Application of binding molecule of actin binding protein LASP1 in prevention and treatment of acute respiratory distress syndrome

Abstract

The invention belongs to the technical field of medicines, and particularly discloses application of binding molecules of actin binding protein LASP1 in prevention and treatment of acute respiratory distress syndrome (ARDS). According to the invention, the LASP1-loaded nanoparticles are constructed, so that the problems that the drug is difficult to penetrate and easy to remove and degrade and the ARDS is difficult to treat are solved. According to the invention, through intranasal instillation of the HA-DA / BP / LASP1 nanoparticles, the level of inflammatory factors can be reduced, the alveolar epithelial barrier structure and function can be repaired, and the death rate of ARDS model mice can be reduced.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of a binding molecule of actin binding protein LASP1 in preventing and treating acute respiratory distress syndrome. Background Art

[0002] Acute respiratory distress syndrome (ARDS) is a common clinical critical illness and the most serious complication of COVID-19. In recent years, although symptomatic supportive treatment strategies such as protective lung ventilation, fluid management, and extracorporeal membrane oxygenation (ECMO) have been successfully applied in clinical practice, these current treatment methods are all "physiological assistance". ARDS remains the main cause of death in critically ill patients, with a mortality rate as high as 35-45%. So far, no specific drug therapy has been proven to be effective for ARDS. Therefore, improving the prognosis of the disease, reducing the mortality rate of ARDS patients, and finding new therapeutic targets and methods are key issues that need to be addressed urgently.

[0003] ARDS is a diffuse lung injury characterized by pulmonary inflammatory storm and destruction of the alveolar-capillary barrier. The alveolar epithelial barrier is an important line of defense against the pulmonary inflammatory storm and is also the main target of invasion by pathogenic factors. The degree of its damage is an important determinant of the severity of the disease. When ARDS occurs, pathogenic microorganisms enter the airway, adhere to the surface of the airway mucosa, break through the mucosal defense and invade the alveolar epithelial cells, inducing mitochondrial depolarization, cytoplasmic Ca 2+ Increased inflammatory factors promote cell damage, which in turn leads to the destruction of the structure and function of the alveolar epithelial barrier. The damaged alveolar epithelial barrier encourages more immune cells to enter the lungs, triggering more uncontrolled inflammatory cell infiltration and inflammatory factor release, forming a vicious cycle between inflammatory cascades and lung barrier damage. At the same time, a large amount of exudate accumulates, causing pulmonary edema, further aggravating the progression of the disease. Therefore, promoting the rapid repair of the damaged alveolar epithelial barrier is an important entry point for the treatment of ARDS.

[0004] The alveolar epithelial barrier is maintained by alveolar epithelial cells and intercellular tight junctions, focal adhesions, gap junctions, and desmosomes. Among them, the state of intercellular tight junctions is an important determinant of the structure and function of the alveolar epithelial barrier. During lung inflammation, the tight junctions between alveolar epithelial cells are destroyed, leading to the formation of a large amount of alveolar exudate and reducing the clearance rate of alveolar exudate; ARDS patients with low clearance rate have a significantly poor prognosis. Therefore, repairing intercellular tight junctions and promoting the recovery of alveolar epithelial barrier structure and function are the key to treating ARDS. At the same time, during the progression of ARDS, airway secretions are viscous, drugs are difficult to penetrate and are easily cleared and degraded, and the effective delivery of drugs is also an urgent problem to be solved.

[0005] LIM and SH3 protein 1 (LASP1) is an F-actin binding protein that was first discovered to be highly expressed in malignant tumor tissues in 1995. Currently, LASP1 has been confirmed to be overexpressed in different types of malignant tumors and is significantly associated with proliferation and invasion. Summary of the invention

[0006] In view of the above-mentioned shortcomings, the present invention provides an application of a binding molecule of actin binding protein LASP1 in preventing and treating acute respiratory distress syndrome. The present invention constructs nanoparticles (HA-DA / BP / LASP1) loaded with LASP1 protein to solve the problem that drugs are difficult to penetrate and are easily cleared and degraded, as well as the problem of treating ARDS. The present invention can downregulate the level of inflammatory factors, repair the structure and function of the alveolar epithelial barrier, and reduce the mortality rate of ARDS model mice by intranasal instillation of HA-DA / BP / LASP1 nanoparticles.

