as a therapeutic drug, and the adopted carrier GDNPs has anti-inflammatory activity, so that when the Cel and the GDNPs are used for treating inflammatory diseases such as colitis and the like, the Cel and the GDNPs can play a multi-component synergistic therapeutic effect, and compared with the single use of the Cel or the GDNPs, the expression of proinflammatory factors (such as TNF-alpha, IL-1beta and IL-6) can be more effectively inhibited, so that the synergistic therapeutic effect is realized; a new strategy is provided for developing a novel natural compound preparation.

Description

Technical Field

[0001] This invention relates to a nanoparticle loaded with triptolide, its preparation method and application, belonging to the field of pharmaceutical technology. Background Technology

[0002] Celastrol (Cel), a pentacyclic triterpenoid compound extracted from the roots of Tripterygium wilfordii Hook. f., a plant in the Celastraceae family, is considered one of the key active ingredients in the pharmacological effects of Tripterygium wilfordii. Clinical studies have shown that celastrol has significant therapeutic potential in a variety of diseases, including atherosclerosis, obesity, arthritis, liver fibrosis, and inflammatory bowel disease, exhibiting broad pharmacological activity.

[0003] However, despite its significant pharmacological activity, triptolide still faces numerous challenges in practical clinical application. First, its extremely poor water solubility results in low oral bioavailability, limiting its efficacy when taken orally. Second, triptolide exhibits certain toxic side effects in vivo; oral administration easily irritates the gastric mucosa, causing gastrointestinal adverse reactions such as nausea and vomiting; long-term or high-dose administration may also cause liver and kidney damage, further restricting its clinical promotion and application prospects.

[0004] To overcome the aforementioned shortcomings, researchers have focused on developing drug delivery systems suitable for triptolide in recent years, aiming to improve its solubility, bioavailability, and reduce toxic side effects. Currently, various synthetic nanocarriers, such as liposomes and polymer nanoparticles, have been attempted for loading and delivering triptolide. However, these synthetic nanocarriers generally suffer from poor biocompatibility, potential in vivo toxicity, and immunogenicity, limiting their further clinical application. For example, Chinese invention patent CN106309364B discloses a triptolide nanosuspension using amphiphilic stabilizers including mPEG-PCL, mPEG-DSPE, benzyl ether 78, SPC, and Tween 80. While this improves efficacy to some extent, the carrier materials still pose a risk of inducing immune responses or tissue accumulation toxicity with long-term use.

[0005] Therefore, developing a triptolide delivery system that combines good biocompatibility, high drug loading efficiency, low toxicity, and suitability for long-term administration remains a key issue that urgently needs to be addressed in this technological field. Summary of the Invention

[0006] To address the aforementioned deficiencies in the prior art, this invention provides nanoparticles loaded with triptolide, their preparation method, and their applications.

[0007] The technical solution of the present invention to solve the above-mentioned technical problems is as follows: One of the objectives of this invention is to provide nanoparticles loaded with triptolide (Cel-GDNPs), comprising ginger exosomes (GDNPs) and triptolide (Cel) encapsulated within the ginger exosomes.

[0008] Furthermore, the encapsulation rate of the triptolide is 65%–75%, and the drug loading is 15%–18%.

[0009] Compared with the prior art, the beneficial effects of the above-mentioned nanoparticles loaded with triptolide of the present invention are as follows: The Cel-GDNPs of this invention use ginger-derived exosomes (GDNPs) as carriers. GDNPs are derived from edible plants and have good biocompatibility and in vivo degradability. Compared with the use of synthetic polymers (such as mPEG-PCL, polylactic acid, etc.) or synthetic lipid materials in the prior art, it fundamentally solves the problems of poor biocompatibility and long-term toxicity of carrier materials, and can significantly reduce the risk of immunogenic reactions and cumulative toxicity.

[0010] This invention utilizes Cel as a therapeutic agent while employing GDNPs, a carrier that possesses anti-inflammatory activity. In treating inflammatory diseases such as colitis, Cel and GDNPs can exert a multi-component synergistic therapeutic effect. Compared to using Cel or GDNPs alone, it can more effectively inhibit the expression of pro-inflammatory factors (such as TNF-α, IL-1β, and IL-6), thereby achieving a synergistic therapeutic effect and providing a new strategy for developing novel natural compound preparations.

