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CN-122005646-A - Lung protecting function exosome and inhalation preparation based on resveratrol-astragalus-ginseng induction and application thereof

CN122005646ACN 122005646 ACN122005646 ACN 122005646ACN-122005646-A

Abstract

The invention discloses a lung protecting function exosome based on resveratrol-astragalus-ginseng induction, an inhalation preparation and application thereof, and belongs to the technical field of biology. The exosomes are obtained by secretion of lung fibroblasts after induction of active ingredients comprising resveratrol, astragalus extract and ginseng extract, wherein the content of astragalus methylglycoside in the astragalus extract is not less than 2.0%, and the total content of ginsenoside Rg1 and Rb1 in the ginseng extract is not less than 5.0%. The invention also provides a dry powder inhalant or an atomized inhalant liquid containing the exosome. The exosome and the preparation thereof exert the lung protection effect through regulating and controlling multiple mechanisms such as inflammation, antioxidant stress, repair promotion and the like, can be used for preventing and/or treating various lung injury diseases such as acute lung injury, pneumonia and the like, and have the advantages of good biocompatibility and strong targeting.

Inventors

  • YANG XIAOCHEN

Assignees

  • 北京圣美细胞生命科学工程研究院有限公司

Dates

Publication Date
20260512
Application Date
20260225

Claims (10)

  1. 1. A functional exosome for lung protection, characterized in that the exosome is obtained by secretion of lung fibroblasts after induction of active ingredients comprising resveratrol, astragalus extract and ginseng extract.
  2. 2. The functional exosome according to claim 1, wherein the final concentration of the active ingredient upon induction is 5-20 μm resveratrol, 10-100 μg/mL of astragalus extract, 10-100 μg/mL of ginseng extract.
  3. 3. The functional exosome of claim 1, wherein the lung fibroblast is human embryonic lung fibroblast MRC-5 or primary lung fibroblast.
  4. 4. A method for producing a functional exosome according to any one of claims 1 to 3, comprising the steps of culturing lung fibroblasts in an induction medium comprising resveratrol, an extract of Astragalus membranaceus and an extract of Panax ginseng at 37℃under 5% CO 2 for 36 to 72 hours, collecting cell supernatants, subjecting the treated supernatants to differential centrifugation and filtration in sequence, subjecting the treated supernatants to ultracentrifugation, collecting precipitates and washing with a buffer solution to obtain the functional exosome.
  5. 5. The method according to claim 4, wherein the differential centrifugation is carried out at 2-8deg.C for 8-15 minutes at 300-500 Xg, at 2000-3000 Xg for 15-25 minutes, and at 10000-12000 Xg for 25-35 minutes.
  6. 6. The method of claim 4, wherein the ultracentrifugation conditions are 4 ℃, 100000-120000 x g centrifugation for 60-90 minutes.
  7. 7. An inhalation formulation comprising the functional exosome of any one of claims 1-3 or prepared by the method of any one of claims 4-6, and a pharmaceutically acceptable carrier.
  8. 8. The inhalation formulation according to claim 7, wherein the inhalation formulation is a dry powder inhaler or an atomized inhalation liquid.
  9. 9. Use of a functional exosome according to any one of claims 1-3 or an inhalation formulation according to any one of claims 7-8 in the manufacture of a medicament for the prevention and/or treatment of a pulmonary injury disorder.
  10. 10. The use according to claim 9, wherein the lung injury disease comprises acute lung injury, pneumonia, pulmonary fibrosis or chronic obstructive pulmonary disease.

Description

Lung protecting function exosome and inhalation preparation based on resveratrol-astragalus-ginseng induction and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to a lung protecting function exosome based on resveratrol-astragalus-ginseng induction, an inhalation preparation and application thereof. Background Pulmonary injury diseases, such as Acute Lung Injury (ALI), pneumonia, chronic Obstructive Pulmonary Disease (COPD), and Pulmonary Fibrosis (PF), are major health problems with high morbidity and mortality worldwide. Their common pathological core involves uncontrolled inflammatory responses, oxidative stress, destruction of alveolar epithelium and vascular endothelial barrier, and subsequent abnormal tissue repair and fibrosis, ultimately leading to progressive loss of lung function. Taking acute lung injury and severe form Acute Respiratory Distress Syndrome (ARDS) as an example, in addition to support treatment such as mechanical ventilation, a specific drug capable of effectively suppressing inflammatory storm and promoting lung tissue repair is still lacking clinically, and patients are poor in prognosis, so that development of novel treatment strategies is urgently needed. In recent years, extracellular vesicles, particularly exosomes, have shown great potential in the field of pulmonary disease treatment as an emerging biologic therapeutic and drug delivery system. The exosomes are nanoscale lipid bilayer vesicles secreted by cells, naturally load bioactive substances such as proteins, nucleic acids and the like, and have low immunogenicity, good biocompatibility and the capability of crossing biological barriers. Research shows that the exosomes derived from the mesenchymal stem cells can show positive effects of protecting alveolar capillary barrier, reducing pulmonary edema and inflammation in an acute lung injury animal model through the mechanisms of delivering anti-inflammatory miRNA, regulating macrophage polarization, reducing oxidative stress and the like. In addition, the exosomes are delivered by an aerosol inhalation mode, so that the local efficient enrichment of the medicine in the lung can be realized, the targeting is obviously improved, the systemic side effect is reduced, and the medicine becomes a very promising administration route. However, the current research and practice of exosomes for lung protection still face several key bottlenecks, namely, firstly, the current research is mostly focused on mesenchymal stem cells or immune cells from healthy donors, and the exosomes have certain anti-inflammatory effects, but the functions of the exosomes are relatively fixed, so that effective regulation and control on complex lung injury microenvironments are difficult, and the cost and quality uniformity control of large-scale preparation are challenging. Second, in many chronic lung diseases, activated lung fibroblasts are core effector cells that cause excessive deposition of extracellular matrix, tissue hardening. Conventional thinking is aimed at inhibiting or eliminating these cells. Third, although natural medicinal components such as ginseng and astragalus have been reported to have anti-inflammatory and antioxidant activities, how to combine them into an induction system capable of precisely regulating the phenotype of lung fibroblasts and inducing them to secrete specific protective exosomes, and to successfully develop exosomes of this origin into stable and inhalable preparations has yet to be explored. Therefore, an integrated lung protection strategy which can utilize key cells in the microenvironment of the lung diseases, obtain the exosomes with enhanced functions through a specific induction system and is suitable for high-efficiency inhalation administration is developed, and the integrated lung protection strategy has important scientific significance and clinical conversion value. Disclosure of Invention In view of the above, the present invention aims to provide a resveratrol-astragalus-ginseng-induced lung protecting function exosome, wherein the functional exosome is obtained by inducing the secretion of lung fibroblasts by a specific combination of traditional Chinese medicine active ingredients, and can realize the targeted delivery of the lung by inhalation route, so as to prevent and/or treat the lung injury diseases. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides a functional exosome for lung protection, which is obtained by secretion of lung fibroblasts after induction of active ingredients comprising resveratrol, astragalus extract and ginseng extract. Preferably, the final concentration of the active ingredients in induction is 5-20 mu M resveratrol, 10-100 mu g/mL radix astragali extract and 10-100 mu g/mL radix Ginseng extract. Preferably, the lung fibroblast is human embryonic lung fibroblast MRC-5