[0007] In order to achieve the above objectives, in a first aspect, the present invention provides use of actin-binding protein LASP1 in the preparation of a drug for preventing and treating acute respiratory distress syndrome.

[0008] In a second aspect, the present invention provides use of a binding molecule of actin-binding protein LASP1 in the preparation of a drug for preventing and treating acute respiratory distress syndrome.

[0009] Furthermore, the actin-binding protein LASP1 binding molecule includes the actin-binding protein LASP1 and a carrier for loading LASP1.

[0010] Furthermore, the carrier is poly-β-amino ester modified by dopamine grafted with hyaluronic acid.

[0011] In a third aspect, the present invention provides a drug for preventing and treating acute respiratory distress syndrome, wherein the drug comprises actin-binding protein LASP1.

[0012] Furthermore, the drug also includes a carrier loaded with LASP1.

[0013] Furthermore, the carrier is poly-β-amino ester modified by dopamine grafted with hyaluronic acid.

[0014] Furthermore, the preparation method of the drug comprises the following steps:

[0015] S1. Dissolve hyaluronic acid in PBS with a pH value of 5.5, then mix with dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride in N2 in the dark, and then dialyze and freeze-dry to obtain dopamine-grafted hyaluronic acid, i.e., HA-DA;

[0016] S2, resuspending the poly-β-amino ester in PBS, adding LASP1 recombinant protein and mixing evenly in an ice bath to obtain a poly-β-amino ester solution loaded with LASP1 recombinant protein, i.e., BP / LASP1 solution;

[0017] S3. Add HA-DA to the BP / LASP1 solution and mix in an ice bath to form binding molecules of the actin-binding protein LASP1.

[0018] Furthermore, the drug administration method includes any one of nasal drip, atomization, oral administration, intravenous injection, and enema.

[0019] Furthermore, the drug is a nanoparticle.

[0020] Beneficial effects of the present invention:

[0021] The present invention provides a binding molecule of actin binding protein LASP1, which is composed of poly-β-amino ester (BP) modified by dopamine grafted hyaluronic acid (HA-DA) as a carrier, has good mucus penetration, and its high specific surface area is conducive to contact with tissues, improves drug efficacy, has good targeting to lung tissue, and can reduce adverse reactions. On this basis, the present invention preliminarily constructs nanoparticles (HA-DA / BP / LASP1) carrying LASP1 protein. Intranasal instillation of the nanoparticles can reduce the mortality of ARDS model mice; alleviate the congestion of the lung tissue of ARDS model mice, reduce the pathological changes such as inflammatory cell infiltration and alveolar septum thickening in the lung tissue of model mice; reduce the wet / dry weight ratio of the lung of ARDS mice; reduce the total protein content in the BALF of ARDS mice; downregulate the level of inflammatory factors in the BALF of ARDS mice; downregulate the level of inflammatory factors in the lung tissue of ARDS mice; improve the tight junctions between alveolar epithelial cells of ARDS mice; reduce the Evans blue (EB) penetration of the lung barrier of ARDS model mice; and upregulate the mRNA levels of molecules related to the lung epithelial barrier in the lung tissue of ARDS model mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The synthesis process of HA-DA / BP / LASP1 nanoparticles of the present invention;

[0023] Figure 2The HA-DA / BP / LASP1 nanoparticles of the present invention improve the mortality rate of ARDS mice, reduce the level of inflammation and improve lung injury; wherein, A is the electron microscope image of HA-DA / BP / LASP1 nanoparticles, scale bar = 100nm; B is HA-DA / BP / LASP1 reducing the mortality rate of lethal ARDS model mice within 168h; C is HA-DA / BP / LASP1 alleviating the congestion phenomenon of lung tissue in ARDS model mice; D is HA-DA / BP / LASP1 alleviating the pathological changes such as inflammatory cell infiltration and alveolar septum thickening in the lung tissue of model mice, scale bar = 20μm; E is HA-DA / BP / LASP1 alleviating the wet / dry weight ratio of ARDS mice lungs; F is HA-DA / BP / LASP1 reducing the total protein content in BALF of ARDS mice; G is HA-DA / BP / LASP1 downregulating the level of inflammatory factors in BALF of ARDS mice; H is HA-DA / BP / LASP1 downregulating the level of inflammatory factors in lung tissue of ARDS mice; Note: ***p<0.001vs NS; ###p<0.001vs LPS+NC; NC: normal mice; LPS+NC: lethal ARDS model mice were treated with blank nanoparticles; LPS+LASP1: lethal ARDS model mice were treated with HA-DA / BP / LASP1 nanoparticles;

