Nano-drug for treating idiopathic pulmonary fibrosis as well as preparation method and application of nano-drug

Abstract

The invention relates to the technical field of biological medicines, in particular to a nano-drug for treating idiopathic pulmonary fibrosis as well as a preparation method and application of the nano-drug. The nano-particles are prepared by loading the TGF-beta1 inhibitor and the RSV by using hemoglobin, so that the TGF-beta1 inhibitor and the RSV are cooperated to effectively treat idiopathic pulmonary fibrosis.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, in particular to a nano drug for treating idiopathic pulmonary fibrosis and a preparation method and application thereof. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease that primarily affects middle-aged and elderly people and is characterized by progressive dyspnea, dry cough, chest discomfort, and finger clubbing.

[0003] The main pathological manifestations of IPF are related to the persistent damage of alveolar epithelium, which leads to the recruitment of downstream effector cells and the secretion of multiple cytokines and fibrogenic factors. For example, TGF-β1 further promotes the occurrence of pulmonary fibrosis by promoting the proliferation and differentiation of fibroblasts, stimulating fibroblasts to synthesize collagen fibers, and inhibiting the degradation of extracellular matrix. Oxidative stress plays a key role in the pathogenesis of IPF. It leads to the imbalance of oxidation / antioxidation in the body, promotes the disorder of cytokine network related to pulmonary fibrosis, causes cell rupture, releases lysosomes, accelerates cell apoptosis, promotes alveolar inflammatory response, aggravates lung damage, and forms pulmonary fibrosis.

[0004] Pirfenidone and nintedanib are currently approved drugs for the treatment of IPF. These two drugs exert anti-inflammatory and anti-fibrotic effects by downregulating TGF-β1 and multiple cytokines. Although these two drugs can relieve lung damage by inhibiting TGF-β1 fibrosis, these drugs cannot improve alveolar cell damage caused by oxidative stress. Summary of the invention

[0005] In order to solve the problems in the above-mentioned background technology, an object of the present invention is to provide a nanomedicine (HbRT) for treating idiopathic pulmonary fibrosis, wherein the nanoparticles include hemoglobin, and the surface of the hemoglobin is loaded with a TGF-β1 inhibitor and RSV, and the TGF-β1 inhibitor is any one or any combination of SB431542, LY2157299, LY2109761, TP0427736, IN1130, and LY3200882.

[0006] TGF-β1 inhibitors can inhibit the expression of TGF-β1 in idiopathic pulmonary fibrosis lesions, thereby inhibiting the expression of fibrotic factors and reducing fibrosis proliferation; while RSV can scavenge free radicals and reduce oxidative stress. The above-mentioned resveratrol includes resveratrol derivatives to fight pulmonary fibrosis from multiple aspects.

[0007] Preferably, the particle size of the nanoparticles is 180-240 nm.

[0008] The second object of the present invention is to provide a method for preparing a nano drug with simple steps, comprising the following steps:

[0009] S1 lysed hemoglobin:

[0010] Dissolve 0.5-3 mg of hemoglobin in 1-3 mL of deionized water, and then perform ultrasonic treatment until the hemoglobin is evenly dispersed to obtain an initial hemoglobin solution;

[0011] The temperature is controlled at 23-27°C, the relative humidity is 40-60%, and the pH value of the initial hemoglobin solution is maintained at 6.8-7.4;

[0012] S2 Preparation of a mixed solution of TGF-β1 inhibitor and RSV:

[0013] A TGF-β1 inhibitor having a concentration of 10 mg / mL and resveratrol (RSV) having a concentration of 40 mg / mL are added to an organic solvent, followed by sonication until the TGF-β1 inhibitor and resveratrol (RSV) are completely dissolved and mixed uniformly to obtain a mixed solution;

[0014] The temperature is controlled at 23-27°C, and the volume ratio of the TGF-β1 inhibitor to RSV and the organic solvent is (2-25) μL: (1-10) μL: (0.2-1.0) mL;

[0015] S3 Preparation of Nanoparticles:

[0016] The mixed solution prepared in step S2 is added dropwise to the hemoglobin solution prepared in step S1 to obtain a base solution, wherein the volume ratio of the mixed solution to the hemoglobin solution is 1:(3-6); the base solution is then subjected to ultrasonic treatment for 5-15 minutes, and then the ultrasonicated base solution is centrifuged at a centrifugal force of 15000g-30000g for 10-30 minutes at a centrifugal temperature of 4-8°C to obtain nanoparticles.

