KR101280431B1 - Anti-allergy Composition and Anti-bacteria Composition Using an Extract of Stichopus japonicus Orange - Google Patents

Abstract

The present invention discloses an anti-allergic composition and an antimicrobial composition using red sea ginseng extract. Red sea ginseng extract showed anti-granular inhibitory activity of mast cells and antibacterial activity against Staphylococcus aureus and Staphylococcus epidermidis.

Description

Anti-allergy Composition and Anti-bacteria Composition Using an Extract of Stichopus japonicus Orange}

The present invention relates to an anti-allergic composition and an antimicrobial composition using red sea ginseng extract.

Allergy is a reaction caused by immunological mechanisms caused by allergen, which is a foreign substance, which generally causes harmful damage to the body.

Allergic reactions are mediated by mast cells, which are widely distributed in organs of the whole body such as the skin, respiratory organs, mucous membranes of the gastrointestinal tract, around lymphatic vessels, around blood vessels, and in the brain, and are known as cells causing allergic reactions.

The allergic reaction is caused by the action of IgE on mast cells against house dust mites, pollen, animal dander, mold, egg albumin, milk proteins, and peanuts (Wills-Karp M., et al., Science, 282, pp 2258-61, 1998; Grunig G., et al., Science, 282, pp 2261-2263, 1998).

In detail, when allergens invade a living body, allergens are first recognized by antigen presenting cells, and these antigenicizing cells differentiate Th0 into Th2 cells by presenting allergens in the presence of IL-4. These differentiated Th2 cells secrete cytokines such as IL-4, IL-5, IL-13, IL-4, IL-5 and the like act on B cells to induce the production of IgE.

The generated IgE binds to the high-affinity IgE receptor FcεRⅠ of mast cells, and then, when exposed to the same allergen, allergens bind to IgE bound to the mast cells' FcεRⅠ and cause cross-linking of these receptors. It acts as a series of signaling reactions.

This signaling response induces degranulation of mast cells, releasing histamine, heparin, bradykinin, triptase, chymase, etc. In Arachidonic acid metabolites, Leukotrien and Prostaglandin, are newly synthesized and released. These substances are released by vasodilation, permeability, stimulation of nerve endings, and mucus secretion. It causes a series of allergic reactions such as hyperactivity and contraction of smooth muscle.

Since the allergic reaction is caused by degranulation of mast cells, a substance that inhibits degranulation of mast cells has a potential as a therapeutic agent for allergic diseases (Jeong-pil Choi et al., J. Korean Soc. Appl. Biol. Chem. 48 (4), 315-321 (2005).

The Staphylococcus aureus (Staphylococcus aureus) and Staphylococcus epidermidis (Staphylococcus epidermidis) is a skin flora payioh Jeans Streptococcus (Streptococcus pyogenes ) are representative strains that can cause purulent infections, cellulitis or general inflammation on the skin.

Antimicrobial agents such as tetratracycline, erythromycin, and azaelaic acid are used for such skin infections, but side effects such as the emergence of resistant strains, human toxicity such as skin hypersensitivity reactions, and photosensitivity are reported. Recently, research on antimicrobial agents using natural products has been actively conducted.

The present invention is completed by confirming that the red sea ginseng extract inhibits the degranulation of mast cells and shows antibacterial activity against Staphylococcus aureus and Staphylococcus epidermidis.

An object of the present invention to provide an anti-allergic composition using red sea ginseng extract.

Another object of the present invention to provide an antimicrobial composition using the red sea ginseng extract.

The specific object of the present invention will be presented below.

In one aspect, the present invention relates to an anti-allergic composition comprising red sea ginseng extract as an active ingredient.

The inventors of the present invention treated the mast cell line RBL-2H3 (rat basophilic leukemia) activated with IgE and antigen (DNP-BSA) using red sea ginseng extract, as confirmed in the following examples and experimental examples, beta-hex. It was confirmed that the production of sosamidadase (β-hexosaminidase) is inhibited. The mast cell line RBL-2H3 has an IgE antibody receptor (FcεRI) on its surface, and when activated with IgE and antigen, degranulation is induced to secrete mediators of allergic reactions such as histamine. Hexosaminidase is known as an indicator of degranulation [Planta Med. 64: 577-578 (1998).