[0024] Figure 3 The HA-DA / BP / LASP1 nanoparticles of the present invention improve the lung epithelial barrier of ARDS mice; wherein, A represents that HA-DA / BP / LASP1 improves the tight junctions between alveolar epithelial cells of ARDS mice, scale bar=1μm; B represents that HA-DA / BP / LASP1 reduces the EB penetration of the lung barrier of ARDS model mice; C represents that HA-DA / BP / LASP1 upregulates the mRNA levels of lung epithelial barrier-related molecules in the lung tissues of ARDS model mice; Note: ***p<0.001vs NS; #p<0.05##p<0.01###p<0.001vs LPS+NC; NC: normal mice; LPS+NC: lethal ARDS model mice were intervened with blank nanoparticles; LPS+LASP1: lethal ARDS model mice were intervened with HA-DA / BP / LASP1 nanoparticles. DETAILED DESCRIPTION

[0025] To make the present invention easier to understand, the present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of the present invention. Unless otherwise defined, the professional terms used below are consistent with the meanings understood by professional and technical personnel in the field; unless otherwise specified, the raw materials and reagents involved in this article can be purchased from the market or prepared by known methods.

[0026] Example 1

[0027] Synthesis of HA-DA / BP / LASP1

[0028] (1) Preparation of HA-DA:

[0029] 500 mg of hyaluronic acid was dissolved in PBS with pH 5.5, and then mixed with 237.1 mg of dopamine hydrochloride and 240 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride in N2 in the dark for 2 h, dialyzed and freeze-dried to obtain HA-DA.

[0030] (2) Preparation of BP:

[0031] 5g of glycerol, 22.6mL of triethylamine and 60mL of dichloromethane were fully mixed in an ice bath, and then 13.7ml of acryloyl chloride was added dropwise, reacted at room temperature for 12h, and triacrylate was collected by filtration and evaporation. Each 1mmol of triacrylate was mixed with 5g of 1,4-butanediol diacrylate and 2.2g of 5-amino 1-pentanol at 90℃ in the dark for 24h. Then 160μL of DMSO and 640ul of 1,3-diaminopropane were added to react at room temperature overnight, and polyβ-amino ester, i.e. BP, was obtained by purification through ether precipitation.

[0032] (3) Preparation of HA-DA / BP / LASP1, the synthesis flow chart is as follows Figure 1 As shown:

[0033] BP was resuspended in PBS, LASP1 recombinant protein was added and mixed in an ice bath for 30 min, and then HA-DA was added to the BP / LASP1 solution and mixed in an ice bath for 30 min to form HA-DA / BP / LASP1.

[0034] The HA-DA / BP / LASP1 prepared above was examined by transmission electron microscopy, and the results are as follows: Figure 2 As shown in A. Figure 2 As shown in A, HA-DA / BP / LASP1 is in the form of nanoparticles.

[0035] Example 2

[0036] Effect of HA-DA / BP / LASP1 on mortality in lethal ARDS model mice

[0037] Control group (LPS+NC group): LPS (sigma L2630, 25 mg / kg) was instilled into normal mice through the trachea to establish a lethal ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later, and the mortality of mice was monitored within 168 hours (7 days).

[0038] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 25 mg / kg) was instilled into normal mice through the trachea to establish a lethal ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled into the nose 2 hours later, and the mortality of mice was monitored within 168 hours (7 days).

[0039] The above results are Figure 2 As shown in B. Figure 2 B shows that HA-DA / BP / LASP1 nanoparticles can improve the mortality rate of ARDS mice.

