Composition for preventing, alleviating, or treating respiratory diseases, comprising atractylodes japonica extract

Abstract

The present invention relates to a composition for treating, preventing, or alleviating respiratory diseases, comprising an Atractylodes japonica extract as a natural extract. The composition of the present invention can exhibit mucolytic and expectorant activity and therapeutic effects on respiratory diseases through two or more combined mechanisms selected from the group consisting of antioxidant activity, anti-inflammatory activity, increased production of substance P and acetylcholine (Ach), and reduced expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA.

Description

Composition for preventing, improving or treating respiratory diseases containing an extract of Saposhnikovia japonica

[0001] This application claims priority to Korean Patent Application No. 10-2024-0074521, filed June 7, 2024, the entire disclosure of which is incorporated herein by reference. The present invention relates to a composition for preventing, improving, or treating respiratory diseases, comprising a natural extract of Saposhnikovia japonica.

[0002] Due to the rapid increase in particulate matter (PM) caused by air pollution, various respiratory diseases are emerging as a serious problem in East Asia, including China, Japan, and Korea. A large amount of PM is generated in the form of yellow dust from the Loess Plateau and deserts of Inner Mongolia and northwestern China, mainly in spring. As these PM pass through inland industrial areas of China, they combine with automobile exhaust and coal gas to form more serious forms of PM. When measured on the ground, most PM are particles with a diameter of about 2.5 μm (PM 2.5), mineral dust, organic pollutants including polycyclic aromatic hydrocarbons, and inorganic pollutants such as sulfate, elemental carbon, nitrate, and ammonium. It is already well known that fine dust can cause serious lung damage and secondary heart damage when inhaled, and asthma is the most representative non-colonial inflammatory airway disease caused by air pollution. According to the WHO announcement in August 2017, the prevalence of asthma is known to be nearly 235 million people worldwide. Accumulated fine dust increases oxidative stress on epithelial cells of the airway and lung tissue, causing local tissue damage and inflammatory responses. The initiation of the inflammatory response can be measured relatively easily by increases in pro-inflammatory cytokines and inflammatory mediators. In particular, with regard to the respiratory system, it has been reported that fine dust is deposited in the lower bronchi and lung parenchyma, causing a decrease in lung function, an increase in the incidence of chronic obstructive pulmonary disease (COPD), asthma, and lung cancer, and an increase in symptoms of the lower respiratory tract and respiratory system. A representative mechanism that has been revealed is that fine dust increases the production of reactive oxygen species, thereby inducing an inflammatory response.

[0003] DEXA is a representative synthetic corticosteroid preparation, and is known to have an anti-inflammatory effect that is approximately 20 to 30 times better than hydrocortisone existing in the body, and approximately 4 to 5 times better than prednisolone, another commonly used synthetic corticosteroid preparation.

[0004] As respiratory disorders caused by fine dust have become a problem, the development of drugs to treat or prevent respiratory damage caused by fine dust has been in progress. In particular, as natural product-derived bioactive substances are known to have relatively few side effects and excellent anti-inflammatory and antioxidant activities, PM 2.5 The development of respiratory protective drugs using natural products is actively underway in induced lung injury models.

[0005] Numerous references and citations are provided throughout this specification. The disclosures of these references are incorporated herein by reference in their entirety to further clarify the state of the art and the scope of the present invention.

[0006] The purpose of the present inventors is to provide a composition for preventing, improving or treating respiratory diseases, which contains an extract of Saposhnikovia japonicus as an active ingredient.

[0007] Another object of the present inventors is to provide a method for producing an extract of Saposhnikovia japonicus that exhibits a preventive, ameliorating or therapeutic effect on respiratory diseases.

[0008] Another object of the present inventors is to provide a method for preventing, improving or treating respiratory diseases using a composition comprising an extract of the plant.

[0009] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0010] One aspect of the present invention is to provide a composition for preventing, improving, or treating respiratory diseases, comprising an extract of Saposhnikovia japonica. The respiratory diseases are at least one selected from the group consisting of bronchitis, bronchial damage, respiratory diseases caused by fine dust, pneumonia, and lung damage, and may include chronic bronchitis, acute bronchitis, bronchiectasis, empyema, chronic obstructive pulmonary disease (COPD), asthma, pneumonia, improvement of bronchial conditions (e.g., coughing, phlegm suppression, etc.), respiratory diseases caused by fine dust, etc.

[0011] The composition of the present invention can exhibit mucolytic expectorant activity through two or more complex mechanisms selected from the group consisting of antioxidant activity; anti-inflammatory activity; increased production of substance P and acetylcholine; and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA, and can also exhibit an effect of preventing, improving, or treating respiratory diseases through the complex mechanisms.

[0012] Another embodiment of the present invention can provide an extract of Saposhnikovia japonicus having two or more complex mechanisms selected from the group consisting of antioxidant activity; anti-inflammatory activity; increased production of substance P and acetylcholine (Ach); and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA.

[0013] Another embodiment of the present invention provides a composition for treating, preventing or improving respiratory diseases, comprising 0.4 to 1.2 mg / g of actrylenolide III and 0.2 to 1.5 mg / g of actrylenolide I, each having two or more complex mechanisms selected from the group consisting of antioxidant activity; anti-inflammatory activity; increasing production of substance P and acetylcholine; and decreasing expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA.

[0014] The above-described use or composition may have a weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) of 1:1 to 1:4 (AI:AIII), preferably 1:1 to 1:2, and more preferably, the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the composition of the present invention may be 1:2±0.5 (AI:AIII).

[0015] Another embodiment of the present invention provides a method for treating a respiratory disease by administering a therapeutically effective amount of a sapling extract to a subject in need thereof. The method can treat any one or more respiratory diseases selected from the group consisting of bronchitis, bronchial damage, respiratory diseases caused by fine dust, pneumonia, and lung damage, and can treat respiratory diseases such as chronic bronchitis, acute bronchitis, bronchiectasis, empyema, chronic obstructive pulmonary disease (COPD), asthma, pneumonia, improvement of bronchial conditions (e.g., cough, phlegm suppression, etc.), respiratory diseases caused by fine dust, pneumonia, and lung damage.

