CN-122012371-A - External vesicle from radix zanthoxyli, preparation method and application thereof in preparation of medicine for preventing and treating acute lung injury

Abstract

The invention belongs to the technical field of biological medicines, and discloses an external vesicle from a radix zanthoxyli, a preparation method and application thereof in preparing a medicine for preventing and treating acute lung injury, wherein the external vesicle with the particle size of 60-100 nm and a lipid bilayer structure is extracted and purified from the root of the radix zanthoxyli for the first time. The external vesicle can be administered in a mode of aerosol inhalation and the like, has excellent curative effects in an acute lung injury model, can obviously relieve pulmonary tissue edema and inflammatory cell infiltration, reduces the level of key pro-inflammatory factors such as IL-6, TNF-alpha, IL-1 beta and the like in serum, promotes macrophage differentiation to an anti-inflammatory M2 type, and has better curative effects than traditional dexamethasone and lower side effects. The invention provides a brand new, efficient and safe candidate drug and natural nanometer treatment strategy for preventing and treating acute lung injury and ARDS.

Inventors

• YUAN XIANG
• LIAO YI
• HUANG BEN

Assignees

• 广西壮族自治区人民医院

Dates

Publication Date

20260512

Application Date

20260409

Priority Date

20260210

Claims (10)

1. 1. An outer vesicle derived from a nitidine, wherein the outer vesicle is a nanoparticle having a lipid bilayer structure extracted from the root of the nitidine.
2. 2. A method of preparing a nitidine-derived outer vesicle as claimed in claim 1, comprising the steps of: (a) Cutting root tissue of radix Zanthoxyli, and performing enzymolysis at 45-55deg.C in solution containing cellulase and pectase; (b) Primary purification, namely centrifuging the enzymolysis liquid and collecting supernatant; (c) Separating outer vesicles, namely sequentially carrying out low-speed centrifugation, high-speed centrifugation and filtration on the supernatant fluid obtained in the step (b) to remove cell fragments and large-particle impurities; (d) And (3) ultracentrifugation enrichment, namely performing ultracentrifugation on the filtrate obtained in the step (c), and collecting precipitate to obtain the outer vesicle from the Zanthoxylum nitidum.
3. 3. The method according to claim 2, wherein in step (a), the enzymatic solution comprises 2-4% (w/v) cellulase, 1-3% (w/v) pectase and 0.5-0.7 mol/L mannitol, and the pH is 5.5-6.0.
4. 4. The method according to claim 2, wherein in the step (c), the condition of the low-speed centrifugation is 1500 to 2500 Xg, the condition of the high-speed centrifugation is 8000 to 12000 Xg, and in the step (d), the condition of the ultracentrifugation is 80,000 to 120,000 Xg.
5. 5. Use of an outer vesicle of needle origin as claimed in claim 1 for the manufacture of a medicament for the prevention and/or treatment of Acute Lung Injury (ALI).
6. 6. The use according to claim 5, wherein the acute lung injury comprises an acute diffuse lung injury condition including Acute Respiratory Distress Syndrome (ARDS).
7. 7. The use according to claim 5, wherein the pharmaceutical dosage form is a pulmonary formulation, preferably the pulmonary formulation is an aerosol inhalation, a dry powder inhalation or an aerosol.
8. 8. A pharmaceutical composition comprising a therapeutically effective amount of an outer vesicle derived from a nitidine according to claim 1 and a pharmaceutically acceptable carrier, preferably for the prevention and/or treatment of acute lung injury.
9. 9. Use of an outer vesicle of needle origin as claimed in claim 1 for the preparation of a formulation for promoting the differentiation of anti-inflammatory macrophages.
10. 10. A method for non-therapeutic purposes comprising contacting a macrophage with an effective amount of an outer vesicle derived from a Zanthoxylum nitidum according to claim 1 under in vitro conditions, thereby detecting its effect on macrophage polarization or inflammatory factor secretion.

