KR20050084725A - Composition comprising an extract of anthriscus sylvesstris for prevention and treatment of inflammatory or allergy diseases - Google Patents

Abstract

The present invention relates to a composition comprising an extract of Anthriscus sylvestris Hoffm. Having anti-inflammatory and anti-allergic activity. Specifically, the extract of the present invention inhibits the production of PAF which causes various inflammations and allergic diseases. In addition, by inhibiting the activity of cyclooxygenase-2 and lipoxygenase (5-LO) to prevent the production of eicosanoid (eicosanoid) and at the same time inhibits the release of histamine (histamin) from mast cells, the present invention The composition containing the extract of Jeonho may be usefully used as a medicine or health functional food for the prevention and treatment of various inflammatory or allergic diseases.

Description

Composition comprising an extract of Anthriscus sylvesstris for prevention and treatment of inflammatory or allergy diseases

The present invention relates to a composition comprising an extract of Anthriscus sylvestris Hoffm. Having anti-inflammatory and anti-allergic activity.

Inflammation is the defense of biological tissue against local injuries. In other words, in response to various harmful stressors, the biodefense reaction to remove the injury caused by the stimulus and restore the original state is an inflammatory response. Stimulation of inflammation includes infection, chemical and physical stimuli, and bio-constitutive factors related to the inflammatory response include low-molecular or polymer chemicals such as free radicals, proteins, sugars, and lipids, plasma, blood cells, and blood vessels. And connective tissue. The process of inflammation can usually be divided into two types: acute and chronic inflammation. Acute inflammation is a short-term reaction within a few days, and plasma components or blood cells are associated with the removal of foreign bodies by posting a microcirculatory system. Chronic inflammation has a long duration, and tissue growth is seen.

Allergic diseases refer to pathological phenomena due to antigen-antibody reactions. Serum diseases, high fever, and bronchial asthma are typical. Nowadays, however, there is a tendency to be considered a disease that can be caused by allergies, which is very similar to the disease in which the antigen-antibody reaction is proved, but the antigen-antibody reaction is not proved, or the involvement of the antigen-antibody reaction is not proved, but the possibility It also includes things. Allergic diseases include organs such as bronchial asthma, hyperthermia, vasomotor neuritis, hypertrophic rhinitis, allergic bronchitis, and Rheoplung syndrome (transient lung infiltration). Diarrhea, allergic stomatitis, enteropolitan purpura, circulatory system such as nodular periarteritis, obstructive endocarditis, angina pectoris, endocarditis, etc. Urticaria in the skin relationship, quinque edema (vascular edema), nodular erythema, purpura The eye and the like are flictene, sympathetic ophthalmitis, allergic conjunctivitis, allergic keratitis, and the like, and diffuse glomerulonephritis and migraine headaches.

In addition, allergic diseases are classified into immediate type (anaphylaxis type) and delayed type (遲發型: tuberculin type). Immediate forms include high fever, bronchial asthma, urticaria, quinque edema, digestive tract allergies, and delayed forms include contact dermatitis and drug allergies. There is some agreement on the therapeutic effect of the reactor and the drug. Currently used drug therapies include antihistamines, corticosteroids, sympathetic nerve excipients such as epinephrine and ephedrine, and parasympathetic antiparalytic agents such as sulfate atropine or lotex, but all are popular treatments rather than radical therapy. Development is urgently needed.

Key mediators to induce inflammatory and allergic diseases include phospholipase A2 and cyclooxygenase, such as prostaglandines, leukotriens, and platelet activating factor (PAF). It is produced from the precursor arachidonic acid by cyclooxygenase) and lipoxygenase.

Leukotriene constitutes a group of locally acting hormones produced in vivo from arachidonic acid, and important leukotrienes include leukotriene B4 (LTB4), leukotriene C4 (LTC4), leukotriene D4 (LTD4) and leukotriene E4 (LTE4). Biosynthesis of these leukotrienes begins by the enzyme 5-lipoxygenase acting on arachidonic acid to produce an epoxide known as leukotriene A4, which is converted to other leukotrienes (LTB4, LTC4, LTD4, LTE4) by a subsequent enzymatic reaction step. Is switched. Leukotriene is involved in allergic and allergic reactions such as asthma, chronic bronchitis and related obstructive airway diseases, allergic rhinitis, contact dermatitis, allergic conjunctivitis, inflammation of arthritis or inflammatory bowel disease, pain, etc. It is known.

Anthriscus sylvestris Hoffm. Of the present invention is a perennial herb that is commonly found in valleys all over the country and is distributed in Japan, Manchuria, and China. Stretches. Flowers bloom in July-August and reddish purple. Many flowers hang on the conical inflorescences at the end of stems, brown hairs on flower shafts, calyxes are divided into 5 pieces, and the fragments are egg-shaped and 5 petals. It is also called horse riding horse or red horse riding horse, and it improves blood circulation, releases blood, releases heat, releases poison, and cramps. It is known to be effective in relieving pain and relieving pain. It is also used to relieve overworked diseases, sore muscles and bone nodes, sore symptoms, bruises, joint pain, postoperative pain, and snake bites (Bae Gi-hwan, Korean medicinal plants, Kyohaksa, p204, 2000).

