Exosome composition and method for treating lung diseases

Abstract

The invention discloses an exosome composition and method for treating lung diseases, and relates to the technical field of medicines.The composition is composed of human alveolar epithelial cell exosome subjected to hypoxia pretreatment, traditional Chinese medicine compound nano-liposome and an atomization stabilizer, the mass ratio of the exosome to the traditional Chinese medicine compound nano-liposome is 1: 1-1: 3, and the mass ratio of the atomization stabilizer to the human alveolar epithelial cell exosome to the traditional Chinese medicine compound nano-liposome is 1: 1-1: 3. The preparation method comprises the steps of cell culture, exosome purification, traditional Chinese medicine liposome preparation and composition mixing treatment, during treatment, an intelligent atomizer can be used for administration, and abdominal respiration training and traditional Chinese medicine iontophoresis therapy are matched. According to the scheme, the targeting property is high, the side effect is small, the inflammatory pathway can be effectively inhibited, the pulmonary epithelium-mesenchymal transition is reversed, the drug delivery efficiency is improved, and the drug delivery system is suitable for treating idiopathic pulmonary fibrosis, acute attack of asthma and the like.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to an exosome composition and method for treating lung diseases. BACKGROUND

[0002] Idiopathic pulmonary fibrosis (IPF) and asthma exacerbation are lung diseases that seriously threaten human health, and their pathogenesis involves complex pathological processes such as lung epithelial cell damage, inflammation pathway activation, and epithelial-mesenchymal transition (EMT). At present, the clinical treatment means are limited, and traditional drugs such as glucocorticoids and immunosuppressants can alleviate symptoms, but have significant systemic side effects, insufficient lung targeting, and limited long-term efficacy. Traditional Chinese medicine has the advantage of multi-target regulation in the treatment of lung diseases, but its effective components have poor water solubility, low lung deposition rate, and insufficient bioavailability, which restricts its clinical application.

[0003] Exosomes as a new type of biological carrier have shown potential in disease treatment due to their natural targeting, low immunogenicity, and good biocompatibility.

[0004] However, the conventional exosome separation and purification method has the problems of low purity and high risk of endotoxin contamination, and the treatment effect of exosomes without pretreatment is limited. At the same time, the existing lung drug delivery system (such as nebulization inhalation) often has uneven particle size and poor stability, resulting in low drug deposition efficiency in alveoli, making it difficult to achieve precise treatment. In addition, single therapy is difficult to regulate multiple pathological pathways at the same time, and there is an urgent need to develop an efficient and low-toxicity combined therapy strategy. SUMMARY

[0005] Therefore, the purpose of the present application is to provide an exosome composition and method for treating lung diseases to solve the technical problems of insufficient targeting, significant side effects, low drug delivery efficiency, and limited multi-pathway regulation in the treatment of existing lung diseases.

[0006] In view of the above problems, the present application provides an exosome composition and method for treating lung diseases.

[0007] In a first aspect of the present application, an exosome composition for treating lung diseases is provided, which is composed of the following components:

[0008] Human alveolar epithelial cell-derived exosomes: concentration of 2x10 9 ~ 8x10 10Particles / mL, secreted after 24 hours of hypoxia pretreatment with 5% oxygen concentration, positive for surface markers CD9, CD81 and Alix expression; human alveolar epithelial cells are primary cells or a safety-verified low-passage immortalized cell line HPAEpiC (passage number ≤ 4), and 0.1-1 mM N-acetylcysteine is added during culture; miR-34a expression can be up-regulated by gene editing.

[0009] Traditional Chinese medicine compound nanoliposomes: a complex containing astragaloside IV, tanshinone IIA and fritillarine, with a mass ratio of 1:0.8:0.5, and a total drug loading of 1.5-6.0 mg / mL; the preparation method is to weigh astragaloside IV, tanshinone IIA and fritillarine monomer standard, mix according to a mass ratio of 1:0.8:0.5, dissolve in an appropriate amount of organic solvent (such as ethanol) to obtain a drug solution, then mix with soybean phospholipid and cholesterol at a ratio of 10:3:1, and prepare liposomes by the film dispersion-high pressure homogenization method, with an encapsulation efficiency of not less than 85%.

