Acute lung injury targeting drug-loaded nano motor and preparation and application thereof

Abstract

The invention provides a preparation method of a drug-loaded nano motor targeting acute lung injury. Researches show that compared with pure tetracycline, the drug-loaded nano motor has a better anti-inflammatory effect. Particularly, the effects of more remarkably improving inflammatory lung injury, more remarkably reducing systemic / local inflammatory response and the like are achieved. Therefore, the motor can improve the curative effect of the medicine and is expected to reduce the side reaction of the medicine.

Description

Technical Field

[0001] The present invention relates to the field of chemical methods, specifically, to the field of synthesizing nanocarriers by chemical methods, and more specifically, to a nanomotor that can be used for drug delivery and targets acute lung injury, a nanomotor loaded with tetracycline, and its preparation and application in targeting acute lung injury. Background Art

[0002] Acute lung injury is one of the important causes of death and disability in critically ill patients. Currently, there is a lack of effective treatment. Excessive inflammatory cell activation and oxidative stress damage are the core pathophysiological mechanisms of acute lung injury.

[0003] Tetracycline (abbreviated TH) is a broad-spectrum antibiotic produced by actinomycetes. Recent studies have found that tetracycline can selectively inhibit the release of IL-1b and IL-18 by inhibiting caspase-1, thereby alleviating inflammatory damage and offering a novel therapeutic approach for acute lung injury. However, systemic, high-dose, and long-term use of tetracycline is associated with severe side effects, including hepatotoxicity, nephrotoxicity, phototoxicity, ototoxicity and bone toxicity, increased intracranial pressure, and dysbiosis, limiting its clinical application, particularly in critically ill patients. For patients with acute lung injury, local administration of tetracycline would significantly reduce side effects and potentially enhance therapeutic efficacy through targeted drug delivery. However, tetracycline is currently not indicated for intratracheal administration. Therefore, optimizing tetracycline delivery and improving its targeting are key research approaches to further improve efficacy. Summary of the Invention

[0004] The present invention aims to overcome the above-mentioned defects and provide a composite nanomotor that can promote the precise delivery of drugs, increase tissue permeability, and improve therapeutic effects.

[0005] The present invention provides a method for preparing a drug-loaded nanomotor targeting acute lung injury, wherein diisopropylamine-diazenediolate (PAPA / NO) is reacted with methacrylic anhydride, methacrylic acid, or methacrylic acid salt to generate methacrylylated PAPA / NO, which is then reacted with N,N'-bisacryloylcystamine to obtain the nanomotor MPN. The synthesis method of methacryloyl PAPA / NO is as follows: diisopropylamine-diazenediolate, water and 1,4-dioxane are mixed, triethylamine is added dropwise, and then methacrylic anhydride or methacrylic acid or methacrylic acid salt is added dropwise while stirring under cooling in an ice-water bath. After the addition is completed, the mixture is stirred at room temperature and reacted overnight. After the reaction is completed, acetone is added to the reaction solution for precipitation to obtain methacryloyl PAPA / NO. The molar ratio of diisopropylamine-diazeniumdiolate to methacrylic anhydride or methacrylic acid or methacrylic acid salt is 1:(1-2); the amount of diisopropylamine-diazeniumdiolate, water and 1,4-dioxane added is 1ml-5ml of water and 0.5ml-2ml of 1,4-dioxane per mmol of diisopropylamine-diazeniumdiolate; the molar amount of triethylamine added is 2-3 times the molar amount of diisopropylamine-diazeniumdiolate; The process of further reacting methacrylylated PAPA / NO with N,N'-bisacryloylcystamine to obtain the nanomotor MPN is as follows: N,N'-bisacryloylcystamine and methacrylylated PAPA / NO are dissolved in a buffer solution and ultrasonically mixed, followed by nitrogen deoxygenation for 30 minutes. Subsequently, an initiator and a catalyst are added under continuous nitrogen protection, and the reaction is stirred at room temperature until the solution becomes turbid. The nanomotor MPN is obtained after centrifugation, washing with water, and freeze-drying. The molar ratio of methacrylated PAPA / NO to N,N'-bisacryloylcystamine is (1-10):1; the initiator is selected from ammonium persulfate; the catalyst is selected from N,N,N',N'-tetramethylethylenediamine; the added amount of the initiator is 1%-10% of the total weight of the system; and the added amount of the catalyst is 5%-15% of the total weight of the system.