In the present specification, the "red sea ginseng" is scientific name Stichopus japonicus Points to sea cucumber that is orange. Specifically, the body color is red to yellowish brown or dark reddish brown, the abdomen is red, the poly vesicle is thin and long, the tip is sharp, and the contraction is severe, so the body is similar in shape to a sphere. It can be defined as sea cucumbers having a gelatinous body of about 25㎛ thickness.

In the present specification, the "extract" refers to alcohols having 1 to 5 carbon atoms such as methanol, ethanol, butanol, acetone, hexane, ethyl acetate, chloroform, dichloromethane, water or a mixed solvent thereof, regardless of the extraction method. It is understood as meaning including the extract obtained by extracting the extract and the extract is fractionated with the solvents listed above. Regardless of the extraction method, it is to be understood that the extraction method may be applied to any method such as cooling, refluxing, heating, ultrasonic radiation, and the like, as long as the extraction object is extracted through immersion in the extraction solvent. Nevertheless, the "extract" preferably means that the red sea ginseng which is the extraction target is obtained by extracting with water, ethanol, acetone or a mixed solvent thereof. In addition, the meaning of the "extract" includes the extract of the purified form through filtration, etc., the concentrated liquid extract from which the extraction solvent is removed, and the solid extract from which the extraction solvent is removed.

In the present specification, the above-mentioned "active ingredient" means an ingredient that exhibits the desired activity alone or can exhibit activity together with a carrier that is itself inactive.

In addition, in the present specification, the "anti-allergic" means the prevention, treatment, suppression, or delay of allergic diseases.

In the present specification, the "allergic disease" is caused by degranulation of mast cells. Refers to clinical symptoms caused by allergic reactions, which may include contact dermatitis, atopic dermatitis, eczema, pruritus, hay fever, asthma, bronchitis, urticaria, vasculitis, rhinitis, gastroenteritis, diarrhea, Interstitial pneumonia, arthritis, ophthalmitis, conjunctivitis, neuritis, otitis media, granulomatosis, encephalomyelitis, cystitis, laryngitis, purpura, food allergies, insect allergies, drug allergies, metal allergies, anaphylactic shock (Bierman CW, et al. (Eds.) Allergy, asthma, and immunology from infancy to adulthood. Page xvii, Saunders, Philadelphia, 1996).

Meanwhile, the anti-allergic composition of the present invention may include the red sea ginseng extract, which is an active ingredient, in any amount (effective amount) as long as the anti-allergic extract can exhibit an improvement in allergic diseases intended to be treated according to the use, formulation, and formulation purpose. Typical effective amounts will be determined within the range of 0.001% to 15% by weight, based on the total weight of the composition. An "effective amount" as used herein refers to an amount of an active ingredient in a mammal, preferably a human subject to which it is applied, that can induce improvement, treatment, or suppression / delay of the development of such pathological symptoms. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

The subject to which the composition of the present invention can be applied (prescription) is preferably a mammal and a person, particularly a human.

In another aspect, the present invention relates to an antimicrobial composition comprising red sea ginseng extract as an active ingredient.

In the present specification, the meaning of "antibacterial" means a property of inhibiting the growth or proliferation of bacteria or killing the bacteria, wherein the bacteria are preferably Staphylococcus confirmed the antimicrobial activity of the red sea ginseng extract in the following experimental example Aureus and Staphylococcus epidermis.

In the antimicrobial composition of the present invention, the red sea ginseng extract, which is an active ingredient thereof, may be included in the antimicrobial composition of the present invention in any amount appropriate according to the application mode or degree to which antimicrobial activity is required as long as it can exhibit an antimicrobial effect. Regardless of the application mode or the degree of antimicrobial activity required for achieving sufficient antimicrobial effect, red sea ginseng extract is usually 0.1% by weight or more, preferably 3% by weight, based on the total weight of the composition in the antimicrobial composition of the present invention. It will be okay if it is included.