[0011] The Cel-GDNPs prepared by this invention have uniform particle size (approximately 200–400 nm), moderate zeta potential, and stable nanostructure, which greatly improves the water dispersibility of Cel and lays a good foundation for subsequent formulation development (such as oral liquids and lyophilized powder injections).

[0012] A second objective of this invention is to provide a method for preparing the triptolide-loaded nanoparticles as described above, comprising the following steps: S1. Extraction and purification of ginger exosomes: Ginger exosomes were extracted and purified from ginger juice by differential centrifugation and ultracentrifugation. S2. Loading of triptolide: The ginger exosomes are mixed with an alcoholic solution containing triptolide and incubated to load the drug. Then, the free triptolide is removed to obtain triptolide-loaded nanoparticles.

[0013] Furthermore, in step S1, the specific operations for extraction and purification include: S11. Wash the ginger, cut it into pieces, juice it, and filter it to obtain ginger juice. S12. The ginger juice obtained in step S11 is centrifuged at a differential speed to remove cell debris and impurities, and the supernatant is collected. S13. Filter the supernatant obtained in step S12 through a membrane to remove residual particles. S14. The filtrate filtered in step S13 is subjected to ultracentrifugation, the precipitate is collected and resuspended in buffer solution to obtain crude ginger exosomes. S15. The crude ginger exosomes obtained in step S14 are purified by sucrose density gradient centrifugation, and the purified ginger exosomes are collected and resuspended.

[0014] Further, in step S12, the differential centrifugation includes centrifugation at 800–1200 × g for 8–12 minutes at 3°C–5°C to remove large particles, centrifugation at 2000–4000 × g for 15–25 minutes to remove small particles, and centrifugation at 8000–12000 × g for 50–80 minutes to remove cell debris.

[0015] Furthermore, in step S14, the conditions for ultracentrifugation include: centrifugation at 80,000 to 120,000 × g for 50 to 70 minutes.

[0016] Further, in step S15, the sucrose density gradient centrifugation purification includes: placing the crudely extracted ginger exosomes in a sucrose density gradient solution with a mass-volume fraction of 15%, 30%, 45%, and 60%, centrifuging at 80,000 to 120,000 × g for 100 to 140 minutes, and collecting the ginger exosomes located in the 30% to 45% sucrose gradient.

[0017] Furthermore, in step S2, the specific operations for loading triptolide include: S21. Dissolve triptolide in an alcohol solvent to prepare a triptolide solution; S22. The purified ginger exosomes obtained in step S1 are mixed with the triptolide solution obtained in step S21 and incubated at 35℃~39℃ for 1~3 hours to load triptolide into the ginger exosomes. S23. The mixture after incubation in step S22 is subjected to ultracentrifugation to remove free triptolide, the precipitate is collected and resuspended to obtain the triptolide-loaded nanoparticles.

[0018] Furthermore, in step S21, the alcohol solvent is anhydrous ethanol.

[0019] Furthermore, in step S22, the volume ratio of ginger exosomes to triptolide solution is 8-10:1, and the ratio of protein concentration of ginger exosomes to mass concentration of triptolide is 3.6:1.

[0020] Furthermore, in step S23, the ultracentrifugation conditions are centrifugation at 80,000–120,000 × g for 50–70 minutes.

[0021] Compared with the prior art, the beneficial effects of the above-mentioned technical solution of the present invention are as follows: This invention employs differential centrifugation combined with sucrose density gradient ultracentrifugation to extract GDNPs, followed by drug loading via an incubation-centrifugation method. This invention utilizes the natural lipid bilayer structure of GDNPs, enabling efficient Cel loading through physical incubation with high encapsulation efficiency. Using inexpensive and readily available ginger as a starting material, this invention extracts nanocarriers through physical methods, avoiding the complex preparation and purification processes of synthesizing polymers or lipid materials. This significantly reduces raw material costs and production energy consumption. The entire process is simple to operate, operates under mild conditions, and requires no complex chemical synthesis or expensive equipment.