Description

External vesicle from radix zanthoxyli, preparation method and application thereof in preparation of medicine for preventing and treating acute lung injury Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to an external vesicle from a Zanthoxylum nitidum, a preparation method and application thereof in preparing a medicine for preventing and treating acute lung injury. Background Acute lung injury (Acute Lung Injury, ALI) and its severe forms acute respiratory distress syndrome (Acute Respiratory Distress Syndrome, ARDS) are one of the major causes of death in critically ill patients, whose central pathological features are alveolar epithelial and capillary endothelial cell injury, massive release of inflammatory factors and inflammatory cell infiltration, leading to a dramatic decline in lung function. Despite the ongoing intensive research into the pathogenesis of ALI/ARDS, clinical treatments still lack efficient, accurate therapeutic approaches. The existing treatment strategies such as mechanical ventilation can cause nonspecific lung tissue injury, and hormone treatment is easy to cause complications such as infection and the like due to immunosuppression, so that balance regulation of 'inflammation fading-tissue repair' is difficult to realize. Radix Zanthoxyli (Zanthoxylum nitidum (Roxb.) DC.) is a plant of Zanthoxylum of Rutaceae, and is used as root drug, and is incorporated into "Chinese pharmacopoeia", which is one of "Gui Ten ingredient" of Guangxi dado medicine, and has effects of promoting blood circulation, removing blood stasis, removing toxic substance, and relieving swelling. Modern pharmacological studies show that the pharmacological activity of radix Zanthoxyli mainly originates from alkaloid components (such as nitidine chloride, chelerythrine, etc.), and has antiinflammatory, antiviral and immunoregulatory effects. However, the clinical application of the radix zanthoxyli as the traditional Chinese medicine faces a plurality of limitations that the traditional dosage form (decoction, pill and the like) has complex components, low bioavailability, obvious first pass effect, difficult full release and absorption of active ingredients, lack of a targeting mechanism, uneven in-vivo distribution of the medicine, difficult accurate action on lesion lung tissues, complex preparation process, large quality difference of different batches and difficult realization of standardized production and quality control. The plant-derived exovesicle (PELNs) is a nanoscale vesicle (30-150 nm) wrapped by a lipid bilayer, carries active ingredients such as nucleic acid, protein and the like, has the advantages of wide sources, low immunogenicity, high biocompatibility, natural targeting and the like, can mediate intercellular communication and accurately regulate and control target tissue functions, and has broad prospects in the fields of anti-inflammatory, anti-tumor and other disease treatment and drug delivery. At present, research on extracting an outer vesicle from a radix zanthoxyli and applying the outer vesicle to acute lung injury prevention and treatment has not been reported, so a method for efficiently extracting the high-purity outer vesicle from the radix zanthoxyli is developed, the application value of the method in acute lung injury treatment is clear, and the method has important clinical significance and application prospect. Disclosure of Invention Aiming at the defects of the prior art, the invention discloses an external vesicle from a Zanthoxylum nitidum, a preparation method and application thereof in preparing a medicine for preventing and treating acute lung injury. The invention comprises the following specific technical scheme: an outer vesicle derived from a nitidine, the outer vesicle being a nanoparticle having a lipid bilayer structure extracted from a nitidine root. The invention creatively extracts and purifies the radix zanthoxyli to obtain the radix zanthoxyli outer vesicle (ZNEXO), which is in a typical round shape or an oval shape, has a complete lipid bilayer structure, has uniform particle size, is in the range of 60-100nm, has stable components and good biocompatibility. In some examples, it was demonstrated that the extraneo-vesicles can be efficiently taken up by target cells such as macrophages, acting to prevent acute lung injury by the following mechanism: 1. significantly reducing inflammatory cell infiltration in the alveolar spaces, improving pulmonary edema and structural destruction; 2. high-efficiency reduction of the levels of proinflammatory factors such as IL-6, TNF-alpha, IL-1 beta and the like in serum; 3. Promoting macrophage differentiation to anti-inflammatory type (M2 type), regulating inflammatory microenvironment balance; 4. the weight recovery speed of the acute lung injury model mice is improved, the lung coefficient is reduced, and the pathological injury of lung tissues is impr