[0010] Atomization stabilizer: composed of 0.5%-2% trehalose by mass volume percentage and lactoferrin-modified chitosan-citric acid nanoparticles with a particle size of 50-200 nm, and the binding efficiency of the nanoparticles to integrin α6β4 on the surface of the exosome membrane is not less than 90%.

[0011] Composition ratio and morphology: the mass ratio of exosomes to traditional Chinese medicine compound nanoliposomes is 1:1-1:3, and a homogeneous suspension with a particle size of not more than 200 nm is formed after ultrasonic emulsification treatment, which is used for outpatient atomization inhalation treatment.

[0012] In the second aspect of the present application, a method for preparing an exosome composition for treating lung diseases is provided, comprising: obtaining and purifying exosomes: expanding human alveolar epithelial cells in a culture system simulating the physiological function of alveoli, and collecting the conditioned medium of the 3rd-4th generation cells; purifying exosomes by size exclusion chromatography combined with affinity chromatography, using anti-CD81 antibody conjugated magnetic beads for affinity chromatography, and using pH 3.0 glycine-HCl buffer as eluent; immediately neutralizing to pH 7.0 with PBS after elution, and the purity of the purified exosomes is not less than 98%, and the endotoxin level is not more than 0.1 EU / mL.

[0013] Mixing and processing of the composition: after mixing the traditional Chinese medicine compound nanoliposomes and the exosomes according to the mass ratio, adding the atomization stabilizer, and processing by a micro-jet nanodispersion system with a pressure of 1500 bar and 3 cycles, a stable system with a Zeta potential of-30 to-15 mV is formed.

[0014] In a third aspect of the present application, there is provided use of the composition in the manufacture of a medicament for treating a lung disease, the medicament being administered by an intelligent nebulizer, the mass median aerodynamic diameter of the nebulized particles being no more than 2.5 μm.

[0015] The use is combined with iontophoresis of traditional Chinese medicine, wherein the traditional Chinese medicine compound nanoliposome is diluted 10 times and then introduced through the Dantian and Feishu points, and the current intensity during the introduction is 0.1-0.3 mA / cm².

[0016] In summary, the present application has the following advantages:

[0017] The present application combines the low-oxygen pretreated human alveolar epithelial cell exosomes with the lactoferritin modified chitosan-citric acid nanoparticles, targets the lung lesion area by integrin α6β4, cooperates with the nebulized particles with a particle size of ≤2.5 μm and the microfluidic stable suspension liquid technology, significantly improves the alveolar deposition efficiency and the drug administration uniformity, and the exosomes (containing up-regulated miR-34a) and the traditional Chinese medicine compound nanoliposome synergistically inhibit the TLR4 / NF-κB pathway activity (inhibition rate ≥75%), down-regulate the α-smooth muscle actin expression by ≥60% and improve the E-cadherin / β-catenin complex stability by 3-5 times, realizes the reversal of the lung epithelial-mesenchymal transition and the multi-pathway inflammation regulation; the human exosomes have low immunogenicity, the traditional Chinese medicine natural ingredients reduce the systemic side effects, the outpatient nebulization combined with abdominal breathing training and iontophoresis at acupoints improves the patient compliance, and the compound enzymolysis combined with ultrahigh pressure extraction, the size exclusion chromatography combined with affinity chromatography and other processes ensure the high yield of the traditional Chinese medicine ingredients and the high purity (≥98%) and low endotoxin (≤0.1 EU / mg) of the exosomes, and the present application has the advantages of high efficiency, low toxicity, precision and safe and controllable preparation. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present application will be clearly and completely described below in combination with the embodiments of the present application. The embodiments described below are exemplary and are used to explain the present application, but cannot be understood as limitations to the present application.

[0019] The embodiments of the present application will be described below according to the overall structure of the present application.

[0020] Embodiment 1: Preparation and performance verification of exosome composition

[0021] Step 1: Preparation of human alveolar epithelial cell exosomes

[0022] The safety-verified low-passage immortalized cell line HPAEpiC (passage number ≤4 passages) is used, and is expanded to the 3rd generation in a culture system simulating the physiological function of alveoli.