[0016] Atractylodes japonica Koidzumi is an herb belonging to the Asteraceae family (Compositae) that is commonly found in Northeast Asia, including the Korean Peninsula. Traditionally, the root has been used for the purposes of tonic, dehumidification, respiratory activation, treatment of eye diseases and colds, and removal of waste products. Up to now, Atractylodes extract has been known to exhibit anti-inflammatory, antioxidant, and anti-obesity effects. Compared to other Atractylodes species (e.g., Atractylodes macrocephala Koidzumi, Atractylodes lancea De Candlle, Atractylodes chinensis Koidzumi), it is more effective in treating, preventing, or improving respiratory diseases, and has a high content of the active ingredients, actylenolide III and actylenolide I.

[0017] The composition of the present invention includes an extract of Saposhnikovia japonicus as an effective ingredient. The term 'extract' used in this specification includes an extraction result obtained by juicing a raw material or treating a raw material with an extraction solvent, or a processed product formulated (e.g., powdered) thereof.

[0018] When the extract used in the composition of the present invention is obtained by treating the raw material with an extraction solvent, various extraction solvents can be used, for example, a polar solvent or a non-polar solvent can be used. Polar solvents include (i) water, (ii) alcohol (preferably, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) DMF (dimethyl-formamide) and (v) DMSO (dimethyl sulfoxide), and non-polar solvents include acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, Chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF can also be used.

[0019] Preferably, the extract used in the present invention is one obtained by extracting using any one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixtures thereof as an extraction solvent.

[0020] In addition, the term 'extract' used in this specification has the meaning commonly used in the art as a crude extract as described above, but in a broad sense, it also includes a fraction obtained by further fractionating an extract. That is, it includes not only an extract obtained by juicing the raw material or using the above-mentioned extraction solvent, but also one obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity), supplementation of a specific component, etc., fractions obtained through various additional purification methods, and products obtained through the addition of a specific component are also included in the extract of the present invention.

[0021] In addition, the extract of the present invention may be obtained by removing the solvent through an additional process, such as filtration, concentration, or drying, or by performing all of filtration, concentration, and drying. Filtration may be performed, for example, using filter paper or a vacuum filter, concentration may be performed using a vacuum concentrator, and drying may be performed using spray drying or freeze drying to obtain a powdered extract.

[0022] In a preferred embodiment, the extract of the present invention may be obtained by first extracting the raw material by grinding it and then extracting it at 85°C±5°C for 6±1 hours using 30 to 50% of the raw material as an extraction solvent in an amount 8 times larger than the raw material, or by mixing the first extract with a second extract obtained by extracting it at 85°C±5°C for 4±1 hours using 30 to 50% of the raw material as an extraction solvent in an amount 7 times larger than the raw material, and then drying the mixture.

[0023] The extract of the present invention may include actrylenolide I (AI) and actrylenolide III (AIII), and in one embodiment, the extract of the present invention may include 0.4 to 1.2 mg / g of actrylenolide III and 0.2 to 1.5 mg / g of actrylenolide I.

[0024] In the composition of the present invention, the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) may be 1:1 to 1:4 (AI:AIII), preferably 1:1 to 1:2, and more preferably, the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the composition of the present invention may be 1:2±0.5 (AI:AIII). The composition of the present invention is characterized in that, by including atactylenolide I and atactylenolide III in the above weight ratio, it exhibits a more remarkable synergistic effect compared to cases where they are included in other weight ratios.

[0025] The above-mentioned extract of the sapling may be included in an amount of 0.1 to 5% by volume based on the total volume of the composition of the present invention, and the composition may exhibit mucolytic expectorant activity through two or more complex mechanisms selected from the group consisting of antioxidant activity, anti-inflammatory activity, increased production of substance P and acetylcholine (Ach), and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA.

[0026] In a preferred embodiment, the respiratory disease may be selected from the group consisting of bronchitis, bronchial damage, respiratory disease due to fine dust, pneumonia and lung damage, and may include chronic bronchitis, acute bronchitis, bronchiectasis, empyema, chronic obstructive pulmonary disease (COPD), asthma, pneumonia, improvement of bronchial condition (e.g., cough, phlegm, etc.), respiratory disease due to fine dust, etc.

[0027] In one embodiment, the composition of the present invention may be a food composition. The food composition of the present invention includes processed forms of all natural materials, such as foods, functional foods, nutritional supplements, health foods, and food additives. The above-mentioned types of food compositions can be prepared in various forms using conventional methods known in the art.

[0028] For example, as a health food, the extract of the present invention itself can be manufactured in the form of tea, juice, and drink and consumed, or can be granulated, encapsulated, or powdered and consumed. In addition, in addition to the extract of the present invention, white peony root, cornelian cherry, scutellaria baicalensis, reishi mushroom, tangerine peel, ginseng root, angelica root, gardenia fruit, astragalus membranaceus, malt, tangerine seed, vitamin C, fructooligosaccharide, stevioside, purified water, maltodextrin, etc. can be further included alone or in mixture within a range that does not inhibit the purpose of the present invention; however, other medicinal ingredients and / or additives that the food composition of the present invention can additionally include are not limited to the above examples.

[0029] For example, the food composition according to the present invention may include water-soluble vitamins such as thiamine (vitamin B1), riboflavin, ascorbic acid, niacin, and vitamin B6; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid; weak acids such as glycolic acid and acetic acid; and amino acids such as eight essential amino acids, threonine, valine, methionine, isoleucine, leucine, phenylalanine, tryptophan, and lysine, as well as aspartic acid, serine, glutamic acid, proline, glycine, alanine, cysteine, tyrosine, histidine, and arginine.

[0030] In another embodiment, the composition of the present invention may be a pharmaceutical composition. The pharmaceutical composition may include the above-described active ingredient and may be formulated into a pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient, or diluent.

[0031] The above “pharmaceutically acceptable” refers to a non-toxic composition that is physiologically acceptable and does not inhibit the action of the active ingredient when administered to humans and does not typically cause allergic reactions such as gastrointestinal upset, dizziness, or similar reactions.

