Application of trace CSE modified polypeptide gold nanoparticles in preparation of medicine for treating chronic obstructive pulmonary disease

Abstract

The invention relates to polypeptide gold nanoparticles modified by trace cigarette extract CSE (0.1%-1%) and application of the polypeptide gold nanoparticles in preparation of drugs for treating chronic obstructive pulmonary diseases. The polypeptide-modified gold nanoparticle is a polypeptide-modified gold nanoparticle as shown in a sequence SEQ ID NO: 1. It is confirmed for the first time that polypeptide (CLPFFD, SEQ ID NO: 1) gold nanoparticles modified by trace CSE (0.1%-1%) can antagonize mtDNA synthesis and inhibit mitochondrial ROS generation so as to inhibit activation of macrophage NLRP3 inflammasome, induce depolarization of macrophage and target lung macrophage in vivo to achieve anti-inflammatory activity, and the polypeptide (CLPFFD, SEQ ID NO: 1) gold nanoparticles can be used for preparing anti-inflammatory drugs. The compound has an obvious treatment effect in a COPD acute exacerbation model and a stable phase model, so that the compound can be used for preparing medicines for treating COPD. The medicine has the advantages of being remarkable in curative effect, capable of acting on specific cells in a targeted mode, convenient to track in vivo and the like, and a new treatment means is provided for COPD.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology. Specifically, it relates to the application of trace CSE (0.1%-1%) modified polypeptide gold nanoparticles in the preparation of drugs for treating chronic obstructive pulmonary disease. Background Art

[0002] Chronic obstructive pulmonary disease (COPD) is a common chronic airway inflammatory disease characterized by persistent respiratory symptoms and airflow limitation, usually associated with airway and / or alveolar abnormalities caused by significant exposure to toxic particles or gases. The pathogenesis of COPD is complex and involves multiple inflammatory pathways, among which the inflammatory response is the main mechanism of COPD development. It has been found that the number of alveolar macrophages in the airways of COPD patients is increased compared with that of healthy people. These cells can secrete various inflammatory mediators, proteases, transforming growth factor-β (TGF-β), reactive oxygen species, etc., and interact with other inflammatory cells to form a complex inflammatory network, inducing chronic inflammatory responses, promoting alveolar wall destruction and fibrosis formation, which play a key role in the progression of COPD. In addition, there are M1 and M2 type polarization activations in the pulmonary macrophages of COPD patients. Among them, M1 type macrophages are involved in the initiation of inflammation and tissue destruction in COPD, while M2 type macrophages are closely related to airway remodeling in COPD. Currently, clinically well-known COPD treatment drugs, such as inhaled corticosteroids combined with long-acting β2-agonists, can partially inhibit the inflammatory response and relieve the clinical symptoms of COPD patients. However, there is still no effective treatment plan that can prevent or reverse the progression of the disease and the symptom burden so far. Summary of the Invention

[0003] The purpose of the present invention is to provide an application of trace CSE (0.1%-1%) modified polypeptide gold nanoparticles for the deficiencies in the prior art.

[0004] On the one hand, the present invention provides an application of trace CSE (0.1%-1%) modified polypeptide gold nanoparticles in inhibiting the activation of the NLRP3 inflammasome signaling pathway by inhibiting mitochondrial DNA synthesis and the generation of mtROS. The polypeptide gold nanoparticles are gold nanoparticles modified with a polypeptide having a sequence as shown in SEQ ID NO:1.

[0005] On the other hand, the present invention provides an application of trace CSE (0.1%-1%) modified polypeptide gold nanoparticles in regulating macrophage polarization in vitro. The polypeptide gold nanoparticles are gold nanoparticles modified with a polypeptide having a sequence as shown in SEQ ID NO:1.

[0006] On the other hand, the present invention provides an application of a polypeptide gold nanoparticle modified with trace CSE (0.1%-1%) in a medicament for treating chronic obstructive pulmonary disease, wherein the polypeptide gold nanoparticle is a gold nanoparticle modified with a polypeptide having the sequence shown in SEQ ID NO: 1.

