KR101474125B1 - Composition comprising complex extract of Astragalus membranaceus BUNGE, Saposhnikovia divaricata Schiskin, Ostericum koreanum Maximowicz, Scutellaria baicalensis Georgi and Atractylodes japonica Koidz for preventing or treating asthma, bronchitis or pneumonia - Google Patents

Abstract

The present invention relates to a composition for the prevention or treatment of asthma, bronchitis or pneumonia, which comprises a complex extract of yellow mugwort, windblown mugwort, golden mugwort, golden mugwort, and alfalfa as an active ingredient. The combined extract of the present invention reduces the number of inflammatory immune cells eosinophils, neutrophils and macrophages, inhibits the synthesis and secretion of immunoglobulin E, an allergic immunity-inducing antibody, and inhibits the allergic immune response-inducing Th2 cytokine IL-4 , Inhibits secretion of IL-5 and IL-13, inhibits the secretion and accumulation of mucin to improve basal membrane damage, inhibits collagen secretion and accumulation, prevents fibrosis and curing of underlying bronchi in lung tissue, Asthma, bronchitis, or pneumonia.

Description

[0001] The present invention relates to a composition for preventing or treating asthma, bronchitis or pneumonia, which comprises a complex extract of Hwanggi, a windbreak, Ganghwam, and Atractylodes japonica Koidz for prevention or treating asthma, bronchitis or pneumonia}

The present invention relates to a composition for the prevention or treatment of asthma, bronchitis or pneumonia, which comprises a complex extract of yellow mugwort, windblown mugwort, golden mugwort, golden mugwort, and alfalfa as an active ingredient.

Bronchitis is a disease that causes damage to the bronchial mucosa due to the stimulation of viruses, bacteria, smoking, air pollution, dust, etc., and causes a lot of inflammation and mucus to cause symptoms such as sputum and cough. Generally, bronchitis is classified into acute bronchitis and chronic bronchitis, and in severe cases, it may be exacerbated by pneumonia or asthma.

Pneumonia is an inflammation of the lung parenchyma, which is called interstitial pneumonia or pulmonary enteritis. Pneumonia is an inflammation of the lungs caused by an infection caused by microorganisms such as bacteria, viruses and fungi. Pneumonia is caused by a variety of symptoms, including coughing, sputum due to the release of inflammatory substances, dyspnea due to impaired breathing function, and pulmonary symptoms that impair the normal functioning of the lungs, as well as digestive symptoms such as nausea, vomiting, diarrhea and headache, fatigue, It is a disease in which systemic symptoms occur throughout the body. Pneumonia should be treated according to the cause, and most of them take antibiotics. However, if the symptoms of pneumonia are severe, even if the appropriate agonist is used, the disease progresses and may die.

Asthma is a disease that causes symptoms such as difficulty breathing, coughing, chest pain and wheezing by causing allergic substances such as house dust mites and ticks to shrink into the human body as they enter the body and cause airway contraction and inflammation. It is a chronic respiratory disease.

In most countries around the world including our country, asthma patients are increasing by about 20 to 50% every 10 years. Studies have shown that worldwide asthma patients reach 150 million people, and about 180,000 people die annually from asthma. Especially, in Korea, the ratio of children aged 6 to 7 years is 13.3%, which is the second highest level among the 8 Asian countries, after Singapore. Asthma is caused by the increase of harmful substances such as air pollution, soot, tobacco smoke, ingestion of contaminated food, activity in enclosed space, and contact with unclean things.

Asthma is often coughing or wheezing as it is simply a cold, so it is often passed without a different prescription, and the symptoms are prolonged as a chronic disease. If the management is not properly done, allergic immune reaction occurs in the airway causing inflammation and death You may.

In addition, lung function may be reduced if asthma is left untreated. There is no cure for asthma yet. Most asthma patients are taking medications to relieve chronic symptoms, to minimize asthma attacks, to maintain normal lung function, and to prevent airway remodeling. Since no special treatment has been developed, antibiotics, anti-inflammatory drugs (Corticosteroids, sodium chromolin / nedocromil, leukotriene modifiers, oral steroids) or bronchodilators (beta adrenergic receptor agonists) that alleviate asthma symptoms.