[0023] 0.5 mM N-acetyl cysteine in culture medium, hypoxic preconditioning condition: 5% oxygen concentration for 24 hours (100 mM cobalt chloride or 50 mM deferoxamine can be used to mimic the effect of hypoxia).

[0024] The conditioned medium was collected and exosomes were purified by size exclusion chromatography (SEC) combined with affinity chromatography using anti-CD81 antibody conjugated magnetic beads.

[0025] Elution buffer: the eluent is glycine-HCl buffer at pH 3.0, immediately neutralized to pH 7.0 with alkaline neutralization buffer (such as 1 M Tris-HCl, pH 8.0-9.0) after elution, and then replaced into PBS buffer system.

[0026] Detection results: exosome purity 98.5% (calculated using nanoparticle tracking analysis (NTA) combined with protein quantification (BCA method)), endotoxin level 0.08 EU / mL, concentration 5 x 10 10 particles / mL (Table 1).

[0027] Step 2: Preparation of traditional Chinese medicine compound nanoliposomes

[0028] The standard substances of astragaloside IV, tanshinone IIA and fritillarine were weighed and mixed according to the mass ratio of 1:0.8:0.5, and then dissolved in an appropriate amount of organic solvent (such as ethanol) to obtain a medicinal liquid.

[0029] The medicinal liquid was mixed with soybean phospholipid and cholesterol at a ratio of 10:3:1, and the liposomes were prepared by the thin film dispersion-high pressure homogenization method.

[0030] Encapsulation efficiency: 92% for astragaloside IV, 88% for tanshinone IIA, and 85% for fritillarine (Table 1).

[0031] Total drug loading: 45 mg / mL.

[0032] Step 3: Composition mixing and atomization treatment

[0033] The exosomes and traditional Chinese medicine liposomes were mixed at a mass ratio of 1:2, 1% trehalose was added (if used for freeze-drying, the concentration of trehalose can be increased to 10%), and lactoferrin modified chitosan-citric acid nanoparticles (particle size 120 nm) were added.

[0034] Microfluidic nanodispersion system (1500 bar, 3 cycles) treatment, obtaining a homogeneous suspension with a particle size of 180 nm and a Zeta potential of -20 mV.

[0035] Table 1: Key parameters of Example 1

[0036] Parameter Result Exosome purity 98.5% Endotoxin level 0.08 EU / mg Traditional Chinese medicine encapsulation rate 92% (astragaloside IV) Nebulized particle size 180 nm TLR4 / NF-κB inhibition rate 78%

[0037] Example 2: Outpatient treatment of idiopathic pulmonary fibrosis (IPF)

[0038] Treatment regimen:

[0039] The composition of Example 1 was administered by a smart nebulizer (MMAD = 2.3 pm), twice a day, 10 minutes each time, for a treatment cycle of 7 days.

[0040] Combined with iontophoresis of traditional Chinese medicine: dilute 10 times of traditional Chinese medicine liposome through the clavicular and lung points (0.2 mA / cm2, once a day).

[0041] Efficacy data (Table 2):

[0042] The expression of a-smooth muscle actin was down-regulated by 65%.

[0043] The stability of E-cadherin / beta-catenin complex was increased by 4.2 times.

[0044] The alveolar deposition efficiency reached 82%.

[0045] Table 2: Efficacy data of Example 2

[0046] Index Result α-SMA down-regulation rate 65% E-cadherin / β-catenin stability 4.2 times Alveolar deposition efficiency 82%

[0047] Example 3: Verification of the synergistic effect of gene-edited exosomes

[0048] Gene editing was performed on HPAEpiC cells to up-regulate the expression of miR-34a (overexpression efficiency ≥ 90%).

[0049] After preparing the exosomes, they were mixed with traditional Chinese medicine liposomes at a ratio of 1:3, and the inhibition rate of TLR4 / NF-κB pathway was detected.

[0050] Results (Table 3): The inhibition rate reached 83%, which was significantly improved compared with unedited exosomes (inhibition rate 68%).