[0007] In the fourth aspect, the present invention provides an application of a polypeptide gold nanoparticle modified with trace CSE (0.1%-1%) in exerting anti-inflammatory activity and promoting protease / antiprotease balance by targeting lung macrophages in vivo, and the polypeptide nanoparticle is a gold nanoparticle modified with a polypeptide having the sequence shown in SEQ ID NO: 1.

[0008] In this article, the polypeptide gold nanoparticle modified with trace CSE (0.1%-1%) and having the sequence shown in SEQ ID NO: 1 is designated as CSE-P12.

[0009] The beneficial effects of the present invention are as follows:

[0010] The present invention for the first time proves that a polypeptide (CLPFFD, SEQ ID NO: 1) gold nanoparticle modified with trace CSE (0.1%-1%) can inhibit the activation of the NLRP3 inflammasome signaling pathway, induce macrophage depolarization, and exert anti-inflammatory effects and promote protease / antiprotease balance by targeting lung macrophages, and has obvious therapeutic effects in both the acute exacerbation model and the stable phase model of COPD. Therefore, it can be used to prepare a medicament for treating COPD. This medicament has the advantages of remarkable curative effect, targetable action on specific cells, and convenient in vivo tracking, etc., providing a new treatment means for COPD.

[0011] The present invention proposes a polypeptide-based nano-drug, which is a new type of drug preparation that has been studied more in recent years. It has a series of advantages such as targetable action on specific cells, increased selectivity of drug delivery, improved treatment efficiency, and reduced side effects.

[0012] When the medicament for treating chronic obstructive pulmonary disease of the present invention is applied, it can not only target specific cells, increase the selectivity of drug delivery, improve the treatment effect, but also reduce side effects. The multiple advantages of this nano-drug provide a brand-new treatment perspective for COPD patients, marking a great progress in the treatment field of respiratory diseases. Its excellent curative effect, precise targeting, and easy in vivo tracking characteristics undoubtedly bring new hope for the treatment of COPD. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figures 1a - 1b For the preparation and characterization of the polypeptide gold nanoparticle CSE-P12 modified with trace CSE;

[0014] Figure 1a For the synthesis and preparation of CSE-P12;

[0015] Figure 1b To analyze the hydrodynamic diameters of naked GNPs, P12, and CSE-P12 by DLS (n = 3);

[0016] Figures 2a - 2c To investigate the inhibitory effect of CSE-P12 on the activation of the NLRP3 inflammasome signaling pathway in mouse bone marrow-derived macrophages (BMDMs).

[0017] Figure 2a To study the effect of CSE-P12 on the components of the NLRP3 inflammasome in bone marrow-derived macrophages (BMDMs) stimulated with LPS for 6 h;

[0018] Figure 2b To examine the expression levels of mature IL-1β and activated caspase-1 proteins in the supernatant after CSE-P12 treatment in bone marrow-derived macrophages (BMDMs) stimulated with LPS for 6 h and then treated with the NLRP3 activator ATP for 1 h;

[0019] Figure 2c To determine the expression level of mature IL-1β in the supernatant after CSE-P12 treatment in bone marrow-derived macrophages (BMDMs) stimulated with LPS for 6 h and then treated with the NLRP3 activator ATP for 1 h;

[0020] Figure 3a 、 Figure 3b 、 Figure 3c To investigate the inhibitory effect of CSE-P12 on mtDNA synthesis, mtROS generation, and the IRF1 / CMPK2 signaling pathway;

[0021] Figure 3a To detect the inhibitory effect of CSE-P12 on mitochondrial DNA synthesis in BMDMs stimulated with LPS for 4 h by RT-qPCR; mitochondrial DNA is represented by D-loop and Non-Numt; nuclear DNA is represented by Tert and B2m.