Recently, efforts have been made to find active ingredients from relatively low-toxic substances derived from natural products. For example, Korean Patent Publication No. 1999-0039985 discloses a novel lignan-based compound, angeloyl grapephytotoxin, isolated from the previous issue. The anti-tumor effect of Angeloyl podophyllotoxin and its extraction method have been disclosed, and Korean Patent Publication No. 2003-0078565 discloses chronic bronchial asthma and heart disease containing mixed extracts of licorice, urea, Schisandra chinensis, Astronomical dong and Jeonho as active ingredients. Therapeutic compositions have been disclosed, but the anti-inflammatory and anti-allergic effects of the composition comprising the extract of Jeonho are not known.

In this regard, the present inventors inhibited the production of prostaglandin D ₂ (PGD ₂ ) and COX-2 in mast cells of rat bone marrow, and at the same time strongly inhibits the production of leukotriene C4 (LTC4) exhibits anti-inflammatory and anti-allergic effects By confirming that the present invention was completed.

An object of the present invention is to provide a pharmaceutical and nutraceutical for the prevention and treatment of inflammatory or allergic diseases containing as an active ingredient extract of Jeonho having excellent anti-inflammatory and anti-allergic activity.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory or allergic diseases containing the extract of Anthriscus sylvestris Hoffm. As an active ingredient.

The extract is water or an organic solvent such as a polar solvent of lower alcohols such as methanol and ethanol, a non-polar solvent of hexane, ethyl acetate, dichloromethane, or a mixed solvent thereof, preferably soluble in a mixed solvent of methanol and dichloromethane. Contains extracts.

In addition, the Jeonho may use an outpost (whole grass), roots, stems or leaves, preferably roots.

The inflammatory or allergic diseases include rheumatoid arthritis, bronchial asthma, acute pain, chronic pain, neuropathic pain, pain such as joint pain, inflammatory bowel disease, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, allergic asthma and the like do.

Hereinafter, the present invention will be described in detail.

Whole extract of the present invention can be obtained as follows.

The extract of Jeonho of the present invention is dried roots, leaves or stems of preferably dried Choho, preferably by cutting the roots of about 1 to 15 times the dry weight, preferably about 2 to 5 times the volume of water or methanol, ethanol, etc. Organic solvents such as polar solvents of lower alcohols, non-polar solvents of hexane, ethyl acetate, dichloromethane or mixed solvents thereof, preferably mixed solvents of methanol and dichloromethane at about 1 day at an extraction temperature of 0 to 70 ° C. The extraction method of the ultrasonic extraction, reflux extraction and the like for 10 days, preferably about 2 to 7 days may be repeated once to 5 times, preferably 2 to 5 times to obtain an organic solvent extract, The extract is suspended in a polar solvent such as water or methanol, and then it is about 1 to 100 times the volume of the suspension, preferably about 1 to 20 times the volume of hexane, ethyl acetate, chloroform, dichloro Nonpolar solvent such as carbon is added, more preferably dichloromethane is added to the polar solvent soluble layer, and the nonpolar solvent soluble layer is fractionated once to 10 times, preferably 2 to 5 times to prepare a nonpolar solvent soluble part and a polar solvent soluble. Wealth can be obtained.

It is also possible to further carry out conventional fractionation processes (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis, 3rd Ed. , Pp 6-7, 1998).

The present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory or allergic diseases containing Jehoho extract obtained by the above manufacturing process as an active ingredient.

The pharmaceutical composition for the prevention and treatment of inflammatory or allergic diseases of the present invention comprises 0.1 to 50% by weight of the extract based on the total weight of the composition.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100mg / kg, preferably 0.001 to 10mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The extract of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The present invention provides a dietary supplement comprising the extract and a food-acceptable food supplement additive exhibiting a prophylactic and improving effect of an inflammatory or allergic disease. Examples of the food to which the extract of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to food or beverages for the purpose of preventing and improving inflammatory or allergic diseases. At this time, the amount of the extract in the food or beverage may be added in 0.01 to 15% by weight of the total food weight, the health beverage composition may be added in a ratio of 0.02 to 5g, preferably 0.3 to 1g based on 100ml. .

The health functional beverage composition of the present invention is not particularly limited to other ingredients except for having the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the extracts of the present invention may contain flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1. Preparation of Jeonho Extract

3 kg of Underground (Anthriscus sylvestris) was purchased from Gyeongdong Market, powdered, 8 liters of a mixed solvent of methanol-dichloromethane (1: 1, v / v) was added and extracted for 1 week at 20 ° C, followed by vacuum filtration. The liquid was recovered. This process was repeated three times to collect the supernatant, and then concentrated under reduced pressure to obtain 260 g of jeonho extract.