[0022] A third objective of this invention is to provide the use of the triptolide-loaded nanoparticles described above in the preparation of medicaments for the prevention and / or treatment of colitis.

[0023] Furthermore, the drug is used to reduce the levels of inflammatory factors TNF-α, IL-1β and / or IL-6 in colonic tissue.

[0024] Compared with the prior art, the beneficial effects of the above-mentioned technical solution of the present invention are as follows: The Cel-GDNPs of this invention can synergistically inhibit the inflammatory cascade response in colonic tissue from multiple targets and pathways. They can significantly inhibit the overexpression of inflammatory factors in the colonic tissue of mice with colitis, improving the therapeutic effect of Cel alone and better alleviating colonic inflammation. Through the encapsulation and delivery of GDNPs, this invention can reduce the exposure and accumulation of Cel in non-target organs (such as the stomach, liver, and kidneys), mitigating the direct irritation of Cel to the gastric mucosa (oral side effects) and reducing the risk of liver and kidney damage due to high doses or long-term use. Therefore, while improving efficacy, it significantly improves the safety window of the medication. Attached Figure Description

[0025] Figure 1 This is a process flow diagram of the extraction and purification process of ginger exosomes in a specific embodiment of the present invention.

[0026] Figure 2 Transmission electron microscope image of ginger exosomes extracted according to a specific embodiment of the present invention.

[0027] Figure 3 The image shows the zeta potential distribution of ginger exosomes extracted in a specific embodiment of the present invention.

[0028] Figure 4This is a particle size distribution diagram of ginger exosomes extracted in a specific embodiment of the present invention.

[0029] Figure 5 Zeta potential distribution of nanoparticles loaded with triptolide prepared according to a specific embodiment of the present invention.

[0030] Figure 6 The particle size distribution diagram of the triptolide-loaded nanoparticles prepared according to a specific embodiment of the present invention.

[0031] Figure 7 This is a comparison of the UV-Vis absorption spectra of triptolide, ginger exosomes, and nanoparticles loaded with triptolide in a specific embodiment of the present invention.

[0032] Figure 8 Comparison of X-ray diffraction patterns of triptolide, Cel-GDNPs, and physical mixtures of Cel and GDNPs in specific embodiments of the present invention.

[0033] Figure 9 This is a laser scanning confocal microscopy (CLSM) image of RAW264.7 macrophages taking up free C6 and C6-GDNPs in a specific embodiment of the present invention.

[0034] Figure 10 This is a laser scanning confocal microscopy (CLSM) image showing the uptake of free C6 and C6-GDNPs by Caco-2 intestinal epithelial cells in a specific embodiment of the present invention.

[0035] Figure 11 This is a statistical chart showing the detection results of TNF-α inflammatory factor content levels in the colon tissue of each group of colitis model mice in a specific embodiment of the present invention.

[0036] Figure 12 This is a statistical chart showing the detection results of IL-1β inflammatory factor levels in the colon tissue of each group of colitis model mice in a specific embodiment of the present invention.

[0037] Figure 13 This is a statistical chart showing the detection results of IL-6 inflammatory factor levels in the colon tissue of each group of colitis model mice in a specific embodiment of the present invention. Detailed Implementation

[0038] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are only for explaining the present invention and are not intended to limit the scope of the present invention.

[0039] I. Extraction and purification of ginger exosomes (GDNPs), see [link to documentation]. Figure 1 This includes the following steps: (1) Weigh 400g of fresh ginger, wash it, cut it into pieces, put it into a juicer, add 800 mL of pre-cooled PBS (phosphate buffer), and extract it repeatedly 4 times until there are no obvious visible fragments. The ginger juice obtained is initially filtered through four layers of sterile gauze to remove coarse residues. The filtrate is collected in a 50 mL centrifuge tube.