[0022] Figure 3b To measure the effect of CSE-P12 on intracellular mitochondrial ROS generation in BMDMs stimulated with LPS for 6 h and then treated with ATP (4 mM) for 1 h using MitoSOX;

[0023] Figure 3c To investigate the inhibitory effect of CSE-P12 on the IRF1 / CMPK2 signaling pathway;

[0024] Figure 4a 、 Figure 4b To investigate the regulatory effect of CSE-P12 on macrophage polarization in vitro;

[0025] Figure 4(a) Mouse bone marrow-derived macrophages (BMDMs) were induced to polarize into M1 by co-stimulation with LPS (10 ng / mL) and IFN-γ (20 ng / mL) for 24 h. CSE-P12 was treated simultaneously with the stimulator, and the effect of CSE-P12 on Nos2 expression was detected by RT-qPCR (n = 3);

[0026] Figure 4b For BMDMs induced to polarize into M2 by co-stimulation with IL-4 (10 ng / mL) and IL-13 (20 ng / mL) for 24 h, CSE-P12 was treated simultaneously with the stimulator, and the effect of CSE-P12 on Arg1 expression was analyzed by RT-qPCR (n = 3);

[0027] Figures 5a - 5f For CSE-P12 to exert an anti-inflammatory effect by targeting lung macrophages in vivo to treat COPD;

[0028] Figure 5a For the effect of CSE-P12 on airway resistance, an index of pulmonary function, in a mouse model of acute exacerbation of COPD induced by LPS combined with cigarette smoke;

[0029] Figure 5b For the effect of CSE-P12 on pulmonary inflammatory cell infiltration in a mouse model of acute exacerbation of COPD induced by LPS combined with cigarette smoke;

[0030] Figure 5c For the effect of CSE-P12 on airway resistance, an index of pulmonary function, in a mouse model of stable COPD induced by cigarette smoke;

[0031] Figure 5d For the effect of CSE-P12 on airway inflammation, alveolar cavity enlargement and airway fibrosis in a mouse model of stable COPD induced by cigarette smoke.

[0032] Figure 5e For the effect of CSE-P12 on the imbalance between protease and anti-protease in the lungs of mice in a mouse model of stable COPD induced by cigarette smoke.

[0033] Figure 5f For the effect of CSE-P12 treatment on the number of inflammatory cells in the bronchoalveolar lavage fluid (BALF) of mice in a model of acute exacerbation of COPD induced by LPS combined with cigarette smoke after macrophages were depleted by clodronate liposomes. Detailed implementation manners

[0034] The following detailed description of the specific implementation manners provided by the present invention will be given in conjunction with the accompanying drawings.

[0035] Example 1

[0036] Preparation of CSE-P12

[0037] 1. Experimental method

[0038] The synthesis of gold nanoparticles was carried out by the method of reducing chloroauric acid with sodium citrate. The specific operation method is as follows: Add 161.2 μL of chloroauric acid (420 mg / mL) to a conical flask containing 200 mL of ultrapure water (18.2 MΩ). After heating to boiling on a magnetic stirrer, quickly add 2 mL of sodium citrate (10%). Continue heating and stirring for 15 minutes, and then stir and cool to room temperature. Polypeptide P12: CLPFFD (SEQ ID NO: 1), with a purity > 95% powder, was dissolved in ultrapure water to prepare a polypeptide solution (1 mM). The polypeptide and the gold nanoparticle solution were mixed at a volume ratio of 1:10 and allowed to stand at room temperature for 24 h to fully combine the polypeptide and the gold nanoparticles. The polypeptide-gold nanoparticle suspension was filtered using a 0.22 μm syringe filter head, then centrifuged (15000 rpm, 30 min), and then washed three times with PBS and concentrated to 100 nM (nanoparticle concentration) for use.

