KR20220078140A - Composition for preventing or improving respiratory disease - Google Patents

Abstract

The present invention relates to a composition for preventing and improving respiratory diseases, golden extract 15 to 25% by weight, gold extract 5 to 15% by weight, oleracea extract 15 to 25% by weight, Sukhumia extract 15 to 25% by weight, Schisandra extract 15 to It is technically characterized to contain 25% by weight and 5 to 15% by weight of cinnamon extract, and has excellent anti-inflammatory effects as well as improvement of respiratory diseases without side effects, and has the advantage of improving immunity.

Description

Composition for preventing or improving respiratory disease

The present invention relates to a composition for preventing and improving respiratory diseases, and more particularly, to a composition for preventing and improving respiratory diseases that has no side effects and has excellent anti-inflammatory effects as well as an improvement effect of respiratory diseases.

The respiratory system of our body is one of the organs that constantly defends against physical and chemical stimuli. When irritants are introduced into the respiratory tract, most irritants are caught in the nose hairs and cannot enter the respiratory tract, but fine irritants passing through are surrounded by mucus, It is expelled upward or out by the movement of microscopic cilia located on the wall, and this is the basic principle of coughing.

As described above, coughing is a defense mechanism that occurs reflexively by irritation of the airway mucosa, and when continuous coughing due to excessive stimulation is induced, the patient's quality of life is reduced and deteriorated. Also, according to the same principle as coughing, when dust or irritants are introduced from the outside, the bronchi of our body exercise muscle together with saliva to push them out. Thick purulent sputum is produced.

As a result, cough and sputum are generated by physical and chemical factors such as cold air, foreign substances including pathogenic microorganisms, air pollutants, and allergens, which can be broadly divided into three categories.

First, if the cough receptors in the larynx, trachea, bronchus, pharynx, sinuses, and diaphragm are stimulated due to physicochemical factors, the stimulus is transmitted to the cough center in the medulla oblongata of the brain and the cough reflex occurs. Mechanisms of antitussives closely related to these irritating cough reflexes are roughly divided into central and peripheral. There are dextromethorphan), noscapine, ephedrine, and zipeprol. These ingredients often cause side effects such as cardiovascular thrombosis, myocardial infarction, respiratory depression, sedation, and gastrointestinal disturbances. there is Peripheral antitussives act on stretch receptors located in the periphery to control cough, and have the advantage of being safe by compensating for the shortcomings of relatively central antitussives. Representative drugs include levopropizine, benzonatate, benzozaine, and benzyl alcohol.

Second, when the parasympathetic nervous system is activated due to physicochemical factors, bronchial smooth muscle contracts, and thus symptoms such as bronchospasm or bronchospasm may appear. At this time, beta 2 agonist (β2-adrenergic receptor agonist) stimulates beta 2 receptor to relax smooth muscle, and xanthine derivative increases cAMP by inhibition of phosphodiesterase or bronchial smooth muscle It acts to directly relax the mucus, dissolves mucus while increasing mucus secretion, and promotes ciliary movement to show antitussive and expectorant activity. Drugs exhibiting this action include beta 2 agonists clenbuterol, bambuterol, formoterol, and the like, and xanthine derivatives, theophylline, aminophylline ( aminophylline), acepifylline, and bamifylline.

In addition, there are mucolytic agents or expectorants that dissolve the produced mucus to reduce the viscosity and promote sputum discharge by promoting ciliary movement of the bronchi. Airway mucus has several important functions in the airways. That is, it protects the airways from particles entering the airways from the outside, maintains the humidity of the inhaled air, and binds the inhaled chemicals or gases to the mucus protein and removes them by ciliary action. In addition, mucus contains substances that play an important role in immune function, such as immunoglobulin A (IgA), lysozyme, lactoferrin, and the like. Most expectorants have been used as folk medicines or empirical prescriptions for thousands of years using plants, and as compounds, cysteine derivatives such as carbocysteine, N-acetylcysteine, and cysteine ethyl ester hydrochloride are expectorants. is being used as The above-described cysteine derivatives are not completely satisfactory when used as an expectorant, and in particular, N-acetyl cysteine and cysteine ethyl ester hydrochloride are both highly toxic and chemically unstable. Therefore, it has been required to develop an expectorant with good physical and chemical stability, fewer side effects, toxicity, and excellent expectorant effect.

Third, inflammatory mediators are released from mast cells due to physicochemical factors. Mast cells contain small cytoplasmic granules that contain inflammatory mediators such as histamine and cytokines. When exposed to an allergen, an antibody called IgE is produced in B cells in the body, which attaches to the surface of mast cells. After that, when exposed to allergens again, chemicals including histamine, a strong inflammatory mediator, are released from mast cells to which IgE is attached, thereby increasing secretions in the airways, exudates caused by inflammation, inhaled foreign substances, and bacteria. leads to Existing drugs such as codeine, which are widely used as antitussives, have excellent sedative effects on breathing and cough, but have the side effect of releasing histamine by inducing degranulation of mast cells.

As described above, existing antitussive expectorants have problems due to side effects, toxicity, and chemical instability of the drug itself. In order to solve this problem, research on antitussive expectorants derived from natural products is being actively conducted in recent years. A representative study related to this includes stimulating the vagus nerve of the airway to promote secretion of the mucous membrane to express expectorant action and at the same time, antitussive action. There are reports on plants containing saponins, alkaloids, etc. that play an auxiliary role, but the results are still insignificant.

Republic of Korea Patent Publication No. 10-1662222 (2016.10.04.) discloses a composition for improving respiratory diseases using herbal medicines.

