KR20020007806A - Composition comprising extract of medicinal herbs for preventing and curing allergy and/or asthma - Google Patents

Abstract

PURPOSE: Provided is a composition for the prevention and treatment of allergy or asthma, containing the extracts of medicinal herbs, thereby the symptoms caused by the allergy or asthma can be effectively inhibited. CONSTITUTION: The composition for the prevention and treatment of allergy or asthma contains an effective amount of at least one extract selected from the group consisting of Moutan Cortex and Acanthopanacis Cortex and a pharmaceutically acceptable carrier, in which the extracting solvent is at least one selected from the purified water, ethanol, methanol, butanol, ethyl acetate and acetone; and the effective amount of the medicinal herb extracts in a person 60 kg heavy is 9 to 27 g/l of Acanthopanacis Cortex and 6 to 12 g/l of Moutan Cortex.

Description

Composition comprising extract of medicinal herbs for preventing and curing allergy and / or asthma}

The present invention relates to a composition for preventing and treating allergies and / or asthma comprising the herbal extract, and more particularly, to an allergy and / or asthma preventing and treating composition comprising the extract of bark or thorns.

In general, allergy is the first term used by von Pirque in 1906 to indicate a hyperimmune reaction. Coombs and Gell are generally classified into four types.

Type I is anaphylatctic (immediate type). The target organs are mainly digestive organs, skin and lungs. Common symptoms include gastrointestinal allergies, gallstones, atrophic dermatitis, allergic rhinitis and bronchial asthma. have. Pathology of type I activates target cells by contacting antigens with IgE antibodies attached to the surface of mast cells and basophils, secreting chemical media such as histamine, serotonin, leukotrienes, and PAF to release blood vessels and smooth muscle. It is known to shrink, which is often combined with type IV (delay type).

The incidence of bronchial asthma among allergic diseases varies according to country, race, and age, but according to the 1991 report of the United Kingdom, about 7% of adults and 13.5% of pediatric age group reported changes in living environment, pollution, It is gradually increasing with the increase of stress.

Bronchial asthma is the most common allergic respiratory disease. Symptoms such as difficulty breathing, coughing, and 'colored' rough breathing (wheezing) are repeated seizures. About 30% of asthma patients and about 80% within 1 year of birth At the age of 4-5, asthma symptoms first appeared in Korea, and the incidence rate of about 10% in Korea, treatment mainly consists of inhalation bronchodilators, oral or injectable bronchodilators (sympathetic stimulants and theophylline drugs), stereo Inhalation, oral, injection, etc.), anti-allergic drugs such as disodium cromoglycate, cromolyn sodium, nedocromil sodium, ketotifen, etc. Treatment and prevention is still one of the important tasks of the medical community, but there is no fundamental treatment. There is a palliative treatment only made it imperative for the development of essential therapies and treatments.

Although there was no term of allergy in traditional medicine, the allergic condition of modern allergy has been continued by using herbal or herbal prescriptions composed of many herbal medicines.

In Korea, Choi, Kami-MakDong-Tang has shown effective treatment for bronchial asthma in clinical trials. (Suk-Bong Choi: An Experimental Study on the Efficacy of Kami-MakDong-Tang, Korean Journal of Oriental Medicine (10) 2, 153 (160,1989), and other literatures reported that Gami-Mak-Dong-Tang was effective in treating clinical symptoms of yeast and bronchial asthma. ), Clinical Research, Korean Journal of Oriental Medicine 8 (2), 32, 1987; Yoseon Yongseon et al .: Literature Review on Allergic Asthma, Korean Journal of Oriental Medicine 11 (1), 39-70, 1990; , Et al .: Clinical study of bronchial asthma, Korean Journal of Oriental Medical Internal Medicine 7 (1), 60-67,1986; ). In addition, Chung Seung-ki and others reported clinical cases of Cheongsangboha-tang and Chung and Kim reported that some herbal remedies were effective for allergic type I and IV allergic reactions. Clinical Review, Papers presented at the 8th Round Korean Medicine Conference, 26-34, 1984; Seung-Gi Jung, Hyung-Koo Lee: Clinical Observation of Gamicheongsangboha-tang in Asthma, Kyunghee Medicine 2 (4), 97- 102, 1986; Jung Seung-gi, Lee Hyung-gu: An Experimental Study on the Effect of Chung Chung-tang on Asthma, Graduate School of Kyung Hee University, 1985; Kim Young-dae, Seung-gi Jung, Hyung-gu Lee A comparative study of the effects of Tang on type I and IV allergic reactions and pulmonary embolism, Kyung Hee Medicine 4 (4), 432-440,1988).