[0039] The preparation of cigarette smoke extract (CSE) was obtained by a modified method using commercially available cigarettes (containing 10 mg of tar and 0.8 mg of nicotine). The specific operation method is as follows: First, two gas sampling bottles were hermetically connected by a rubber tube, and a cigarette with the filter tip cut off was fixed on one side of the first gas sampling bottle using a rubber tube. 10 mL of RPMI 1640 medium was added to this gas sampling bottle, and a 50 mL syringe was connected to the other side of the second gas sampling bottle through a rubber tube. After the sealed connection was completed, the cigarette was lit, and the 50 mL syringe was used to quickly draw air. The cigarette smoke was dissolved in 10 mL of RPMI 1640 medium to form 100% CSE of cigarette smoke extract. Then it was filtered and diluted 100 times and fully mixed and incubated with the polypeptide-gold nanoparticle complex (P12) for 1 h, and finally became CSE-P12 containing 0.1% - 1% CSE.

[0040] 2. Experimental results

[0041] In this application, an anti-inflammatory nanoparticle CSE-P12 of a trace CSE-modified polypeptide-gold nanoparticle complex was designed. One cigarette was lit and the gas was quickly drawn out using a syringe, and the cigarette gas was dissolved in 10 mL of cell culture medium to form cigarette smoke extract (100% CSE); it was filtered and diluted to 0.1% - 1% CSE and then fully mixed with the polypeptide-gold nanoparticle complex P12, and finally became CSE-P12. After the preparation of the trace CSE-modified polypeptide-gold nanoparticles, their physicochemical properties were detected (Figure 1).

[0042] Example 2

[0043] CSE-P12 inhibits the activation of the NLRP3 inflammasome signaling pathway

[0044] 1. Experimental methods

[0045] In in vitro experiments, mouse bone marrow cells were obtained from wild-type mice (8 - 10 weeks old, C57BL / 6, SPF grade). The cells were cultured in IMDM medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and 20 ng / mL M-CSF for 7 days to obtain bone marrow-derived macrophages (BMDMs). On the 7th day, the macrophages were collected and seeded in 24-well plates (5×10 5 cells / well). The cells were treated with LPS (100 ng / mL) alone or in combination with P12 (100 nM), CSE, and CSE-P12 (100 nM) for 6 h. Protein immunoblotting was used to detect the expression of NLRP3, ASC, pro-IL-1β, and pro-caspase-1, the components of the NLRP3 inflammasome in the cells, to evaluate the effect of CSE-P12 on the initial stage of NLRP3 inflammasome activation. After stimulating BMDMs with LPS (100 ng / mL) for 6 h, the NLRP3 inflammasome activator ATP (4 mM, 1 h) was added to treat the cells, and then the supernatant was collected, freeze-dried and concentrated for protein. Protein immunoblotting was used to detect the expression of mature IL-1β and activated caspase-1 in the supernatant, and ELISA was used to analyze the secretion level of IL-1β in the supernatant to clarify the inhibitory effect of CSE-P12 on NLRP3 inflammasome activation in Mφ.

[0046] 2. Experimental results

[0047] To verify the effect of CSE-P12 on NLRP3 inflammasome activation, we investigated the effect of CSE-P12 on the activation process of NLRP3, the processing of pro-IL-1β, and the NLRP3 inflammasome in LPS-primed macrophages. The experimental results showed that CSE-P12 could significantly reduce the expression of NLRP3, ASC, pro-IL-1β, and pro-caspase-1, the components of the NLRP3 inflammasome, while P12 and CSE alone had no significant inhibitory effect ( Figure 2a ). More importantly, CSE-P12 inhibited the release of activated caspase-1 and mature IL-1β proteins in macrophages induced by the NLRP3 agonist ATP ( Figure 2b ). In addition, enzyme-linked immunosorbent assay was used to detect the inhibitory effect of each group of nanoparticles on the production of IL-1β in LPS-primed macrophages induced by the NLRP3 agonist ATP, and it was found that CSE-P12 could significantly reduce the secretion of IL-1β ( Figure 2c ).