The composition for improving respiratory diseases using the herbal medicine has the advantage of being effective in the prevention of respiratory diseases without side effects, but has a disadvantage in that the anti-inflammatory effect is insufficient.

KR 10-1662222 B1 2016.10.04.

It is an object of the present invention to provide a composition for preventing and improving respiratory diseases, which has excellent anti-inflammatory effects as well as an improvement effect on respiratory diseases without side effects.

Another object of the present invention is to provide a composition for preventing and improving respiratory diseases, which can improve immunity.

In order to achieve the above object, the present invention provides the following means.

The present invention is, golden extract 15-25% by weight, gold and silver extract 5-15% by weight, oleracea extract 15-25% by weight, Sukhumia extract 15-25% by weight, Schisandra extract 15-25% by weight, and cinnamon extract 5-15% by weight %, which provides a composition for preventing and improving respiratory diseases.

The golden extract is obtained by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of gold and extracting at 75 to 85° C. for 3 to 4 hours, and the gold and silver extract is 70% (v/v) to 1 part by weight of gold and silver coins. 20 parts by weight of ethanol is added and extracted for 3 to 4 hours at 75-85° C., and the extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of A. oleander and extracting at 75-85° C. for 3 to 4 hours. and 20 parts by weight of 70% (v/v) ethanol is added to 1 part by weight of Sukhumia extract, and extracted at 75-85° C. for 3 to 4 hours, and the Schisandra extract is 70% (v / v) 20 parts by weight of ethanol is added and extracted at 75-85° C. for 3-4 hours, and the cinnamon extract is obtained by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of cinnamon and adding 20 parts by weight of 70% (v/v) ethanol at 75-85° C. for 3-4 hours extract while

15 to 25% by weight of the golden extract, 5 to 15% by weight of the ginseng extract, 15 to 25% by weight of the oleander extract, 15 to 25% by weight of the sagebrush extract, 15 to 25% by weight of the Schisandra extract and 5 to 15% by weight of the cinnamon extract 1 to 5 parts by weight of an immune enhancer is additionally included in 100 parts by weight of a mixture, wherein the immunity enhancer contains 30% by weight of red ginseng concentrate, 30% by weight of Angelica asiatica extract, 20% by weight of chlorella and 20% by weight of leaf extract, the red ginseng concentrate 300-400 parts by weight of citrus juice is added to 100 parts by weight of silver red ginseng, heated at 80-85°C for 20-24 hours, centrifuged, filtered, and concentrated under reduced pressure, After extraction using an ultrasonic extractor in a water bath 3 times each hour, the leaflet extract is filtered after adding 500 parts by weight of 100% methanol to 100 parts by weight of the leaflets, immersing them at room temperature for 12 hours.

15 to 25% by weight of the golden extract, 5 to 15% by weight of the ginseng extract, 15 to 25% by weight of the oleander extract, 15 to 25% by weight of the sagebrush extract, 15 to 25% by weight of the Schisandra extract and 5 to 15% by weight of the cinnamon extract 0.1 to 1 part by weight of a taste enhancer is additionally included in 100 parts by weight of a mixture, wherein the taste enhancer contains 50% by weight of Goji berry extract, 35% by weight of maekmundong extract, 10% by weight of stevia extract and 5% by weight of fructooligosaccharide, Goji berry extract is added 300-400 parts by weight of water to 100 parts by weight of Goji berry, heated at 95-105° C. for 1-2 hours, and filtered, and the maekmun-dong extract is 300-400 parts by weight of water added to 100 parts by weight of maekmun-dong and 105-110 After heating at ℃ for 5-7 hours and filtering, the stevia extract is filtered after extraction by adding 600-700 parts by weight of purified water to 100 parts by weight of stevia and heating at 95-100 ℃ for 3-4 hours.

The composition for preventing and improving respiratory diseases according to the present invention has the advantage of having excellent anti-inflammatory effects as well as improving effects of respiratory diseases without side effects.

In addition, the composition for preventing and improving respiratory diseases of the present invention has the advantage of improving immunity.

1 is a graph showing the NO production inhibitory effect and cytotoxicity test results of the composition for preventing and improving respiratory diseases prepared in Example 1. FIG.
2 is a graph showing the cytokine inhibitory effect of the composition for preventing and improving respiratory diseases prepared in Example 1.
3 is a graph showing the mitogen-induced spleen cell proliferation of the composition for preventing and improving respiratory diseases prepared in Example 1. FIG.
4 is a graph showing the activation of NK cells of the composition for preventing and improving respiratory diseases prepared in Example 1.
5 is a diagram illustrating a bronchial asthma Mouse model.
6 is a graph showing the effect of the composition for preventing and improving respiratory diseases prepared in Example 1 on changes in airway hypersensitivity in OVA-induced bronchial asthma.
7 is a graph showing the effect of the composition for preventing and improving respiratory diseases prepared in Example 1 on the histology of lung tissue in OVA-induced bronchial asthma.

Hereinafter, the present invention will be described in detail.

First, a composition for preventing and improving respiratory diseases according to the present invention will be described.

The composition for preventing and improving respiratory diseases of the present invention,

Contains 15 to 25% by weight of golden extract, 5 to 15% by weight of ginseng extract, 15 to 25% by weight of oleracea extract, 15 to 25% by weight of Sulfuricaceae extract, 15 to 25% by weight of Schisandra extract and 5 to 15% by weight of cinnamon extract. .

The golden extract is preferably extracted for 3 to 4 hours at 75 ~ 85 ℃ by adding 20 parts by weight of 70% (v / v) ethanol to 1 part by weight of gold.