[0032] Examples of the carrier, excipient or diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, the pharmaceutical composition may further include a filler, an anticoagulant, a lubricant, a wetting agent, a fragrance, an emulsifier or a preservative.

[0033] The term "pharmaceutically effective amount" means an amount that produces a response greater than that of a negative control, and preferably an amount sufficient to produce an effect in the prevention and / or treatment of respiratory diseases.

[0034] Additionally, the pharmaceutical composition of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to a mammal. The formulation may be, for example, a formulation selected from the group consisting of gels, pastes, ointments, powders, emulsions, sprays, and aerosols, but is not limited thereto.

[0035] The pharmaceutical composition of the present invention may be administered orally or parenterally, and the present invention is not limited thereto. When oral administration is included, the composition may be prepared by conventional means with pharmaceutically acceptable excipients, and may be formulated in a form suitable for sustained release, controlled release, sustained release, etc. to smoothly achieve the intended purpose. In the case of parenteral administration, various routes may be included, such as transdermal, nasal, intraperitoneal, subcutaneous, intramuscular, and intravenous, and these may also be prepared by conventional means with pharmaceutically acceptable excipients.

[0036] The preferred dosage of the pharmaceutical composition of the present invention may vary depending on the patient's condition, the degree of the disease, the form of the drug, the route of administration, and the period of administration, and therefore the concentration and dosage of the composition may be appropriately selected by a person skilled in the art.

[0037] In addition, the pharmaceutical composition of the present invention can be administered in combination with a known compound having a preventive and / or therapeutic effect on respiratory diseases.

[0038] Additionally, the composition of the present invention may be a composition for animals as well as humans.

[0039] Preventing, ameliorating, or treating respiratory disease involves, for example, preventing the full development of a respiratory disease in an individual at risk for the disease. "Treatment" refers to a therapeutic intervention that improves the signs or symptoms of a disease or pathological condition after they have already developed. The term "amelioration" refers to a reduction in the severity of some or all clinical symptoms of the disease, a delay in the onset of clinical symptoms, or a slow progression of the disease. "Prevention" can be understood as reducing the risk of developing pathological symptoms in an individual who does not exhibit signs of the disease or who exhibits only early signs.

[0040] The selection of a specific effective dosage can be determined by those skilled in the art in consideration of several factors, including the disease to be treated or prevented, related symptoms, the subject's medical history, the subject's physical condition such as age, weight, and / or immune status, the administered composition, and other factors known to those skilled in the art. The dosage to be used in a preparation comprising a composition containing the extract of the present invention may vary depending on the subject's age, weight, sex, dosage form, health condition, and degree of disease, and may be administered once or several times a day at regular intervals at the discretion of a physician or pharmacist. For example, the daily dosage may be 0.001 to 2 g / kg, preferably 0.5 to 500 mg / kg, based on the content of the active ingredient. The above dosage is an example of an average case, and the dosage may be higher or lower depending on individual differences.

[0041]

[0042] Another embodiment of the present invention provides a method for producing an extract of Saposhnikovia japonicus having an excellent effect in treating, preventing or improving respiratory diseases.

[0043] S1) A step of crushing the shovel to prepare the shovel crushed material;

[0044] S2) A step of preparing a first extract, a step of preparing a first extract using ethanol, preferably 7 to 10 times, preferably 8 times, the total weight of the above-mentioned crushed sapling, as an extraction solvent, preferably 30 to 50% (w / w) of ethanol;

[0045] S3) A step of preparing a secondary extract, wherein the secondary extract is prepared using 30 to 50% (w / w) of alcohol as an extraction solvent, preferably 5 to 10 times, more preferably 6 to 9 times, and even more preferably 7 times the total weight of the primary extract;

[0046] S4) A step of concentrating the secondary extract at a concentration temperature of 50 to 70°C, preferably 60 to 65°C, to achieve 15 to 20 brix; and

[0047] S5) A step for manufacturing a dry powder includes a step of spray drying the concentrated secondary extract under conditions of an inlet temperature of 180-200°C, preferably 190°C, and an outlet temperature of 70-90°C, preferably 80°C. For example, spray drying can be performed using a Yamato spray dryer ADL311SA, a Mini spray dryer S-300, etc.

[0048] The step of preparing the first extract of the above step S2) is to extract at 60-100°C, preferably 70-90°C, for 3 hours or more, preferably 4 hours or more, and more preferably at 85°C±5°C for 6±1 hours.

[0049] The step of preparing the secondary extract of the above S3) step is to extract at 60-100°C, preferably 70-90°C, for 2 hours or more, preferably 3 hours or more, and to extract at 85°C±5°C for 4±1 hours.

[0050] The extract of the sapling manufactured by the above manufacturing method has an excellent effect in treating respiratory diseases, and the contents of actrylenolide III and actrylenolide I in the extract of the sapling may be 0.4 to 1.2 mg / g and 0.2 to 1.5 mg / g, respectively.

[0051] Through one embodiment, a method for producing an extract of Saposhnikovia japonica having a content of actrylenolide III and actrylenolide I of 0.4 to 1.2 mg / g and 0.2 to 1.5 mg / g in the extract of Saposhnikovia japonica can be provided.

[0052] In another embodiment, the use of an extract of the plant having a content of actrylenolide III and actrylenolide I of 0.4 to 1.2 mg / g and 0.2 to 1.5 mg / g for treating, preventing, or improving respiratory diseases may be provided.

[0053] Another embodiment of the present invention provides a method for producing an extract of Saposhnikovia japonicus having mucolytic expectorant activity through two or more complex mechanisms selected from the group consisting of antioxidant activity, anti-inflammatory activity, increased production of substance P and acetylcholine, and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA, wherein the extract has an Actrylenolide III content of 0.4 to 1.2 mg / g and an Actrylenolide I content of 0.2 to 1.5 mg / g through two-step extraction.

[0054] All ingredients described in the present invention preferably do not exceed the maximum usage levels stipulated in relevant laws and regulations of Korea, China, the United States, Europe, Japan, etc. That is, preferably, the food or pharmaceutical composition according to the present invention contains the ingredients according to the present invention within the content limits permitted by relevant laws and regulations of each country.