[0006] Furthermore, the present invention provides a method for preparing a drug-loaded nanomotor targeting acute lung injury, which has the following characteristics: the reaction process of methacrylylated PAPA / NO and N,N'-bisacryloylcystamine is carried out in a phosphate buffer system.

[0007] Furthermore, the present invention also provides a tetracycline-loaded nanomotor, which is characterized in that tetracycline is loaded on the nanomotor MPN prepared according to the above method.

[0008] The preparation method is as follows: disperse the nanomotor MPN in a buffer solution containing tetracycline, stir at room temperature for 10-24 hours, and then collect by centrifugation to obtain the MPN@TH nanomotor; Among them, the mass ratio of nanomotor MPN to tetracycline is 1:(0.5-1).

[0009] In addition, the present invention also suggests the application of tetracycline-loaded nanomotors in the preparation of anti-inflammatory drugs.

[0010] In addition, the present invention also suggests the use of tetracycline-loaded nanomotors in the preparation of drugs for treating, alleviating and preventing inflammatory lung injury, reducing systemic / local inflammatory responses, etc.

[0011] In addition, the present invention also suggests the application of tetracycline-loaded nanomotors in the preparation of preparations for inhibiting inflammatory factors and INF-β signaling pathways.

[0012] The effects and effects of the present invention: The present invention uses diisopropylamine-diazenediolate PAPA / NO to react with methacrylic anhydride or methacrylic acid or methacrylic acid salt to generate methacrylylated PAPA / NO, which is then reacted with N,N'-bisacryloylcystamine to obtain a new drug-loaded nanomotor MPN (synthetic route is as follows Figure 6 (As shown). PAPA / NO, a diisopropylamine-diazenediolate salt, was first used to develop drug-loaded nanomotors. The present invention's research revealed that these drug-loaded nanomotors can be used to carry anti-inflammatory drugs, such as tetracycline. The present invention also discovered that when tetracycline is loaded onto this drug-loaded nanomotor MPN, it can achieve a superior anti-inflammatory effect compared to tetracycline alone. Specifically, the tetracycline-loaded nanomotor MPN exhibited significantly greater anti-inflammatory effects than tetracycline, including significantly improved improvements in inflammatory lung injury and reduced systemic and local inflammatory responses. Therefore, the present invention's research demonstrates that these nanomotors have the potential to enhance drug efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Characterization of MPN nanomotors and MPN@TH nanomotors (a) Zeta potential; (b) dynamic light scattering (DLS) particle size distribution results; (c) cumulative release curve of TH from MPN@TH in PBS within 24 hours; (d) cumulative release curve of TH from PB@TH in PBS within 24 hours; (e) comparison of drug loading between MPN@TH and PB@TH.

[0014] Figure 2 Therapeutic effect of MPN@TH on inflammatory lung injury in ALI mice (a) Representative images of H&E-stained lung tissue from ALI mice. (b) Intergroup differences in lung injury scores, including total injury score, alveolar neutrophil infiltration score, interstitial neutrophil infiltration score, hyaline membrane score, protein fragmentation score, and alveolar membrane thickening score. TH, PB@TH, and MPN@TH interventions all significantly ameliorated LPS-induced lung injury, with MPN@TH demonstrating a significantly superior therapeutic effect compared to TH alone and non-motor-loaded PB@TH (statistical analysis performed using the Kruskal-Wallis test and Dunn's multiple comparison method. Data are expressed as mean ± SD. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001).

[0015] Figure 3 MPN@TH alleviates systemic inflammatory response in ALI mice (a) Routine blood test results of ALI mice; (b) Heat map analysis of multiple inflammatory factors in the serum of ALI mice; (c) Quantitative results of representative cytokine ELISA (statistical analysis was performed using the Kruskal-Wallis test and Dunn's multiple comparison method; data are expressed as mean ± SD. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001).

[0016] Figure 4 MPN@TH alleviates local inflammatory response in ALI mice (a) Changes in the number of neutrophils in bronchoalveolar lavage fluid; (b) Changes in the number of macrophages in bronchoalveolar lavage fluid; (c) Changes in the mean fluorescence intensity (MFI) of CD86 on macrophages in bronchoalveolar lavage fluid; (d) Changes in the mean fluorescence intensity (MFI) of CD206 on macrophages in bronchoalveolar lavage fluid; (e) Immunofluorescence analysis revealed changes in the expression of IL-1β (green fluorescence), IL-6 (red fluorescence), and TNFα (pink fluorescence) in lung tissue. Statistical analysis was performed using the Kruskal-Wallis test and Dunn's multiple comparison method. Data are expressed as mean ± standard deviation. * indicates P < 0.05, ** indicates P < 0.01, and *** indicates P < 0.001.