However, since the above administration preparation can temporarily improve the symptoms, it is difficult to cure it and there is no change in the severity, so it can not be said to be a fundamental treatment method, and thus development of a therapeutic agent therefor is urgently required.

On the other hand, herbal medicines have been used to treat diseases and protect the body, and they are currently being used for the prevention and treatment of various diseases. Since herbal medicines are derived from natural resources, they have multiple pharmacological mechanisms and can produce complex effects. In addition, the herbal medicines contain various pharmacologically active substances and physiologically active substances, and are deeply involved in maintenance of the homeostasis and immune response of the human body, and it is possible to cure diseases. Recently, as side effects of biochemical pharmaceuticals manufactured by chemical synthesis have been controversial in the media and media, there is a great interest in treating diseases from safer natural resources, i.e., medicinal herbs.

Astragalus Root (Astragalus Root, Scientific name: Astragalus membranaceus Bunge) is used as a medicinal herb by drying roots. It is flavorful, warm in nature, and works in the spleen and lungs. Hwanggi has the effect of seeing the flag and tightening the body, stopping sweating, warming the body, helping the diuretic effect and removing the tumor. It also cleanses the skin and removes toxins to renew the skin. Therefore, Hwanggi is not a solid body, weakness of the weak person, weak aura, difficult to behave symptoms, poor urine symptoms, skin type, lack of energy and blood, swelling and malformation in the depression, Or all kinds of surgical and dermatological diseases are known to be used for the treatment of symptoms that do not get well.

Saposhnikovia Root (Saposhnikovia divoricata Schischkin ) is a dried plant root which is used as a medicinal herb. Its taste is temperate and warm in nature, and acts on liver, spleen and bladder. Windfight is an anti-inflammatory drug that treats the symptoms caused by external stimuli by releasing the fragility of the skin, relieving the wind, eliminating the joint pain, neuralgia and pain caused by the stagnation of moisture and stopping the seizure. It has been commonly used. Windshields can be used irrespective of the type of symptoms such as fever, fever, blistering, and demonstration, and have been used to treat chills, headache or body aches, fever or fever caused by cold winds, sore throat, and sore throat . Winds are used to cure cold and windy winds and limbs and to treat sickness. It is also used for the treatment of heavy joints, painful bones, symptoms of bones, limb muscles, twitching and twitching Is used. In addition, the windshield has the effect of eliminating the winds and stop the itching, improving the windshield, treating skin pruritus, etc., entering into the ligament attached to the liver, eliminating the winds and relieving the mysterious stretch of the limbs. It is known to be used to treat backward swelling, axillary spasm, and other symptoms.

Ostericum Root (Scientific name: Ostericum koreanum Maximowicz) is used as a herb medicine by drying the roots of plants. It is tasted, warm in nature, and works on bladder and nerve. Ganghwam has the effect of eliminating the cold, removing the wind, removing moisture and stopping the pain, treating the headache caused by cold or windy external winds, body pain, and the joints caused by cold winds and humid air It is a medicinal herb that is commonly used in cold and joint related diseases because it has the effect of treating pain.

Scutellaria Root (Scientific name: Scutellaria baicalensis Georgi) is a herb that is used as a medicinal herb by removing roots from a plant, and is used in the taste, texture, and function in the lungs, feces, stomach, large intestine and small intestine. Gold has the effect of cooling the heat, drying the moisture, and removing the poison by burning the heat. In addition, gold is known to have the effect of stabilizing hemostasis and pregnancy. Gold is a fever that is caused by moisture, hot and warm, and the chest is blocked and full and stuffy due to symptoms of fever and sickness due to chest exhaustion, humid heat, while symptoms such as unaffected symptoms, jaundice, It is known to be used to treat symptoms of fever and thirst in the chest, symptoms of fever with high fever in the blood, swelling and swelling in the blood, symptoms of uncomfortable movements of pregnant women, uterine bleeding, hematuria and the like.

Atractylodes japonica Koidz or Atractylodes macrocephala Koidz (= Atractylis ovata Thunb.)) Is used to dry rootstocks, to taste, to wear, to warm, to spleen Lt; / RTI > It makes the spleen strong and refreshing, and it dries the moisture and makes the urine go well. In addition, it is known that it has the effect of stopping perspiration and stabilizing the fetus of pregnant women. It is known to be used for the treatment of unstable symptoms, diarrhea, sputum, asthma, symptoms of dizziness when eating food, sweat, hyperhidrosis, and pregnancy and pregnancy.

