Pharmaceutical composition for preventing, treating or ameliorating respiratory diseases, comprising agrimonia pilosa extract as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for preventing, treating or ameliorating respiratory diseases, comprising an Agrimonia pilosa extract as an active ingredient. The pharmaceutical composition can alleviate inflammatory responses occurring in lung tissues and reduce allergic responses, and thus can be effectively used for preventing, ameliorating or treating various respiratory diseases, particularly lung diseases caused by house dust mites.

Description

Pharmaceutical composition for preventing, treating or improving respiratory diseases containing dragon's-eye extract as an active ingredient

[0001] The present invention relates to a pharmaceutical composition for preventing, treating or improving respiratory diseases, comprising an extract of dragon's moss as an active ingredient.

[0002] This invention was made possible with the support of the Korean government under the Ministry of SMEs and Startups' Project No. S3282568 in 2024, "Efficacy and mechanism of action of functional raw materials for respiratory health from dragon's eye extract" (Research period: January 1, 2024 - June 30, 2024).

[0003] House dust mites (HDMs) are known to be the leading cause of allergic respiratory diseases, with 79% of patients with allergic rhinitis and asthma being sensitized to them. Allergic rhinitis, which causes chronic cough, nasal congestion, and a runny nose, can also lead to chronic fatigue, decreased exercise capacity, and difficulty concentrating, interfering with daily activities such as work and study. If left untreated, it can lead to other allergic diseases, such as allergic asthma.

[0004] Respiratory diseases caused by house dust mites require long-term rest and medication to improve symptoms and recover, resulting in significant socioeconomic costs. For example, current treatments for allergic rhinitis include nasal steroids, oral steroids, and antihistamines, which are used to reduce inflammation in the nasal mucosa. However, long-term use of steroids can cause many problems, including decreased immunity, endocrine disruption, slowed growth, and weight gain. Antihistamines, on the other hand, only provide temporary symptom relief rather than a fundamental treatment, and their side effects, such as drowsiness, decreased concentration, and constipation, can cause inconvenience in other aspects of daily life.

[0005] Agrimonia pilosa, also known as "Seonhakcho" or "Jipsinnamul," is a plant known for its diverse physiological functions. Its abundant polyphenol content has been studied for its antioxidant, tumor-suppressing, antiviral, and antibacterial activities. Furthermore, administration of Agrimonia pilosa extract to diabetic rats has been shown to improve blood sugar levels. Furthermore, Agrimonia pilosa has been reported to have hemostatic and vasoconstrictive effects. However, there has been no report on whether Agrimonia pilosa can help improve respiratory diseases caused by house dust mites.

[0006] Accordingly, the inventors of the present invention conducted research to improve respiratory diseases using dragon's claw and completed the present invention.

[0007] One object of the present invention is to provide a pharmaceutical composition for preventing or treating respiratory diseases, which contains an extract of dragon's moss as an active ingredient.

[0008] Another object of the present invention is to provide a method for preventing or treating respiratory diseases, comprising administering the composition to a subject.

[0009] Another object of the present invention is to provide a health functional food for preventing or improving respiratory diseases, which contains a dragon's-eye extract as an active ingredient.

[0010] One aspect of the present invention provides a pharmaceutical composition for preventing or treating respiratory diseases, comprising a dragon's-eye extract as an active ingredient.

[0011] According to one specific example of the present invention, the extract may be a hot water extract.

[0012] According to one specific example of the present invention, the respiratory disease may be a lung disease caused by house dust mites.

[0013] According to one specific example of the present invention, the lung disease may be an inflammatory lung disease or an allergic lung disease.

[0014] Another aspect of the present invention provides a method for preventing or treating a respiratory disease comprising administering the composition to a subject.

[0015] Another aspect of the present invention provides a health functional food for preventing or improving respiratory diseases, which contains a dragon's-eye extract as an active ingredient.

[0016] According to a pharmaceutical composition for preventing, treating or improving respiratory diseases containing a dragon's-eye extract as an active ingredient, it can not only alleviate inflammatory reactions occurring in lung tissue but also improve allergic reactions, and thus can be effectively utilized for preventing, improving or treating various respiratory diseases, particularly lung diseases caused by house dust mites.

[0017] Figure 1 is a graph showing the body weight (A) and body weight change (B) by test group according to administration of dragon's-eye extract in an HDM lung disease model.