The gold (herbal medicine name: Scutellaria Root, scientific name: Scutellaria baicalensis Georgi) is used as a herbal medicine by drying the root from which the bark of the plant has been removed. Gold has the effect of cooling heat, drying moisture, and detoxifying by burning heat. In addition, gold is known to have the effect of hemostasis and stabilizing the fetus of pregnant women. Gold is a fever caused by moisture, a symptom of vomiting and sickness due to hot and warm chest congestion, a chest full and stuffy due to humid heat, while not sick, jaundice, a symptom of coughing due to fever in the lungs, body heat It is known to be used to treat symptoms of feeling hot and thirsty in the chest, symptoms of nosebleeds due to high blood temperature, boils and swelling, symptoms of unstable fetal movement during pregnancy, uterine bleeding, and hematuria.

It is preferable to extract 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of gold and silver coins, and extract the extract at 75 to 85° C. for 3 to 4 hours.

The gold and silver flower (Lonicerae Flos) is a flower bud or just started blooming of the honeysuckle ( Lonicera japonica Thunberg ) of the Caprifoliaceae family. In honeysuckle, a stem or a stem to which leaves are attached is called honeysuckle. In Korea, it grows well on roadsides, at the foot of mountains, and near streams all over the country. In June or July, the white flowers later turn into yellow flowers, so they are called gold and silver flowers. In early summer, the flower buds form before they bloom, and they are collected and dried in the shade for use. Flower buds are a mixture of small rod-shaped buds and lip-shaped flowers, and are about 15-35 mm long, with the upper part being about 3 mm in diameter and the lower part being about 1.5 mm in diameter. The outer surface is yellowish white or greenish white, and the longer it is stored, the darker the color. If you look through a magnifying glass, light brown hairs are dense, and the calyx is green and the tip is divided into 5 parts. There are 5 stamens, yellow, and 1 pistil, and there is no hair in the ovary. The taste is cigarette, bitter, and has a characteristic odour. The main active substances of gold and silver coins include phenolic compounds with various functional groups, iridolis, and flavonoids. According to the study on physiologically active substances, substances with excellent physiological activity and content among the contained natural substances include chlorogenic acid, 3,5-dicaffeoylquinic acid among phenolic compounds, swerosids and (E)-aldosecologanin in iridolis, and Among flavonoids, luteolin-7-O-glucoside has been reported. Gold and silver coins have the weakness of removing heat from the body and eliminating the action of toxic substances, and are widely used in oriental medicine for boils, syphilis, gonorrhea, hemorrhoids, and vasculitis. Pharmacologically, relatively strong anti-inflammatory action has been reported on Shigella, Typhoid, Paratyphoid, Escherichia coli, Pertussis, Diphtheria, Mycobacterium tuberculosis, Staphylococcus, Hemolytic Streptococcus, etc. It has been used as a neutralizing drug.

It is preferable that 20 parts by weight of 70% (v/v) ethanol is added to 1 part by weight of the oleracea extract, and extracted at 75-85° C. for 3-4 hours.

The sagebrush is also called sagebrush and the scientific name is Acanthopanax sessiliflorus , which is a deciduous shrub belonging to the Araliaceae family taxonomically, and the root bark of the genus is used medicinally. Acanthoside B and D (eleutheroside E) of the lignan family contained in sagebrush are effective in tonicity, carbohydrate and fat metabolism, alcohol detoxification, appetite enhancement, blood circulation, heat retention, improvement of constipation, and fatty liver. Acantosides A and C are effective in bone marrow regulation, white blood cell increase (immunity, resistance improvement), allergic disease improvement, and rapid recovery after cancer treatment.

It is preferable to extract 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of Sukhumus extract and extract at 75-85° C. for 3 to 4 hours.

The Sukjihwang is a processed root of Rehmannia glutinosa Liboschitz var. purpurea Makino or R. glutinosa Liboschitz. is known Symptoms such as back pain, dizziness, dizziness, tinnitus, hearing loss, periodic fever, and night sweats appear according to the nutritional deficiency of the liver and heart. Sukjihwang is known to replenish worms and fill bones, and is known to relieve cough, wheezing, and asthma caused by nutritional deficiency of the kidneys.

It is preferable to extract the Schisandra extract by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of Omija and extracting it at 75 to 85° C. for 3 to 4 hours.

The Schisandra chinensis is a fruit of the Schisandra tree, which is a creeping plant belonging to the Magnolia family. It is called Schisandra because it has five tastes, such as acidity of the skin, sweetness of the flesh, spicy and bitterness of the seeds, and saltiness of the whole. Schisandra is favorably used for food and medicinal purposes, and the main constituents of Schisandra include about 30 types of lignans with a dibenzo[a,c]cyclooctadiene skeleton. Active substances include schizandrin, gomisin, deoxyschizandrin, schizandrol, and the like. As the pharmacological action of Schisandra, antioxidant, antibacterial, antithrombotic, anti-obesity, anti-inflammatory, anticancer, and hepatoprotective effects have been reported.

The cinnamon extract is preferably extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of cinnamon and extracting at 75-85° C. for 3-4 hours.