[0048] Example 3

[0049] CSE-P12 inhibits mtDNA synthesis and mtROS production

[0050] 1. Experimental methods

[0051] To further explore whether CSE-P12 inhibits the activation of NLRP3 inflammasome by improving mitochondrial function, the extracted and induced differentiated BMDMs were seeded in 24-well plates (5×10 5 cells / well). They were treated with LPS (100 ng / mL) alone or in combination with P12 (100 nM), CSE, CSE-P12 (100 nM) for 4 h, and then the total cellular DNA was extracted using a blood and tissue extraction kit. Specific primers for the mitochondrial D-loop (displacement loop, D-loop) and a specific mitochondrial DNA region that is not inserted into nuclear DNA (non-nuclear mitochondrial DNA segment, Non-Numt) were used to quantify mitochondrial DNA by qPCR. Nuclear DNA encoding Tert and B2m was used for normalization. BMDMs were treated with P12 (100 nM), CSE, CSE-P12 (100 nM) and LPS (100 ng / mL) for 6 h, and then stimulated with the NLRP3 inflammasome activator ATP (4 mM) for 30 min. Then 4 μM of MitoSOX was added to incubate the cells for 20 min. After washing twice with PBS, the cells were resuspended in PBS, and then cells from different groups (with the same number of cells) were added to a 96-well plate for fluorescence reading to minimize differences between groups. The fluorescence intensity was measured at 510 / 580 nm using a microplate reader. In addition, after BMDMs were treated with LPS (100 ng / mL) in combination with P12 (100 nM), CSE, CSE-P12 (100 nM) for 6 h, the supernatant was discarded, and cellular proteins were extracted. The expression of key proteins IRF1 and CMPK2 that regulate DNA replication was detected by Western blotting.

[0052] 2. Experimental results

[0053] Using the qPCR method to detect the production of mitochondrial DNA, it was found that CSE-P12 could significantly inhibit LPS-induced mtDNA synthesis in macrophages ( Figure 3a ); and the production of mtROS was detected using the MitoSOX superoxide indicator. The results showed that P12, CSE and CSE-P12 could all inhibit LPS+ATP-induced mtROS production, and CSE-P12 had a better inhibitory effect ( Figure 3b ). Verification by Western blotting found that CSE-P12 could inhibit the expression of IRF1 and CMPK2 in LPS-induced BMDMs ( Figure 3c) This result suggests that CSE-P12 may inhibit the generation of mtDNA by regulating the IRF1 / CMPK2 signaling pathway, inhibit the production of mtROS, and thus inhibit the activation of the NLRP3 inflammasome.

[0054] Example 4

[0055] CSE-P12 Regulates Macrophage Polarization in Vitro

[0056] 1. Experimental Method

[0057] The extracted and induced differentiated BMDMs were seeded in 12-well plates (1×10 6 cells / well). BMDMs were co-stimulated with LPS (10 ng / mL) and IFN-γ (20 ng / mL) or IL-4 (10 ng / mL) and IL-13 (20 ng / mL) to induce M1 or M2 polarization. These cells were co-treated with CSE-P12 (100 nM) and the stimulants for 24 h. After 24 h, the supernatant was discarded, and cell RNA was extracted by the Trizol method for analysis of the expression of M1 or M2 macrophage-specific genes.

[0058] 2. Experimental Results

[0059] BMDMs were co-stimulated with LPS and IFN-γ to induce M1 polarization. We found that CSE-P12 significantly decreased the expression of the M1-type specific marker gene Nos2. Under the stimulation of IL-4 and IL-13 co-induced M2 polarization in BMDMs, CSE-P12 was also able to inhibit the expression of the M2-type specific marker gene Arg1 ( Figure 4a and b).