[0051] Table 3: Comparison of the effects of gene-edited exosomes

[0052] Group TLR4 / NF-κB inhibition rate Unedited exosome group 68% Gene-edited exosome group 83%

[0053] Example 4: Comparison of the effects of exosomes from primary cells and immortalized cell lines

[0054] Step 1: Preparation of primary cell exosomes

[0055] Primary human alveolar epithelial cells were isolated from healthy donor lung tissue and cultured in a three-dimensional gas-liquid interface to the 3rd passage.

[0056] The hypoxic pretreatment conditions were the same as in Example 1, and the purity of the purified exosomes was 97.8% and the endotoxin was 0.09 EU / mg.

[0057] Step 2: Efficacy comparison

[0058] The TLR4 / NF-κB inhibition rate was detected when the primary cell exosome group and the immortalized cell line (HPAEpiC) exosome group were mixed with traditional Chinese medicine liposomes, respectively.

[0059] Results (Table 4): The inhibition rate of the primary cell group was 81%, and that of the HPAEpiC group was 78%, with no significant difference (P>0.05).

[0060] Table 4: Inhibition rate comparison of different cell sources

[0061] Group TLR4 / NF-κB inhibition rate Primary cell exosome group 81% HPAEpiC exosome group 78%

[0062] Example 5: Optimization of aerosol stabilizer ratio

[0063] Step 1: Stability test of different trehalose concentrations

[0064] 0.5%, 1%, and 2% trehalose were respectively combined with chitosan-sodium citrate nanoparticles (particle size 120 nm).

[0065] The particle size change of the suspension stored at 25°C for 7 days was detected (Table 5).

[0066] Table 5: Effect of trehalose concentration on stability

[0067] Trehalose concentration Initial particle size (nm) Particle size after 7 days (nm) 0.5% 180 210 1% 175 185 2% 170 195

[0068] Conclusion: The 1% trehalose group has the best stability, with a particle size change of ≤5%.

[0069] Example 6: Effect of treatment cycle length on efficacy

[0070] Step 1: Different cycle treatment of IPF model mice

[0071] The treatment cycle was 5 days and 10 days, respectively, and the other conditions were the same as in Example 2.

[0072] The proportion of pulmonary fibrosis area was detected (Table 6).

[0073] Table 6: Treatment cycle and fibrosis inhibition effect

[0074] Treatment cycle Fibrosis area ratio 5 days 15.2% 10 days 8.7%

[0075] Conclusion: The 10-day course is significantly better than the 5-day course (P<0.01).

[0076] Comparative Example 1: Exosomes without hypoxic pretreatment

[0077] HPAEpiC cells were cultured under normoxic conditions (21% oxygen) to prepare exosomes.

[0078] After mixing with traditional Chinese medicine liposomes, the down-regulation rate of α-SMA was detected.

[0079] Results (Table 7): The down-regulation rate of α-SMA was only 38%, which was significantly lower than that of the hypoxic preconditioning group (65%).

[0080] Table 7: Comparison of the effects of hypoxic preconditioning

[0081] Group α-SMA down-regulation rate Non-hypoxic pretreatment group 38% Hypoxic pretreatment group 65%

[0082] Comparative Example 2: Single Traditional Chinese Medicine Component Liposomes

[0083] Liposomes were prepared only using astragaloside A (drug loading amount 15 mg / mL) and mixed with exosomes.

[0084] The inhibition rate of TLR4 / NF-κB was detected.

[0085] Results (Table 8): The inhibition rate was 42%, which was significantly lower than that of the compound traditional Chinese medicine group (78%).

[0086] Table 8: Comparison of the effects of compound and single traditional Chinese medicine

[0087] Group TLR4 / NF-κB inhibition rate Single astragaloside IV group 42% Compound traditional Chinese medicine group 78%

[0088] Comparative Example 3: Unmodified Chitosan Nanoparticles

[0089] Unmodified chitosan-citric acid nanoparticles (without lactoferrin binding) were used.

[0090] The binding efficiency with exosome membrane integrin α6β4 was detected.

[0091] Results (Table 9): The binding efficiency was only 52%, which was significantly lower than that of the modified group (92%).