[0040] (2) Perform a series of differential centrifugations on the filtrate to gradually remove impurities: ① Centrifuge at 1,000 × g for 10 minutes at 4℃ to remove larger fragments and collect the supernatant; ② At 4°C, the supernatant was centrifuged at 3,000 × g for 20 minutes to remove smaller fragments, and the supernatant was collected again; ③ At 4℃, the above supernatant was centrifuged at 10000 × g for 60 minutes to remove cell debris and other impurities.

[0041] (3) The supernatant after centrifugation in step (2) is filtered sequentially through microporous membranes with pore sizes of 0.88 μm, 0.45 μm and 0.22 μm to further remove residual microparticles.

[0042] (4) Take the filtrate from step (3) into centrifuge tubes, weigh them to balance the mass, and ensure that the mass difference between each tube does not exceed 0.01 g. After balancing, centrifuge at 4°C and 100,000 × g for 60 min in an ultracentrifuge. Discard the supernatant and resuspend the precipitate in 2 mL of PBS to obtain crude GDNPs.

[0043] (5) Purification of crude GDNPs was performed using a sucrose density gradient centrifugation method: In an ultracentrifuge tube, 2 mL, 3 mL, 3 mL, and 2 mL of 60%, 45%, 30%, and 15% (w / v) sucrose solutions were layered sequentially from bottom to top. The crude GDNP sample was slowly added to the top layer of the gradient solution. The tube was centrifuged at 100,000 × g for 120 minutes at 4°C. After centrifugation, exosomes located in the 30%–45% sucrose layer were aspirated. The exosomes were resuspended in an equal volume of PBS and centrifuged again at 100,000 × g for 60 minutes at 4°C to remove excess sucrose solution, yielding the purified GDNPs.

[0044] (6) The protein concentration of purified GDNPs was determined to be 4 mg / mL using the BCA protein quantification kit. The concentration was adjusted to 1.0 mg / mL with PBS. After aliquoting, the GDNPs were stored in a -80℃ freezer for later use, avoiding repeated freeze-thaw cycles.

[0045] Result characterization: Transmission electron microscopy (TEM) revealed that the extracted GDNPs were cup-shaped or round with disc-shaped concave structures and intact morphology (see [link to TEM]). Figure 2 The particle size distribution and zeta potential of GDNPs were determined using a Malvern laser particle size analyzer, and the results showed that the particle size range of GDNPs was 100–200 nm (see [link to study]). Figure 4 The zeta potential is approximately -15 mV to -5 mV (see [reference]). Figure 3 ).

[0046] II. The preparation method of triptolide-loaded nanoparticles (Cel-GDNPs) is as follows: (1) Weigh 10.0 mg of triptolide (Cel) and dissolve it in 10 mL of anhydrous ethanol until it is fully dissolved, so as to prepare a Cel ethanol solution with a concentration of 1 mg / mL for subsequent use. (2) Dilute the 1.0 mg / mL GDNPs solution prepared in step one above to a protein concentration of 0.1 mg / mL, and dilute the 1.0 mg / mL Cel ethanol solution to 0.25 mg / mL. Take 9 mL of the 0.1 mg / mL GDNPs solution and slowly add 1 mL of the 0.25 mg / mL Cel ethanol solution dropwise under gentle vortex stirring (i.e., the volume ratio of GDNPs solution to Cel solution = 9:1). Under this ratio, the protein concentration of GDNPs to the mass concentration of Cel is 3.6:1.

[0047] (3) Place the mixed solution in a 37°C constant temperature water bath shaker and incubate for 2 hours to allow Cel to be fully loaded into GDNPs.

[0048] (4) After incubation, transfer the mixture to a centrifuge tube and centrifuge at 10,000 × g for 10 min at 4 °C. Transfer the supernatant to an ultracentrifuge tube and centrifuge at 100,000 × g for 60 min at 4 °C. Discard the supernatant and resuspend the precipitate in PBS to obtain purified Cel-GDNP nanoparticles.

[0049] The effects of different concentration ratios of Cel and GDNPs on encapsulation efficiency (EE%) and drug loading (DL%) were studied through single-factor experiments.