The inventors of the present invention have been studying a therapeutic agent for asthma, bronchitis or pneumonia while reducing the number of inflammatory immune cells eosinophils, neutrophils and macrophages, and the synthesis of immunoglobulin E And suppresses the secretion of IL-4, IL-5 and IL-13 which are Th2 cytokines and suppresses the secretion and accumulation of mucin to improve basement membrane damage and suppress collagen secretion and accumulation The present inventors have completed the present invention by confirming that they have excellent therapeutic effects on asthma, bronchitis or pneumonia by preventing fibrosis and curing of the lower part of the bronchus in lung tissue.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of asthma, bronchitis or pneumonia which contains a combination extract of yellow mugwort, windblown mugwort, golden mugwort, golden mugwort,

Another object of the present invention is to provide a food composition for preventing or ameliorating asthma, bronchitis or pneumonia, which contains as an active ingredient a complex extract of yellow mugwort, windblown mugwort, golden mugwort, and golden mugwort.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating asthma, bronchitis or pneumonia, which comprises a combined extract of yellow mugwort, windblown mugwort, golden mugwort, and golden mugwort as an active ingredient.

Further, the present invention provides a food composition for preventing or ameliorating asthma, bronchitis or pneumonia, which contains a combination extract of hwangggi, windblown, ganghwam, gold and alfalfa as an effective ingredient.

The combined extract of Hwanggi, Kwangwoon, Kwangmyung, Gwangmyeong, Gwangmyeong, and Baekwang according to the present invention reduces the number of eosinophils, neutrophils and macrophages which are inflammatory immune cells, inhibits the synthesis and secretion of immunoglobulin E as an allergic immune response inducing antibody, Inhibition of the secretion of IL-4, IL-5 and IL-13, the Th2 cytokines that induce the immune response, and inhibition of mucin secretion and accumulation to improve basement membrane damage and inhibit collagen secretion and accumulation, By preventing fibrosis and hardening of the lower bronchus, it has excellent therapeutic effect on asthma, bronchitis or pneumonia.

1 is a graph showing the effect of reducing the number of inflammatory cells in bronchoalveolar lavage fluid in ovalbumin-induced asthma mice of the combination extract of the present invention ( ^* P <0.05, ^** P <0.01 and ^*** P <0.001) (a) and negative control group (b)).
FIG. 2 is a graph showing the effect of inhibiting the secretion of ovalbumin-specific IgE in serum of an egg albumin-induced asthma mouse of the combination extract of the present invention ( ^* P <0.05, ^** P <0.01 and ^*** P < (a) and negative control group (b)).
FIG. 3 is a graph showing the effect of inhibiting ovalbumin-specific IgE secretion in the bronchoalveolar lavage fluid of the combination extract of the present invention in ovalbumin-induced asthma mice ( ^* P <0.05, ^** P <0.01 and ^*** P <0.001. Normal group (a) and negative control group (b)).
FIG. 4 is a graph showing the inhibitory effect of IL-4 on the secretion of ovalbumin-induced asthma in combination extract of the present invention ( ^* P <0.05, ^** P <0.01 and ^*** P <0.001) a), negative control (b) and positive control (c).
FIG. 5 is a graph showing the inhibitory effect of IL-5 on serum secretion in ovalbumin-induced asthma mice of the combination extract of the present invention ( ^* P <0.05, ^** P <0.01 and ^*** P < a), negative control (b) and positive control (c).
FIG. 6 is a graph showing the inhibitory effect of IL-13 on serum secretion in ovalbumin-induced asthma mice of the combination extract of the present invention ( ^* P <0.05, ^** P <0.01 and ^*** P < a), negative control (b) and positive control (c).
7 shows the effect of the combined extract of the present invention on the damage of lung tissue in an egg albumin induced asthmatic mouse (H & E staining; purple and red color, × 200)
8 is a graph showing the effect of inhibiting goblet cell proliferation and mucin secretion in the basal membrane of lung tissue in ovalbumin-induced asthmatic mice of the combination extract of the present invention (PAS staining; purple, × 200)
FIG. 9 is a graph showing collagen accumulation and fibrosis inhibitory effects of pulmonary tissues in an egg albumin-induced asthmatic mouse of the multiple extract of the present invention (PAS staining: blue, × 200)

The present invention provides a composition for the prevention or treatment of asthma, bronchitis or pneumonia, which comprises a combined extract of yellow mugwort, windblown mugwort, golden mugwort, and golden mugwort as an active ingredient.