[0017] Figure 5 .Transcriptome sequencing revealed that MPN@TH has a therapeutic effect on ALI mice by inhibiting inflammatory response, neutrophil chemotaxis, and interferon and other cytokine activation mechanisms.

[0018] Figure 6 .The synthetic route using methacrylic anhydride as an example, where PAPA / NO: diisopropylamine-diazenediol salt, TEA: triethylamine, MAC: methacrylic anhydride, AS: ammonium sulfate, TMEDA: N,N,N',N'-tetramethylethylenediamine, MPN: nanomotor, MPN@TH: tetracycline-loaded nanomotor. DETAILED DESCRIPTION

[0019] The present invention is susceptible to various modifications and embodiments, and thus specific embodiments are illustrated and described in the accompanying drawings. However, this is not intended to limit the present invention to specific embodiments, but rather should be understood to encompass all modifications, equivalents, and even substitutes that fall within the spirit and technical scope of the present invention.

[0020] Example 1. Synthesis of MPN nanomotor: Diisopropylamine-diazenediolate reacts with methacrylic anhydride, methacrylic acid, or methacrylic acid salt to generate methacrylylated PAPA / NO, which is then reacted with N,N'-bisacryloylcystamine to obtain the nanomotor MPN. The synthesis method of methacryloyl PAPA / NO comprises: mixing diisopropylamine-diazenediolate, water and 1,4-dioxane, adding triethylamine TEA dropwise to the mixture, then adding methacrylic anhydride or methacrylic acid or methacrylic acid salt dropwise while stirring in an ice-water bath, stirring at room temperature overnight after the addition is completed, and adding acetone as a precipitation agent to the reaction solution to precipitate to obtain methacryloyl PAPA / NO after the reaction is completed; The molar ratio of diisopropylamine-diazeniumdiolate to methacrylic anhydride MAC or methacrylic acid MMA or methacrylic acid salt is 1:1-2; the added amounts of diisopropylamine-diazeniumdiolate, water and 1,4-dioxane are 1-5 ml of water and 0.5-2 ml of 1,4-dioxane per mmol of diisopropylamine-diazeniumdiolate; and the added molar amount of triethylamine is 2-3 times the molar amount of diisopropylamine-diazeniumdiolate.

[0021] The process of further reacting methacrylylated PAPA / NO with N,N'-bisacryloylcystamine to obtain the nanomotor MPN is as follows: N,N'-bisacryloylcystamine and methacrylylated PAPA / NO are dissolved in a buffer solution (any commonly used buffer solution can be used), ultrasonically mixed, and then deoxygenated with nitrogen for 30 minutes. Subsequently, an initiator and a catalyst are added under continuous nitrogen protection, and the reaction is stirred at room temperature until the solution becomes turbid. After post-treatment, the nanomotor MPN is obtained; wherein the initiator is selected from ammonium persulfate AS; and the catalyst is selected from N,N,N',N'-tetramethylethylenediamine TMEDA. The molar ratio of methacrylylated PAPA / NO to N,N'-bisacryloylcystamine is 1-10:1; the added amount of the initiator is 1-10% of the total weight of the system; and the added amount of the catalyst is 5-15% of the total weight of the system.

[0022] The best test case is as follows: 1. Synthesis of Tetracycline-Loaded Nanomotors (MPN@TH) 1.1. Synthesis of diisopropylamine-diazenediolate derivatives First, diisopropylamine-diazeniumdiol salt (PAPA / NO) (1.94 g, 12 mmol, Mcclin Reagent, HY-134636) was dissolved in a mixed solution of distilled water (20.0 mL) and 1,4-dioxane (8.5 mL, National Pharmaceutical Group Chemical Reagent Co., Ltd.). Then, triethylamine (4.5 mL, 32.3 mmol, Aldrich Chemical Company) was added dropwise to the mixture. After the reaction system was cooled in an ice water bath, methacrylic anhydride (3.0 mL, 18.9 mmol, Aldrich Chemical Company) was slowly added dropwise under stirring, and the dropwise addition time was controlled within 10 minutes. The ice water bath was removed, and the reaction was stirred at room temperature overnight. The final product was obtained by twice reprecipitation by adding 400.0 mL of acetone to the reaction solution and finally drying under vacuum at 60°C. The final product was named methacrylated PAPA / NO, abbreviated as MPN.