[0055]

[0056] One embodiment of the present invention provides a method for treating a respiratory disease by administering an effective amount of a sapling extract to a subject in need thereof.

[0057] The above method may be such that the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the extract of the present invention may be 1:1 to 1:4 (AI:AIII), preferably 1:1 to 1:2, and more preferably, the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the composition of the present invention may be 1:2±0.5 (AI:AIII).

[0058] Another embodiment of the present invention provides an extract of Saposhnikovia japonicus, or a composition comprising the same, for use in a medicine or health functional food for treating, preventing or improving respiratory diseases.

[0059]

[0060] The composition of the present invention can exhibit mucolytic expectorant activity through two or more complex mechanisms selected from the group consisting of antioxidant activity, anti-inflammatory activity, increased production of substance P and acetylcholine (Ach), and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA, and can also exhibit an effect of preventing, improving, or treating respiratory diseases through the complex mechanism.

[0061] Figure 1 shows a photograph of a shovel extract obtained as reddish brown powders according to one embodiment.

[0062] Figure 2 shows the results of analyzing the Actractylenolide III content in the extract of the plant using the HPLC method.

[0063] Figure 3 shows the results of analyzing the Actractylenolide I content in the extract of the sapling using the HPLC method.

[0064] Figure 4 shows the results of measuring the amount of nitric oxide produced by measuring absorbance.

[0065] Figure 5 shows the results of measuring PGE2 content by measuring absorbance.

[0066] Figure 6 shows the results of determining whether pulmonary edema was suppressed based on the macroscopic autopsy findings of the lungs. A = intact vehicle, B = PM 2.5 control, C=DEXA (0.75mg / kg), D=AJ400 (400mg / kg of injection), E=AJ200 (200mg / kg of injection), F=AJ100 (100mg / kg of injection)

[0067] Figure 7 shows the results of measuring the amount of mucus secretion using Phenol Red O staining.

[0068] Figure 8 shows the results of measuring substance P and ACh contents in lung tissue.

[0069] Figure 9 shows the results of measuring the expression of MUC5AC and MUC5B mRNA related to mucus production in lung tissue.

[0070] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0071]

[0072] Example

[0073] I. Preparation of the extract of the shovel and confirmation of its optimal composition

[0074] 1. Preparation of Atractylodis Rhizoma (root of Atractylodes japonica Koidzumi) extract

[0075] 500 g of Atractylodis Rhizoma (root of Atractylodes japonica Koidzumi) purchased from a herbal medicine store was prepared and ground, and then extracted at 85℃ for 6 hours using 30% (w / w) ethanol (8 times the volume of the original material) as an extraction solvent, followed by filtration to obtain a first extract. Next, 7 times the volume of 30% (w / w) ethanol compared to the first extract was used, extracted at 85℃ for 4 hours, followed by filtration to obtain a second extract. Afterwards, the extract was concentrated at 60-65℃ to 15-20 brix, and spray dried under the conditions of inlet 190℃ and outlet 80℃ to prepare a reddish brown powders of Atractylodis Rhizoma extract (Fig. 1).

[0076]

[0077] 2. Confirmation of the content of effective ingredients in the extract of the shovel

[0078] For the analysis of Atractylenolide III contained in the extract of Saposhnikovia japonica, 1.74 mg of the standard Atractylenolide III was precisely weighed and dissolved in methanol (MeOH) to make exactly 50 mL. 10 mL of this solution was dissolved in methanol and diluted to 25 mL, and this solution was used as the standard solution. Approximately 1500 mg of the original sample was diluted to 50 mL with 50% methanol. Approximately 1000 mg of the powder of Saposhnikovia japonica extract was precisely weighed, suspended in 10 mL of H2O, diluted to 50 mL with methanol, ultrasonically extracted for 30 minutes, and then filtered to use as a sample. The HPLC conditions for the analysis of Atractylenolide III are shown in Table 1 below.

[0079]

[0080] In addition, for the analysis of Atractylenolide I contained in the extract of Saposhnikovia japonica, 3.02 mg of the standard Atractylenolide I was precisely weighed and dissolved in methanol (MeOH) to make exactly 50 mL, and 2 mL of this solution was dissolved in methanol to make 20 mL, which was used as the standard solution. The original sample was approximately 1500 mg and 50 mL was made by adding 50% methanol. Approximately 1000 mg of the powder of Saposhnikovia japonica extract was precisely weighed, suspended in 10 mL of H2O, and then methanol was added to make 50 mL, ultrasonic extraction was performed for 30 minutes, and then filtered to use as a sample. The HPLC conditions for the analysis of Atractylenolide I are shown in Table 2 below.

[0081]

[0082] High speed liquid chromatography (HPLC) analysis was performed under the conditions shown in Tables 1 and 2 above, and the results are shown in Figures 2 (Actractylenolide III) and 3 (Actractylenolide I), respectively.

[0083] As a result of the experiment, it was confirmed that the extract of the sapling manufactured in the above 1 contains actrylenolide III in a content of 0.4 to 1.2 mg / g and actrylenolide I in a content of 0.2 to 1.5 mg / g.

[0084]

[0085] 3. Efficacy evaluation according to the content ratio of active ingredients

[0086] In order to evaluate the efficacy of the optimal mixing ratio of atactylenolide I and atactylenolide III, which are the active ingredients of the extract of Saposhnikovia japonica, the anti-inflammatory activity and NO production inhibition ability and PGE2 production inhibition ability for each single treatment and their combined ratio were confirmed as follows.

[0087]

[0088] a. Cell culture

[0089] The mouse-derived macrophage cell line (Raw264.7) was obtained from ATCC (Rockville, MD, USA). Raw264.7 cells were cultured in DMEM medium containing 10% FBS, 50 units / mL penicillin, and 50 μg / mL streptomycin. All cells were cultured at 37°C in 5% CO2.