The composition comprises a pharmaceutical composition and a food composition.

Hereinafter, the present invention will be described in detail.

The complex extract of the present invention can be obtained by a conventional extraction method, and commercially available ones can be purchased and used. Typical extraction methods include, but are not limited to, ultrasonic extraction, filtration, and reflux extraction.

The complex extract of the present invention can be obtained through the following steps. First of all, after washing the hwanggi, wind wind, ganghwam, gold and algae, they are dried, crushed and mixed. The medicinal herb may be cultivated, marketed, etc. without limitation. Water or an organic solvent is added to the mixture in an amount of about 10 to 30 times, and the mixture is then extracted, filtered, concentrated under reduced pressure, and lyophilized to obtain a complex extract.

In the present invention, the water used for hot water extraction of the mixed medicinal materials according to the present invention refers to water suitable for drinking standards on the basis of the Ministry of Health and generally refers to water such as purified water, sterilized water and anion water, tap water, Etc. may be used. However, in the present invention, the solvent used for extracting the active ingredient of the mixed medicinal herb is not limited to water. If an apparatus for removing the harmful substance is used, an organic solvent such as methanol, ethanol, acetone, Can be used. That is, in the present invention, in order to prevent or treat asthma, bronchitis or pneumonia in addition to water, it is possible to extract the active ingredient of the medicinal material thoroughly while extracting it using an organic solvent if it is completely harmless to the human body, have.

The composition of the present invention preferably contains 10 to 30% by weight of sulfur, 10 to 20% by weight of windshield, 10 to 20% by weight of hardness, 10 to 20% by weight of gold and 20 to 50% by weight of flour.

If the above-mentioned maximum use amount is exceeded for each of the above-mentioned medicinal herbs, the other necessary components can not be used sufficiently, and if the use amount is less than the minimum usage amount, the effective effect of each medicinal herb is reduced. Therefore, it is preferable that the combined extracts are used within the use ranges for each herbal medicine described above to exhibit the maximum effect on asthma, bronchitis or pneumonia.

The combined extract of Hwanggi, Kwangwoon, Kwangmyung, Gwangmyeong, Gwangmyeong, and Baekwang according to the present invention reduces the number of eosinophils, neutrophils and macrophages which are inflammatory immune cells, inhibits the synthesis and secretion of immunoglobulin E as an allergic immune response inducing antibody, Inhibition of the secretion of IL-4, IL-5 and IL-13, the Th2 cytokines that induce the immune response, and inhibition of mucin secretion and accumulation to improve basement membrane damage and inhibit collagen secretion and accumulation, By preventing fibrosis and hardening of the lower bronchus, it has excellent therapeutic effect on asthma, bronchitis or pneumonia.

Preferably, the asthma is bronchial asthma.

The bronchitis includes, but is not limited to, acute bronchitis, chronic bronchitis, allergic bronchitis, infectious bronchitis, traumatic bronchitis, catarrhal bronchitis, purulent bronchitis, obstructive bronchitis, ulcerative bronchitis and invasive bronchitis.

The pneumonia includes, but is not limited to, bacterial pneumonia, mycoplasma pneumonia, pneumonia caused by viruses, and pneumonia caused by rickettsia.

The composition of the present invention may contain at least one known active ingredient having a therapeutic effect of asthma, bronchitis or pneumonia in combination with the combined extract.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the combined extract of the present invention may be administered in an amount of 100 mg / kg to 500 mg / kg per day, and may be administered once or several times a day.

The pharmaceutical composition of the present invention may be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of asthma, bronchitis or pneumonia.

In the present invention, the term "health functional food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease and aging inhibition.