[0017] Preferably, the organic solvent is dimethyl sulfoxide.

[0018] The third object of the present invention is to provide an application of nanomedicine in the preparation of anti-fibrosis preparations.

[0019] The dosage form of the preparation includes tablets, powders, suspensions, granules, capsules, injections, sprays, solutions, enemas, emulsions, films, suppositories, patches, nasal drops or pills.

[0020] Preferably, the preparation is in the form of a spray. Forming the nano drug into a spray facilitates non-invasive inhalation of the drug into the lungs, helps the drug to accumulate at the wound site, and improves the therapeutic effect.

[0021] A fourth object of the present invention is to provide a drug delivery system.

[0022] In summary, the beneficial effects of the present invention are:

[0023] TGF-β1 inhibitors can inhibit the expression of TGF-β1, thereby inhibiting the expression of fibrosis factors and reducing fibrosis.

[0024] RSV can scavenge free radicals and reduce oxidative stress.

[0025] The HbRT of the present application can be inhaled into the lungs non-invasively, exerting a synergistic effect to combat pulmonary fibrosis while improving alveolar cell damage caused by oxidative stress.

[0026] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention, it can be implemented in accordance with the contents of the specification. In order to make the above and other purposes, features and advantages of the present invention more obvious and easy to understand, the following specifically cites a preferred embodiment and describes it in detail with the accompanying drawings as follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 : Transmission electron microscopy images and particle size distribution of HbRT;

[0028] Figure 2 : HbRT particle size and polydispersity index monitoring;

[0029] Figure 3 : Paramagnetic spectra of hydroxyl radicals and nitroxide radicals;

[0030] Figure 4 : Confocal imaging of intracellular reactive oxygen species levels;

[0031] Figure 5 : Quantitative detection of TGF-β1 expression at the cellular level;

[0032] Figure 6 :Flow cytometry detection of cell fibrosis indicators α-SMA and Col-Ⅰ expression;

[0033] Figure 7 : Fluorescence imaging of idiopathic pulmonary fibrosis model mice after drug administration;

[0034] Figure 8 : Imaging of HE, α-SMA and Col-Ⅰ expression in mouse lung tissue after drug treatment;

[0035] Fig. 9 : Pulmonary function test of mice after drug treatment. DETAILED DESCRIPTION

[0036] In order to make the content of the present invention more clearly understood, the present invention is further described below based on specific embodiments in conjunction with the accompanying drawings.

[0037] Resveratrol (RSV) of the present embodiment is a non-flavonoid polyphenol compound, and its chemical name is E-3,5,4'-trihydroxystilbene. Its chemical structure contains three phenolic hydroxyl groups, especially the 4-parahydroxyl group, which is the main source of its antioxidant capacity. Resveratrol is present in the dry root bark of Moraceae plants. Resveratrol has an anti-fibrotic effect, can inhibit the proliferation of fibroblasts, and reduce the generation of collagen fibers. In terms of antioxidant, resveratrol can inhibit the generation of free radicals, reduce oxidative stress, and thus has the effect of reducing alveolar damage.

[0038] In the treatment of IPF, it is of great significance to select effective drugs and increase the cumulative amount of drugs in the fibrosis site to improve the effective treatment effect. The present invention uses hemoglobin to load TGF-β1 inhibitors and RSV to prepare nanoparticles, thereby achieving effective treatment of idiopathic pulmonary fibrosis with TGF-β1 inhibitors and RSV.

[0039] The experimental steps of the present invention include:

[0040] S1 lysed hemoglobin:

[0041] Dissolve 0.5-3 mg of hemoglobin in 1-3 mL of deionized water, and then perform ultrasonic treatment until the hemoglobin is evenly dispersed to obtain an initial hemoglobin solution;

[0042] The temperature is controlled at 23-27°C, the relative humidity is 40-60%, and the pH value of the initial hemoglobin solution is maintained at 6.8-7.4;

[0043] Preferably, the concentration of the prepared hemoglobin is 0.75 mg / mL, the temperature is controlled at 25°C, the relative humidity is 50%, and the pH value of the initial hemoglobin solution is maintained at 7;

[0044] S2 prepares a mixed solution of TGF-β1 inhibitor and resveratrol (RSV):