[0090]

[0091] B. Measurement of nitric oxide production

[0092] Raw264.7 cells were treated with 0.1 mg / mL dexamethasone or 200 μg / mL of atractylenolides I and III, which were identified as the active ingredients or marker components of Sapju. In addition, atractylenolides I and III were mixed in ratios of 4:1, 2:1, 1:1, 1:2, and 4:1, and each was added at the same concentration of 200 μg / mL for treatment. After 1 hour of treatment, 1 μg / mL of LPS was treated for 18 hours, and the culture medium was recovered. An equal volume of Griess reagent was added, and the absorbance was measured at 540 nm using an automated microplate reader to measure the amount of nitric oxide produced. The results are shown in Fig. 4.

[0093] The experimental results showed that, compared to the single treatment groups of atactylenolides I and III, the complex treatment group of the two components showed a superior NO production reduction effect in all weight ratios, confirming the synergistic effect of the combination of the two components. In particular, the complex treatment group in which atactylenolides I (AI) and atactylenolides III (AIII) were mixed in weight ratios of 1:1, 1:2, and 1:4 (AI:AIII) showed a significantly superior NO production reduction effect in cells compared to the cases in which they were included in other weight ratios, and the complex treatment group mixed in a weight ratio of 1:2 (AI:AIII) was confirmed to show the best effect (Fig. 4).

[0094]

[0095] D. Measurement of PGE2 content

[0096] The PGE2 content in the cell culture medium was measured using the Prostaglandin E2 Parameter Assay Kit. Briefly, a 96-well plate coated with goat anti-mouse PGE2 antibody was reacted with cell culture medium and primary antibody, and then additionally reacted with horseradish peroxidase-conjugated PGE2. The 96 wells were washed, reacted with 3,3',5,5'-tetramethylbenzidine (TMB) to develop color, and the reaction was stopped with sulfuric acid solution. The absorbance of the reaction solution was measured at a wavelength of 450 nm using an automated microplate reader, and the results are shown in Figure 5.

[0097] The experimental results showed that, compared to the single treatment groups of atractylenolides I and III, the complex treatment group of the two components showed a superior PGE2 inhibition effect in all weight ratios, confirming the synergistic effect of the combination of the two components. In particular, the complex treatment group in which atractylenolides I and atractylenolides III were mixed in weight ratios of 1:1, 1:2, and 1:4 (AI:AIII) showed a significantly superior PGE2 inhibition effect in cells compared to the cases in which they were included in other weight ratios, and the complex treatment group mixed in a weight ratio of 1:2 (AI:AIII) was confirmed to show the best anti-inflammatory activity through PGE2 inhibition (Fig. 5).

[0098] Thus, further in vivo efficacy evaluation was performed using the 1:2 complex (a composition including the extract of Saposhnikovia japonicus adjusted so that AI and AIII are included in a weight ratio of 1:2 (AI:AIII)) (hereinafter referred to as AJ), which was confirmed to have the most remarkable effect in oxidative stress inhibition activity and anti-inflammatory activity.

[0099]

[0100] II. In vivo efficacy evaluation

[0101] Experimental animals and experimental methods

[0102] A total of 88 SPF / VAF Inbred Balb / cAnNCrlOri male mice (OrientBio, Seungnam, Korea) were acclimatized for 7 days and then PM 2.5 Body weight measured 1 day before the first intranasal injection and administration of the experimental substance (normal medium control group average 20.80±0.79 g, 19.40 to 21.80 g and PM 2.5 Based on the average lung damage-induced experimental group of 20.83±1.23 g, 18.60 ~ 23.80 g), 10 animals per group were selected and used in the experiment. This animal experiment was conducted with prior approval from the Animal Experiment Ethics Committee of Daegu Haany University (Approval No. DHU2022-015, February 22, 2022). All experimental animals fasted for 18 hours before the start of experimental substance administration and the final necropsy (drinking was not restricted at this time).

[0103] Military separation (6 groups total; 10 animals per group):

[0104] Normal medium control group: Oral administration of sterile distilled water and intranasal injection of 0.1 ml / kg of normal saline solution

[0105] - PM 2.5 Control group: Oral administration of sterile distilled water and PM 2.5 1 mg / kg intranasal injection lung injury vehicle control

[0106] - Oral administration of DEXA 0.75 mg / kg (11.40 mg / kg as DEXA-water soluble) and PM 2.5 1 mg / kg intranasal injection control drug group

[0107] - AJ 400 mg / kg oral administration and PM 2.5 1 mg / kg intranasal injection high-dose experimental drug group

[0108] - AJ 200 mg / kg oral administration and PM 2.5 1 mg / kg intranasal injection intermediate dose experimental drug group

[0109] - AJ 100 mg / kg oral administration and PM 2.5 1 mg / kg intranasal injection low-dose experimental drug group

[0110]

[0111] Causes of lung damage:

[0112] PM 2.5 The test substance was suspended in saline at a concentration of 10 mg / ml and administered intranasally twice at a dose of 0.1 ml / kg (1 mg / kg) at 48-hour intervals (Day 0 and Day 2) 1 hour before oral administration to induce subacute lung injury. In the normal medium control group, PM was administered to apply the same correction and administration stress. 2.5 Instead of the suspension, the same volume of medium (saline solution) was injected intranasally using the same method. PM in the suspension 2.5 To prevent excessive aggregation of molecules, sonication was performed using an ultrasonicator (Model 5210, Branson, St. Louise, MO, USA) for 30 minutes before each intranasal injection.

[0113]

[0114] Administration of experimental substances:

[0115] The experimental material AJ was dissolved in sterile distilled water at concentrations of 40, 20, and 10 mg / ml, and then administered orally by force once daily for 10 days at a dose of 10 ml / kg (400, 200, and 100 mg / kg), using a metal sonde attached to a 1 ml syringe. DEXA-water soluble (Sigma-Aldrich, St. Louis, MO, USA) was also dissolved in sterile distilled water at a concentration of 1.14 mg / ml (0.075 mg / ml as DEXA itself), and administered orally by force once daily for 10 days at a dose of 10 ml / kg (11.40 mg / kg, 0.75 mg / kg as DEXA itself). Normal medium and PM 2.5In the control group, in order to apply the same correction and administration stress, the same volume of medium, sterile distilled water, was administered orally in the same manner instead of the test substance or DEXA.