In Japan, Jiangsu has examined the effects of 32 kinds of herbal medicines, such as hyssop, on chemical mediators, antihistamines and guinea pigs, and the smooth muscles of the bronchioles (江 田昭英, 勝 田榮 二, 渡邊 茂 勝: 生 藥) Shibo-tang combined with Soshiho-Tang and Ha-Ha-Hak-Bak-Tang, PCA contact dermatitis and atopic asthma suppression. It has been reported to be effective, and in addition, 11 drugs such as "siho, licorice", which are recognized to feed back the IgE antigen by acting in the first stage of the allergic type I reaction and promoting the production of IgG antibodies, act in the second stage. We report 10 drugs, including gold, that have the effect of inhibiting the release of a chemical transfer material from mast cells involved in antigen-antibody reactions. Allergic Type IV is effective in delaying hypersensitivity reactions such as Candida and Aspergillus. As a result of 48 hours of PCA response by DNA-As-IgE, ginseng, pepper, bokyeongpi, licorice, half, jujube, leaflet are effective. However, it was reported that Chinese pepper was the most effective (江 田昭英 等: 和 漢 藥 の 抗 ア レ キ, Japanese pharmacological magazine 80, 31-32, 1982; 江 田昭英 江: Effects of various herbal medicines on reaginic antibody, Japanese pharmacological magazine 69, 889, 1973).絲 et al. Have reported six drugs, such as acting in the third step, antagonizing the chemicals released from mast cells or suppressing the inflammatory response. (絲川 秀 治 等: to histamine and barium hydrochloride Plant components that antagonize harvesting tube contraction (3rd Natural Products Development and Application Symposium Collection), 49, 1980).

In China, Kwak reported that 400 cases of dam margin and eczema were effective in Changchang, Hyeong-ae, Red Peony, Housewives, Baek-sun-pi, Baek-jap, and Seon-gyo. (郭 辨證: 淺談 蕁痲疹 的 辨證 施治, 浙江 中醫 雜誌 10, 412-414, 1981), Yu-Yeong has effective effects on contact dermatitis, and nutrient tonics such as clear heat detoxification drugs (Rhubarb, yellow and white, spectators, and silkworm) and Ganoderma lucidum are positive for HBsAg in chronically active hepatitis. He reported that the patient was converted to negative, and the most famous Beijing Medical Center in China divides asthma into ischemia and emphysema in this hospital. Boheungtang gamibang and Geumgwangi-hwan are used for emphysema, and Sagan Ma Huangtang and Ma-haenggam-tang (demonstration).麻 杏 甘 石 湯), Samjatang-hapyijintang, Samjajingi-tang, etc. were added or subtracted.

Therefore, the inventors of the present invention are currently increasing and allergic diseases need to be developed for the prevention and treatment of allergies and asthma using domestic herbs for allergic diseases worldwide.

The present invention solves the above problems, and drawn out by the above necessity, an object of the present invention is to provide a composition for the prevention and treatment of allergy and / or asthma comprising the herbal extract.

In order to achieve the above object, the present invention provides a composition for the prevention and treatment of allergies and / or asthma containing an effective amount of one or more extracts selected from the group consisting of bark and thorny scabies with a pharmaceutically acceptable carrier. to provide.

As the herbal extract used in the present invention, it is preferable to use the extract as an extract solvent, preferably by adding one solvent selected from an extracting solvent, preferably purified water, ethanol, methanol, butanol, ethyl acetate, and acetone.