[0045] A TGF-β1 inhibitor having a concentration of 10 mg / mL and resveratrol (RSV) having a concentration of 40 mg / mL are added to an organic solvent, followed by sonication until the TGF-β1 inhibitor and resveratrol (RSV) are completely dissolved and mixed uniformly to obtain a mixed solution;

[0046] The temperature is controlled at 23-27°C, and the volume ratio of the TGF-β1 inhibitor to RSV and the organic solvent is (2-25) μL: (1-10) μL: (0.2-1.0) mL;

[0047] Preferably, the temperature is controlled at 25°C, the volume ratio of TGF-β1 inhibitor to resveratrol (RSV) and organic solvent is 20 μL: 5 μL: 1 mL, and the ultrasonic time is 10 min;

[0048] S3 Preparation of HbRT nanoparticles:

[0049] The mixed solution prepared in step S2 is added dropwise to the hemoglobin solution prepared in step S1 to obtain a base solution, wherein the volume ratio of the mixed solution to the hemoglobin solution is 1:(3-6); the base solution is then subjected to ultrasonic treatment for 5-15 minutes, and then the ultrasonicated base solution is centrifuged at a centrifugal force of 15000g-30000g for 10-30 minutes at a centrifugal temperature of 4-8°C to obtain nanoparticles.

[0050] Preferably, the ultrasonic time is 10 min, the centrifugal temperature is controlled at 4° C., the centrifugal force is 16,000, and the centrifugal time is 20 min.

[0051] Preferably, the HbRT nanoparticles of the present invention are used to prepare spray-type medicines.

[0052] The hemoglobin-loaded transforming growth factor-β1 (TGF-β1) and RSV nanoparticles (HbRT) of the present invention, that is, hemoglobin has the function of loading drugs, and can effectively deliver drugs to fibrotic lesions after inhalation, thereby solving the problem that oral drugs cannot accumulate in fibrotic lesions due to the need to be transported through the blood circulation.

[0053] In the present application, the TGF-β1 inhibitor in HbRT can inhibit the proliferation of fibroblasts and the synthesis of collagen, reduce the degree of pulmonary fibrosis, inhibit inflammatory response and oxidative stress response, and further alleviate the deterioration of pulmonary fibrosis.

[0054] At the same time, resveratrol in HbRT can scavenge free radicals, inhibit inflammatory responses, and reduce oxidative stress responses, thereby alleviating damage to lung cells and slowing the progression of pulmonary fibrosis.

[0055] In conclusion, the HbRT delivery system can inhibit the inflammatory response in the lesions, inhibit the release of TGF-β1 factor, scavenge ROS and exert anti-oxidative stress effect, so as to achieve the purpose of non-invasive and efficient delivery of drugs to fibrotic lesions, and provide a new idea for the treatment of pulmonary fibrosis.

[0056] The hemoglobin of the present invention can also be loaded with anti-fibrosis drugs such as pirfenidone and nintedanib by the same method as above.

[0057] Embodiment 1

[0058] S1 lysed hemoglobin:

[0059] 2.25 mg of hemoglobin was dissolved in 3 ml of deionized water, followed by sonication to obtain an initial hemoglobin solution;

[0060] The temperature was controlled at 25°C, the relative humidity was 50%, and the pH value of the initial hemoglobin solution was maintained at 7.

[0061] S2 prepares a mixed solution of TGF-β1 inhibitor and resveratrol (RSV):

[0062] A TGF-β1 inhibitor SB431542 at a concentration of 10 mg / mL and RSV at a concentration of 40 mg / mL were added to a dimethyl sulfoxide (DMSO) solvent, followed by sonication until the TGF-β1 inhibitor and RSV were completely dissolved to obtain a mixed solution;

[0063] The temperature was controlled at 25°C, the volume of TGF-β1 inhibitor was 20 μL, the volume of RSV was 5 μL, the volume of organic solvent was 1.0 mL, and the ultrasonic time was 10 min;

[0064] S3 Preparation of Nanoparticles:

[0065] The mixed solution prepared in step S2 is added dropwise to the hemoglobin solution prepared in step S1 to obtain a base solution, which is then ultrasonicated for 10 minutes. The ultrasonicated base solution is then centrifuged at a centrifugal force of 16,000 g for 20 minutes with the centrifugal temperature controlled at 4°C to obtain nanoparticles (HbRT).