[0116]

[0117] Experimental results

[0118] 1. Macroscopic autopsy findings and changes in lung weight

[0119] PM 2.5 In the control group, lung enlargement with marked local congestion was observed, and a significant (p<0.01) increase in the area of ​​visual congestion and absolute and relative lung weights was observed compared to the normal medium control group. On the other hand, PM 2.5 Compared to the control group, a significant (p<0.01) decrease in the lung macrocongested area, absolute and relative weight was observed in a dose-dependent manner in all three AJ 400, 200 and 100 mg / kg groups, respectively. However, all three AJ groups showed a relatively lower PM compared to the DEXA 0.75 mg / kg group. 2.5 Inhibitory activity against induced pulmonary congestion, enlargement, and increase in absolute and relative weight was recognized (Fig. 6).

[0120] The area of ​​visible congestion in the lungs is PM 2.5 In the control group, a change of 2471.35% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -86.27, -76.32, -60.03, and -40.16%, respectively. The absolute lung weight was PM 2.5 In the control group, a change of 47.17% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5Compared to the control group, the changes were -31.16, -19.71, -15.30, and -11.78%, respectively. The relative lung weight was PM 2.5 In the control group, a change of 44.70% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, changes of -22.23, -19.88, -14.70, and -12.26% were observed, respectively.

[0121]

[0122] 2. Changes in cytokine levels in lung tissue - TNF-α, IL-6, CXCL1, and CXCL2

[0123] PM 2.5 In the control group, a significant (p<0.01) increase in the contents of cytokines - TNF-α, IL-6, CXCL1, and CXCL2 - in lung tissue was observed compared to the normal medium control group. Meanwhile, PM 2.5 Compared to the control group, a significant (p<0.01 or p<0.05) decrease in the contents of TNF-α, IL-6, CXCL1, and CXCL2 in lung tissue was observed in a dose-dependent manner in all three doses of AJ 400, 200, and 100 mg / kg groups, respectively. However, all three doses of AJ groups showed relatively lower PM compared to the DEXA 0.75 mg / kg group. 2.5 The inhibitory activity on the increase in the contents of TNF-α, IL-6, CXCL1, and CXCL2 in induced lung tissue was recognized (Table 3: Activity of the injection to improve lung inflammation).

[0124]

[0125]

[0126] TNF-α content in lung tissue PM 2.5 In the control group, a change of 659.00% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively.2.5 Compared to the control group, the changes were -69.17, -60.59, -47.96, and -36.71%, respectively. IL-6 content in lung tissue was higher in PM 2.5 In the control group, a change of 1242.14% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -82.11, -66.59, -50.00, and -35.41%, respectively. CXCL1 content in lung tissue was higher in PM 2.5 In the control group, a change of 912.47% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -69.36, -59.24, -50.73, and -40.13%, respectively. CXCL2 content in lung tissue was higher in PM 2.5 In the control group, a change of 1012.01% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -69.07, -59.91, -46.11, and -31.90%, respectively.

[0127]

[0128] 3. Changes in lipid peroxidation and antioxidant defense systems in lung tissue

[0129] PM 2.5 In the control group, a significant increase (p<0.01) in lipid peroxidation (MDA content) and ROS content in lung tissue was observed compared to the normal medium control group, along with a decrease in GSH content, SOD, and CAT activity. Meanwhile, PM 2.5Compared to the control group, a significant decrease (p<0.01 or p<0.05) in lipid peroxidation and ROS content in lung tissue, and an increase in GSH content, SOD and CAT activity were observed in all three doses of AJ 400, 200 and 100 mg / kg administration groups in a dose-dependent manner, but all three doses of AJ administration groups showed relatively low antioxidant activity compared to the DEXA 0.75 mg / kg administration group. 2.5 It was shown in mice with induced lung injury (Table 4: Antioxidant activity of the sap).

[0130]

[0131]

[0132] Lipid peroxidation in lung tissue PM 2.5 In the control group, a change of 230.65% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -51.07, -41.36, -34.42, and -27.23%, respectively. The ROS content in lung tissue was PM 2.5 In the control group, a change of 395.30% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were -69.04, -50.18, -38.85, and -24.55%, respectively. GSH content in lung tissue was higher in PM 2.5 In the control group, a change of -86.90% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were 176.15, 129.40, 87.98, and 55.93%, respectively. SOD activity in lung tissue was higher in PM 2.5In the control group, a change of -78.31% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were 167.08, 122.08, 91.81, and 53.89%, respectively. CAT activity in lung tissue was higher in PM 2.5 In the control group, a change of -87.34% was observed compared to the normal medium control group, but in the DEXA 0.75 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were 261.62, 184.85, 139.39, and 77.78%, respectively.

[0133]

[0134] 4. Changes in organ pH-sensitive phenol red secretion

[0135] As a positive control group, AX (ambroxol hydrochloride) 250 mg / kg was used. Ambroxol is a drug that promotes mucus secretion, thins sputum, and helps with sputum discharge.

[0136] PM 2.5 In the control group, a significant (p<0.01) increase in OD value and phenol red secretion of tracheal washing fluid was observed compared to the normal medium control group, and PM 2.5 Compared to the control group, a significant (p<0.01) increase in tracheal phenol red secretion was observed in a dose-dependent manner in all three dose groups of AJ 400, 200, and 100 mg / kg, and in particular, AJ 400 mg / kg showed an increase in OD value and phenol red secretion of tracheal lavage fluid comparable to that of AX 250 mg / kg. 2.5 It was shown in mice with induced subacute lung injury (Fig. 7).

[0137] OD value of tracheal washing fluid, phenol red secretion amount PM 2.5In the control group, a change of 31.83% was observed compared to the normal medium control group, and in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, changes of 30.50, 23.80, 28.32, and 15.51% were observed, respectively.

[0138] In this way, the administration of ambroxol and sap extract as positive control drugs showed a significant increase in PM as measured by Phenol Red O staining. 2.5 It was confirmed that mucus secretion was increased more significantly than in the administration group. This can be considered to be due to the action of AJ as an expectorant, which promotes mucus secretion, removes clumped sputum, and acts as a lubricant against airway irritants.