[0018] Figure 2 is a graph showing the lung tissue weight (A) and the relative weight of lung tissue to body weight (B) by test group according to the administration of dragon's-eye extract in the HDM lung disease model.

[0019] Figure 3 is a photograph (A) showing lung tissue by test group in the HDM lung disease model and a graph (B) showing the results of evaluating lung tissue inflammation using a 5-point score.

[0020] Figure 4 is a graph showing the concentrations of blood ALT (A) and AST (B) in an HDM lung disease model.

[0021] Figure 5 is a graph showing the cytoplasmic staining of immune cells present in bronchoalveolar lavage fluid by test group in the HDM lung disease model (A), the total number of immune cells (B), and the number of immune cells by type (C).

[0022] Figure 6 is a graph showing the relative gene expression levels of TNF-α (A), IL-6 (B), and IL-1β (C), which are inflammation indicators in lung tissue, by test group in the HDM lung disease model.

[0023] Figure 7 is a graph showing the concentration of IgE, an indicator of allergic reaction in the blood, by test group in the HDM lung disease model.

[0024] Figure 8 shows immunoblotting results (A) and a graph (B) comparing the gene expression levels of NF-κB and IκB-α in the lungs of each test group in a DM lung disease model.

[0025] One aspect of the present invention provides a pharmaceutical composition for preventing or treating respiratory diseases, comprising a dragon's-eye extract as an active ingredient.

[0026] The composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the composition of the present invention are those commonly used in the manufacture of pharmaceuticals, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above ingredients, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweetening agents, flavoring agents, emulsifiers, suspending agents, preservatives, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington: the science and practice of pharmacy 22nd edition (2013).

[0027] A pharmaceutical composition according to one embodiment of the present invention may be administered together with a substance that exhibits a preventive or therapeutic effect on one or more respiratory diseases.

[0028] In addition, the pharmaceutical composition according to one embodiment of the present invention may be used alone or in combination with methods using procedures, hormone therapy, drug therapy, and / or biological response modifiers for the prevention or treatment of respiratory diseases.

[0029] The composition of the present invention may include various bases and / or additives necessary and appropriate for the formulation of the dosage form, and may be manufactured by further including known compounds such as nonionic surfactants, silicone polymers, pigments, fragrances, preservatives, bactericides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softeners, antioxidants, free radical scavengers, opacifiers, stabilizers, emollients, silicones, α-hydroxy acids, antifoaming agents, moisturizers, vitamins, insect repellents, fragrances, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, alkalizing or acidifying agents, or colorants, within a range that does not reduce the effectiveness thereof.

[0030] The appropriate dosage of the composition of the present invention may be prescribed in various ways depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. The dosage of the composition of the present invention may be 0.001 to 1000 mg / kg for adults.

[0031] The composition of the present invention can be administered orally.

[0032] The composition of the present invention can be administered in various dosage forms when administered orally, such as tablets, pills, hard / soft capsules, liquids, suspensions, emulsifiers, syrups, granules, elixirs, troches, etc., and may further include various excipients, for example, wetting agents, sweeteners, fragrances, preservatives, etc. Specifically, when the composition of the present invention is formulated into an oral administration dosage form, it may further include appropriate carriers, excipients, and diluents commonly used in the manufacture thereof. Examples of the carrier, excipient and diluent may include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and / or mineral oil. In addition, the formulation may be prepared by including diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants and surfactants commonly used in formulations, and in addition to the excipients, a lubricant such as magnesium stearate or talc may be further included.

[0033] According to one specific example of the present invention, the respiratory disease may be an inflammatory respiratory disease or an allergic respiratory disease.

[0034] According to one specific example of the present invention, the respiratory disease may be asthma, chronic obstructive pulmonary disease, bronchitis, pharyngitis, tonsillitis, laryngitis, sinusitis or pneumonia.

[0035] According to one specific example of the present invention, the allergic respiratory disease may be allergic rhinitis or allergic asthma.

[0036] The dragon's mantle extract of the present invention can not only alleviate inflammatory reactions in the lungs but also improve allergic reactions, and thus can be effectively used in the prevention or treatment of various respiratory diseases, particularly lung diseases caused by house dust mites.

[0037] According to one specific example of the present invention, the extract may be a hot water extract.

