Application of caffeic acid phenethyl ester in preparation of medicine for treating acute lung injury caused by erosive toxic agent

Abstract

The invention relates to an application of caffeic acid phenethyl ester (CAPE) in preparation of a medicine for treating acute lung injury caused by erosive toxic agents. Aiming at an erosive toxic agent 2-chloroethyl ethyl sulfide (CEES) induced mouse acute lung injury model, the experimental result shows that the CAPE can obviously relieve pathological injury of lung tissues and recover lung function indexes including ventilation per minute, inspiration time, expiration time and relaxation time through an antioxidant effect, and can effectively reduce the level of reactive oxygen species (ROS) and the content of malondialdehyde (MDA) in the lung tissues. The invention shows that the CAPE as a potential therapeutic drug has a remarkable protective effect on the acute lung injury induced by the erosive toxic agent, provides a new candidate drug and a prevention and treatment strategy for treating the erosive toxic agent poisoning, and has important clinical application prospects and social values.

Description

Technical Field

[0001] The invention relates to the field of medical drugs, and in particular to application of caffeic acid phenethyl ester in preparing a drug for acute lung injury caused by erosive poisons. Background Art

[0002] Sulfur Mustard (SM), commonly known as mustard gas, has a chemical name of dichlorodiethyl sulfide and is known as the "king of poisons". It can not only easily penetrate the skin, eyes and respiratory mucosa, causing systemic poisoning, but also because of its lipophilicity, it can be quickly absorbed by the human body, causing direct invasion and damage to multiple organs, tissues and cells. In the First World War, mustard gas was first used on the battlefield on a large scale, causing millions of casualties, and its horrific influence is still remembered by the world.

[0003] Acute lung injury caused by sulfur mustard is one of the important causes of death from poisoning. When sulfur mustard enters the human body through the respiratory tract, it quickly triggers a severe inflammatory response, leading to pathological features such as epithelial necrosis, tracheal ulcers, and tracheal mucosal shedding. These pathological changes not only seriously affect the normal function of the lungs, but may also lead to acute respiratory distress syndrome, thereby endangering the patient's life. Therefore, the development of effective preventive and therapeutic drugs for acute lung injury caused by sulfur mustard is of great strategic significance for safeguarding national public security and citizen health.

[0004] However, the poisoning mechanism of sulfur mustard is complicated, and current treatment methods are still ineffective. A large number of studies have shown that oxidative stress damage induced by reactive oxygen species (ROS) is one of the important toxic mechanisms of sulfur mustard poisoning. Sulfur mustard damages biological macromolecules such as DNA and proteins through alkylation in the body, resulting in the generation of a large amount of ROS. These ROS then trigger peroxidation reactions of lipids, proteins, and carbohydrates, seriously damaging the structure and function of lung tissue. Therefore, finding potential drugs that can effectively inhibit oxidative stress has become a new challenge for the treatment of sulfur mustard poisoning.

[0005] 2-Chloroethyl ethyl sulfide (CEES), as a sulfur mustard mimetic, is often used in the medical field to study the specific mechanism of sulfur mustard injury and screen therapeutic drugs. Similar to sulfur mustard, CEES can also construct an acute lung injury model through aerosol inhalation to simulate the pathological process of sulfur mustard poisoning. The establishment of this model provides an important experimental platform for in-depth research on the mechanism of sulfur mustard poisoning and the development of new therapeutic drugs.

[0006] Caffeic acid phenethyl ester (CAPE) is a bioactive ingredient extracted from propolis, which has a wide range of biological characteristics, including antioxidant, immunomodulatory and anti-tumor activities. In recent years, CAPE has shown strong clinical application prospects in the treatment of burns, tumors, cardiovascular, diabetes, local trauma and skin diseases. However, there are relatively few studies on whether CAPE has a protective effect on sulfur mustard-induced lung injury.

[0007] In order to solve the above problems, the applicant proposed the use of caffeic acid phenethyl ester in the preparation of drugs for acute lung injury caused by erosive toxic agents. Summary of the invention

[0008] The purpose of the present invention is to provide an application of caffeic acid phenethyl ester in the preparation of a drug for treating acute lung injury caused by erosive poisons, so as to solve the problems in the prior art.

[0009] To achieve the above object, the present invention provides the following technical solution: a compound for protecting lung damage caused by erosive toxic agents, wherein the compound is caffeic acid phenethyl ester.

[0010] Optionally, the blister agent is sulfur mustard or its analog 2-chloroethyl ethyl sulfide.