[0023] 1.2. Synthesis of MPN nanomotor First, N,N'-bisacryloyl cystamine (BAC, 2.00 mg, 0.0077 mmol, Shanghai Mcclin Biochemical Technology Co., Ltd.) and MPN (9.25 mg, 0.0285 mmol) were dissolved in phosphate buffered saline (PBS, 2.0 mL) and ultrasonically mixed. The solution was deoxygenated with nitrogen for 30 minutes. Then, initiator ammonium persulfate (APS, 4% of the total system, 0.45 mg, Shanghai Mcclin Biochemical Technology Co., Ltd.) and N,N,N',N'-tetramethyl ethylenediamine (TEMED, 8% of the total system, 0.90 mg, Shanghai Aldrich Biochemical Technology Co., Ltd.) were added under continuous nitrogen protection. The reaction was stirred at room temperature for 2 hours until the solution became turbid. The product was centrifuged (8000 rpm, 10 minutes), washed with water twice, and freeze-dried to obtain a white solid.

[0024] 1.3. Synthesis of non-nanomotor PB BAC (12.00 mg, 0.0462 mmol) was dissolved in PBS (2.0 mL) and ultrasonically mixed. The solution was deoxygenated with nitrogen for 30 minutes. Initiator APS (4% of the total system, 0.48 mg) and TEMED (8% of the total system, 0.96 mg) were added under continuous nitrogen protection. The reaction was stirred at room temperature for 2 hours until turbidity appeared. The product was centrifuged (8000 rpm, 10 minutes), washed with water twice, and freeze-dried to obtain a white solid, which was recorded as PB non-nanomotor. PB (20 mg) was dispersed in a PBS solution containing TH (5 mL, 2.5 mg / mL) and stirred at room temperature for 12 hours. The white solid was collected by centrifugation and named PB@TH non-nanomotor.

[0025] Example 2. Preparation method of tetracycline-loaded nanomotor: The nanomotor MPN was dispersed in a buffer solution containing tetracycline, stirred at room temperature for 10-24 hours, and then collected by centrifugation to obtain the MPN@TH nanomotor; wherein the mass ratio of the nanomotor MPN to tetracycline was 1:0.5-1.

[0026] The best test case is as follows: Preparation of MPN@TH nanomotors MPN (20 mg) was dispersed in PBS solution containing TH (5 mL, 2.5 mg / mL) and stirred at room temperature for 12 hours. The white solid was collected after centrifugation and named MPN@TH nanomotors.

[0027] Example 3. Results and Characterization 1. Characterization of Nanomotors Nanoparticle (NP) size was analyzed using an HT7800 transmission electron microscope (Hitachi, Japan). Zeta potential and particle size distribution were determined using a Nano-Z Zetasizer (Malvern Instruments, UK). Proton nuclear magnetic resonance (H-NMR) spectra of the samples were acquired using a Bruker Avance 400 NMR spectrometer. Absorbance at 360 nm was measured using an automated microplate reader (Tecan (Shanghai) Laboratory Equipment Co., Ltd., Switzerland). Fourier transform infrared (FT-IR) spectra were acquired using a Cary 5000 FT-IR spectrometer (Varian, USA). The motion trajectories of the nanomotors in the cellular environment were recorded using a fluorescence microscope (MF53-N, Guangzhou Mingmei Optoelectronics Technology Co., Ltd.). Chemical structures were drawn using ChemDraw Ultra 7.0 software.

[0028] RAW 264.7 macrophages (5 × 10 4 cells / mL) were inoculated into culture dishes and cultured overnight. After 12 hours of stimulation with lipopolysaccharide (LPS, Shanghai Yisheng Biotechnology Co., Ltd.), Cy5-labeled MPN@TH and PB@TH (200 μg / mL) were added. Unstimulated macrophages were added to the control group samples simultaneously. The movement trajectory was tracked using an inverted fluorescence microscope (×100 objective lens), and the image sequence was analyzed using Image J software (v2.0.0). The average velocity of the particles was calculated by averaging the velocity within the time interval. Ten nanoparticles were tracked in each group to determine the average velocity and mean square displacement (MSD) value. The particle motion type was classified by MSD fitting. Results are shown in Figure 1 .