[0050] Dilute the GDNPs solution to 0.1 mg / mL and the Cel ethanol solution to 0.1 mg / mL. Take 9 mL of the 0.1 mg / mL GDNPs solution and slowly add 1 mL of the 0.1 mg / mL Cel ethanol solution dropwise while gently vortexing (i.e., the GDNPs solution:Cel solution volume ratio = 9:1). At this ratio, the protein concentration of GDNPs to the mass concentration of Cel is 9:1, and the operation procedure is the same as above.

[0051] Dilute the GDNPs solution to 0.1 mg / mL and the Cel ethanol solution to 0.5 mg / mL. Take 9 mL of the 0.1 mg / mL GDNPs solution and slowly add 1 mL of the 0.1 mg / mL Cel ethanol solution dropwise while gently vortexing (i.e., GDNPs solution:Cel solution volume ratio = 9:1). At this ratio, the protein concentration of GDNPs to the mass concentration of Cel is 1.8:1, and the operation procedure is the same as above.

[0052] The highest encapsulation efficiency was achieved when the ratio of GDNP protein concentration to Cel mass concentration was 3.6:1. Specific results are shown in Table 1 below. Table 1. Encapsulation efficiency and drug loading of nanoparticles with different concentration ratios of Cel and GDNPs.

[0053] III. Characterization and Drug Loading Performance Evaluation of Cel-GDNPs: (1) Physicochemical characterization: The particle size and zeta potential of Cel-GDNPs were measured using a Malvern laser particle size analyzer. The results showed that the average particle size of Cel-GDNPs was 200–400 nm (see [reference needed]). Figure 6 The zeta potential is approximately -10 mV to -5 mV (see [reference]). Figure 5 UV-Vis spectrophotometric scanning showed that Cel-GDNPs exhibited a distinct absorption peak at the characteristic absorption wavelength of Cel, while blank GDNPs showed no absorption peak at this location, indicating that Cel was successfully loaded (see [link to relevant documentation]). Figure 7 Cel-GDNPs were freeze-dried into powder for X-ray diffraction pattern testing. The results showed that the crystal form diffraction peaks of Cel disappeared after loading, indicating that the crystal form of Cel changed and it exists in an amorphous or molecular state in GDNPs (see [link to relevant documentation]). Figure 8 ).

[0054] (2) Determination of drug loading and encapsulation efficiency: The drug loading efficiency of Cel-GDNPs was determined by high performance liquid chromatography combined with ultracentrifugation. Specifically, the Cel-GDNPs precipitate obtained after centrifugation in step two was resuspended in 1.0 mL of PBS. 10 μL of Cel-GDNPs nanoparticles were placed in a test tube, and then 990 μL of anhydrous ethanol was added to disrupt the exosome membrane. The mixture was filtered through a 0.22 μm microporous membrane, and the concentration of Cel in Cel-GDNPs was determined by HPLC. The encapsulation efficiency (EE%) and drug loading (DL%) of Cel-GDNPs were calculated according to the following formulas.

[0055] EE%=(M 包 / M 总 ) * 100%; DL%=[M包 / (M) 总 +M 载 )*100%; in, M 包 The quality of triptolide encapsulated in ginger exosomes; M 总 Total mass of triptolide added during the preparation of drug-loaded exosomes; M 载 The quality of ginger exosomes; Chromatographic conditions: Column: C18, 4.6*250 mm, 5 µm; column temperature: 25 ℃; mobile phase: acetonitrile: 0.1% phosphoric acid water (80:20, v / v); flow rate: 1 mL·min -1 The detection wavelength was 425 nm and the injection volume was 10 μL.

[0056] The results showed that the encapsulation efficiency (EE%) of Cel in Cel-GDNPs was 70.8% and the drug loading (DL%) was 16.4%, indicating that Cel-GDNPs have a high drug loading capacity.

[0057] IV. Cellular uptake of Cel-GDNPs assay: To visually observe cellular uptake, C6-labeled GDNPs (C6-GDNPs) were prepared using the hydrophobic fluorescent probe coumarin 6 (C6) instead of Cel, following the same method as described in this invention.