In addition, the composition of the present invention may further include conventionally used excipients, disintegrants, sweeteners, lubricants, flavoring agents, etc., tablets, capsules, powders, granules, suspensions, emulsifiers, It may be formulated as a pharmaceutical formulation for single or multiple administration, such as a syrup, liquid or parenteral formulation.

The composition of the present invention can be parenterally administered or orally as desired, and the effective amount of the daily herbal extract for 60 kg body weight of the present invention is in the range of 125 mg / kg-500 mg / kg in the People's Republic of China and experimentally Considering the effective effect, the dose of prickly pear skin is 9g-27g and the amount of bark skin is 6-12g. Administration may be from one to several times a day. The dosage level for a particular patient may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion, severity of disease, and the like.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited to these examples.

Example 1: Preparation of an extract having anti-allergic effect using the herbal medicine

The Danmi herb used in the present invention is Mokpipi and psoriasis, which was purchased from Gyeongdong Market. The preparation method of extract from each herbal medicine is as follows. 300 g of powdered herbal medicines were soaked in 1 liter of ethanol for 1 week, and then concentrated under reduced pressure using a rotary evaporator (VI 461) and lyophilized. Each herb extract was stored at 4 ℃, was dissolved in DMSO in the experiment and used to dissolve in PBS or RPMI-1640 medium.

Example 2: Each sweet medicine and prescription in vitro Histamine glass experiment

Animals for histamine release experiments from mast cells were used as male Spraque-Dawley rats in the Korea Research Institute of Chemical Technology. In order to separate mast cells from rats, the rats were killed by ether, followed by tyrode solution (NaCl 137 mM, KCl 2.7 mM, CaCl ₂ .2H ₂ O 1 mM, NaH ₂ PO ₄ 0.4 mM, NaHCO ₃ 12 mM, glucose). 5.5 ml BSA) was injected into the abdominal cavity of the rat, and the abdominal wall was lightly massaged for 2 minutes, and then a small incision was made in the abdominal wall. The washing solution was centrifuged at 3000 rpm for 5 minutes to remove the supernatant and washed twice more using a tyrode solution containing no BSA. The mast cell suspension resuspended with tyrode solution containing no BSA was adjusted to 1 X 10 ⁶ / ml / well, dispensed into each well, stabilized at 37 ° C for 10 minutes, and then histamine glass compound 48/80 (Sigma). Co., 10 ㎍ / ㎖) 250µl and 250µl of herbal prescription solution adjusted to various concentrations were added to adjust the total amount of 2.5ml by the addition of the Tyrode solution and incubated for 20 minutes. By cooling the test tube with ice, the histamine free reaction was stopped and centrifuged at 3000 rpm for 5 minutes to separate the supernatant and pellets. Histamine was extracted through the extraction of the organic layer-water layer using the supernatant according to the Shore method. The amount of released histamine was stopped by adding 100 µl of 0.2% o-phthalehyde, incubating for 5 minutes, and then adding 140 µl of 0.5NH ₂ SO _4. The concentration of the reaction product was measured at the induced wavelength of 360 nm and the measured wavelength of 440 nm. It was. Histamine standard solution was obtained by using 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1 ㎍ / mL standard curve, and the quantitative and relative inhibitory concentration of histamine by substance by substituting the fluorescence intensity of the experimental group into this standard curve. Was calculated.

Experimental results on the inhibition of histamine induced by degranulation-inducing substance (compound48 / 80) in rat abdominal cells showed that staphylococcal and thorny skin had a statistically significant inhibitory effect on histamine release compared to the control group. Table 1).

TABLE 1 Effect of Bark and Thorn on the Histamine Release from Peritoneal Mast Cells Induced by Degranulation 48/80 (10µg / ml)

group Concentration (µg / ml) % Against control Neck skin 500 6.14 ± 0.39 ^*** Go away 500 15.75 ± 1.02 ^***

: ( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 3: Relaxation Test for Extraction Bronchoconstriction

SD was inhaled under anesthesia with CO ₂ gas and bleeding through the abdominal dome vein. The bronchus was carefully dissected and cut to include 4-5 cartilage rings of the bronchus per sample, opening the rings to connect two specimens with silk. Both sides were fixed with cell pins and suspended in a 15 ml tissue bath containing Krebs-henseleit solution (pH 7.4, 37 ° C.). Stabilization was maintained by maintaining the basic tension to 1g and then induced contraction with acetylcholine (ED ₅₀ : 10 ^-4 M). After shrinkage was stabilized, the sample was added cumulatively to determine the extent to which the test material reduced the shrinkage. The change in tension was measured with an isometric transducer and the results are as follows.