[0092] Table 9: Comparison of the effects of nanoparticle modification

[0093] Group Binding efficiency Unmodified nanoparticle group 52% Lactoferrin-modified group 92%

[0094] Comparative Example 4: Difference in efficacy between only nebulization and combined iontophoresis

[0095] Step 1: Group treatment

[0096] Group 1: Example 1 composition only nebulization (treatment period 7 days).

[0097] Group 2: Nebulization + traditional Chinese medicine iontophoresis (same as Example 2).

[0098] The E-cadherin / β-catenin stability was detected (Table 10).

[0099] Table 10: Comparison of the effects of combined therapy

[0100] Group Complex stability (times) Nebulization only group 3.1 Nebulization + ion introduction group 4.2

[0101] Conclusion: The combination therapy significantly improved stability (P<0.05).

[0102] Comparative Example 5: Gene Editing Effects of Different miRNAs

[0103] Step 1: Editing exosomes of miR-34a and miR-21

[0104] HPAEpiC cells were edited with miR-34a and miR-21 respectively (overexpression efficiency ≥90%).

[0105] The TLR4 / NF-κB inhibition rate was measured (Table 11).

[0106] Table 11: Inhibition rate of different miRNA editing

[0107] Group TLR4 / NF-κB inhibition rate miR-34a editing group 83% miR-21 editing group 62%

[0108] Conclusion: miR-34a is a more effective target (P<0.01).

[0109] Comparative Example 6: Effect of changes in ultra-high pressure extraction conditions on the yield of traditional Chinese medicine

[0110] Step 1: Extraction under different pressures and times

[0111] Extraction conditions:

[0112] Group A: 500 MPa, 10 minutes (Example 1 conditions).

[0113] Group B: 300 MPa, 15 minutes.

[0114] Group C: 700 MPa, 5 minutes.

[0115] The yield of astragaloside A was determined (Table 12).

[0116] Table 12: Relationship between ultra-high pressure parameters and yield

[0117] Group Pressure (MPa) Time (min) Yield (%) A 500 10 92% B 300 15 78% C 700 5 85%

[0118] Conclusion: 500 MPa × 10 minutes is the optimal condition.

[0119] Overall Comparison Table

[0120] Group / parameter Exosome purity (%) TLR4 / NF-κB inhibition rate (%) α-SMA down-regulation rate (%) Binding efficiency (%) Fibrosis area (%) Example 1 98.5 78 65 92 - Example 2 97.8 81 68 90 8.7 (10 days) Example 3 (gene editing) 98.2 83 70 93 - Example 4 (primary cell exosome) 97.8 81 67 91 - Example 5 (1% trehalose optimization) 98.0 79 66 92 - Example 6 (10-day treatment) 98.1 80 69 90 8.7 (10 days) Comparative Example 1 (non-hypoxic) 97.5 45 38 50 22.5 Comparative Example 2 (single traditional Chinese medicine) 98.0 42 30 88 18.9 Comparative Example 3 (unmodified nanoparticle) 98.3 70 55 52 15.8 Comparative Example 4 (nebulization only) 98.1 75 60 89 12.4 Comparative Example 5 (miR-21 editing) 97.9 62 48 90 - Comparative Example 6 (300 MPa x 15 min) 98.0 65 50 85 14.3

[0121] Hypoxia pretreatment enhances exosome activity:

[0122] Hypoxia (5% O2, 24 hours) significantly enhanced exosome function, with the downregulation rate of α-SMA increasing from 38% (non-hypoxia group) to 65%, and the inhibition rate of TLR4 / NF-κB increasing from 45% to 78%.

[0123] Gene editing of miR-34a further enhanced the anti-inflammatory effect, with an inhibition rate of 83%, which was 22% higher than the unedited group (68%) (Example 3 vs Comparative Example 1).

[0124] Synergistic Effect of Compound Traditional Chinese Medicine:

[0125] The combined inhibition rate of astragaloside A, tanshinone IIA, and fritillary acetylcholine A (78%) was significantly higher than that of the single component (42%), indicating a synergistic effect of multi-target regulation (Example 1 vs. Comparative Example 2).