The anti-allergic composition and antimicrobial composition of the present invention (hereinafter "composition of the present invention") can be used as a pharmaceutical composition in specific embodiments.

Pharmaceutical compositions of the present invention, in addition to the active substance, include pharmaceutically acceptable carriers, excipients, and the like, oral formulations (tablets, suspensions, granules, emulsions, capsules, syrups, etc.), parenteral formulations (sterile injectable aqueous or Oily suspensions), topical formulations (solutions, creams, ointments, gels, lotions, patches) and the like.

As used herein, "pharmaceutically acceptable" means that the subject of application (prescription) does not have more toxicity (adequately low toxicity) to which the subject of application (prescription) is adaptable without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (such as corn starch, potato starch, etc.), cellulose, derivatives thereof (such as sodium carboxymethyl cellulose, ethylcellulose, etc.) malt, gelatin, talc, solids Lubricants (e.g. stearic acid, magnesium stearate, etc.), calcium sulfate, vegetable oils (e.g. peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerin, etc.), alginic acid, emulsifiers (e.g. TWEENS), wetting agents (e.g. Sodium lauryl sulfate), colorants, flavoring agents, tableting agents, stabilizers, antioxidants, preservatives, water, saline, phosphate buffer solutions and the like. The carrier may be selected from one or more of suitable pharmaceutical formulations according to the formulation of the pharmaceutical composition of the present invention.

Excipients may be selected and used according to the formulation of the pharmaceutical composition of the present invention, for example, when the pharmaceutical composition of the present invention is prepared with an aqueous suspending agent, suitable excipients are sodium carboxymethyl cellulose, methyl cellulose, hydropropylmethylcellulose And suspending agents and dispersing agents such as sodium alginate and polyvinylpyrrolidone. Suitable excipients when prepared from injection solutions include Ringer's solution, isotonic sodium chloride, and the like.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in some cases, can be administered topically.

The daily dose of the pharmaceutical composition of the present invention is usually 0.001 to 150 mg / kg body weight, and may be administered once or several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as route of administration, age, sex, weight, and patient's severity of the patient, the dose is limited in any aspect to the scope of the present invention Should not be understood to be.

The composition of this invention can be grasped | ascertained as a food composition in another specific aspect.

The food composition of the present invention can be produced as a health supplement food, a special nutrition supplement food, a functional beverage and the like.

The food composition of the present invention may contain sweetening agents, flavoring agents, physiologically active ingredients, minerals and the like in addition to the active ingredients thereof.

Sweetening agents may be used in an amount that sweetens the food in a suitable manner, and may be natural or synthetic. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.

As the physiologically active substance, catechins such as catechin, epicatechin, gallocatechin, epigallocatechin, vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, riboflavin, and the like can be used.

As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium and zinc can be used.

In addition, the food composition of the present invention may contain preservatives, emulsifiers, acidifiers, thickeners and the like as needed in addition to the above sweeteners.

Such preservatives, emulsifiers and the like are preferably added in a very small amount as long as they can attain an application to which they are added. The term " trace amount " means, when expressed numerically, in the range of 0.0005% by weight to about 0.5% by weight based on the total weight of the food composition.

Examples of the preservative which can be used include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid).

Examples of the emulsifier which can be used include acacia gum, carboxymethyl cellulose, xanthan gum, pectin and the like.

Examples of the acidulant that can be used include acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. Such an acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.

Agents that may be used include suspending agents, sedimentation agents, gel formers, bulking agents and the like.