The direct relaxation effect on bronchial contractile material in vitro was investigated. Acetylcholine was used as a bronchial contraction inducer. As a result of measuring the effect of the test substance on the acetylcholine-induced contraction, the relaxation effect was not clear up to 3.5 mg / ml in both neck and bark, but significant relaxation was observed at 5 mg / ml (Table 2). .

Table 2 Relaxation Effects of Bark Bark and Prickly Pear Extract on Acetylcholine E ₅₀ Pretreated Bronchial Contraction

group density Shrink % Acetylcholine ED ₅₀ 10 ^-4 M 100 Neck skin 0.5mg / ml 96.75 ± 2.93 1mg / ml 99.33 ± 4.84 3.5mg / ml 94.75 ± 4.03 5mg / ml 48.25 ± 7.53 ^** Go away 0.5mg / ml 97.33 ± 2.67 1mg / ml 113.67 ± 6.84 3.5mg / ml 118.33 ± 6.12 5mg / ml 42.67 ± 6.35 ^***

( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 4 Measurement of IgE Production in vitro

Isolation of mesentric lymphocytes and spleen lymphocytes from balb / c 1 × 10⁶After each aliquot of 24 plates at cells / ml, ConA (2.5 μg / ml) and LPS (10 μg / ml) were added as active sources and incubated with samples adjusted to various concentrations for 24 hours. The amount of IgE secreted into the culture was measured by the sandwich ELISA method. IgE antibodies were diluted 1000-fold with 50 mM carbonate-bicarbonate buffer (pH 9.6) and dispensed into 100 μl in 96 well plates and incubated at 37 ° C. for 1 hour. After washing three times with PBS (PBS-Tween) containing 0.05% Tween 20, and blocked for 3 hours at 4 ℃ using 3% BSA (washing three times with PBS-Tween). Then, 100 μl of lymphocyte culture supernatant was added, incubated at 37 ° C. for 2 hours, washed, and then diluted 2000 times with a solution of biotin bound to the coated antibody and the IgE antibody having different epitopes. Incubate for 2 hours at 100 ml of avidin-HRP solution diluted 3x after 3 washes. Incubate at 37 ° C. for 30 minutes. After washing each well of the plate three times with PBS-Tween, substrate solution [0.006% H₂O₂After dissolving OPD (orthophenylenediamine) as a substrate in 0.2M citric acid buffer, pH 4.0), add 100µl to each well, and develop for 30 minutes.Stop color development with 4N sulfuric acid solution and use 490nm with ELISA reader. Absorbance is measured at.

As a result of measurement of IgE production in vitro using single medicinal herb extracts, the present inventors showed that the endothelial and spinal cord of the present invention reduced the production of IgE from the lymphocytes of the neck and spine of the mesentic lymph nodes (MLN) and splenic lymphocytes stimulated with ConA and LPS. Table 3.