[0038] The extract of the dragon's moss included as an active ingredient in the composition according to the present invention can be obtained as follows. The dragon's moss, specifically the dragon's moss leaves, are washed with water to remove foreign substances, dried in the shade, and an appropriate amount of solvent is added to the dragon's moss so that it is completely immersed. At this time, the dragon's moss can be used as is in its dried state or ground and used in powder form. Dragon's claw can be extracted using a conventional extraction solvent, preferably (a) anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, etc.), (b) a mixed solvent of the lower alcohols and water, (c) acetone, (d) ethyl acetate, (e) chloroform, (f) 1,3-butylene glycol, (g) hexane, (h) diethyl ether, (i) butyl acetate (j) chloroform-methanol, or (k) water, and more preferably, it can be extracted with water. The extraction can be performed by soaking or heating at room temperature, and hot water extraction is most preferable.

[0039] According to one specific example of the present invention, the respiratory disease may be a lung disease caused by house dust mites.

[0040] House dust mites are so small that they cannot be easily observed with the naked eye. However, if the dead bodies or excrement of house dust mites enter the respiratory tract, lung diseases such as inflammatory lung disease or allergic lung disease can be induced. For example, in people who are allergic to house dust mites, inflammation and constriction of the airway of the lungs can occur, and various reactions such as an increase in immune cells and / or an increase in IgE in the blood due to an inflammatory reaction in the lung tissue can occur. Since the dragon's-foot-and-mouth disease extract of the present invention is excellent in improving such reactions induced by house dust mites in the lungs, it can be usefully utilized in the prevention or treatment of respiratory diseases caused by house dust mites, particularly inflammatory lung disease or allergic lung disease.

[0041] According to one specific example of the present invention, the lung disease may be an inflammatory lung disease or an allergic lung disease.

[0042] Another aspect of the present invention provides a method for preventing or treating a respiratory disease comprising administering the composition to a subject.

[0043] The pharmaceutical composition comprising the dragon's-eye extract of the present invention may be administered orally in an amount effective for the treatment or prevention of a subject or patient, depending on the intended purpose. It should be understood that the dosage for a specific subject or patient should be determined based on various related factors such as the patient's weight, age, race, sex, health condition, diet, administration time, administration method, and disease severity, and may be appropriately increased or decreased by a specialist. For example, a physician may start the dosage of the pharmaceutical composition of the present invention at a level lower than that required to achieve the intended therapeutic effect, and gradually increase the dosage until the intended effect is achieved, and may easily determine and prescribe the dosage as needed.

[0044] Another aspect of the present invention provides a health functional food for preventing or improving respiratory diseases, which contains a dragon's-eye extract as an active ingredient.

[0045] To avoid excessive complexity due to unnecessary repetition in this specification, description of common items is omitted.

[0046] When the dragon's mantle extract according to one specific example of the present invention is manufactured into a health functional food, it may include ingredients that are typically added during the manufacture of health functional foods, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

[0047] For example, when the health functional food of the present invention is manufactured as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, Eucommia ulmoides extract, jujube extract, and / or licorice extract may be additionally included in addition to the composition of the present invention.

[0048] In addition, the health functional food of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0049] These components may be used independently or in combination, and the proportion of these additives may be selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the health functional food of the present invention, but is not limited thereto.

[0050] The present invention will be described in more detail below through one or more examples. However, these examples are intended to exemplify the present invention and the scope of the present invention is not limited to these examples.

[0051]

[0052] Example 1. Preparation of dragon's claw extract

[0053] The leaves of Agrimonia pilosa were purchased, thoroughly shaken to remove foreign substances, and soaked in purified water 20 times the volume of the raw material for 6 hours at 90±2℃, and extracted with normal hot water under conditions of more than the standard 0.1 Brix. The extract was filtered using a 50μm filter and passed through a magnetic bar to remove metals, impurities, and foreign substances. The extract was concentrated at a steam temperature of 80 to 90℃ (internal temperature 40 to 60℃) under a vacuum of 10 or less until the Brix was more than 19 Brix, and then sterilized at 90±5℃ for more than 30 minutes. For freeze-drying, the concentrate was poured into a freeze-drying tray, frozen at -45 to -25℃ for 20 hours, dried for 52 hours, and freeze-dried for a total of 72 hours. After that, it was ground in a mixer to prepare Agrimonia pilosa freeze-dried powder (APFD).