The composition of the present invention may be added to a health functional food for the purpose of preventing or improving asthma, bronchitis or pneumonia. When the compound extract of the present invention is used as a food additive, the compound extract may be directly added or used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the complex extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, preferred examples, experimental examples, and formulation examples are provided to facilitate understanding of the present invention. However, the following examples, experimental examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples, experimental examples and preparation examples.

[Example 1: preparation of complex extract]

In order to prepare the complex extract of the present invention, the origin of Korean herb, windblown, ganghwam, gold and liquor purchased from Kwang Myung Dang Pharmaceutical, a Korean traditional medicine manufacturer, was washed and washed with water. The dried yellow, wind, wind, yellow, and golden leaves were crushed and mixed according to the weights shown in Table 1 below.

Herbal medicine A good name Weight used (g) Hwanggi Astragalus Root ( Astragalus membranaceus ) 6 Wind wind Saposhnikovia Root ( Saposhnikovia divaricata ) 4 Ganghwa Ostericum Root ( Ostericum koreanum ) 4 Gold Scutellaria Root ( Scutellaria baicalensis ) 4 Boiling Atractylodes Rhizome White ( Stractylodes lancea ) 8 Yield (%) 42%

As shown in Table 1, 20 times volume of purified water was added to the mixed herbal medicines prepared by mixing 6 g of sulfur, 4 g of windblown, 4 g of gangbyeol, 4 g of gold, 4 g of gold and 8 g of flour, and the mixture was heated at 95 to 100 ° C for 4 hours And heated and extracted with hot water. The hot-water extract was filtered through a filter (50 mu m and 1 mu m cartilage), the filtrate was introduced into a concentrator and then concentrated under reduced pressure at a temperature of 60 DEG C and 700 mmHg for 15 hours. The concentrate was then lyophilized to produce a complex extract. The yield of the obtained complex extract was 42%, which was used as a drug for evaluation of efficacy while being stored at 4 ° C in a refrigerator.

[Example 2: Preparation of ovalbumin-sensitized allergic asthma animal model and administration group setting]

1. Production of an albumin induced asthma animal model

20 mg of ovalbumin (OVA) dissolved in 2 mg of aluminum hydroxide in 8-week-old BALB / c mice was intraperitoneally administered to the mice at 0, 7, 14, and 21 days Allergic immune response was induced. Mice in which systemic allergy immunoreactivity was induced were inoculated into a standard size acrylic box once a day from the 21st to 27th day of OVA peritoneal administration (for a total of 7 days), and 1% albumin was inhaled for 20 minutes with a Nebulizer Sensitized albumin induced asthmatic mouse model.

2. Administration group setting

In order to examine the preventive or therapeutic effect of the combined extract according to the present invention on asthma, bronchitis or pneumonia, the administration group was set up as shown in Table 2 below.

Normal group Saline-treated group Negative control group OVA treated group (egg albumin induced asthmatic mouse) Experimental group A OVA + compound extract 100 mg / kg group Experimental group B OVA + compound extract 200 mg / kg group Positive control group OVA + Montelukast 10 mg / kg group

As shown in Table 2, normal group to which physiological saline was administered; A negative control in which OVA was administered to induce albumin asthma; In order to induce albumin asthma, OVA was administered and the dose of Compound A (100 mg / kg of body weight of the compound extract) and B (200 mg / mouse of the combined extract sample, ); The administration group was set as OVA to induce albumin asthma and a positive control group to which montelukast, a leukotriene antagonist, was administered. The administration of the combined extract and leukotriene antagonist was performed in the same manner as in the last administration of ovalbumin , And was orally administered once daily once daily for 7 days of inhalation sensitization.

2. Statistical Test

All results are the average standard error of the triplicate experiments; showed a (standard deviation SD), statistical analysis was determined to be significant with a p-value less than .05 by Student t -test black in the GraphPad program.

[Experimental Example 1: Measurement of the effect of decreasing the number of inflammatory immune cells in bronchoalveolar lavage fluid of ovalbumin induced asthmatic mice against the compound extract of the present invention]

In order to determine the effect of reducing the number of inflammatory immune cells in the bronchoalveolar lavage fluid of the ovalbumin-induced asthmatic mouse to the compound extract of the present invention, the following experiment was conducted.