[Table 3] Effects of Bark Bark Migogapi on IgE Production

accelerant group Concentration (µg / ml) % Against control Mesenteric lymph nodes ConA (2.5 μg / ml) Neck skin 125 74.48 ± 0.56 *** 250 71.69 ± 1.20 *** 500 81.66 ± 1.22 *** 1000 83.50 ± 0.93 *** Go away 125 74.96 ± 1.11 *** 250 76.38 ± 1.46 *** 500 75.85 ± 0.99 *** 1000 76.74 ± 0.76 *** LPS (10 μg / ml) Neck skin 125 82.32 ± 1.42 *** 250 83.23 ± 2.27 *** 500 92.22 ± 2.90 * 1000 82.75 ± 1.38 *** Go away 125 74.85 ± 1.45 *** 250 76.97 ± 3.33 *** 500 77.40 ± 0.94 *** 1000 85.54 ± 1.25 ** Spleen lymphocyte ConA (2.5 μg / ml) Neck skin 125 77.87 ± 1.81 *** 250 78.78 ± 0.39 *** 500 88.33 ± 1.69 ** 1000 83.86 ± 1.24 *** Go away 125 73.83 ± 0.96 *** 250 74.55 ± 0.35 *** 500 77.03 ± 1.56 *** 1000 84.34 ± 0.15 *** LPS (10 μg / ml) Neck skin 125 88.36 ± 1.06 ** 250 86.34 ± 1.10 ** 500 85.06 ± 1.44 ** 1000 83.18 ± 1.60 *** Go away 125 83.24 ± 0.99 *** 250 82.84 ± 0.88 *** 500 86.68 ± 1.79 ** 1000 92.19 ± 1.60

( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 5 Response to Vascular Permeability by Chemical Mediator in Animal Experiments with SD

The following experiments were conducted to investigate the activity of bark and thorns on the response to immediate allergy. Five SD rats were divided into 1 group and the control group and the experimental group, and the experimental group was orally administered with the same amount of saline. After 30 minutes, 1 ml of 1% Evans blue saline was injected intravenously into each animal, and 0.1 ml of saline containing histamine (10 ug) was immediately injected intradermally into the entire embryo. After 30 minutes, the blood was bleeded, the skin was peeled off, and the leaked pigment amount of the clean part was measured according to the method of Katayama et al. That is, after cleansing the cleansing area, the skin plate was dissolved in 1.2N KOH for 24 hours, 0.6NH ₃ PO ₄ and acetone were mixed in a ratio of 5:13, and Evans blue was extracted for 24 hours to absorb absorbance at 620 nm. Was measured. The response to vascular permeability by the sample was calculated by applying the OD value of the experimental group from a search line previously prepared with Evans blue standard solution.

As a result, in the control group, the residual amount of evans blue in the tissue was 26.98 ± 2.82µg / ml, and the oral administration of neck skin was 15.79 ± 2.31µg / ml. The response to permeability was shown to be significantly reduced compared to the control (Table 4).

[Table 4] Effects of Bark Skin and Prickly Pear on Response to Vascular Permeability by Chemical Intermediates

group Volume (mg / kg / day) Die evacuation (μg / mL) Control - 26.98 ± 2.82 Neck skin 500 15.79 ± 2.31 ^* Go away 500 23.94 ± 1.11

( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 6: Delayed Allergic Dermatitis Response

Picryl chloride (PC, SIGMA) was used as an antigen for experiments on the delayed allergic dermatitis response. That is, 7% PC ethanol solution was used as a sensitizing antigen, and 1% PC olive solution was used as a trigger antigen. Induction of connective dermatitis by PC was performed according to the method of Asherson and Ptak. In summary, 10 animals were divided into the experimental group and the control group, and the experimental group was orally administered once daily for 3 days before the experiment. On the day before the experiment, each group of mice was sensitized by applying 0.1 ml of 7% PC ethanol solution to the abdomen. After 7 days of sensitization, 0.02 ml of 1% PC olive solution was applied to both ears of the mouse to induce skin irritation.

Experimental results showed that the dilatation was 0.030 ± 0.010mm in the control group, 0.010 ± 0.003mm in the group of pelvic dermis and 0.016 ± 0.002mm in the group of thorny skin, and each sample was skin-induced by picryl chloride Inhibitory activity was investigated (Table 5).

Table 5 Effects of MDP and KOK Extracts on Delayed Allergic Dermatitis

group Volume (mg / kg / day) Ear swelling (mm) Control 0.030 ± 0.010 MDP 500 0.010 ± 0.003 KOK 500 0.016 ± 0.002

^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 7: Delayed Allergic Foot Edema Reaction

Facet cells were used as antigens for the delayed allergic foot edema reaction. Five mice per group were sensitized by intravenously administering 10 ⁶ cells of red blood cells intravenously, and 4 days later, 2.5 x 10 ⁶ red blood cells per mouse were injected bilaterally subcutaneously to induce inflammation. In order to investigate the inhibitory activity of the inflammation-induced samples, the test substance was administered orally once a day for 3 days before the test, and twice orally, immediately before and 6 hours after the onset of inflammation. Inflammatory inhibitory activity by the sample was measured by comparing the difference in the thickness of the foot before and after inflammation, measured by the dial gauge.