The cinnamon ( Cinnamomum verum ) is mainly produced in southern China and northern Vietnam. The shape is semi-tubular, semi-regular, or a tubular shell that is rolled to one side, mostly in the form of pieces, and the size varies. The outer surface is dark reddish-brown, and the inner surface is flat, but it is also reddish-brown and easy to fracture. It has a peculiar smell and the taste is slightly sweet and very sweet at first, and then it becomes mucous and astringent. Cinam aldehyde is the main ingredient, and in addition, there are cinnamyl acetate, phenylpropyl acetate, cinnamic acid, salicylaldehyde, etc., and synzeilanol exists as diterpenoids. In addition, there are campen, cineol, and eugenol as essential oils. Cinnamon has been widely used in oriental medicine and folk remedies because of its various effects and scents, and has been used for a long time as a material for tea, beverages, or herbal medicines. Known pharmacological actions include blood pressure enhancement, peripheral vasodilation, and blood circulation. In oriental medicine, it is used for colds and pain relief.

The composition for preventing and improving respiratory diseases according to the present invention has the advantage of having excellent anti-inflammatory effects as well as improving effects of respiratory diseases without side effects.

In addition, the composition for preventing and improving respiratory diseases of the present invention has the advantage of improving immunity.

The composition for preventing and improving respiratory diseases according to the present invention

Golden extract 15~25% by weight, gold ginseng extract 5~15% by weight, oleracea extract 15~25% by weight, Sukhumus extract 15~25% by weight, Schisandra extract 15~25% by weight, and cinnamon extract 5~15% by weight are mixed. 1 to 5 parts by weight of an immune enhancer may be additionally included in 100 parts by weight of the mixture.

The composition for preventing and improving respiratory diseases according to the present invention can further enhance immunity by additionally including an immune enhancing agent.

The immunity enhancer includes 30% by weight of red ginseng concentrate, 30% by weight of Angelica asiatica extract, 20% by weight of chlorella and 20% by weight of leaf extract.

The red ginseng (Panax ginseng C.A. Meyer) is a dried ginseng that is steamed and dried without peeling it after selecting 4 to 6 year old fresh ginseng, a perennial grass belonging to the Orchidaceae family. In oriental medicine, it has been used as a representative herbal medicine that has the effect of promoting health and preventing diseases, such as slightly warm in nature, slightly sweet in taste, and slightly bitter in nature, and enters the rain, lungs, and heart to replenish energy and make extracts. The active ingredients of red ginseng are more concentrated in the process of drying after steaming, and physiologically active ingredients effective for the human body such as saponins are produced, which improves digestion and absorption, and helps to enhance the immune system and strengthen functions.

The red ginseng concentrate can be prepared by adding 300 to 400 parts by weight of citrus juice to 100 parts by weight of red ginseng, heating it at 80 to 85° C. for 20 to 24 hours, and then centrifuging, filtration and concentration under reduced pressure.

The Angelica gigas (Angelica gigas NAKAI), Angelica sinensis DIELS, Angelica acutiloba (A. acutiloba KITAG.), which are perennial herbs belonging to the Umbelliferae, and dried roots of related plants As that, it is known that it has the effects of bohyeolhwahyeol, sciatica, and lubricating organs. The root has sedative, antispasmodic, and blood pressure lowering effects. When an aqueous solution of root essential oil is used for arthritis, it is said to have analgesic and anti-inflammatory effects. Used for crumbs, etc.

The Angelica asiatica extract can be prepared by filtration after extracting Angelica quince with 85% methanol using an ultrasonic extractor on a water bath 3 times for 1 hour each.

The chlorella refers to a living organism that survives in various ecosystems such as freshwater, seawater, air, and soil, and has chlorophyll to obtain energy necessary for photosynthesis from sunlight and to obtain a carbon source from inorganic substances such as carbon dioxide to grow independently. Chlorella has no flagella and has no motility, and the cell consists of a single nucleus and cup-shaped chloroplasts, and the grown cells proliferate rapidly through specific cell division, and usually 4 daughter cells form endospores, Pyrenoidosa), Bulgaris, Ellipsoidea, etc. Currently, about 10 species are known. Chlorella not only contains all 40 nutrients including the 5 major nutrients, but also has excellent photosynthetic ability, so it contains 10 times more chlorophyll than general vegetables, and contains a large amount of minerals and vitamins that are lacking in modern people. It balances the body's nutrition and has excellent effects on strengthening immunity, excreting heavy metals, and inhibiting the absorption of dioxins. In addition, as an alkaline food, chlorella has a useful function of maintaining the balance of ion balance in the body by changing the acidic constitution caused by excessive consumption of meat or grains to an alkaline constitution.

The leaflet (Perilla Leaf) is a leaf of Perilla frutescens var. acuta KUDO. as an annual herbaceous plant belonging to the family Lamiaceae.

The leaflet extract can be prepared by adding 500 parts by weight of 100% methanol to 100 parts by weight of the leaflets, immersing them at room temperature for 12 hours, and then filtering.

The composition for preventing and improving respiratory diseases according to the present invention

Golden extract 15~25% by weight, gold ginseng extract 5~15% by weight, oleracea extract 15~25% by weight, Sukhumus extract 15~25% by weight, Schisandra extract 15~25% by weight, and cinnamon extract 5~15% by weight are mixed. 0.1 to 1 part by weight of a taste enhancer may be additionally included in 100 parts by weight of the mixture.

The composition for preventing and improving respiratory diseases according to the present invention has the advantage of facilitating ingestion by additionally including a taste enhancer.

The taste enhancer includes 50% by weight of Goji berry extract, 35% by weight of maekmundong extract, 10% by weight of stevia extract, and 5% by weight of fructooligosaccharide.