Bone marrow bronchoalveolar labile fluid (BALF) was obtained by repeatedly inserting and inhaling a DMEM culture medium containing 10% FBS with a syringe at 37 ° C in the airway of the asthma-induced mouse, . Various immune cells contained in the obtained alveolar lavage fluid were stained on a slide glass at 1000 rpm using a cytospin chamber, stained with Diff-Quik, and the number of macrophages, lymphocytes, neutrophils and eosinophils was counted 400-fold optical microscope (Nicon, Japan) and the number of cells was calculated. The results are shown in Fig.

As shown in FIG. 1, the number of inflammatory immune cells in the bronchoalveolar lavage fluid of asthma-induced negative control group was significantly increased compared to that of the normal group, and inflammatory immune cells such as macrophages, neutrophils, and eosinophils Of the total number.

On the other hand, the number of immune cells in the bronchoalveolar lavage fluid of the experimental groups A (100 mg / kg) and B (200 mg / kg) administered with the compound extract of the present invention decreased depending on the administration dose and significantly decreased . In particular, it was confirmed that the number of neutrophils and eosinophils was significantly reduced.

Therefore, the combined extract of the present invention significantly reduces the number of eosinophils and inflammatory immune cells that play a key role in the pathogenesis of asthma, bronchitis or pneumonia, thereby significantly inhibiting the production of allergic inducers and inflammatory substances It was confirmed that the effect of strongly controlling the allergic reaction and the inflammatory reaction was excellent.

[Experimental Example 2: Measurement of secretion inhibitory effect of ovalbumin-specific IgE and inflammatory cytokines (IL-4, IL-5, IL-13) in serum and bronchoalveolar lavage fluid of ovalbumin-induced asthmatic mice against compound extracts]

The secretion inhibitory effect of ovalbumin-specific immunoglobulin E (IgE) and inflammatory cytokines (IL-4, IL-5, IL-13) in serum and bronchoalveolar lavage fluid of ovalbumin induced asthmatic mice to the compound extract of the present invention For the measurement, the following experiment was carried out.

1. Obtain serum and bronchoalveolar lavage fluid from asthma-induced mice

The serum of the asthma-induced mouse was intraperitoneally administered with 10% chloral hydrate to the mouse in which the test was completed on the next day (day 28) after the inhalation sensitization of ovalbumin in Example 1, Blood was collected by cardiac blood collection and centrifuged at 3000 rpm for 10 minutes. Bronchoalveolar lavage fluid of asthma-induced mice was obtained using the same method as described in Experimental Example 1 above.

2. Measurement of immunoglobulin E and cytokine inhibitory activity

(IgE) and cytokines (IL-4, IL-5 and IL-13), which are allergic response factors or inflammation-inducing immunity factors, from the serum of the obtained mice and the supernatant of bronchoalveolar lavage fluid The ELISA kit was used for this purpose. Each antibody was diluted in a coating buffer, coated on a microwell, and allowed to react overnight at 4 ° C. Each well was washed three times with buffer solution for washing, and then 100 쨉 l of serum and bronchoalveolar lavage fluid were dispensed. After incubation for 1 hour at room temperature, the cells were washed twice with a buffer solution for washing and 100 쨉 l of avidin-HRP-conjugated antibody was dispensed into each well and left at room temperature for 1 hour. After washing again, 100 .mu.l of TMB substrate was added, reacted for 30 minutes in a dark state, reacted with 50 .mu.l of the reaction completion solution, and the absorbance at 450 nm was measured with an ELISA meter.

3. Results

end. IgE measurement results

The concentrations of IgE, an allergic immunity-inducing antibody, in the serum of the egg albumin-induced asthmatic mouse and the bronchoalveolar lavage fluid of the compound extract of the present invention were measured, and the results are shown in FIGS.

As shown in FIGS. 2 and 3, the concentrations of IgE in serum (FIG. 2) and bronchial alveolar lavage fluid (FIG. 3) in mice in which asthma was induced by ovalbumin sensitization as a negative control group were measured, And it was confirmed that it was significantly increased.