[0054]

[0055] Example 2. Preparation of a house dust mite lung disease model, composition of test groups, and test items.

[0056] 2-1. Preparation of experimental animals

[0057] Six-week-old female BALB / c mice were provided by the Central Laboratory Animal (Seoul, Korea) and were housed in the experimental animal room of Chonbuk National University Hospital under the following conditions: temperature 23±3℃, relative humidity 55±15%, ventilation rate 10–20 times / hr, lighting time 12 h (lights on 8:00 AM–8:00 PM), and illumination intensity 150–300 LUX. During the rearing period, environmental conditions such as temperature / humidity, ventilation rate, and illumination intensity of the animal room were regularly measured. Food was provided ad libitum by the Central Laboratory Animal (7 Baumoe-ro 7-gil, Seocho-gu, Seoul). Purified water was provided ad libitum from polycarbonate drinking bottles. Wooden bedding was provided by Coretech Co., Ltd. (Gyeonggi-do, Korea). Mice were housed in polycarbonate cages for rodents (W235 × L380 × H175 mm) with a population of 5 or fewer per cage during the acclimatization, administration, and blood collection periods, and cages, bedding, and water bottles were changed at least once a week.

[0058]

[0059] 2-2. Manufacturing of a house dust mite lung disease model

[0060] The mice prepared in Example 2-1 were acclimatized for one week and then infected with house dust mites (HDM) to create a house dust mite lung disease model.

[0061] Specifically, the body weights of the mice were measured and randomly distributed to each group so that the average body weights were distributed as evenly as possible according to the ranked body weights. Then, to prepare the HDM lung disease model, lyophilized house dust mites (NC9756554, GREER Laboratory) were dissolved in saline at a concentration of 2 μg / μL and prepared. After anesthetizing the mice, HDM was administered at a dose of 100 μg / 50 μL per mouse in volume mode (Respiratory rate 80 bpm, Tidal column 0.18 mL) using a small animal ventilator (SAR-1000, CWE Inc.) at one-week intervals for a total of four times. The normal control group was prepared by administering 50 μL of saline using the same equipment.

[0062]

[0063] 2-3. Composition of test group for administration of dragon's-eye extract

[0064] For the HDM lung disease model prepared in Example 2-2, a test group was formed as shown in Table 1, and the extract of the dragon's mantle prepared in Example 1 was suspended in distilled water and administered intragastrically to the administration group every day at 10:00 AM.

[0065]

[0066] Classification Number of animals Group of administered substances Single dose (mpk) Route of administration Number of administrations G115 Normal control group (Ctrl) Distilled water - intragastric administration 1 time / day G215 Negative control group (DW) Distilled water - intragastric administration 1 time / day G315 Low concentration administration group (L) Dragon's moss extract 25 Intragastric administration 1 time / day G415 Medium concentration administration group (M) Dragon's moss extract 50 Intragastric administration 1 time / day G515 High concentration administration group (H) Dragon's moss extract 100 Intragastric administration 1 time / day

[0067]

[0068] 2-4. Observation and autopsy examination items

[0069] During the acclimatization, administration, and observation periods, the type of general symptoms, including death, the date of onset, and the severity of symptoms were observed once a day and recorded for each individual. Body weight was measured for each individual using an electronic scale when separating the groups, once a week during the administration period, and on the day of necropsy, and recorded. Feed intake was measured before the start of administration and once a week after the start of administration. The measurement method was to measure the amount fed on the day after feeding the feed and the remaining amount on the next day, and divide the difference by the number of mice per cage to calculate the average feed intake (g / mouse / day).

[0070] Meanwhile, for autopsy, mice were anesthetized with ketamine, blood was collected, and euthanized. After that, a macroscopic examination was performed on the external surface and all orifices, cranial cavity, thoracic and abdominal cavities and their contents. In addition, after a macroscopic examination was performed on all mice during autopsy, the organ weight was measured by removing the lungs from the mice whose abdomens were cut, washing them with saline solution, and removing moisture with filter paper.

[0071]

[0072] Example 3. Confirmation of the improving effect of dragon's mantle extract on lung disease induced by HDM.