The goji berries have traditionally been known for their efficacy in preventing diseases (liver, kidney, blood pressure improvement, diabetes, etc.) and restoring energy (dizziness, neurological diseases, etc.), and the main active ingredients include betaine and zeaxanthin. ), β-sitosterol, physalien, and choline, among which betaine and zeaxanthin are known as the most representative components. Betaine is water-soluble and is present in tea or hot water using water as a solvent, and zeaxanthin is fat-soluble and is a major functional ingredient extracted in an organic solvent such as ethanol. The clinical efficacy of Goji berry can be found in various herbal literatures such as Bonchogangmok and Donguibogam, and it is known that it is mainly effective for adult diseases such as hypertension, headache, paralysis, neurological diseases, diabetes, etc. has been reported

The Goji berry extract can be prepared by adding 300 to 400 parts by weight of water to 100 parts by weight of Goji berry, heating at 95 to 105° C. for 1 to 2 hours, and then filtering.

The maekmundong has been used by adding it as a part of herbal medicines rather than being used alone as medicinal substances. Representative examples of these include saengmaeksan, ongyeongtang, and licorice broth. The main pharmacological components of maekmundong include steroid-based saponins opiopogenine A, B, methylophiopogonone A, B, opiopogonanone A, methylophiopoginanone A Homo-isoflavonoids such as , B, and other polysaccharides such as β-sitosterol, stigmasterol, β-sitosterol glucoside, and oligosaccharides are contained in the tuber root by more than 60%. Paekmundong tuberous root has anticancer activity, and its main component is saponin having the structure of spicatoside A and B. In addition, when the intake of maekmundong extract is increased in rats fed a high-cholesterol diet, lipid excretion is increased and lipids are decreased, and the concentration of corticosterone in the blood of mice induced by starvation stress is decreased, thereby reducing starvation stress.

The maekmundong extract can be prepared by adding 300 to 400 parts by weight of water to 100 parts by weight of maekmundong, heating at 105 to 110° C. for 5 to 7 hours, and then filtering.

The stevia is a perennial plant belonging to the family Asteraceae. It grows wild in the mountainous areas of the borders of Paraguay, Argentina, and Brazil in South America. The leaf is lanceolate, has fine sawtooth, is curved, has no petiole, and is attached to the stem in opposite directions. The stem grows about 70-100 cm, and each node has a leaf, and a branch sprouts from the leaf axil. During the winter, the stem loses its function, and a new branch grows from the hidden eye close to the root to form a new stem every year. In the root, the development of the original root is not clear and there are many side roots and membrane roots, and thick roots develop at the end of growth to have a storage function. Flowers bloom in clusters of 5-6 common flowers in one cluster, showing self-incompatibility. As it is a single plant, flowering is promoted under single conditions. Stevia leaves contain 6-7% of stevioside compared to the weight of the whole plant as a major sweetening component, and the content varies greatly depending on the individual. The stevioside is 250 times sweeter than sugar, and as a non-carcinogenic or calorie-free sugar substitute sweetener, it is used in seafood, pickled vegetables, desserts, beverages and confectionery, etc., and is used in adult diseases such as obesity, diabetes, heart disease, hypertension, and hyperlipidemia and as a hypoglycemic sweetener for caries patients.

The stevia extract is obtained by adding 600 to 700 parts by weight of purified water to 100 parts by weight of stevia, extracting by heating at 95 to 100° C. for 3 to 4 hours, and then filtration.

The fructooligosaccharide has a sweetening quality very similar to that of sugar, has a sweetness level of 60% of that of sugar, and has health functional properties, so it is attracting attention as a sugar substitute. Fructooligosaccharide refers to a mixture of components in which 1 to 3 fructose are bonded to sugar using a fructose transferase using sugar as a substrate. Fructooligosaccharide has the effect of intestinal function that helps the growth of beneficial bacteria in the intestine and suppresses harmful bacteria, and helps to facilitate bowel movements.

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

A composition for the prevention and improvement of respiratory diseases was prepared by mixing 20% by weight of golden extract, 10% by weight of gold extract, 20% by weight of oleracea extract, 20% by weight of saccharin extract, 20% by weight of Schisandra extract and 10% by weight of cinnamon extract.

After drying the gold extract, 20ℓ of 70% (v/v) is added to 1 kg of the dried gold, extracted 3 times at 80° C. for 3 hours, filtered to remove impurities, and then freeze dry (PVTFD20R, Ilshin Lab (Note) ), Korea) was obtained by freeze-drying.

After drying the gold and silver coins, 20 liters of 70% (v/v) was added to 1 kg of the dried gold and silver coins, extracted three times at 80 ° C. for 3 hours, filtered to remove impurities, and then freeze dry (PVTFD20R, Ilshin Lab (Co.) ), Korea) was obtained by freeze-drying.

After drying the Orchis spp., 20ℓ of 70% (v/v) was added to 1 kg of the dried sageum, extracted three times at 80° C. for 3 hours, filtered to remove impurities, and then Freeze dry (PVTFD20R, Ilshin Lab (Note) ), Korea) was obtained by freeze-drying.

After drying the extract, 70% (v/v) 20ℓ was added to 1 kg of dried sukhumus, extracted three times at 80° C. for 3 hours, filtered to remove impurities, and then freeze dry (PVTFD20R, Ilshin Lab (Note) ), Korea) was obtained by freeze-drying.

After drying the Schisandra extract, 70% (v/v) 20ℓ of the dried Schisandra 1 kg was added, extracted 3 times at 80° C. for 3 hours, filtered to remove impurities, and then freeze dry (PVTFD20R, Ilshin Lab (Note) ), Korea) was obtained by freeze-drying.