[0139]

[0140] 5. Changes in substance P and ACh content in lung tissue

[0141] Intestinal secretion in the respiratory system is mainly regulated by ACh and substance P. Stimulation / excitation of the vagus nerve is known to induce local secretion of ACh and result in large amounts of intestinal secretion, and substance P is also a representative intestinal secretion-promoting factor in the respiratory system.

[0142] PM 2.5 In the control group, a significant increase (p<0.01) in substance P and ACh contents in lung tissue was observed compared to the normal medium control group. PM 2.5 Compared to the control group, a significant increase (p<0.01) in the substance P and ACh contents in the lung tissue was observed in a dose-dependent manner in all three doses of AJ 400, 200, and 100 mg / kg, respectively. In particular, AJ 400 mg / kg showed an increase in the substance P and ACh contents in the lung tissue comparable to AX 250 mg / kg. 2.5 It was shown in mice with induced subacute lung injury (Fig. 8).

[0143] The content of substance P in lung tissue is PM 2.5 In the control group, a change of 124.55% was observed compared to the normal medium control group, and in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were 137.62, 143.31, 84.10, and 59.81%, respectively. The ACh content in lung tissue was PM 2.5 In the control group, a change of 116.24% was observed compared to the normal medium control group, and in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5 Compared to the control group, changes of 114.84, 117.71, 68.81, and 43.40% were observed, respectively.

[0144] In this way, as part of the inflammatory response due to the invasion of external pollutants, the production of viscous mucus increased due to the increase in the content of substance P and Ach, which promote serous production, and the increase in the expression of MUC5AC and MUC5B, but the expectorant ambroxol and sphagnum mosquitoes increased the production of PM. 2.5 It was confirmed that the secretion of intestinal fluid was promoted by increasing the content of substance P and Ach, which are involved in more intestinal fluid production.

[0145]

[0146] 6. Changes in MUC5AC and MUC5B mRNA expression related to mucus production in lung tissue

[0147] When an infection (fine dust, bacteria, etc.) occurs in the respiratory tract, the respiratory system secretes mucus on the surface to provide a first line of defense and expel infectious agents that have invaded from the outside. When an external infectious agent (fine dust, etc.) invades, inflammation is induced, and the expression of proteins such as MUC5AC and MUC5B, which are involved in the synthesis of viscous mucin components, increases, thereby increasing mucin synthesis, resulting in the production of sticky, viscous phlegm.

[0148] PM 2.5 In the control group, a significant increase (p<0.01) in the expression of MUC5AC and MUC5B mRNA related to mucus production in lung tissue was observed compared to the normal medium control group, but in PM 2.5 Compared to the control group, a significant (p<0.01) decrease in the expression of MUC5AC and MUC5B mRNA in lung tissue was observed in all three dose groups of AJ 400, 200, and 100 mg / kg in a dose-dependent manner, and in particular, AJ 400 mg / kg showed comparable PM to AX 250 mg / kg. 2.5 Each showed inhibitory activity on the increase in MUC5AC and MUC5B mRNA expression related to mucus production in induced lung tissue (Fig. 9).

[0149] MUC5AC mRNA expression in lung tissue is higher in PM 2.5 In the control group, a change of 477.29% was observed compared to the normal medium control group, but in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5 Compared to the control group, the changes were -48.96, -52.94, -38.41, and -29.94%, respectively. MUC5B mRNA expression in lung tissue was higher in PM 2.5 In the control group, a change of 189.01% was observed compared to the normal medium control group, but in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5Compared to the control group, the changes were -36.61, -37.19, -28.03, and -21.05%, respectively.

[0150] In this way, it was confirmed that the expression of mucin-producing proteins with viscosity was suppressed by AJ administration, similar to the positive control drug, ambroxol (Fig. 9).

[0151] In summary of the above, FIGS. 8 and 9, it was confirmed that the positive control drug and AJ secrete a lot of mucus (FIG. 8), but block the synthesis of highly viscous mucin (FIG. 9), and have expectorant activity by thinning the mucus through increased secretion of thin intestinal fluid, etc.

[0152]

[0153] 7. Histopathological changes in the lung PM 2.5

[0154] In the control group, significant sarcomatous lesions - thickening of the alveolar septum due to inflammatory cell infiltration, thickening of the secondary bronchial mucosa, and increase in PAS-positive mucus-producing cells were observed. Compared to the normal medium control group, a significant (p<0.01) increase in the mean alveolar septum and secondary bronchial mucosa thickness, number of peri-alveolar infiltrating inflammatory cells, and number of PAS-positive mucus-producing cells in the secondary bronchial mucosa, and a decrease in ASA related to these were recognized, respectively. However, compared to the PM2.5 control group, a significant (p<0.01) increase in ASA, decrease in the mean alveolar septum thickness, and number of peri-alveolar infiltrating inflammatory cells were recognized in a dose-dependent manner in all three doses of AJ 400, 200, and 100 mg / kg, respectively, and in particular, AJ 400 mg / kg showed comparable PM2.5 levels to AX 250 mg / kg. 2.5 It showed inhibitory activity on induced alveolar septal hypertrophy and inflammatory cell infiltration and the reduction of ASA associated with it. In addition, as part of expectorant activity, PM 2.5Compared to the control group, a significant increase in the average thickness of the secondary bronchial mucosa and the number of PAS-positive mucus-producing cells was also observed in a dose-dependent manner in all three AJ 400, 200, and 100 mg / kg groups, and in particular, AJ 400 mg / kg showed a comparable increase in the thickness of the secondary bronchial mucosa and the number of PAS-positive mucus-producing cells to AX 250 mg / kg. 2.5 It was shown in mice with induced subacute lung injury (Table 5: Respiratory improvement activity of the sap (ASA, Alveolar septal thickness, SB thickness)).