[0040] Example 3

[0041] Effects of HA-DA / BP / LASP1 on lung pathological morphology in ARDS mice

[0042] Normal control group (NS group): Lung tissues of normal mice were obtained 24 hours later, and gross images of the tissues were taken. After paraffin embedding, the slices were sliced ​​and HE staining was performed to detect the pathological morphology of the lung tissues.

[0043] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. Lung tissues were obtained 24 hours later, and gross images of the tissues were taken. After paraffin embedding, the sections were sliced ​​and stained with HE to detect the pathological morphology of the lung tissues.

[0044] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. Lung tissues were obtained 24 hours later, and gross images of the tissues were taken. The sections were embedded in paraffin and stained with HE to detect the pathological morphology of the lung tissues.

[0045] The above results are Figure 2 C- Figure 2 D. Figure 2C- Figure 2 D shows that HA-DA / BP / LASP1 alleviates the congestion of lung tissue in ARDS model mice, and reduces pathological changes such as inflammatory cell infiltration and alveolar septum thickening in the lung tissue of model mice.

[0046] Example 4

[0047] Effects of HA-DA / BP / LASP1 on pulmonary edema in ARDS mice

[0048] Normal control group (NS group): lung tissues of normal mice were taken after 24 hours, the wet weight of tissues was measured, and the dry weight of tissues was measured after drying at 65°C for 48 hours, and the lung wet / dry weight ratio was calculated.

[0049] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. Lung tissue was collected 24 hours later, the wet weight of the tissue was weighed, and the dry weight of the tissue was weighed after drying at 65°C for 48 hours, and the lung wet / dry weight ratio was calculated.

[0050] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled into the nose 2 hours later. Lung tissue was collected 24 hours later, and the wet weight of the tissue was weighed. The dry weight of the tissue was weighed after drying at 65°C for 48 hours, and the lung wet / dry weight ratio was calculated.

[0051] The above results are Figure 2 E. Figure 2 E shows that HA-DA / BP / LASP1 reduces the lung wet / dry weight ratio of ARDS mice.

[0052] Example 5

[0053] Effects of HA-DA / BP / LASP1 on total protein in BALF of ARDS mice

[0054] Normal control group (NS group): 24 hours later, normal mice were perfused with 1 ml of pre-cooled PBS through the trachea, and the BALF was obtained by repeated perfusion three times.

[0055] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. 1 ml of pre-cooled PBS was instilled through the trachea 24 hours later. BALF was obtained by repeated lavage 3 times.

[0056] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. 1 ml of pre-cooled PBS was instilled through the trachea 24 hours later. The BALF was obtained by repeated lavage 3 times.

[0057] The total protein concentration in BALF of the control group and the experimental group was determined by BCA method. Figure 2 F. Figure 2 F shows that HA-DA / BP / LASP1 reduces the total protein content in BALF of ARDS mice.

[0058] Example 6

[0059] Effects of HA-DA / BP / LASP1 on inflammatory factors in BALF of ARDS mice

[0060] Normal control group (NS group): 24 hours later, normal mice were perfused with 1 ml of pre-cooled PBS through the trachea, and the BALF was obtained by repeated perfusion three times.

[0061] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. 1 ml of pre-cooled PBS was instilled through the trachea 24 hours later. BALF was obtained by repeated lavage 3 times.

[0062] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. 1 ml of pre-cooled PBS was instilled through the trachea 24 hours later. The BALF was obtained by repeated lavage 3 times.

[0063] The inflammatory factor concentrations in the control group and experimental group were detected by ELISA kit. The results are as follows: Figure 2 G. By Figure 2 G shows that HA-DA / BP / LASP1 downregulates the levels of inflammatory factors in BALF of ARDS mice.

[0064] Example 7

[0065] Effects of HA-DA / BP / LASP1 on inflammatory factors in lung tissue of ARDS mice

[0066] Normal control group (NS group): Lung tissues were collected from normal mice 24 hours later, RNA was extracted by trizol method, reverse transcribed into cDNA, and the expression of inflammatory factors was detected by qPCR using corresponding gene primers.