[0058] Preparation of C6 solution: Weigh 10 mg of C6 powder in the dark and dissolve it in 10 mL of anhydrous ethanol until fully dissolved to prepare a C6 ethanol solution with a concentration of 1 mg / mL. Then, use PBS to serially dilute to 10 μg / mL to obtain the C6 solution that will be diluted with DMEM basal medium.

[0059] Preparation of C6-GDNPs solution: The preparation method for Cel-GDNPs was the same as described above, except that 1 mg / mL of C6 ethanol solution was used instead of 1 mg / mL of Cel ethanol solution. The prepared C6-GDNPs were dissolved in PBS to a concentration of 10 μg / mL. RAW264.7 cells (mouse macrophages) and Caco-2 cells (human colon adenocarcinoma cells) in logarithmic growth phase were then cultured at a concentration of 2 × 10⁻⁶ cells / mL. 5Cells were seeded at the specified density in glass-bottomed culture dishes, with 2 mL of DMEM complete medium added to each dish. The dishes were incubated overnight at 37°C with 5% CO2. Two cell lines were established: a control group (C6 group) and an experimental group (C6-GDNPs group). Once the cells adhered and reached 80% confluence, 2 mL of C6 solution diluted with DMEM basal medium (C6 concentration 1 μg / mL) was added to the C6 group, and C6-GDNPs solution (C6 concentration 1 μg / mL) was added to the C6-GDNPs group. The cells were incubated at 37°C for 1 h. After incubation, the medium was discarded, and the cells were washed with PBS. 1 mL of 4% paraformaldehyde fixative was added to each dish, and the cells were fixed at room temperature for 10 min. After fixation, the cells were washed three times with PBS, and then 4',6-diamidinyl-2-phenylindole (DAPI) staining solution was added, and the cells were stained at room temperature for 10 min. After staining, the cells were washed three times with PBS, and the uptake of C6 by RAW264.7 cells and Caco-2 cells was observed using a laser scanning confocal microscope (CLSM, Ex = 488 nm).

[0060] The results are as follows Figure 9 and Figure 10 As shown, green represents C6-labeled GDNPs, and blue represents the nuclei of RAW264.7 and Caco-2 cells stained with DAPI. The results showed that no significant green fluorescence was observed in either RAW264.7 or Caco-2 cells in the C6 group, while significant green fluorescence was observed in both RAW264.7 and Caco-2 cells in the C6-GDNPs group. This indicates that compared to the free C6 group, C6-GDNPs can be taken up more efficiently by RAW264.7 and Caco-2 cells, thereby improving Cel utilization efficiency, reducing the dosage, and lowering the probability of toxic side effects caused by high-dose administration.

[0061] V. In vivo anti-inflammatory activity study of Cel-GDNPs: (1) Animal model establishment and drug administration: 5-6 week old male C57BL / 6 mice (18g-22g) were randomly divided into 4 groups: control group, DSS group, DSS+Cel group and DSS+Cel+GDNPs group, with 5 mice in each group. The control group was given normal drinking water and 200 μL of sterile water was administered by gavage daily. The drinking water of the DSS group, DSS+Cel group and DSS+Cel+GDNPs group was replaced with drinking water containing 3% (w / v) sodium dextran sulfate (DSS) for 7 consecutive days to induce an acute ulcerative colitis model. At the same time as the model was established, the DSS group was administered 200 μL of sterile water by gavage daily, the DSS+Cel group was administered 200 μL of Cel solution daily, and the DSS+Cel+GDNPs group was administered 200 μL of Cel-GDNPs solution daily. The dosage was 3 mg / kg.

[0062] (2) Sample collection and index detection: On the 8th day of the experiment, the mice were sacrificed and colon tissues from the same location were taken. The contents of inflammatory factors (TNF-α, IL-1β and IL-6) in the colon tissues of each group were measured by ELISA (enzyme-linked immunosorbent assay).

[0063] The results are as follows Figures 11-13 As shown, compared with the DSS group and the DSS+Cel group, the content of inflammatory factors in the DSS+Cel+GDNPs group was significantly reduced, indicating that the Cel-GDNPs nanoparticles of the present invention can more significantly reduce the expression of inflammatory factors through synergistic effects, and more effectively inhibit the local inflammatory response in the colon of colitis mice, thus exhibiting significant anti-colitis efficacy.