[0073] 3-1. Method of analyzing treatment effects

[0074] 3-1-1. Blood biochemical analysis

[0075] After the end of the test, mice that had been fasted for more than 14 hours were anesthetized with ketamine, and 1 ml of blood was collected through cardiac puncture. The collected blood was centrifuged (3000 rpm, 10 minutes) to obtain serum, which was then transferred to EP tubes and stored at -80℃. Biochemical analysis was performed on triglycerides, total cholesterol, AST (Aspartate aminotransferase), and ALT (Alanine aminotransferase). For this purpose, a triglyceride measuring reagent (Asan Pharmaceutical Co., Ltd., AM-157S-K), a cholesterol measuring reagent (Asan Pharmaceutical Co., Ltd., AM 202-K), a GOT reagent (Asan Pharmaceutical Co., Ltd., AM-103K), and a GPT reagent (Asan Pharmaceutical Co., Ltd., AM-102K) were used. Blood biochemical tests were analyzed according to the guidelines of the Jeonbuk National University Hospital Effectiveness Evaluation Center SOP.

[0076]

[0077] 3-1-2. Lung Tissue Pathology Analysis

[0078] Lung tissue was extracted and fixed, and paraffin blocks were made for histopathological analysis. These were sectioned at 4 μm thickness, stained, and interpreted. Histopathological examinations were performed using hematoxylin & eosin staining (H&E), and lesions and inflammation progression in lung tissue were evaluated by a pathologist based on a 5-point score.

[0079]

[0080] 3-1-3. Measurement of IgE in blood

[0081] Approximately 300 to 400 μL of blood was collected through the orbital vein of mice and centrifuged for 15 minutes to separate plasma. The plasma was collected and stored at -80°C. The plasma was collected the day after the third HDM injection. IgE levels were measured using an ELISA method using the collected plasma.

[0082]

[0083] 3-1-4. Analysis of cells in bronchoalveolar lavage fluid

[0084] The recovered bronchoalveolar lavage fluid was centrifuged and the supernatant was stored at -20°C for cytokine analysis. After lysis of red blood cells in the cell pellet, the number of lymphocytes, eosinophils, neutrophils, and macrophages was measured using a particle counter.

[0085]

[0086] 3-1-5. Cytokine RNA expression analysis

[0087] RNA was isolated from lung tissue using Tripure Isolation Reagent (Roche). 5 μg of total RNA was synthesized into cDNA using a High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). Real-time PCR was performed with 1 μL of the synthesized cDNA, along with 1 μL of Taqman primer, 10 μL of Taqman Universal Master Mix II (Thermo Fisher Scientific), and 8 μL of triple-distilled water. The TaqMan gene used in the quantitative polymerase reaction was ordered from Applied Biosystems. The real-time PCR reaction conditions were 50°C for 2 minutes, 95°C for 10 minutes, and 40 cycles of denaturation at 95°C for 15 seconds and annealing at 60°C for 15 seconds.

[0088]

[0089] 3-1-6. Western blot analysis

[0090] Lung tissues were washed twice with PBS, and proteins were extracted using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo, Cat#: 78835). The extracted proteins were quantified after measuring the concentration using Bradford reagent (Bio-Rad, Hercules, CA, USA). The quantified proteins were mixed with sample buffer, heated at 95°C for 5 minutes, and separated by 10–12% SDS-PAGE. After separation, the proteins were transferred to PVDF membranes using a semi-dry transfer system at 15 V for 60 minutes through SDS-PAGE, reacted with blocking buffer (5% Skim milk in 1X TBST) for more than 1 hour, reacted with primary antibodies overnight at 4°C, and then washed five times with 1X TBST at 7-minute intervals. After reacting with the secondary antibody at room temperature for more than 1 hour, the membrane was washed 5 times with 1X TBST at 7-minute intervals, developed with ECL reagent, and exposed to an X-ray film.

[0091]

[0092] 3-1-7. Analysis and Analysis Program

[0093] For the results of this study, parametric or non-parametric multiple comparison procedures were used. In the case of parametric multiple comparisons, normality of the data was assumed and a parametric one-way ANOVA was used for the test. If the ANOVA results were significant, a post hoc test was performed using Dunnett's multiple comparison test. In the case of non-parametric multiple comparisons, the Kruskal-Wallis' H-test was used for the test. If the results were significant, a post hoc test was performed using Dunn's multiple comparison test. Statistical analysis was performed using Prism 7.04 (GraphPad Software Inc., San Diego, CA, USA), and a p-value less than 0.05 was considered statistically significant.