[0067] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later, RNA was extracted by the trizol method, and the expression of inflammatory factors was detected by qPCR using the corresponding gene primers after reverse transcription into cDNA.

[0068] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later, RNA was extracted by the trizol method, and the corresponding gene primers were used to detect the expression of inflammatory factors by qPCR after reverse transcription into cDNA.

[0069] The results of the experimental and control groups are as follows Figure 2 H. Figure 2 It can be seen that HA-DA / BP / LASP1 downregulates the levels of inflammatory factors in the lung tissue of ARDS mice.

[0070] Example 8

[0071] Effects of HA-DA / BP / LASP1 on tight junctions between alveolar epithelial cells in ARDS mice

[0072] Normal control group (NS group): Lung tissues of normal mice were taken after 24 hours and quickly placed in electron microscope fixative (Wuhan Saiweier G1102). The lungs were cut into pieces no larger than 2 mm using a sharp blade in the fixative. 3 The tissue blocks were placed in a new fixative and fixed in the dark for 2 hours at room temperature. They were then transported and sent for examination using ice packs. Transmission electron microscopy was used to analyze the morphological changes of the tight junctions between alveolar epithelial cells.

[0073] Control group (LPS+NC group): Normal mice were instilled with LPS (sigma L2630, 5 mg / kg) through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later and quickly placed in electron microscopy fixative (Wuhan Saiweier G1102). The lung tissues were cut into pieces no larger than 2 mm in the fixative using a sharp blade. 3The tissue blocks were placed in a new fixative and fixed in the dark for 2 hours at room temperature. They were then transported and sent for examination using ice packs. Transmission electron microscopy was used to analyze the morphological changes of the tight junctions between alveolar epithelial cells.

[0074] Experimental group (LPS+LASP1 group): Normal mice were instilled with LPS (sigma L2630, 5 mg / kg) through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later and quickly placed in electron microscopy fixative (Wuhan Saiweier G1102). The lung tissues were cut into pieces no larger than 2 mm in the fixative using a sharp blade. 3 The tissue blocks were placed in a new fixative and fixed in the dark for 2 hours at room temperature. They were then transported and sent for examination using ice packs. Transmission electron microscopy was used to analyze the morphological changes of the tight junctions between alveolar epithelial cells.

[0075] The results of the experimental and control groups are as follows Figure 3 As shown in A. Figure 3 A shows that HA-DA / BP / LASP1 improves the tight junctions between alveolar epithelial cells in ARDS mice.

[0076] Example 9

[0077] Effects of HA-DA / BP / LASP1 on lung tissue permeability in ARDS mice

[0078] Normal control group (NS group): normal mice were injected with 2% EB (0.2 ml) through the tail vein 22 hours later, and 5 ml of normal saline was lavaged through the heart 2 hours later. Lung tissue was obtained and photographed, and then EB was extracted from the tissue by formamide method for quantitative analysis.

[0079] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. 2% EB (0.2 ml) was injected through the tail vein 22 hours later. 5 ml of normal saline was lavaged through the heart 2 hours later, lung tissue was obtained, photographed, and then tissue EB was extracted by the formamide method for quantitative analysis.

[0080] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. 2% EB (0.2 ml) was injected through the tail vein 22 hours later. 5 ml of normal saline was lavaged through the heart 2 hours later, lung tissue was obtained, photographed, and then tissue EB was extracted by the formamide method for quantitative analysis.

[0081] The results of the experimental and control groups are as follows Figure 3 As shown in B. Figure 3 B shows that HA-DA / BP / LASP1 reduces EB penetration in the lung barrier of ARDS model mice.

[0082] Example 10

[0083] Effects of HA-DA / BP / LASP1 on lung barrier-related molecules in ARDS mice

[0084] Normal control group (NS group): Lung tissues were collected from normal mice 24 hours later, RNA was extracted by the trizol method, and the expression of barrier-related molecules was detected by qPCR using the corresponding gene primers after reverse transcription into cDNA.

[0085] Control group (LPS+NC group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP nanoparticles (blank nanoparticles) were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later, RNA was extracted by the trizol method, and the expression of barrier-related molecules was detected by qPCR using the corresponding gene primers after reverse transcription into cDNA.