[0064] Note: Figures 11-13 In the table, *P<0.05, **P<0.01, and ****P<0.0001 indicate statistically significant differences compared to the model control group.

[0065] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A nanoparticle loaded with triptolide, characterized in that, It includes ginger exosomes and triptolide encapsulated within the ginger exosomes.

2. The nanoparticles loaded with triptolide according to claim 1, characterized in that, The encapsulation rate of triptolide is 65%–75%, and the drug loading is 15%–18%.

3. A method for preparing nanoparticles loaded with triptolide as described in claim 1 or 2, characterized in that, Includes the following steps: S1. Extraction and purification of ginger exosomes: Ginger exosomes were extracted and purified from ginger juice by differential centrifugation and ultracentrifugation. S2. Loading of triptolide: The ginger exosomes are mixed with an alcoholic solution containing triptolide and incubated to load the drug. Then, the free triptolide is removed to obtain triptolide-loaded nanoparticles.

4. The method for preparing nanoparticles loaded with triptolide according to claim 3, characterized in that, In step S1, the specific operations for extraction and purification include: S11. Wash the ginger, cut it into pieces, juice it, and filter it to obtain ginger juice. S12. The ginger juice obtained in step S11 is centrifuged at a differential speed to remove cell debris and impurities, and the supernatant is collected. S13. Filter the supernatant obtained in step S12 through a membrane to remove residual particles. S14. The filtrate filtered in step S13 is subjected to ultracentrifugation, the precipitate is collected and resuspended in buffer solution to obtain crude ginger exosomes. S15. The crude ginger exosomes obtained in step S14 are purified by sucrose density gradient centrifugation, and the purified ginger exosomes are collected and resuspended.

5. The method for preparing nanoparticles loaded with triptolide according to claim 4, characterized in that, In step S12, the differential centrifugation includes centrifuging at 800–1200 × g for 8–12 minutes at 3°C–5°C to remove large particles, centrifuging at 2000–4000 × g for 15–25 minutes to remove small particles, and centrifuging at 8000–12000 × g for 50–80 minutes to remove cell debris. In step S14, the conditions for ultracentrifugation include centrifugation at 80,000–120,000 × g for 50–70 minutes.

6. The method for preparing nanoparticles loaded with triptolide according to claim 4, characterized in that, In step S15, the sucrose density gradient centrifugation purification includes: placing the crudely extracted ginger exosomes in a sucrose density gradient solution with a mass-volume fraction of 15%, 30%, 45%, and 60%, centrifuging at 80,000 to 120,000 × g for 100 to 140 minutes, and collecting the ginger exosomes located in the 30% to 45% sucrose gradient.

7. The method for preparing triptolide-loaded nanoparticles according to claim 3, characterized in that, In step S2, the specific operations for loading triptolide include: S21. Dissolve triptolide in an alcohol solvent to prepare a triptolide solution; S22. The purified ginger exosomes obtained in step S1 are mixed with the triptolide solution obtained in step S21 and incubated at 35℃~39℃ for 1~3 hours to load triptolide into the ginger exosomes. S23. The mixture after incubation in step S22 is subjected to ultracentrifugation to remove free triptolide, the precipitate is collected and resuspended to obtain the triptolide-loaded nanoparticles.

8. The method for preparing triptolide-loaded nanoparticles according to claim 7, characterized in that, In step S21, the alcohol solvent is anhydrous ethanol; in step S22, the volume ratio of ginger exosomes to triptolide solution is 8-10:1, and the ratio of protein concentration of ginger exosomes to mass concentration of triptolide is 3.6:1; in step S23, the ultracentrifugation conditions are centrifugation at 80,000-120,000 × g for 50-70 minutes.

9. The use of the triptolide-loaded nanoparticles as described in claim 1 or 2 in the preparation of medicaments for the prevention and / or treatment of colitis.

10. The application according to claim 9, characterized in that, The drug is used to reduce the levels of inflammatory factors TNF-α, IL-1β and / or IL-6 in colon tissue.

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