[0060] Example 5

[0061] CSE-P12 Targets Lung Macrophages to Exert Anti-Inflammatory Activity and Treat Chronic Obstructive Pulmonary Disease 1. Experimental Method

[0062] Establish a mouse model of acute exacerbation of COPD induced by LPS combined with cigarette smoke and a mouse model of stable COPD induced by cigarette smoke alone: Select C57BL / 6 wild-type mice (6 - 8 weeks old), and replicate the COPD mouse model by the method of mainstream cigarette smoke inhalation (commercially available Marlboro cigarettes, tar content 10 mg / cigarette, 5 cigarettes / time, four times / day, with an interval of 30 min, 5 days / week). For the acute exacerbation model of COPD, not only should the smoke exposure last for 4 weeks, but also LPS (O111:B4, 10 mg / kg) should be instilled into the airway on the 21st day. The mouse model of stable COPD is established by continuous cigarette smoke exposure alone for 6 months. The mice will be raised in a clean environment at the SPF level, and their body weights will be measured weekly and their health status will be recorded. Before cigarette smoke exposure, intratracheal instillation (500 nM, 50 μL) will be administered using a laryngoscope. An in vivo animal pulmonary function testing analyzer will be used. In the way of sealing the posterior end of the nose, using the bony prominence behind the animal's ear, and fixing the animal's position with a U-shaped clip, the pulmonary function-related indexes will be recorded by measuring the airflow at the nose and chest of the mouse, including the raw inspiratory resistance (RAW), respiratory frequency (F), minute ventilation volume (MV), and 50% volume expiratory flow (EF50).

[0063] After the experiment, the neck and chest will be dissected, the trachea and the right lung lobe will be ligated, a puncture needle will be inserted into the upper end of the trachea, and 0.4 mL of PBS will be injected into the left lung with a syringe and left to stay for about 30 s for unilateral lavage, usually repeated twice. The retrieved lavage fluid will be collected into a 1.5 mL EP tube (placed in an ice bath), and about 0.65 mL of lung lavage fluid can be recovered; after measuring the volume of the specimen, it will be centrifuged at 4°C (1500 r / min) for 10 min, the supernatant will be aliquoted and stored at -80°C, and the cell sediment will be resuspended in 1 or 0.5 mL of PBS for direct counting and smear of nucleated cells; wait for the slide to dry naturally, and then fix it in 10% neutral formaldehyde solution for more than 10 minutes for routine HE staining. The BALF of the mice will be collected, and the total number of infiltrating cells and the numbers of neutrophils, macrophages, and lymphocytes will be detected by cell counting in the BALF. The lower right lobe of the lung will be fixed with 4% paraformaldehyde for histological examination with H&E staining and Masson staining. The evaluation of COPD histopathology is based on the following features: airway inflammation, alveolar septal intercept, and peribronchial collagen deposition. The protein expression levels of matrix metalloproteinase MMP12 and tissue inhibitor of metalloproteinase TIMP1 in the lung tissue will be detected by Western blotting, and finally, the improvement effect of CSE-P12 intervention on emphysema in COPD mice will be systematically evaluated.

[0064] To evaluate the anti-inflammatory activity of CSE-P12 in vivo through macrophages in a COPD model, after clearing pulmonary macrophages in mice using clodronate liposomes, a mouse model of acute exacerbation of COPD induced by LPS combined with cigarette smoke was constructed. 72 h before the start of the experiment, the mice were anesthetized by intraperitoneal injection of 2.5% avertin. Under the assistance of a laryngoscope, a pipette was used to inject clodronate liposomes (75 μL, 5 mg / mL). The group without macrophage clearance was replaced with blank liposomes, and the treatment method was the same as above. 72 h after clearing pulmonary macrophages in the mice, the mice were anesthetized, and nanoparticles CSE-P12 (500 nM, 50 μL) were instilled into the trachea by the method of drug administration through a laryngoscope. The CS+LPS model group was given PBS in the same way. After that, a smoking experiment was carried out, 5 days a week, 10 cigarettes each time, twice a day for continuous smoking for 4 weeks. On the 19th day, clodronate liposomes were also given to clear pulmonary macrophages in the mice. 72 h later, CSE-P12 (500 nM, 50 μL) was given in the same way as before, and LPS (10 mg / kg) was instilled into the trachea through a laryngoscope for model establishment. After continuous smoking for 48 h, the experiment was ended, and the mice were dissected according to the above sample collection protocol.