[0011] Optionally, it is used to mitigate lung tissue damage caused by blistering agents.

[0012] Alternatively, it reduces the oxidative stress level in lung tissue by inhibiting the generation of reactive oxygen species in lung tissue and reducing the content of lipid oxidation products such as malondialdehyde (MDA).

[0013] Optionally, it is administered by intraperitoneal injection, and the recommended dosage is 10 mg per kilogram of body weight.

[0014] Optionally, when used, caffeic acid phenethyl ester is administered within a period of time after exposure to the blister agent and is administered repeatedly to maintain its effective concentration in the body.

[0015] Optionally, the period of time after exposure to the blistering agent specifically refers to administration of caffeic acid phenethyl ester at 1 hour, 25 hours and 49 hours after exposure.

[0016] A method for protecting against lung damage caused by erosive agents, comprising administering the caffeic acid phenethyl ester compound according to any one of claims 1 to 7 to a person who may be exposed to the erosive agent.

[0017] Optionally, the method also includes monitoring lung function-related parameters, lung tissue morphological changes, antioxidant indices, and oxidative stress levels of personnel exposed to erosive agents to evaluate the protective effect.

[0018] Alternatively, this approach is suitable for persons who may be at risk for lung injury from exposure to blister agents.

[0019] Beneficial effects: 1. Effectively relieve acute lung injury:

[0020] By constructing a mouse acute lung injury model, the present invention confirmed that CAPE has a significant protective effect on acute lung injury induced by 2-chloroethyl ethyl sulfide (CEES). The lung function indicators of mice in the CAPE treatment group were significantly restored, including minute ventilation, inspiratory time, expiratory time and relaxation time, and the degree of pathological damage to lung tissue was also significantly reduced.

[0021] 2. Significant antioxidant effect:

[0022] The present invention found that CAPE effectively alleviated CEES-induced oxidative stress damage through antioxidant effects. CAPE can significantly reduce the level of reactive oxygen species (ROS) and malondialdehyde (MDA) content in lung tissue, which indicates that CAPE has a strong antioxidant capacity and can remove excess ROS in the body, thereby reducing oxidative damage to lung tissue.

[0023] 3. Provide new ideas for the treatment of erosive toxic agents:

[0024] As one of the earliest weapons used, the in-depth study of the poisoning mechanism and the formulation of effective prevention and treatment strategies of erosive agents have always attracted much attention. The present invention provides a new treatment idea for acute lung injury caused by erosive agents, that is, using the antioxidant effect of CAPE for intervention treatment. This not only brings more effective treatment hope to patients, but also lays the foundation for intervention measures for poisoning treatment of erosive agents and research on potential therapeutic drugs.

[0025] 4. Has broad application prospects:

[0026] Caffeic acid phenethyl ester (CAPE) is a bioactive ingredient extracted from propolis, which has a wide range of biological characteristics, including antioxidant, immunomodulatory and anti-tumor activities. The present invention not only confirms the therapeutic effect of CAPE in acute lung injury caused by erosive agents, but also provides new possibilities for its application in the treatment of burns, tumors, cardiovascular, diabetes, local trauma and skin diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 is the chemical structural formula of caffeic acid phenethyl ester;

[0028] Figure 2 This is a diagram showing the effect of CAPE intervention on lung injury in CEES-exposed mice;

[0029] Figure 3 The figure shows the effect of CAPE treatment on the pathological morphology of lung tissue in mice exposed to CEES;

[0030] Figure 4 This is a graph showing the effect of CAPE intervention on the lung function of mice exposed to CEES;

[0031] Figure 5 This is the effect of CAPE intervention on oxidative stress in lung tissue of CEES-exposed mice;

[0032] Figure 6 The effect of CAPE treatment on the antioxidant function of lung tissue in CEES-exposed mice DETAILED DESCRIPTION

[0033] The following describes the preferred embodiments of the present invention with reference to the drawings in the specification, so that the technical content is clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0034] Example 1

[0035] This technical solution revolves around four core links: selection and preparation of CAPE, design of dosing regimen, comprehensive evaluation of protective effects, and subsequent application and promotion strategy. It aims to build a complete protection system from source to application to ensure that CAPE can be efficiently and safely used in the protection against lung injury caused by erosive agents.

[0036] CAPE is mainly contained in natural products such as propolis, and its extraction method must take into account both efficiency and purity. This plan tends to adopt a green and environmentally friendly natural extraction method, by precisely controlling the extraction conditions (such as temperature, pH value, solvent selection, etc.), to ensure that the active ingredients of CAPE are retained to the maximum extent while avoiding harmful residues.