On the other hand, the IgE concentrations in serum and bronchoalveolar lavage fluid of asthma-induced mice in the experimental group A (100 mg / kg) and B (200 mg / kg) administered with the compound extract of the present invention were measured, IgE concentration was decreased, and it was confirmed that the amount of IgE was significantly decreased in the negative control group. In particular, the effect of reducing the IgE concentration in experimental group B was confirmed to be superior to that of montelukast, a positive control leukotriene antagonist.

Therefore, it was confirmed that the combined extract of the present invention is excellent in suppressing the synthesis and secretion of IgE, which is a major factor in inducing an allergic immune response in asthma, bronchitis or pneumonia.

I. Measurement results of cytokine

The concentrations of IL-4, IL-5 and IL-13, which are Th2 cytokines inducing allergic immunity in the serum of ovalbumin-induced asthma mice, were measured for the combined extract of the present invention. Respectively.

As shown in FIG. 4 and FIG. 6, the concentrations of IL-4, IL-5 and IL-13 in the serum of mice in which asthma was induced by ovalbumin sensitization as a negative control group were measured. Which was significantly increased after the induction.

On the other hand, the concentrations of IL-4, IL-5 and IL-13 in the serum of asthma-induced mice of the group A (100 mg / kg) and B , Dose-dependently decreased, and it was confirmed that it was significantly decreased in the negative control group. In particular, the reduction of IL-4 and IL-5 levels in serum of asthma-induced mice in experimental group B was found to be superior to montelukast, a leukotriene antagonist, a positive control.

Therefore, it was confirmed that the combined extract of the present invention is excellent in suppressing the secretion of IL-4, IL-5 and IL-13, which are Th2 cytokines secreted from T lymphocytes important for controlling allergic immune responses in asthma, bronchitis or pneumonia Respectively.

[Experimental Example 3: Evaluation of bronchial and lung tissue damage improvement effects of ovalbumin-induced asthma mice on the combined extracts]

In order to measure the effect of the combined extract of the present invention on the lung tissue damage improving effect of the ovalbumin induced asthmatic mouse, the ovalbumin-induced asthmatic mice that had been tested on the next day (28 days) And then fixed on 4% paraformaldehyde to prepare a tissue slice. The tissue slice was cut to 5 to 6 탆 and adhered to the slide to prepare a tissue specimen.

1. Observation of improvement effect on bronchial and lung tissue damage

In order to examine the effect of the combined extract according to the present invention on the damage of the bronchial and lung tissues of the ovalbumin-induced asthmatic mouse, the paraffin tissue specimen was prepared and stained with H & E (Hematoxylin and Eosin) epithelium) damage and hypertrophy, pulmonary vascular occlusion, airway obstruction, infiltration of inflammatory cells were observed under a microscope, and the results are shown in FIG.

As shown in FIG. 7, asthma induced bronchial and lung damage was observed in ovalbumin-sensitized mice. As a result, morphological damage of bronchial airways, endothelial damage and thickening of the basement membrane, and pulmonary vascular occlusion Airway obstruction, massive influx of inflammatory cells, and inflammatory responses were observed, indicating that the inflammatory damage of the bronchial and lung tissues worsened.

On the other hand, observing the effects of the group A (100 mg / kg) and the group B (200 mg / kg) administered with the complex extract of the present invention to improve bronchial and lung tissue damage, Negative control group. In particular, it was observed that the effect of improving the bronchial and lung tissue damage of the experimental group B was remarkably excellent and restored similarly to the normal group.

Therefore, it has been confirmed that the combined extract of the present invention is excellent in the effect of protecting bronchial airway and lung tissue by improving and restoring structural damage of bronchial and lung tissues in asthma, bronchitis or pneumonia.

2. Observation of mucin secretion in basolateral bronchial and lung tissue

In order to examine the effect of the combined extract of the present invention on the inhibition of basement membrane damage in bronchial and lung tissues of ovalbumin-induced asthmatic mice, periodic acid-Schiff (PAS) staining was performed on the tissue samples , And goblet cell proliferation were observed under a microscope. The results are shown in FIG.

As shown in FIG. 8, the bronchial airway and the basal membrane of the lung tissue of the asthma-induced mice were observed by ovalbumin sensitization as a negative control group. As a result, goblet cells secreting mucin strongly stained with PAS in the endothelial cell layer ), And it was confirmed that the basement membrane damage was large.