[0029] 2. Nanomotor validation in a mouse model: Alleviating acute lung injury in mice Male C57BL / 6J mice aged 8-12 weeks (purchased from Saiye Biolabs) were used. Brief description of the acute lung injury (ALI) model: After anesthesia with an intraperitoneal injection of 80 mg / kg sodium pentobarbital, 70 μL of PBS containing 160 μg of LPS was intratracheally instilled to induce lung injury. Subsequently, 80 μL of PBS containing TH (1 μg / g), PB@TH (4.7 μg / g, including 1 μg / g of TH), or MPN@TH (5.1 μg / g, including 1 μg / g of TH) was instilled to evaluate the efficacy of ALI treatment. A sham-operated group was instilled with an equal volume of PBS as a control. Mice were sacrificed 24 hours after the intervention, and peripheral blood, lung tissue, and bronchoalveolar lavage fluid (BALF) were collected for subsequent analysis.

[0030] To evaluate lung injury scores and local inflammatory responses, fixed sections of the left upper lung of 3-4 mice were obtained from each group. Five pathological features (alveolar neutrophil infiltration, interstitial neutrophil infiltration, hyaline membrane formation, alveolar protein exudates, and alveolar septal thickening) were scored using HE staining. The severity of the injury was scored as 0, 1, or 2 points, and the weighted average was used to obtain a final score of 0-1. Results are available at Figure 2 , indicating the therapeutic effect of MPN@TH on inflammatory lung injury in ALI mice.

[0031] 3. Nanomotors alleviate systemic inflammatory responses in mice with acute lung injury Routine blood testing: 24 hours after model establishment, anticoagulated blood samples were collected via cardiac puncture and analyzed by Wuhan Service Biotechnology Co., Ltd. (official website: https: / / www.servicebio.cn / ). Four mice were included in each experimental group.

[0032] Serum multiplex ELISA assay: 24 mice after modeling, 500 μL of whole blood samples were collected from each mouse and separated by centrifugation (5000 rpm, 10 min). The resulting protein samples were used in a multiplex ELISA assay (Boster, Catalog No. MEK1003, 13-factor combined assay) to assess systemic inflammation. Each experimental group included 3 mice. Results are available at [link missing]. Figure 3 , indicating that MPN@TH alleviates the systemic inflammatory response in ALI mice.

[0033] 4. Nanomotors alleviate local inflammatory responses in the lungs of mice with acute lung injury Immunofluorescence staining of lung tissue: Local inflammation analysis was performed by immunofluorescence staining to detect the expression of proinflammatory cytokines and inflammatory cell activation. The primary antibodies used included IL-1β (Abcam, TA5103, 1:200), IL-6 (Abcam, TD6087, 1:200), TNFα (Abcam, PY19810S, 1:500), F4 / 80 (Abcam, ab300421, 1:100), LY6G (Proteintech, 65078-1-Ig, 1:100), and CCR7 (Proteintech, 25898-1-AP, 1:200).

[0034] Bronchoalveolar lavage fluid flow cytometric analysis: Bronchoalveolar lavage fluid (BALF) was collected from mice and centrifuged at 350 g for 5 minutes to harvest immune cells. After washing with PBS, the cells were resuspended in 50 μL of buffer and stained for the following markers: live / dead markers and immune cell activation markers. Data were acquired using a BD FACSCanto II flow cytometer and analyzed using FlowJo software. After excluding doublets and dead cells, neutrophils were identified as CD45+CD11b+Ly6G+ cells and macrophages as CD45+CD11b+Ly6G- cells. Macrophage polarization was analyzed by assessing the mean fluorescence intensity (MFI) of CD86 and CD206. Three to four mice were included in each experiment.

[0035] Lung tissue multiplex ELISA assay: Total protein was extracted from lung lobe tissue and the total protein concentration in lung tissue was determined using a BCA protein quantification kit. The extracted protein was then analyzed using a multiplex ELISA kit (Boster, Cat. No. MEK1003, 13-factor assay) to assess the level of local inflammation. Three mice were included in each experimental group. Results are shown in the table. Figure 4 , indicating that MPN@TH alleviated the local inflammatory response in ALI mice.

[0036] 5. Transcriptome sequencing reveals the therapeutic mechanism of nanomotors Total RNA from the left lower lobe of the mouse lung was extracted using Trizol and sequenced. Differential expression analysis was performed using the DESeq algorithm. A false discovery rate-corrected P value < 0.05 and a log2FC > 1 or < -1 were set as the screening thresholds for differentially expressed genes (DEGs). Gene ontology (GO) and gene set enrichment analysis (GSEA) were further performed on the DEGs to explore their functions and biological significance. Figure 5 Transcriptome sequencing revealed that MPN@TH had a therapeutic effect on ALI mice by inhibiting inflammatory response, neutrophil chemotaxis, and interferon and other cytokine activation mechanisms.