[0094]

[0095] 3-2. Evaluation of the effects of dragon's-eye extract administration on body weight and lung tissue weight changes in the HDM lung disease model.

[0096] According to Example 2, the body weight was measured and recorded once a week from the time of administering the extract of dragon's moss to the HDM lung disease model, and the change in body weight due to administration was compared and analyzed. The body weight at the time of starting administration was measured and the change in body weight during the test was calculated. As a result, there was no significant difference in the change in body weight among the normal control group, negative control group, and test group, confirming that administration of the dragon's moss extract did not affect the body weight of the mice (Fig. 1).

[0097] Meanwhile, after the test, the mice were sacrificed, dissected, and their lungs were removed. The removed tissues were observed for any unusual features, and their weights were measured to confirm that there were no unusual features in the tissues. Since the weight of the tissues may vary depending on the body weight of the mouse, the weight of the lung tissues was calculated as a percentage of the body weight and compared and analyzed to exclude this. As a result, there was no statistically significant difference between the groups in both the weight of the lung tissues and the relative weight of the tissues to the body weight, confirming that administration of the dragon's-ear extract did not affect the weight of the lung tissues of the mice (Fig. 2).

[0098]

[0099] 3-3. Confirmation of the effect of improving lung tissue inflammation following administration of dragon's-eye extract in the HDM lung disease model.

[0100] The degree of inflammation in the mouse lung tissues extracted in Example 3-2 was compared.

[0101] Specifically, the left lobe was fixed in a formalin solution using the method of Example 3-1-2, paraffin sections were created, and stained using the H&E staining method. The stained lung tissue slides were then referred to a specialized pathologist, and after establishing suitable criteria for testing based on inflammatory cell infiltration, they were interpreted and compared for analysis.

[0102] As a result, it was found that the inflammation score in lung tissue was increased due to HDM-induced lung disease, whereas infiltration of inflammatory cells in tissue was suppressed in the group administered with the extract of Yong-A-Cho for 4 weeks (Fig. 3).

[0103] Through these results, it was confirmed that the extract of dragon's claw can improve inflammation of lung tissue in HDM-induced lung disease.

[0104]

[0105] 3-4. Confirmation of the improvement in blood liver function parameters following administration of dragon's claw extract in the HDM lung disease model.

[0106] Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are enzymes that normally exist primarily in hepatocytes and perform their functions. However, when liver toxicity causes cell death, they are released into the bloodstream. Therefore, blood ALT and AST concentrations are used in the diagnosis of liver toxicity.

[0107] Therefore, the amount of liver enzymes in the blood was quantified using a biochemical analysis technique in an animal model of lung disease induced by HDM. Specifically, blood biochemical analysis was performed using the method of Example 3-1-1 to compare and analyze the serum concentrations of ALT and AST, which are enzymes in the body known to be related to liver toxicity and obesity.

[0108] As a result, it was confirmed that there was no significant difference in the concentrations of blood ALT and AST among all groups (Fig. 4), and it was confirmed that the dragon's mantle extract according to the present invention did not exhibit liver toxicity.

[0109]

[0110] 3-5. Confirmation of the effect of administration of dragon's-eye extract on improving allergic inflammation in the bronchial tubes in the HDM lung disease model.

[0111] HDM-induced lung disease is a type of allergic airway inflammation. Therefore, to assess the improvement of allergic inflammation in the bronchial airways, we compared and analyzed the expression of inflammatory cells in bronchoalveolar lavage fluid (BALF) and inflammatory cytokines in lung tissue.

[0112] Specifically, immune cells present in BALF were counted and compared by dividing them into neutrophils, eosinophils, lymphocytes, or macrophages according to cytoplasmic staining patterns and nuclear morphology using the method of Example 3-1-4.

[0113] As a result, all types of classified immune cells were confirmed to be significantly increased in the disease-induced group compared to the normal control group. On the other hand, the number of neutrophils, eosinophils, and lymphocytes in BALF was confirmed to be decreased in the group administered with the extract of Yong-A-Cho (Fig. 5).