[0066] Experimental Analysis 1

[0067] HbRT structure detection

[0068] Take 10 μL of HbRT prepared in Example 1, disperse it in 1 mL of deionized water to obtain a diluted HbRT solution, take 10 μL of the diluted HbRT solution and drop it on the amorphous carbon film copper mesh, and observe it by transmission electron microscope after drying. The results are as follows Figure 1 As shown in (A), HbRT nanoparticles are spherical in structure, well dispersed and have relatively uniform particle size.

[0069] 10 μL of the HbRT nanoparticle solution prepared in Example 1 was dispersed in 1 mL of deionized water, and the hydrated particle size was detected by dynamic light scattering. Figure 1 As shown in (B), the hydrated particle size of HbRT is 200 nm.

[0070] Experimental Analysis II

[0071] HbRT stability test.

[0072] The HbRT prepared in Example 1 was dispersed into 1 mL of PBS and 10% FBS solution, respectively, and allowed to stand, with the volume of HbRT being 10 μL. Samples were then taken at 0, 2, 4, 6, and 8 days, and the particle size and polydispersity index (PDI) stability of HbRT were detected by dynamic light scattering.

[0073] The results are as follows Figure 2 As shown, the particle size and PDI of HbRT did not change significantly during the 7-day test period, and HbRT had good stability.

[0074] Experimental Analysis 3

[0075] Test of HbRT's ability to scavenge reactive oxygen free radicals.

[0076] The reactive oxygen radical scavenging ability of HbRT and an equal amount of RSV was characterized by electron spin resonance (ESR) spectroscopy. Hydroxyl radicals (·OH) were produced by Fenton reaction (the reactants were 70 μL of 0.735 mM FeCl2 and 25 μL of 0.315 mM H2O2) and captured by DMPO. Nitrogen oxide radicals (·NO) were produced by 70 μL of 1.125 mM SNAP under ultraviolet irradiation and captured by DEPMPO. 200 μg / mL HbRT and an equal amount of RSV were added to the above mixture, and the changes in the amplitude of the ESR spectra of DMPO-·OH adduct and DEPMPO-·NO adduct indicated the scavenging effect. Ctrl was the untreated control group and RSV was the positive control group.

[0077] The results are as follows Figure 3 As shown, HbRT can scavenge ·OH and ·NO, and its scavenging efficiency is not significantly different from that of free RSV.

[0078] Experimental Analysis 4

[0079] Detection of free radical scavenging ability at the cellular level.

[0080] Human embryonic lung fibroblasts (MRC5 cells) were placed in a constant temperature incubator at 37°C and 5% CO2 concentration. When the cells grew to 80%, they were passaged and inoculated in confocal dishes. The number of cells inoculated was 10,000 / well. After adherence, a TGF-β1 inducer with a concentration of 10 ng / mL was added. After 24 hours of culture, different groups of samples and an equal volume of 200 μmol / L hydrogen peroxide were added, followed by 10 mM free radical scavenger DCFH-DA. After 30 minutes, the cells were washed and confocal imaging was performed. The green fluorescence is the level of reactive oxygen in the cells. The higher the fluorescence intensity, the higher the level of reactive oxygen, indicating that the effect of producing free radicals is better. The different groups of samples are: control group (Ctrl), PBS, HbR, HbT and HbRT groups, with 3 replicates in each group.

[0081] The samples in the PBS, HbR, HbT and HbRT groups were equal amounts of PBS, hemoglobin nanoparticles loaded only with the free radical scavenger RSV, hemoglobin nanoparticles loaded only with the TGF-β1 inhibitor SB431542, and hemoglobin nanoparticles loaded with both RSV and SB431542, respectively. The control group (Ctrl) did not undergo the treatment step of adding TGF-β1 inducer, and the other steps were the same.

[0082] The results are as follows Figure 4 As shown, HbRT nanoparticles have good ROS scavenging efficiency and can effectively inhibit cellular oxidative stress response.

[0083] Experimental analyses three and four reflect the excellent free radical scavenging ability of HbRT.

[0084] Experimental Analysis 5

[0085] Detection of HbRT inhibiting TGF-β1 expression in vitro.