[0037] After extraction, CAPE needs to go through a multi-step purification process, including but not limited to column chromatography, thin layer chromatography and other high-performance chromatography techniques to remove impurities and improve purity. Subsequently, high-performance liquid chromatography (HPLC) is used for purity testing to ensure that the product meets high standards. At the same time, advanced technologies such as mass spectrometry (MS) and nuclear magnetic resonance (NMR) are used to accurately identify the chemical structure of CAPE to ensure the accuracy of its molecular structure.

[0038] According to the physical and chemical properties of CAPE, suitable preparation forms are designed, such as oral solution, injection or solid tablets, to meet the needs of different routes of administration. During the preparation process, stabilizers and preservatives are added to ensure the stability and safety of CAPE during long-term storage and transportation. At the same time, a strict quality control system is implemented, including sterility testing, content determination, etc., to ensure that each batch of products meets the pharmaceutical production specifications. CAPE preparations need to be stored in a specific environment, such as low temperature and away from light, to maintain their biological activity.

[0039] Based on the pharmacological properties of CAPE and the exposure characteristics of erosive agents, this protocol recommends intraperitoneal injection as the preferred route of administration to ensure that CAPE quickly enters the blood circulation and reaches the lung tissue directly. Based on previous animal experiments and clinical data support, 10 mg per kilogram of body weight was determined as the initial recommended dose, which not only ensures the protective effect but also avoids unnecessary side effects.

[0040] The timing and frequency of administration are crucial to the protective effect. Combining the mechanism of action of erosive agents and the pharmacokinetic characteristics of CAPE, a flexible dosing regimen was designed, that is, CAPE was administered at key time points after exposure (such as 1 hour, 25 hours, and 49 hours) to achieve continuous and effective protection. At the same time, the frequency and dosage of administration were adjusted in time according to individual differences and actual conditions to achieve the best protective effect.

[0041] Taking individual differences into consideration, this program encourages research on personalized dosing regimens. By monitoring the subjects' physiological indicators, pharmacokinetic parameters, etc., the most appropriate dosing regimen can be tailored for each subject to further improve the targetedness and effectiveness of protection.

[0042] Advanced lung function testing equipment is used to conduct real-time and dynamic monitoring of the subjects' lung function parameters, including but not limited to vital capacity, respiratory rate, blood oxygen saturation, airway resistance, etc., to comprehensively evaluate the protective effect of CAPE on lung function.

[0043] Through histological staining, immunohistochemistry, electron microscopy and other technical means, we conducted in-depth morphological analysis of lung tissue, observed alveolar structure, pulmonary interstitial changes, cell morphology and inflammatory cell infiltration, and quantitatively evaluated the repair effect of CAPE on lung tissue damage.

[0044] In order to explore the protective mechanism of caffeic acid phenethyl ester (CAPE) against erosive agent-induced acute lung injury, we constructed a complete oxidative stress detection system, which covers the determination of multiple key indicators and aims to comprehensively evaluate the inhibitory effect of CAPE on oxidative stress.

[0045] Detection process: Lung tissue ROS detection: Dihydroethidium (DHE) is a superoxide anion fluorescence detection probe. It dehydrogenates under the action of intracellular superoxide anions to form ethidium bromide. Ethidium bromide can bind to RNA or DNA to produce red fluorescence, and the fluorescence intensity is proportional to the ROS level. After the lung tissue was dissected from mice, it was quickly frozen in liquid nitrogen, embedded and fixed with OTG embedding agent, and then immediately cut into frozen sections with a thickness of about 5um. The nuclear fluorescent dye Hoechst (10μmol / L) and the reactive oxygen fluorescent probe DHE (10μmol / L) were stained, and the sections were observed and photographed under a microscope after sealing with glycerol, and the average fluorescence intensity was finally calculated.

[0046] Detection of MDA, SOD and CAT activity in lung tissue. First, we accurately weighed 50 mg of mouse lung tissue, cut it into pieces and placed it in a centrifuge tube. Subsequently, 450 μL of pre-cooled saline was added and mechanically homogenized using a high-throughput tissue grinder under ice bath conditions. The homogenizer operates at a frequency of 50 Hz, running for 30 seconds each time, then stopping for 30 seconds, and repeating this six times to ensure that the lung tissue is fully ground into a uniform slurry. Next, the homogenate was centrifuged at 3000 rpm for 10 minutes to separate the supernatant. Strictly following the instructions of the lung tissue MDA, CAT and SOD content assay kit, we determined the content of the above indicators. At the same time, in order to accurately evaluate the protein concentration of lung tissue, we also used the BCA kit for related measurements.