After drying the cinnamon extract, 20 liters of 70% (v/v) was added to 1 kg of the dried cinnamon, extracted three times at 80 ° C. for 3 hours, filtered to remove impurities, and then Freeze dry (PVTFD20R, Ilshin Lab (Note) ), Korea) was obtained by freeze-drying.

[Experimental Example 1]

(1) Cell culture and differentiation induction

The RAW 264.7 cell line used in the experiment was purchased from the Korea Cell Line Bank (KCLB, Seoul, Korea) and used. Dulbecco's modified Eagel's medium, DMEM (Thermo, Waltham, MA, USA) was added with 10% FBS (fetal bovine serum) and 1% penicilin/streptomycin. Incubated and maintained at 37° C., 5% CO ₂ conditions.

(2) Cytotoxicity test

MTS assay was performed to determine whether the composition for preventing and improving respiratory diseases prepared in Example 1 is toxic to macrophages. RAW 264.7 was dispensed in a 96-well plate at 5×10 ³ cells/well, and DMEM containing 10% NBCS was added to make a final volume of 100 μL per well. After culturing for 24 hours, the extracts were treated at each concentration (0, 100, 200, 300, 400, 500 μg/ml) and incubated for 48 hours. After that, 1/10 times the volume of each cell culture solution was added to the MTS lysate, and after incubation at 37°C for 2 hours, the absorbance was measured at 490 nm using an ELISA microplate reader (Infinite 200 pro, TECAN, Austria).

(3) NO (nitric oxide) concentration measurement

In order to measure the NO production inhibitory effect of the composition for preventing and improving respiratory diseases prepared in Example 1 in RAW 264.7 cells activated with LPS, several concentrations were pre-treated and then LPS (500 ng/ml) was treated for 24 hours. did Then, it was performed according to the manufacturer's protocol using the griess reagent system (Promega, Fitchburg, WI, USA). The RAW 264.7 cell line was aliquoted to a concentration of 2×10 ⁵ cells/ml in a 96 well plate, and incubated for 24 hours at 37°C and 5% incubator, and the extract was applied at a concentration of 100, 200, 300, 400 and 500 μg/ml Treated for 24 hours. After that, the cell culture medium and griess reagent were mixed in a 1:1 ratio and reacted for 10 minutes, and then absorbance was measured at 540 nm using a microplate reader Infinite® 200 PRO (TECAN, Mannedorf, Switzerland), as shown in FIG. .

The results obtained in this experiment were expressed as mean ± standard deviation (mean ± S.D.), and the SPSS (18.0, Statistical Package for Social Science Inc., Chicago, IL, USA) statistical package program was used to test the significance between the experimental groups. Therefore, one-way ANOVA was performed for animal experiments. In case of significance, Duncan's multiple range test was performed at the level of p<0.05.

According to FIG. 1, it can be seen that the composition for preventing and improving respiratory diseases prepared in Example 1 is excellent in the inhibitory effect of NO (nitric oxide) production.

NO is a free radical with a small molecular weight, and plays an important role in the human body, such as controlling nerve transmission and blood pressure. In addition, since it has cytotoxicity, it is known as a mediator involved in cytotoxicity in the host defense mechanism when an inflammatory response occurs. Therefore, it can be seen that the inhibition of NO can suppress the inflammatory response, and the composition for preventing and improving respiratory diseases prepared in Example 1 has anti-inflammatory activity.

[Experimental Example 2]

ELISA assay for TNF-α, IL-6, IL-1β was performed to find out how the composition for preventing and improving respiratory diseases prepared in Example 1, which had a high NO production inhibitory effect, affects the expression of inflammatory mediators. proceeded. The extracts were pre-treated at a concentration of 50, 100, 250 and 500 μg/ml, respectively, and then LPS was treated at a concentration of 500 ng/ml for 24 hours.

The secretion of TNF-α and interleukin-6 in the cell culture was measured using an ELISA kit (BD Bioscience, San Diego, CA, USA). Each of the capture antibodies (anti-mouse TNF-α and interleukin-6) was dispensed into a 96-well plate and coated at 4°C overnight. This was washed with washing buffer (phosphate-buffered saline (PBS) containing 0.05% Tween-20), blocked with PBS containing 10% FBS for 1 hour, and washed with washing buffer. In each microplate, 100 μL of the supernatant of the same cell culture medium as in which nitric oxide (II) was measured was dispensed and reacted for 2 hours at room temperature, followed by washing with washing buffer. Each diluted biotinylated anti-mouse TNF-α and interleukin-6 detection antibody and streptavidin-horseradish peroxidase conjugate solution were added and reacted at room temperature for 1 hour. After washing the reacted plate with washing buffer, tetramethylbenzidine (TMB) substrate solution was added to develop color for 30 minutes in the dark, 1M phosphoric acid was added to stop the color reaction, and absorbance was measured at 450 nm, as shown in FIG. 2 .

According to Figure 2, the concentration of TNF-α was increased to a concentration of 979.06 ± 3.98 pg / ㎖ in the LPS-only treatment group compared to the untreated group, but when 500 μg / ㎖ of OJ-E70 was treated 650.17 ± 3.25 pg / ml. (A)

In the IL-6 ELISA assay, in the LPS-only group, IL-6 production was increased to 1990.38 ± 10.79 pg/ml, but decreased to a concentration of 1804.13 ± 27.46 pg/ml when 500 μg/ml of OJ-E70 was treated ( B)

Next, as a result of confirming the production of IL-1β, the production amount of the LPS-only group increased to 288.28 ± 14.84 pg/ml, but when 500 μg/ml of AY-E70 was treated, it was reduced to a concentration of 220.38 ± 3.67 pg/ml. .(C)

Taken together, the decrease in IL-6 and IL-1β increased by 500 ng/ml LPS treatment was insignificant, but the production of TNF-α was reduced when AY-E70 at a concentration of 500 μg/ml was treated. was reduced visibly.