[0086] MRC5 cells were seeded in 96-well plates, with 5000 cells / well. After adherence, 10ul of 10ng / mL TGF-β1 inducer was added. After 24h of culture, the cell culture medium was removed, washed 3 times with PBS, and different groups of samples were added. After 24h, the cell supernatant was collected, and the expression of TGF-β1 in the cell supernatant of different groups was detected by enzyme-linked immunosorbent assay (ELISA). The different groups of samples were: control group (Ctrl), PBS, HbR, HbT and HbRT groups, with 3 replicates in each group.

[0087] The samples in the PBS, HbR, HbT and HbRT groups were equal amounts of PBS, hemoglobin nanoparticles loaded only with the free radical scavenger RSV, hemoglobin nanoparticles loaded only with the TGF-β1 inhibitor SB431542, and hemoglobin nanoparticles loaded with both RSV and SB431542, respectively. The control group (Ctrl) did not undergo the treatment step of adding TGF-β1 inducer, and the other steps were the same.

[0088] The results are as follows Figure 5 As shown, the expression of TGF-β1 in cells after HbRT intervention was significantly decreased, indicating that HbRT can effectively inhibit the expression of TGF-β1 in cells.

[0089] Experimental Analysis VI

[0090] In vitro anti-fibrosis assay of HbRT.

[0091] MRC5 cells were seeded in 24-well plates, with 10,000 cells per well. After adherence, 10ul of 10ng / mL TGF-β1 inducer was added. After 24 hours of culture, different groups of samples were added. After 12 hours, the cells were washed 3 times with PBS, and the cell fibrosis indicators α-SMA and Col-Ⅰ were fluorescently stained. The expression of cell fibrosis indicators α-SMA and Col-Ⅰ in different groups was detected by flow cytometry. The different groups of samples were: blank control group (Ctrl), PBS, HbR, HbT and HbRT groups, with 3 replicates in each group.

[0092] The samples in the PBS, HbR, HbT and HbRT groups were equal amounts of PBS, hemoglobin nanoparticles loaded only with the free radical scavenger RSV, hemoglobin nanoparticles loaded only with the TGF-β1 inhibitor SB431542, and hemoglobin nanoparticles loaded with both RSV and SB431542, respectively. The control group (Ctrl) did not undergo the treatment step of adding TGF-β1 inducer, and the other steps were the same.

[0093] The results are as follows Figure 6 As shown in (A), compared with the PBS group, the α-SMA signal in cells after HbRT intervention was weakened; Figure 6 As shown in (B), compared with the PBS group, the Col-Ⅰ signal in the cells treated with HbRT was weakened. Therefore, HbRT can effectively inhibit the fibrotic process of fibroblasts.

[0094] Experimental Analysis VII

[0095] C57BL / 6 mice were raised under specific pathogen-free conditions with a humidity of 50±10%, a temperature of 20±2°C, and a light-dark cycle of 12h. When the mice were 8 weeks old and weighed 20-24g, 1.5U / kg of bleomycin was administered by aerosol to establish an IPF model. Seven days after modeling, the mice were randomly divided into two groups and 200μg HbRT was administered to each mouse by tail vein (iv) and aerosol administration (Inhalation). Small animal imaging was performed at different time points after administration (0, 3, 6, 9, 12, 24, 48, 72, 96, 120h).

[0096] The results are as follows Figure 7 As shown in (A), the fluorescence intensity of the lung tissue of mice in the aerosol administration (Inhalation) group was stronger than that in the tail vein (iv) injection group. Figure 7 As shown in (B), compared with the tail vein (iv) injection group, the fluorescence retention time in the lung tissue of mice in the aerosol administration (Inhalation) group was longer, indicating that HbRT can be efficiently retained in the IPF lesion site after aerosol administration.

[0097] Experimental Analysis 8

[0098] Evaluation of the antifibrotic effect of HbRT in vivo.

[0099] IPF mice were randomly divided into 4 groups, 6 mice in each group, divided into: PBS group, HbR group, HbT group and HbRT group. The corresponding samples were given for treatment on days 5, 9, 13 and 17 after modeling. At the same time, healthy mice (Healthy) were given PBS as the control group. The mice were killed on the 22nd day, and the lung tissues were taken. After fixation in 4% paraformaldehyde, they were gradient dehydrated according to standard procedures and embedded in paraffin. The tissues were cut into 10 μm sections for HE staining. At the same time, the fibrosis indicators α-SMA and Col-Ⅰ of the lung tissue were fluorescently stained, and the lung fibrosis indicators α-SMA and col-Ⅰ were detected by immunofluorescence.