[0126] The ultra-high pressure extraction process (500 MPa × 10 minutes) optimized the yield of traditional Chinese medicine to 92%, which is 18% higher than that under non-standard conditions (300 MPa × 15 minutes) (Example 1 vs Comparative Example 6).

[0127] Targeted delivery system optimization:

[0128] The binding efficiency of lactoferrin-modified chitosan-citric acid nanoparticles to exosomal membrane integrin α6β4 reached 92%, which was nearly 80% higher than that of the unmodified group (52%), significantly improving lung targeting (Example 1 vs. Comparative Example 3).

[0129] After optimizing the atomized particle size (≤200 nm) and aerodynamic diameter (MMAD≤2.5 μm), the alveolar deposition efficiency reached 82%, reducing drug waste (Example 2).

[0130] Optimization of preparation process and treatment plan

[0131] High purity and low toxicity:

[0132] Size exclusion chromatography combined with anti-CD81 antibody affinity chromatography ensured exosome purity ≥98% and endotoxin ≤0.1 EU / mg, meeting clinical safety requirements (Example 1).

[0133] The immortalized cell line (HPAEpiC) has similar efficacy to primary cell exosomes (78% vs. 81%), but the former is less expensive and suitable for large-scale production (Example 4).

[0134] Treatment cycle and combination therapy:

[0135] A 10-day treatment course significantly reduced the area of ​​pulmonary fibrosis compared to a 5-day treatment course (8.7% vs 15.2%), indicating that extending the treatment period can enhance the therapeutic effect (Example 6).

[0136] The combined therapy of nebulized inhalation and acupoint iontophoresis increased the stability of the E-cadherin / β-catenin complex by 4.2 times, which was superior to the single nebulization group (3.1 times), highlighting the synergistic effect of multi-pathway treatment (Example 2 vs. Comparative Example 4).

[0137] Atomization stability optimization:

[0138] The combination of 1% trehalose and modified nanoparticles resulted in a particle size change of ≤5% after 7 days of suspension storage, which is superior to the 0.5% or 2% group, ensuring the long-term stability of the drug (Example 5).

[0139] Breakthrough in existing technologies

[0140] Multi-pathway joint regulation:

[0141] Simultaneously inhibiting the TLR4 / NF-κB inflammatory pathway (inhibition rate ≥75%) and reversing epithelial-mesenchymal transition (α-SMA downregulation ≥60%), it overcomes the limitations of traditional single therapy.

[0142] The synergistic effect of gene editing technology (miR-34a) and traditional Chinese medicine compound formulas provides a new strategy for precision treatment.

[0143] Safety and controllability:

[0144] Human exosomes have low immunogenicity and are combined with natural Chinese medicine ingredients to reduce systemic side effects.

[0145] The combined enzymatic hydrolysis-ultra-high pressure extraction and microfluidic dispersion processes ensure the activity of the components and the uniformity of the formulation (encapsulation rate ≥85%, particle size ≤200 nm).

[0146] It is suitable for refractory diseases such as idiopathic pulmonary fibrosis (IPF) and acute asthma exacerbations. The outpatient nebulization combined with iontophoresis program is convenient, efficient and suitable for widespread application.

[0147] This invention, through hypoxia pretreatment of exosomes, synergistic effects of compound traditional Chinese medicine, optimized targeted delivery systems, and gene editing technology, demonstrates outstanding performance in inhibiting inflammation, reversing fibrosis, and improving drug delivery efficiency. The tabular data fully supports its high efficiency, safety, and controllability, providing an innovative solution for the precision treatment of lung diseases.

[0148] Although embodiments of the present invention have been shown and described, these specific embodiments are merely explanations of the invention and are not intended to limit it. The specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. After reading this specification, those skilled in the art may make modifications, substitutions, and variations to the embodiments as needed without departing from the principles and spirit of the invention, but such modifications, substitutions, and variations are protected by patent law as long as they are within the scope of the claims of the present invention.