[Experimental Example 3]

After cervical dislocation of the experimental animal, the spleen was removed aseptically, and then lightly crushed with a sterile glass rod to release it. The separated cell suspension was passed through a 200 mesh stainless steel sieve in RPMI 1640 medium (Hyclone, Logan, UT, USA) to release the cells. The separated cell suspension was passed through 100 μm cell strainers (BD Falcon, Palo Alto, CA, USA), washed twice with culture medium, centrifuged at 3,000 rpm for 10 minutes, and red blood cell lysing buffer (Hybri-MaxTM, Sigma). -Aldrich) for 5 minutes to remove red blood cells. Splenocytes from which red blood cells were removed were again dispersed in RPMI 1640 culture medium, stained with trypan blue solution, and the number of cells was measured using a hemocytometer. It was dispensed in a 24-well plate at 3.0×106 cells/mL. Con A (concanavalin A, 7.5 μg/mL) and lipopolysaccharide (30 μg/mL) were added as mitogens to each group, and incubated at 37° C., 5% CO ₂ for 72 hours in an incubator. Cell proliferation ability was measured by CCK-8 assay. Thus, it is shown in FIG. 3 .

In the case of non-activated lymphocytes, cell proliferation occurs at the stage of activation by various stimuli such as various antigens, mitogens, and cytokines, and the degree of activation can be easily confirmed by the mitogen-stimulated response. Therefore, it is widely used as a method to search for new immunomodulators. To determine whether OJ administration affects the proliferation effect of splenocytes, the degree of lymphocyte proliferation was measured using Con A, a mitogen for T cells, and lipopolysaccharide, a mitogen for B cells, in splenocytes isolated from the spleen. One result was as shown in Fig. 3A.

The T lymphocyte proliferation rate was significantly increased in all groups administered with the positive control group (PC) and the composition for preventing and improving respiratory diseases (GHW) prepared in Example 1, compared to the normal control group (CON), and the proliferative ability of B lymphocytes Also, in all groups administered at 100 mg/kg and 200 mg/kg of the composition for preventing and improving respiratory diseases (GHW) prepared in Example 1, the activity was significantly higher than that of the normal control group and the positive control group (Fig. 3 B).

These results show that administration of the composition for prevention and improvement of respiratory diseases prepared in Example 1 has mitogen activity to proliferate splenocytes, and the number of immune cells that induce immune responses to antigens when exposed to external antigens It can be confirmed that effective defense against external infection can be induced by increasing it.

[Experimental Example 4]

For the cytotoxicity of natural killer cells, a method was used to measure lactate dehydrogenase (LDH) released from tumor cells that are destroyed by natural killer cells, Yac-1 cells. Yac-1 (targetcell) cells were adjusted to be 1×10 ⁴ cells/well in a 96-well U-bottom culture plate, and then cultured with isolated splenocytes (effector cells). The effector-to-target cell ratio was 10: The number of cells was adjusted to be 1, 5:1, or 2.5:1. The amount of lactate dehydrogenase released from target cells by the killing ability of natural killer cells after incubation for 4 hours in an incubator at 37°C, 5% CO ₂ was measured using EZ-LDH Cell Cytotoxicity Assay Kit (DoGen, Seoul, Korea). was measured and shown in FIG. 4 .

Natural killer cells are mainly present in the blood and are the cells responsible for intrinsic immunity, and are very important for the initial immune defense of the body in cancer or viral infections. It has cellular immune activity that kills non-self cells such as infected cells or cancer cells, so when it comes into contact with a target substance, it is activated and destroys the target cell. It also prevents pathogens such as viruses from entering the body and entering host cells for replication, or selectively finds and removes infiltrating cells.

Activation of natural killer cells by the composition for preventing and improving respiratory diseases prepared in Example 1 was performed by culturing Yac-1 cells, known to be sensitive to natural killer cells and splenocytes isolated from the mouse spleen, together. -1 It was confirmed by measuring the degree of apoptosis of cells.

In the splenocytes of mice orally administered the composition for preventing and improving respiratory diseases prepared in Example 1 at a concentration of 200 mg/kg BW, the ratio of effector cells to target cells was significant at 10:1 compared to the normal control group (CON). was increased (p<0.05) and was found to be at a level similar to that of the positive control group (PC). This shows that the degree of apoptosis of YAC-1 cells was improved by 81.2 and 63.1%, respectively, in the positive control group and the group administered with the composition for preventing and improving respiratory diseases (GHW, 200 mg/kg) prepared in Example 1 compared to the normal control group. (refer to Fig. 4)

Therefore, it was confirmed that the composition for preventing and improving respiratory diseases prepared in Example 1 affects the activity of NK cells, and the higher the treatment concentration, the higher the activity of natural killer cells.

[Experimental Example 5]

In order to confirm the effect in the bronchial asthma model of the composition for preventing and improving respiratory diseases prepared in Example 1, which could significantly inhibit the production of TNF-alpha and IL-5 in splenocytes, 100 μg of OVA and 1 mg Alum was mixed with 0.2 ml of PBS (phosphate buffered saline, pH 7.4) and injected intraperitoneally. After primary systemic sensitization, immune boosting was induced through secondary sensitization in the same way on the 15th day, and then, A was diluted by concentration with physiological saline and orally administered from the 18th to the 24th for a week. In addition, allergic bronchial asthma was induced by spraying 5% OVA in PBS for 30 minutes for 3 consecutive days on days 22, 23, and 24. Mice were analyzed 24 hours after the last OVA spray (see FIG. 5 ).