[0100] The results are as follows Figure 8 As shown, compared with the PBS control group, the fibrosis pathology in the HbRT group was significantly alleviated, and the expression of fibrosis indicators α-SMA and col-Ⅰ was significantly decreased, indicating that HbRT can effectively inhibit the process of bleomycin-induced lung fibrosis.

[0101] Experimental Analysis IX

[0102] Lung function test of mice after HbRT treatment.

[0103] IPF mice were randomly divided into 4 groups, 6 mice in each group, divided into PBS group, HbR group, HbT group and HbRT group. The corresponding samples were given for treatment on days 5, 9, 13 and 17 after modeling. At the same time, healthy mice (Healthy) were given PBS as a control group. On the 21st day, the lung function indicators of mice were detected: peak expiratory flow (PEF), forced vital capacity (FVC), lung resistance (RL), and dynamic lung compliance (Cdyn).

[0104] The results are as follows Fig. 9 As shown, compared with the PBS control group, the lung function of mice in the HbRT group was significantly improved, indicating that HbRT can effectively repair the lung function of mice with bleomycin-induced pulmonary fibrosis.

[0105] The above embodiments are only preferred implementation modes of the present invention and cannot be used to limit the protection scope of the present invention. Any non-substantial changes and modifications made by those skilled in the art on the basis of the invention shall fall within the protection scope of the present invention.

Claims

1. A nanomedicine for treating idiopathic pulmonary fibrosis, characterized in that: The nanoparticles include hemoglobin, the surface of which is loaded with a transforming growth factor-β1 (TGF-β1) inhibitor and resveratrol (RSV), wherein the transforming growth factor-β1 (TGF-β1) inhibitor is any one or any combination of SB431542, LY2157299, LY2109761, TP0427736, IN1130, and LY3200882.

2. A nanomedicine for treating idiopathic pulmonary fibrosis according to claim 1, characterized in that: The particle size of the nanoparticles is 180-240 nm.

3. A method for preparing a nanomedicine for treating idiopathic pulmonary fibrosis according to any one of claims 1 or 2, characterized in that: The following steps are involved: S1 lysed hemoglobin: Dissolve 0.5-3 mg of hemoglobin in 1-3 mL of deionized water, and then perform ultrasonic treatment until the hemoglobin is evenly dispersed to obtain an initial hemoglobin solution; The temperature is controlled at 23-27°C, the relative humidity is 40-60%, and the pH value of the initial hemoglobin solution is maintained at 6.8-7.4; S2 Preparation of a mixed solution of TGF-β1 inhibitor and RSV: A TGF-β1 inhibitor having a concentration of 10 mg / mL and resveratrol (RSV) having a concentration of 40 mg / mL are added to an organic solvent, followed by sonication until the TGF-β1 inhibitor and resveratrol (RSV) are completely dissolved and mixed uniformly to obtain a mixed solution; The temperature is controlled at 23-27°C, and the volume ratio of the TGF-β1 inhibitor to RSV and the organic solvent is (2-25) μL: (1-10) μL: (0.2-1.0) mL; S3 Preparation of Nanoparticles: The mixed solution prepared in step S2 is added dropwise to the hemoglobin solution prepared in step S1 to obtain a base solution, wherein the volume ratio of the mixed solution to the hemoglobin solution is 1:(3-6); the base solution is then subjected to ultrasonic treatment for 5-15 minutes, and then the ultrasonicated base solution is centrifuged at a centrifugal force of 15000g-30000g for 10-30 minutes at a centrifugal temperature of 4-8°C to obtain nanoparticles.

4. The method for preparing a nano drug for treating idiopathic pulmonary fibrosis according to claim 3, characterized in that: The organic solvent is dimethyl sulfoxide.

5. An application of a nano drug in the preparation of an anti-fibrosis preparation, characterized in that: Use the nano drug as described in any one of claims 1-2.

6. The use of the nano drug according to claim 5 in the preparation of an anti-fibrosis preparation, characterized in that: The dosage form of the preparation includes tablets, powders, suspensions, granules, capsules, injections, sprays, solutions, enemas, emulsions, films, suppositories, patches, nasal drops or pills.

7. A drug delivery system, characterized in that Use the nano drug as described in any one of claims 1-2.

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