[0114] Through these results, it was confirmed that the extract of Yong-A-Cho could improve allergic inflammation in the bronchi in HDM-induced lung disease.

[0115]

[0116] 3-6. Confirmation of the effect of improving inflammation in lung tissue following administration of dragon's-eye extract in the HDM lung disease model.

[0117] The anti-inflammatory efficacy of the extract of Yong-A-Cho was verified in the HDM lung disease model using the method of Example 3-1-5.

[0118] As a result, it was found that TNF-α, IL-6, and IL-1β increased in the negative control group, whereas TNF-α, IL-6, and IL-1β decreased in a concentration-dependent manner in the group administered with the extract of dragon's ginseng (Fig. 6).

[0119] Through these results, it was confirmed that the extract of Yong-A-Cho could improve inflammation in lung tissue in HDM-induced lung disease.

[0120]

[0121] 3-7. Confirmation of the inhibitory effect of dragon's-eye extract on the increase in blood IgE concentration in the HDM lung disease model.

[0122] Allergic rhinitis is a common upper respiratory allergy. Its prevalence has been steadily increasing recently, driven by environmental factors such as fine dust and air pollution. Allergic rhinitis is well known to be a disease driven by a typical Th2 immune response. When exposed simultaneously to fine dust and allergens, it stimulates the existing Th2 immune response, leading to increased IgE production and eosinophils.

[0123] Therefore, the IgE concentration in the blood of mice was evaluated using the method of Example 3-1-3.

[0124] As a result, it was found that the blood IgE concentration increased in the HDM lung disease model, whereas the increase in the blood IgE concentration was suppressed in the group administered with the dragon's-eye extract (Fig. 7).

[0125] Through these results, it was confirmed that the extract of Yong-A-Cho can alleviate allergic symptoms such as allergic rhinitis in lung diseases induced by HDM.

[0126]

[0127] 3-8. Confirmation of the inhibitory effect of NF-kB expression following administration of dragon's-eye extract in the HDM lung disease model.

[0128] Pathological mechanisms associated with lung disease are known to involve signaling pathways such as NF-kB, MAPK, PTEN / PI3K / AKT, and STAT6. Among these, NF-kB signaling is known to be activated when epithelial cells in the airway are continuously exposed to oxidative stress or allergens. When signaling is activated, NF-kB undergoes phosphorylation and translocates into the nucleus, inducing gene expression of inflammatory mediators. This, in turn, is known to activate inflammatory cells in the alveoli.

[0129] Therefore, biomarkers that are the pathological mechanism in the HDM lung disease model were compared using the Western blot method of Example 3-1-6.

[0130] As a result, it was confirmed that in the normal control group, there was almost no phosphorylated NF-kB in the nucleus of the cells, and in the lung disease model induced by HDM, the amount of phosphorylated p-NF-kB in the nucleus was confirmed to increase. On the other hand, when the extract of Yong-A-Cho was administered after inducing lung disease by HDM, it was confirmed that the amount of phosphorylated NF-kB in the nucleus was decreased (Fig. 8).

[0131] Through these results, it was confirmed that the extract of Yong-A-Cho could show an improvement effect on lung disease induced by HDM by inhibiting NF-kB signaling.

[0132]

[0133] The present invention has been described above, focusing on specific embodiments thereof. Those skilled in the art will appreciate that the present invention can be implemented in modified forms without departing from its essential characteristics. Therefore, the disclosed embodiments should be considered illustrative rather than limiting. The scope of the present invention is set forth in the claims, not the foregoing description, and all differences within the scope equivalent thereto should be construed as being encompassed by the present invention.

Claims

1. A pharmaceutical composition for preventing or treating respiratory diseases, containing an extract of dragon's moss as an active ingredient.

2. A pharmaceutical composition for preventing or treating respiratory diseases, wherein the extract in paragraph 1 is a hot water extract.

3. A pharmaceutical composition for preventing or treating a respiratory disease, wherein the respiratory disease in claim 1 is a lung disease caused by house dust mites.

4. A pharmaceutical composition for preventing or treating respiratory disease, wherein the lung disease in claim 3 is an inflammatory lung disease or an allergic lung disease.

5. A method for preventing or treating a respiratory disease, comprising administering the composition of claim 1 to a subject.

6. Health functional food for preventing or improving respiratory diseases containing dragon's-eye extract as an active ingredient.

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