[0155]

[0156] ASA is an indirect indicator of the gas exchange capacity of the lungs, and a decrease in ASA indicates a decline in lung function due to a decrease in the gas exchange surface area of ​​the lungs. PM 2.5 A decrease in ASA was confirmed due to inflammation, and ASA recovery was confirmed by administration of positive control drugs and sap injection, confirming that lung function was protected. In addition, it was confirmed that changes in the thickness of the alveolar septum were induced by changes due to inflammation, which resulted in a decrease in ASA, and this was significantly improved.

[0157] The average ASA of lung tissue is PM 2.5 In the control group, a change of -47.85% was observed compared to the normal medium control group, but in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5 Compared to the control group, the changes were 64.31, 65.75, 55.30, and 45.47%, respectively. The mean alveolar septal thickness was PM 2.5 In the control group, a change of 709.96% was observed compared to the normal medium control group, but in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5Compared to the control group, the changes were -59.93, -61.09, -47.81, and -36.40%, respectively. The average thickness of the secondary bronchial mucosa was PM 2.5 In the control group, a change of 34.19% was observed compared to the normal medium control group, and in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, the changes were 41.48, 45.20, 32.06, and 18.44%, respectively. The average number of peri-alveolar infiltrating inflammatory cells was PM 2.5 In the control group, a change of 756.75% was observed compared to the normal medium control group, but in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM 2.5 Compared to the control group, the changes were -63.34, -64.71, -53.22, and -38.49%, respectively. The average number of PAS-positive mucus-producing cells in the secondary bronchial mucosa was PM 2.5 In the control group, a change of 71.25% was observed compared to the normal medium control group, and in the AX 250 mg / kg, AJ 400, 200 and 100 mg / kg administration groups, PM was observed, respectively. 2.5 Compared to the control group, changes of 99.03, 101.95, 63.50, and 37.71% were observed, respectively.

[0158] Thus, it was confirmed that the extract of the sapling protected lung function through the recovery of ASA and significantly improved the thickness changes of the alveolar septum and the decrease in ASA due to inflammation.

[0159]

[0160] The extract of the sapling has a restorative effect on lung function and has therapeutic and improvement effects on respiratory diseases, including those of the lungs and bronchial tubes. Therefore, it can be used as a composition or pharmaceutical product for the treatment, prevention, or improvement of respiratory diseases.

Claims

1. A composition for preventing, improving or treating respiratory diseases containing an extract of Saposhnikovia japonica.

2. A composition according to claim 1, characterized in that the extract is extracted using any one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixtures thereof as an extraction solvent.

3. In the first paragraph, the extract is a composition characterized in that the first extraction is performed at 85°C±5°C for 6±1 hours using 30 to 50% of the raw material weight as an extraction solvent after crushing the raw material, or the first extract is mixed with a second extract obtained by extracting at 85°C±5°C for 4±1 hours using 30 to 50% of the raw material weight as an extraction solvent, and the first extract is dried.

4. A composition according to claim 1, characterized in that the respiratory disease is selected from the group consisting of chronic bronchitis, acute bronchitis, bronchiectasis, empyema, chronic obstructive pulmonary disease (COPD), asthma, pneumonia, cough and phlegm suppression, respiratory disease caused by fine dust, pneumonia, and lung damage.

5. In the first paragraph, the composition is characterized in that it exhibits mucolytic expectorant activity through two or more complex mechanisms selected from the group consisting of antioxidant activity, anti-inflammatory activity, increased production of substance P and acetylcholine (Ach), and decreased expression of MUC5AC (Mucin-5AC) and MUC5B (Mucin-5B) mRNA.

6. A composition according to claim 1, characterized in that the extract of the sapling contains 0.4 to 1.2 mg / g of actrylenolide III and 0.2 to 1.5 mg / g of actrylenolide I.

7. A composition according to claim 6, characterized in that the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the composition is 1:1 to 1:4 (AI:AIII).

8. A composition according to claim 6, characterized in that the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the composition is 1:2±0.5 (AI:AIII).

9. A composition according to any one of claims 1 to 8, characterized in that the composition is a food composition for preventing or improving respiratory diseases.

10. A composition according to any one of claims 1 to 8, characterized in that the composition is a pharmaceutical composition.

11. Health functional food containing the composition according to Article 9.

12. A composition according to any one of claims 1 to 8, characterized in that the composition is a pharmaceutical composition for animals.

13. A method for manufacturing an extract of the shovel, S1) A step of crushing the shovel to prepare the shovel crushed material; S2) A step for preparing a first extract, a step for preparing a first extract using 7 to 10 times the total weight of the above crushed shovel as an extraction solvent; S3) A step for preparing a secondary extract, wherein the secondary extract is prepared using 5 to 10 times the total weight of the primary extract as an extraction solvent; and S4) A method for producing an extract of the sapling, comprising a step of concentrating the secondary extract at a concentration temperature of 50 to 70°C and concentrating it to 15 to 20 brix.

14. In paragraph 13, The above step S2) is a method for producing an extract of Saposhnikovia japonica, characterized in that the first extract is prepared by extracting at 85°C±5°C for 6±1 hours using 30 to 50% (w / w) of distilled alcohol as an extraction solvent.

15. In paragraph 13, The above S3) step is a method for producing an extract of Saposhnikovia japonica, characterized in that a secondary extract is prepared by extracting at 85°C±5°C for 6±1 hours using 30 to 50% (w / w) of distilled alcohol as an extraction solvent.

16. A method for producing an extract of Saposhnikovia japonica, characterized in that the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the extract of Saposhnikovia japonica is 1:1 to 1:4 (AI:AIII), in any one of the 13th to 15th clauses.

17. A method for producing an extract of Saposhnikovia japonicus, wherein the method is for producing an extract of Saposhnikovia japonicus for treating, preventing or improving respiratory diseases, in any one of the 13th to 15th clauses.

18. A method for treating respiratory diseases by administering an effective amount of an extract of the plant to an individual in need thereof.

19. In the 18th paragraph, the method is a method in which the weight ratio of atactylenolide I (AI) to atactylenolide III (AIII) in the extract of the plant is 1:1 to 1:4 (AI:AIII).

20. A composition comprising an extract of Saposhnikovia japonica for use in a medicine or health functional food for treating, preventing or improving respiratory diseases.

Images