[0047] Through this series of sophisticated detection steps, we were able to deeply reveal the protective mechanism of CAPE against erosive agent-induced acute lung injury, providing strong scientific support for the application of CAPE in related fields.

[0048] The results of the test are as follows Figure 5 and 6As shown in the results, compared with the blank control group, the MDA content and ROS level in the lung tissue of mice in the CAPE control group did not change significantly (P<0.05), but the MDA content and ROS level in the lung tissue of mice in the CEES poisoning group increased significantly (P<0.05). After CAPE treatment, the above index changes were significantly reversed (P<0.05); in addition, compared with the blank control group, the CAT and SOD content in the lung tissue of mice in the CAPE control group did not change significantly (P<0.05), but after CEES poisoning, the above indexes increased significantly (P<0.05). After CAPE treatment, the increase of CAT and SOD content in lung tissue induced by CEES was reversed. Conclusion: CAPE can inhibit the excessive generation of ROS and MDA, thereby alleviating CEES-induced oxidative stress damage in the lung tissue of mice; in addition, the CAT and SOD content in the lung tissue increased significantly, indicating that at this dose, the organism produced a stress antioxidant protection response, protecting its own state by upregulating the expression of antioxidant genes. After CAPE treatment, the CAT and SOD content in the lung tissue decreased significantly. The above results show that CAPE can remove excessive ROS produced by CEES and exert an antioxidant effect.

[0049] The above evaluation indicators are comprehensively analyzed, and statistical methods are used to evaluate the protective effect of CAPE. A feedback mechanism is established to adjust the dosing regimen in a timely manner according to the evaluation results to form a closed-loop optimization process. At the same time, the subjective feelings, clinical symptoms and quality of life evaluation of the subjects are combined to provide a more comprehensive evaluation of the protective effect.

[0050] Based on the previous research results, a detailed clinical application guide was developed, including indications, usage and dosage, precautions, etc., to provide guidance for the clinical application of CAPE in the protection of lung injury caused by erosive agents. At the same time, the application of CAPE was promoted through academic conferences and professional training to improve the medical staff's awareness and acceptance of this protective measure.

[0051] This technical solution aims to build a scientific and effective protection system to provide an innovative solution to this problem by comprehensively and systematically planning the application of CAPE in the protection of lung injury caused by erosive agents. Through the selection of raw materials, optimization of dosing regimens, comprehensive evaluation of effects and continuous research and promotion, we are confident that we can make CAPE a shining pearl in the field of protection against erosive agents and contribute to the health of mankind.

[0052] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the attached claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0053] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A compound for protecting against lung damage caused by erosive toxic agents, characterized in that: The compound is caffeic acid phenethyl ester.

2. The compound according to claim 1, characterized in that The erosive agent is sulfur mustard or its analog 2-chloroethyl ethyl sulfide.

3. The compound according to claim 1, characterized in that It is used to reduce lung tissue damage caused by blistering agents.

4. The compound according to claim 1, characterized in that It reduces the oxidative stress level of lung tissue by inhibiting the generation of reactive oxygen species in lung tissue and reducing the content of lipid oxidation products such as malondialdehyde (MDA).

5. The compound according to claim 1, characterized in that It is administered by intraperitoneal injection, and the recommended dose is 10 mg per kilogram of body weight.

6. The compound according to claim 1, characterized in that When used, caffeic acid phenethyl ester must be administered within a certain period of time after exposure to blister agents and administered repeatedly to maintain its effective concentration in the body.

7. The compound according to claim 6, characterized in that The period of time after exposure to the blister agent specifically refers to administration of caffeic acid phenethyl ester at 1 hour, 25 hours and 49 hours after exposure.

8. A method for protecting against lung damage caused by corrosive toxic agents, characterized in that: The method comprises administering the caffeic acid phenethyl ester compound of any one of claims 1 to 7 to a person who may be exposed to a blistering agent.

9. The method according to claim 8, characterized in that The method also includes monitoring lung function-related parameters, lung tissue morphological changes, antioxidant indicators, and oxidative stress levels of personnel exposed to erosive agents to evaluate the protective effect.

10. The method according to claim 8, characterized in that This approach is intended for persons who may be at risk for lung injury from exposure to blister agents.

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