On the other hand, as a result of observing damage of bronchial airway and basal membrane of lung tissue of asthma-induced mice of Experimental group A (100 mg / kg) and B (200 mg / kg) administered with the compound extract of the present invention, And basal membrane damage were improved, and it was confirmed that the negative control was significantly reduced. Especially, in experimental group B, PAS staining was not observed similar to normal group, and it was confirmed that airway and basement membrane damage were hardly found.

Accordingly, the combined extract of the present invention is effective in preventing damage to the basement membrane in the bronchial and lung tissues and preventing airway obstruction by inhibiting the secretion and accumulation of mucin upon proliferation of goblet cells in asthma, bronchitis or pneumonia Respectively.

3. Observation of fibrosis in lung tissue

In order to examine the effect of the combined extracts of the present invention on the damage caused by the fibrosis generated in the lower bronchus in the lung tissue of the ovalbumin-induced asthmatic mouse according to the present invention, the tissue specimen was stained with Masson's trichrome and then collagen secretion and accumulation The degree of fibrosis was observed under a microscope, and the results are shown in FIG.

As shown in FIG. 9, in the lung tissue of the asthma-induced mouse, the fibrogenesis was observed in the lower bronchus of the mouse, and the accumulation of collagen strongly stained in Masson's trichrome in the lower bronchus (blue) The resulting fibrosis was observed.

On the other hand, fibrosis produced in the lower bronchus was observed in the lung tissues of mice of the group A (100 mg / kg) and B (200 mg / kg) administered with the compound extract of the present invention, , And the negative control group was significantly reduced. In particular, in experimental group B, collagen secretion and fibrosis were inhibited by not being stained with Masson's trichrome almost in the same manner as the normal group.

Accordingly, it has been confirmed that the combined extract of the present invention is excellent in improving damage of lung tissue by inhibiting fibrosis and sclerosis of lung tissue due to accumulation of collagen in asthma, bronchitis or pneumonia.

Hereinafter, formulation examples of the composition of the present invention will be described, but the present invention is not intended to be limited but is specifically described.

[Preparation Example 1: Preparation of pharmaceutical preparation]

1. Manufacturing of powder

Compound extract 20 ml

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2. Preparation of tablets

Compound extract 10 ml

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

Compound extract 10 ml

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

4. Preparation of injections

Compound extract 10 ml

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Manufacture of liquids

Compound extract 20 ml

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

[Formulation example 2. Preparation of food preparation]

1. Manufacture of health food

Compound extract 100 ml

Vitamin mixture quantity

Vitamin A Acetate 70 g

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Manufacture of health drinks

Compound extract 100 ml

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

Although the composition ratio is relatively mixed with a component suitable for a favorite beverage as a preferred embodiment, the blending ratio may be arbitrarily varied depending on the regional or national preference such as the demand class, demand country, use purpose, and the like.

Claims (8)

As an active ingredient, a complex extract comprising 10 to 30% by weight of a sulfur-containing group, 10 to 20% by weight of a windshield, 10 to 20% by weight of a hardy oil, 10 to 20% by weight of gold, and 20 to 50% Or &lt; / RTI &gt;
delete The pharmaceutical composition for preventing or treating asthma, bronchitis or pneumonia according to claim 1, wherein the combined extract is a complex hot-water extract.
delete The method of claim 1, wherein the bronchitis is selected from the group consisting of acute bronchitis, chronic bronchitis, allergic bronchitis, infectious bronchitis, traumatic bronchitis, quartz bronchitis, purulent bronchitis, obstructive bronchitis, ulcerative bronchitis and invasive bronchitis Or a pharmaceutically acceptable salt or solvate thereof, for the prophylaxis or treatment of asthma, bronchitis or pneumonia.
The pharmaceutical composition according to claim 1, wherein the pneumonia is at least one pneumonia selected from the group consisting of bacterial pneumonia, mycoplasma pneumonia, viral pneumonia, and pneumonia caused by rickettsia. Gt;
Bronchitis or bronchial asthma comprising as an active ingredient a complex extract comprising 10 to 30% by weight of sulfur, 10 to 20% by weight of windshield, 10 to 20% by weight of hardness, 10 to 20% by weight of gold and 20 to 50% Food composition for improving pneumonia.
delete

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