[0037] The effects and effects of this embodiment are as follows: This study demonstrated that drug-loaded nanomotors exhibited superior anti-inflammatory effects compared to tetracycline alone. Specifically, they significantly improved inflammatory lung injury and reduced systemic and local inflammatory responses. This demonstrates that these motors can enhance drug efficacy and potentially reduce side effects.

[0038] While the above description focuses on the embodiments, this is merely illustrative and does not limit the present invention. Persons skilled in the art will readily appreciate that various modifications and applications not illustrated above are possible without departing from the essential characteristics of the embodiments. For example, the various components specifically illustrated in the embodiments may be implemented with modifications. Furthermore, any differences associated with such modifications and applications should be construed as being within the scope of the present invention as defined in the appended claims.

Claims

1. A method for preparing drug-loaded nanomotors targeting acute lung injury, characterized by: Diisopropylamine-diazenediolate reacts with methacrylic anhydride, methacrylic acid, or methacrylic acid salt to generate methacrylylated PAPA / NO, which is then reacted with N,N'-bisacryloylcystamine to obtain the nanomotor MPN. The synthesis method of the methacryloylated PAPA / NO is as follows: diisopropylamine-diazenediolate, water and 1,4-dioxane are mixed, triethylamine is added dropwise, and then methacrylic anhydride or methacrylic acid or methacrylic acid salt is added dropwise while stirring under cooling in an ice-water bath. After the addition is completed, the mixture is stirred at room temperature and reacted overnight. After the reaction is completed, acetone is added to the reaction solution for precipitation to obtain the methacryloylated PAPA / NO; The molar ratio of the diisopropylamine-diazeniumdiolate to methacrylic anhydride or methacrylic acid or methacrylic acid salt is 1:(1-2); the added amounts of the diisopropylamine-diazeniumdiolate, water and 1,4-dioxane are 1ml-5ml of water and 0.5ml-2ml of 1,4-dioxane per mmol of diisopropylamine-diazeniumdiolate; the added molar amount of the triethylamine is 2-3 times the molar amount of the diisopropylamine-diazeniumdiolate; The process of further reacting methacrylylated PAPA / NO with N,N'-bisacryloylcystamine to obtain the nanomotor MPN is as follows: N,N'-bisacryloylcystamine and methacrylylated PAPA / NO are dissolved in a buffer solution and ultrasonically mixed, followed by nitrogen deoxygenation for 30 minutes, followed by addition of an initiator and a catalyst under continuous nitrogen protection, and reacting at room temperature with stirring until the solution becomes turbid, followed by centrifugation, washing with water, and freeze-drying to obtain the nanomotor MPN; The molar ratio of the methacrylated PAPA / NO to N,N'-bisacryloylcystamine is (1-10):1; the initiator is selected from ammonium persulfate; the catalyst is selected from N,N,N',N'-tetramethylethylenediamine; the added amount of the initiator is 1%-10% of the total weight of the system; and the added amount of the catalyst is 5%-15% of the total weight of the system.

2. The method for preparing a drug-loaded nanomotor targeting acute lung injury according to claim 1, wherein: The reaction process of the methacrylylated PAPA / NO and N,N'-bisacryloylcystamine is carried out in a phosphate buffer system.

3. A tetracycline-loaded nanomotor, characterized in that: Tetracycline is loaded on the nanomotor MPN obtained by any one of the preparation methods of claims 1-2.

4. The method for preparing a tetracycline-loaded nanomotor according to claim 3, wherein: The nanomotor MPN was dispersed in a buffer solution containing tetracycline, stirred at room temperature for 10-24 hours, and then collected by centrifugation to obtain the MPN@TH nanomotor. The mass ratio of the nanomotor MPN to tetracycline is 1:(0.5-1).

5. Use of the tetracycline-loaded nanomotor according to claim 3 in the preparation of anti-inflammatory drugs.

6. Use of the tetracycline-loaded nanomotor according to claim 3 in the preparation of a drug for treating, alleviating and preventing inflammatory lung injury and reducing systemic / local inflammatory response.

7. Use of the tetracycline-loaded nanomotor according to claim 3 in preparing a preparation for inhibiting inflammatory factors and INF-β signaling pathway.

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