Changes in airway hypersensitivity caused by allergic hypersensitivity reactions were observed by applying the composition for preventing and improving respiratory diseases prepared in Example 1, which showed a significant effect on the expression of TNF-α and IL-5 in mouse splenocytes, to a bronchial asthma model. In order to confirm the concentration of methacholine in the normal group (Normal), the asthma-inducing group (ovalbumin, OVA), and the group treated with the composition for prevention and improvement of respiratory diseases prepared in Example 1 (GHW 0.05, 0.1, 0.5 g/kg treatment group) The airway resistance value (Penh) was investigated by exposure to the star, and is shown in FIG. 6 .

According to FIG. 6, the Penh value of the composition treatment group (GHW) for preventing and improving respiratory diseases prepared in Example 1 decreased in a concentration-dependent manner at the concentrations of 0.5 g/kg and 0.1 g/kg, thereby confirming a significant effect. .

[Experimental Example 6]

In order to confirm the effect of the composition for preventing and improving respiratory diseases prepared in Example 1 on the lung tissue of bronchial asthma-induced mice, H&E staining of the lung tissue was observed 24 hours after the last spraying of OVA, and the results are shown in FIG. it was

According to FIG. 7 , in the normal group, the airway wall is thin and clear, and infiltration of inflammatory cells is not seen, but in the asthma-induced group (OVA), the airway stenosis and the thickness of the airway wall are increased, and the deposition of inflammatory cells is significantly increased. You can check what has been done.

In the 0.5 g/kg group (GHW) of the composition for preventing and improving respiratory diseases prepared in Example 1, the airway stenosis and the thickness of the airway wall were reduced compared to the asthma-inducing group, and the respiratory disease prevention and improvement prepared in Example 1 In the 0.1 g/kg group (GHW), the airway wall became thinner, the damaged alveolar morphology returned to a normal level, and the infiltration of inflammatory cells was also reduced to the level of the normal group compared to the asthma-inducing group.

Claims (4)

15 to 25% by weight of golden extract, 5 to 15% by weight of ginseng extract, 15 to 25% by weight of oleracea extract, 15 to 25% by weight of sorghum extract, 15 to 25% by weight of Schisandra extract and 5 to 15% by weight of cinnamon extract ,
A composition for preventing and improving respiratory diseases.
The method of claim 1,
The golden extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of gold and extracting at 75-85° C. for 3-4 hours,
The gold and silver flower extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of gold and silver coins and extracting at 75 to 85° C. for 3 to 4 hours,
The extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of the oleander and extracting it at 75-85° C. for 3-4 hours,
The extract is obtained by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of Sukhumang and extracting it at 75-85° C. for 3 to 4 hours,
The Schisandra extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of Schisandra and extracting it at 75-85° C. for 3-4 hours,
The cinnamon extract is extracted by adding 20 parts by weight of 70% (v/v) ethanol to 1 part by weight of cinnamon and extracting at 75-85° C. for 3-4 hours,
A composition for preventing and improving respiratory diseases.
The method of claim 1,
15 to 25% by weight of the golden extract, 5 to 15% by weight of the ginseng extract, 15 to 25% by weight of the oleander extract, 15 to 25% by weight of the sagebrush extract, 15 to 25% by weight of the Schisandra extract and 5 to 15% by weight of the cinnamon extract 1 to 5 parts by weight of an immunity enhancer is additionally included in 100 parts by weight of one mixture,
The immunity enhancer contains 30% by weight of red ginseng concentrate, 30% by weight of Angelica asiatica extract, 20% by weight of chlorella and 20% by weight of leaf extract,
The red ginseng concentrate is obtained by adding 300 to 400 parts by weight of citrus juice to 100 parts by weight of red ginseng, heating it at 80 to 85° C. for 20 to 24 hours, centrifugation, filtration, and concentration under reduced pressure,
The Angelica asiatica extract is filtered after extracting Angelica quince with 85% methanol using an ultrasonic extractor on a water bath 3 times for 1 hour each,
The leaflet extract is filtered after adding 500 parts by weight of 100% methanol to 100 parts by weight of the leaflets and immersing it at room temperature for 12 hours,
A composition for preventing and improving respiratory diseases.
The method of claim 1,
15 to 25% by weight of the golden extract, 5 to 15% by weight of the ginseng extract, 15 to 25% by weight of the oleander extract, 15 to 25% by weight of the sagebrush extract, 15 to 25% by weight of the Schisandra extract and 5 to 15% by weight of the cinnamon extract 0.1 to 1 part by weight of a taste enhancer is additionally included in 100 parts by weight of a mixture,
The taste enhancer contains 50% by weight of goji berry extract, 35% by weight of maekmundong extract, 10% by weight of stevia extract, and 5% by weight of fructooligosaccharide,
The Goji berry extract is filtered after adding 300 to 400 parts by weight of water to 100 parts by weight of Goji berry and heating at 95 to 105° C. for 1 to 2 hours,
The maekmundong extract is filtered after adding 300 to 400 parts by weight of water to 100 parts by weight of maekmundong and heating at 105 to 110° C. for 5 to 7 hours,
The stevia extract is filtered after extraction by adding 600 to 700 parts by weight of purified water to 100 parts by weight of stevia and heating at 95 to 100° C. for 3 to 4 hours,
A composition for preventing and improving respiratory diseases.

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