Claims

1. An exosome composition for treating lung diseases, characterized in that, It consists of the following components: a) Exosomes derived from human alveolar epithelial cells, at a concentration of 2×10⁻⁶. 9 ~8×10 10 The exosomes were secreted after hypoxia pretreatment (5% oxygen concentration for 24 hours) and were positive for surface markers CD9, CD81 and Alix. b. A compound nanoliposome containing traditional Chinese medicine, comprising a complex of astragaloside A, tanshinone IIA and fritillary acetylcholine A, with a mass ratio of 1:0.8:0.5 and a total drug loading of 1.5–6.0 mg / mL; C. Atomization stabilizer, composed of 0.5% to 2% by mass volume of trehalose and chitosan-citric acid nanoparticles with a particle size of 50 to 200 nm; The mass ratio of the exosomes to the traditional Chinese medicine compound nanoliposomes is 1:1 to 1:3, and the composition is ultrasonically emulsified to form a homogeneous suspension with a particle size not exceeding 200 nm, which is used for outpatient nebulized inhalation therapy for idiopathic pulmonary fibrosis or acute asthma attacks.

2. The composition according to claim 1, characterized in that: The traditional Chinese medicine compound nanoliposomes were prepared by the following method: S1: Weigh out the standards of astragaloside A, tanshinone IIA and fritillary acetylcholine A monomer, mix them in a mass ratio of 1:0.8:0.5, dissolve them in an appropriate amount of organic solvent to obtain the drug solution; S2: The drug solution is mixed with soybean phospholipids and cholesterol in a ratio of 10:3:1, and liposomes are prepared by thin film dispersion-high pressure homogenization, with an encapsulation rate of not less than 85%.

3. The composition according to claim 1, characterized in that: The human alveolar epithelial cells are primary cells or the low-passage immortalized cell line HPAEpiC with verified safety, with ≤8 passages, and 0.1-1 mM of N-acetylcysteine ​​is added during culture.

4. The composition according to claim 1, characterized in that: The chitosan-citric acid nanoparticles in the atomization stabilizer are modified with lactoferrin, and their binding efficiency with integrin α6β4 on the surface of exosome membrane is not less than 90%.

5. A method for preparing the composition of claim 1, characterized in that, include: S1: Expand human alveolar epithelial cells in a culture system that simulates alveolar physiological function, and collect the conditioned medium of cells from the 3rd to 5th generation. S2: Exosomes were purified by size exclusion chromatography combined with affinity chromatography to a purity of not less than 98% and an endotoxin level of not more than 0.1 EU / mg; S3: After mixing the traditional Chinese medicine compound nanoliposomes and exosomes in a certain mass ratio, an atomization stabilizer was added, and the mixture was treated by a microfluidic nanodispersion system at a pressure of 1500 bar for 3 cycles to form a stable system with a zeta potential of -30 to -15 mV.

6. The method according to claim 5, characterized in that: In step S2, affinity chromatography uses anti-CD81 antibody-conjugated magnetic beads, with glycine-HCl buffer at pH 3.0 as the elution buffer, and the solution is immediately neutralized to pH 7.0 with PBS after elution.

7. The use of the composition according to any one of claims 1-4 in the preparation of a medicament for treating lung diseases, characterized in that, The drug is administered via a smart nebulizer, wherein the mass median aerodynamic diameter of the atomized particles does not exceed 2.5 μm.

8. The use according to claim 7, characterized in that, The application is combined with traditional Chinese medicine iontophoresis therapy, wherein the traditional Chinese medicine compound nanoliposomes are diluted 10 times and introduced through the Tanzhong and Feishu acupoints, with the current intensity during introduction being 0.1 to 0.3 mA / cm².

9. The use of the composition according to claim 1 in the preparation of a drug for reversing pulmonary epithelial-mesenchymal transition, characterized in that, The composition downregulates α-smooth muscle actin expression by at least 60% and increases the stability of the E-cadherin / β-catenin complex by 3 to 5 times.

10. The composition according to claim 1, characterized in that: The exosomes were gene-edited to upregulate miR-34a expression and synergistically inhibited TLR4 / NF-κB pathway activity with traditional Chinese medicine compound nanoliposomes, with a pathway inhibition rate of not less than 75%.