[0065] 2. Experimental results

[0066] Using a mouse model of acute exacerbation induced by LPS combined with cigarette smoke, we verified that CSE-P12 could effectively reduce the airway resistance (Raw) and inflammatory cell infiltration in the pulmonary function measurement indexes of mice caused by CS+LPS ( Figure 5a and b). In a COPD stable-phase model induced by cigarette smoke alone for 6 months, through the analysis of mouse airway resistance, H&E staining and Masson staining, it was confirmed that CSE-P12 could effectively improve CS-induced airway obstruction, airway inflammation, increased alveolar septal intercept and peribronchial collagen deposition ( Figure 5c and Figure 5d ). Thus, it can be seen that CSE-P12 has a significant therapeutic effect in the acute exacerbation model and stable-phase mouse model of COPD. In addition, Western blot experiments also verified that CSE-P12 treatment could significantly inhibit the up-regulation of Arg1 and MMP12 and the degradation of tissue inhibitor of metalloproteinase 1 (TIMP1) in lung tissue ( Figure 5e ). These results confirmed that CSE-P12 could inhibit the protease / antiprotease imbalance in lung tissue, thereby reducing the formation of emphysema and alleviating the pathogenesis of COPD.

[0067] An acute exacerbation model of COPD was constructed by depleting pulmonary macrophages, and BALF was collected to evaluate the infiltration of inflammatory cells. The results showed that in the case of macrophage depletion, CSE-P12 treatment could not reduce the total cell count induced by CS+LPS stimulation, while in the presence of pulmonary macrophages, the protective effect of CSE-P12 still existed. These results demonstrated the importance of pulmonary macrophages in the anti-inflammatory effect of CSE-P12 in the LPS combined with CS-induced acute exacerbation model of COPD( Figure 5f ).

[0068] The above shows and describes the basic principles, main features and advantages of the present invention. Each component mentioned in the present invention is a common technology in the existing field. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and the description in the specification only illustrate the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of protection claimed by the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for treating chronic obstructive pulmonary disease by using a peptide gold nanoparticle modified with trace amounts of CSE, characterized in that The polypeptide gold nanoparticles are gold nanoparticles modified with a polypeptide having a sequence as shown in SEQ ID NO: 1, and the polypeptide gold nanoparticles are used to inhibit the activation of NLRP3 inflammasomes.

2. The use of a trace amount of CSE-modified polypeptide gold nanoparticles in a drug for treating chronic obstructive pulmonary disease according to claim 1, characterized in that The application of polypeptide gold nanoparticles inhibits the activation of NLRP3 inflammasome signaling pathway by inhibiting mitochondrial DNA synthesis and the generation of mtROS.

3. The use of a trace amount of CSE-modified polypeptide gold nanoparticles in a drug for treating chronic obstructive pulmonary disease according to claim 1, characterized in that The application of polypeptide gold nanoparticles inhibits mtDNA synthesis by antagonizing the IRF1 / CMPK2 signaling axis, inhibits the production of mitochondrial mtROS, and thus prevents the activation of macrophage NLRP3 inflammasome.

4. The use of a trace amount of CSE-modified polypeptide gold nanoparticles in a drug for treating chronic obstructive pulmonary disease according to claim 1, characterized in that The polypeptide gold nanoparticles simultaneously inhibit the polarization of macrophages to M1 type and M2 type, and are used in the preparation of drugs for depolarizing macrophages.

5. The use of a trace amount of CSE-modified polypeptide gold nanoparticles in a drug for treating chronic obstructive pulmonary disease according to claim 4, characterized in that Application of peptide gold nanoparticles to exert anti-inflammatory activity in vivo and promote protease / antiprotease balance by targeting lung macrophages.

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