[0086] Experimental group (LPS+LASP1 group): LPS (sigma L2630, 5 mg / kg) was instilled into normal mice through the trachea to establish the ARDS mouse model. HA-DA / BP / LASP1 nanoparticles were instilled through the nose 2 hours later. Lung tissues were collected 24 hours later, RNA was extracted by the trizol method, and the expression of barrier-related molecules was detected by qPCR using the corresponding gene primers after reverse transcription into cDNA.

[0087] The results of the experimental and control groups are as follows Figure 3 C. Figure 3 C shows that HA-DA / BP / LASP1 upregulates the mRNA levels of lung epithelial barrier-related molecules in the lung tissues of ARDS model mice.

[0088] Comparative Example 1

[0089] Synthesis of HA-DA / BP nanoparticles (blank nanoparticles)

[0090] (1) Preparation of HA-DA:

[0091] 500 mg of hyaluronic acid was dissolved in PBS with pH 5.5, and then mixed with 237.1 mg of dopamine hydrochloride and 240 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride in N2 in the dark for 2 h, dialyzed and freeze-dried to obtain HA-DA.

[0092] (2) Preparation of BP:

[0093] 5g of glycerol, 22.6mL of triethylamine and 60mL of dichloromethane were fully mixed in an ice bath, and then 13.7ml of acryloyl chloride was added dropwise, reacted at room temperature for 12h, and triacrylate was collected by filtration and evaporation. Each 1mmol of triacrylate was mixed with 5g of 1,4-butanediol diacrylate and 2.2g of 5-amino 1-pentanol at 90℃ in the dark for 24h. Then 160μL of DMSO and 640ul of 1,3-diaminopropane were added to react at room temperature overnight, and polyβ-amino ester, i.e. BP, was obtained by purification through ether precipitation.

[0094] (3) Preparation of HA-DA / BP:

[0095] BP was resuspended in PBS and placed on ice for 30 min, and then HA-DA was added to the BP solution and mixed on ice for 30 min to form HA-DA / BP nanoparticles.

[0096] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with the art within the technical scope disclosed in the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims.

Claims

1. Application of actin-binding protein LASP1 in the preparation of drugs for preventing and treating acute respiratory distress syndrome.

2. Application of binding molecules of actin-binding protein LASP1 in the preparation of drugs for preventing and treating acute respiratory distress syndrome.

3. The use according to claim 2, characterized in that: The actin-binding protein LASP1 binding molecule comprises the actin-binding protein LASP1 and a carrier for loading LASP1.

4. The use according to claim 3, characterized in that: The carrier is poly-β-amino ester modified by dopamine grafted with hyaluronic acid.

5. A drug for preventing and treating acute respiratory distress syndrome, characterized in that: The drugs include the actin-binding protein LASP1.

6. The drug for preventing and treating acute respiratory distress syndrome according to claim 5, characterized in that The drug also includes a carrier loaded with LASP1.

7. The drug for preventing and treating acute respiratory distress syndrome according to claim 6, characterized in that The carrier is poly-β-amino ester modified by dopamine grafted with hyaluronic acid.

8. The drug for preventing and treating acute respiratory distress syndrome according to claim 7, characterized in that: The preparation method of the drug comprises the following steps: S1. Dissolve hyaluronic acid in PBS with a pH value of 5.5, then mix with dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride in N2 in the dark, and then dialyze and freeze-dry to obtain dopamine-grafted hyaluronic acid, i.e., HA-DA; S2, resuspending the poly-β-amino ester in PBS, adding LASP1 recombinant protein and mixing evenly in an ice bath to obtain a poly-β-amino ester solution loaded with LASP1 recombinant protein, i.e., BP / LASP1 solution; S3. Add HA-DA to the BP / LASP1 solution and mix in an ice bath to form binding molecules of the actin-binding protein LASP1.

9. The drug for preventing and treating acute respiratory distress syndrome according to claim 5, characterized in that: The drug administration method includes any one of nasal drip, atomization, oral administration, intravenous injection, and enema.

10. The drug for preventing and treating acute respiratory distress syndrome according to claim 5, characterized in that: The drug is a nanoparticle.

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