Experimental results showed that the foot edema rate of the control group was 4.80 ± 0.59, whereas that of the endothelial group was 3.17 ± 0.59, and that of the pelvic vertebrae group was 3.16 ± 0.42, which was significantly suppressed compared to the control group (p <0.05, Table 6).

TABLE 6 Effect of MDP and KOK extracts on delayed allergic foot edema reaction

group Volume (mg / kg / day) FPSI Control 4.80 ± 0.59 Neck skin 500 3.17 ± 0.59 Go away 500 3.16 ± 0.42 ^*

( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

Example 8 Measurement of Lung Throbarbituric Acid (TBA)

10 animals are assigned to each group and divided into normal group, control group and experimental group. Induced pulmonary edema in rats was injected very slowly through the vein of 0.1 ml per 100 g of the experimental animals according to the method of small intestine. Samples of the experimental group were orally administered 2ml 3 hours after 2% ethanol-xylene administration, and left at room temperature for 3 hours. Lung extraction from rats was performed after stunned by the laryngeal dislocation method under anesthesia, and was used for the experiment after removing the bronchus of both lungs. 0.5 g of the deceased lung was taken, and 5 ml of 0.05M phosphate buffer (pH7.4) was used for homogenization. After homogenization, 200 ul of the homogenate was added, and 200 ul of the 8.1% sodium dodecyl sulfate (SDS) solution was added to the homogenate. After mixing for 2 seconds, 1.5 ml of 20% acetic acid was added again and mixed vigorously for 5 seconds. Then, 1 ml of 1.2% TBA solution was added to each of the tubes, covered with glass beads, boiled in a water bath for 30 minutes, and then cooled at room temperature for 30 minutes. After that, only the supernatant was collected by centrifugation at 2000 rpm for 10 minutes, and the lung TBA value was measured by measuring the absorbance at 532 nm.

As a result of the experiment, after measuring lung edema with 2% xylene ethanol, the TBA reaction product contained in the lung was measured to be 3.10 ± 0.08µg / ml in the control group. As shown by the two samples, the results showed that the activity of significantly reducing the lung TBA (Table 7).

Table 7 Effect of MDP and KOK on Pulmonary TBA

group Volume (mg / kg / day) Lung Weight / Body Weight TBA (μg / ml) normal - 0.053 ± 0.00002 2.45 ± 0.08 Control - 0.0067 ± 0.0007 3.10 ± 0.09 MDP 500 0.0065 ± 0.0002 2.70 ± 0.15 ^* KOK 500 0.0064 ± 0.0002 2.34 ± 0.11 ^**

( ^* : P <0.05, ^** : P <0.01, ^*** : P <0.001)

As described in the above examples, the endothelial and spinal cords had antihistamine free inhibitory activity that effectively inhibits histamine from the peritoneal mast cells of the mouse, thereby inhibiting bronchial contraction relaxation and IgE production, which are symptoms induced by asthma and allergy. It showed effective activity in the back.

Claims (3)

A composition for the prevention and treatment of allergies and / or asthma, comprising, as an active ingredient, an effective amount of at least one extract selected from the group consisting of bark and thorny scabies with a pharmaceutically acceptable carrier. The allergy and / or asthma prevention according to claim 1, wherein the extracting solvent of the extract is an extracting solvent, preferably one solvent selected from purified water, preferably ethanol, methanol, butanol, ethyl acetate and acetone. And therapeutic compositions. According to claim 1, wherein the effective amount of the herbal extract is allergic and / or asthma, characterized in that the dose of thorny skin is 60gg 9g-27g / 1 day, the capacity of the neck skin 6-12g / 1day Prophylactic and therapeutic compositions.