Example 2. Preparation of Whole Lake Dichloromethane Fraction

260 g of the extract of Jeonho obtained in Example 1 was suspended in 5 L of distilled water, and then dissolved in an equal amount of dichloromethane. Then, only the components dissolved in the dichloromethane layer were separated and dried in vacuo. This process was repeated three times to give 89 g of dichloromethane fractions.

Experimental Example 1. Prostaglandin D ₂  Measure impact on biosynthesis

In order to examine the effect on the production of prostaglandin D ₂ (PGD ₂ ) of the extract of the present invention, the following experiment was performed.

That is, mouse bone marrow-derived mast cells (BMMC) from BALB / C mice were obtained from Murakami et al. (Murakami et al., J. Biol. Chem., 269 , pp22269-22275). , 1994), isolated from bone marrow, 50% WEHI-3 medium (conditioned medium, 10) containing culture supernatant of IL-3 producing cells, WEHI-3 cells (provided by Professor Kudo Ichiro, School of Pharmacy, Showa University, Japan). Cultured in% FCS). After 3 weeks of incubation, mast cells were treated with SCF / LPS / IL-10 mixed stimulant (Sigma) for 30 minutes, and PGD ₂ of supernatant after cell stimulation was analyzed using EG (Enzyme) using PGD ₂ assay kit (Cayman). measured by linked immunoassay. At this time, in order to suppress the production of PGD ₂ produced by COX-1 (cyclooxygenase-1), BMMC, which had been pretreated with 10 μg / ml of aspirin 1 hour before, was used for BMMC. After pre-treatment for a minute, the amount of PGD ₂ produced by adding a stimulant was measured. In addition, the production of PGD ₂ by COX-1 was measured after 1 hour after treatment with SCF / LPS / IL-10 mixed stimulant (Sigma, USA).

As a result, it was confirmed that Jehoho extract of the present invention significantly inhibited PGD ₂ production in a dose-dependent manner.

Experimental Example 2. Determination of the effect on the production of leukotriene

In order to determine the effect on leukotriene C4 (LTC4) production of Jeonho extract of the present invention was carried out as follows.

That is, mouse bone marrow-derived mast cells (BMMC, mouse bone marrow-derived mast cells) were cultured as in Experimental Example 1. After 3 weeks of culture, mast cells were treated with SCF / LPS mixed stimulant (Sigma, USA) for 30 minutes, and then LTC4 production amount of supernatant after cell stimulation using LTC4 assay kit (Cayman) and EIA (Enzyme linked immunoassay). Was measured. Jeonho extract was pre-treated for 15 minutes in advance and measured the amount of LTC4 produced by adding a stimulant.

As a result, it was confirmed that the Jehoho extract of the present invention significantly inhibited the production of LTC4 in a dose-dependent manner.

One example of the formulation of the pharmaceutical composition comprising the extract of the present invention, but the present invention is not intended to limit it, but is intended to explain in detail.

Formulation Example 1 Preparation of Powder

20 mg of the extract of Example 1

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

10 mg of extract of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

10 mg of the extract of Example 2

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

10 mg of extract of Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

20 mg of the extract of Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding lemon flavor appropriately, mixing the above components, adding purified water, adjusting the whole to 100 ml by adding purified water, and then filling into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

1000 mg of extract of Example 2

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic 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 mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

1000 mg of jeonho extract of Example 1

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is mixed with a relatively suitable component for a preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the extract of the present invention is a lysoPAF-acetyltransferase for synthesizing PAF from lyso PAF, which is a precursor of platelet activating factors that cause various inflammation or allergic diseases. ) Inhibits the activity of the enzyme, inhibits the production of PAF, and inhibits the activities of cyclooxygenase-2 and lipoxygenase (5-LO), showing excellent anti-inflammatory and anti-allergic activity. The composition can be usefully used as a pharmaceutical and nutraceutical for the prevention and treatment of inflammatory or allergic diseases.

Claims (6)

A pharmaceutical composition for the prevention and treatment of inflammatory or allergic diseases containing the extract of Anthriscus sylvestris Hoffm. As an active ingredient. The pharmaceutical composition according to claim 1, wherein the extract is an extract available in a mixed solvent of methanol and dichloromethane. The pharmaceutical composition according to claim 1, wherein an outpost, root, stem or leaf of Jeonho is used. The method of claim 1, wherein the inflammatory or allergic disease is rheumatoid arthritis, bronchial asthma, acute pain, chronic pain, neuropathic pain, pain such as joint pain, inflammatory bowel disease, allergic rhinitis, allergic conjunctivitis, allergic dermatitis or A pharmaceutical composition that is allergic to asthma. A health functional food comprising the Jeonho extract of claim 1 and a foodstuff acceptable food supplement additive, which exhibit a prophylactic effect of inflammation or allergic disease. The health functional food of Claim 5 which is a health drink.