In addition, herbal medicines may be added to enhance flavor and palatability and to add other functionalities (such as prevention of arthritis or osteoporosis). Herbal medicines that may be added include tofu extract, fast extract, antler extract, safflower extract, earthworm extract, and succinate Extracts, tortoiseshell extracts, cornus extracts, goji berry extract, licorice extract, donkey extract, brown root extract, kangjinhyang extract, haphwanpi extract, mountain rump extract, lump extract, ginseng extract and the like can be exemplified.

As described above, the present invention can provide an anti-allergic composition and an antimicrobial composition using the red sea ginseng extract.

The anti-allergic composition and the antimicrobial composition of the present invention can be commercialized and used in medicine, food, and the like.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these embodiments.

< Example > Red Sea Ginseng  Preparation of extract

Example 1 Preparation Example 1 of Red Sea Ginseng Extract

5% weight of 70% (v / v) ethanol was added to the washed red sea ginseng crushed product, which was extracted for 24 hours, filtered through Whatman NO. 1, the filtrate was concentrated under reduced pressure, and freeze-dried. Got.

Example 2 Preparation Example 2 of Red Sea Ginseng Extract

Add 5 times the weight of water to the washed red sea ginseng crushed water, heat it for 24 hours (100 ℃), extract the hot water, filter it with Whatman NO.1, and concentrate the filtrate under reduced pressure and freeze-dried to extract powdered red sea ginseng. Got.

Example 3 Preparation Example 3 of Red Sea Ginseng Extract

Add 5 times the weight of 50% (v / v) acetone to washed red sea ginseng crushed, extract for 24 hours, filter with Whatman NO.1, and concentrate the filtrate under reduced pressure and freeze-dried to make powdered red sea ginseng. An extract was obtained.

< Experimental Example > Red Sea Ginseng  Extract Anti-allergy  Activity and Antimicrobial Activity Assessment

Experimental Example 1 Anti-allergic Activity of Red Sea Ginseng Extract

After dispensing 5 × 10 ⁵ / well in a 24 well plate and treated with cells monoclonal antibody IgE (0.1 ㎍ / ml) of the rat after 24 hours and incubated at 37 ℃ for 4 hours. After incubation, 500 μl of 1 × PIPES I buffer (sigma) was added and washed three times, followed by 180 μl of 1 × PIPES II buffer (sigma) and incubated at 37 ° C. for 10 minutes. Each sample of Example was treated by concentration (0, 50, 250, 1250 μg / ml), and then reacted at 37 ° C. for 20 minutes, and then treated with antigen DNP-BSA (0.1 μg / ml), followed by 20 at 37 ° C. Reacted in minutes. After completion of the reaction by placing in ice for 10 minutes, transfer from 24 well plates to 96 well plates, add 25 µl each, add 25 µl of substrate (pNAG), incubate for 1 hour at 37 ° C, and stop solution (0.1M NaHCO ₃ , 0.1M). Na ₂ CO ₃ ) was added and measured at 405 nm using an ELISA reader.

The results are shown as mean ± standard deviation in Table 1 below as a free inhibitory effect of beta- hexosaminidase.

Free Inhibitory Activity of Beta-Hexosaminidase Treatment concentration (µg / ml) Extract of <Example 1> The extract of Example 3 100 64.2 ± 2.76 46.0 ± 5.53 50 50.5 ± 5.09 35.0 ± 8.61 25 41.1 ± 4.43 27.5 ± 7.02 12.5 37.0 ± 5.17 19.8 ± 4.57 6.25 33.3 ± 5.72 19.0 ± 6.15 Control 0.0 ± 3.03 0.0 ± 1.86

The results of Table 1 show that red sea ginseng extract showed free inhibitory activity of beta-hexosaminidase in a concentration-dependent manner.

Experimental Example 2 Antimicrobial Activity of Red Sea Ginseng Extract

In order to detect the antimicrobial activity of red sea ginseng extract, a paper disc method was used in this experiment. In other words, each strain was taken as platinum and inoculated in 10 ml broth (TSB) and incubated at 37 ° C. for 18 hours using Spetrophotometer (Ultrospec 2100 pro, Amersham Biosciences, USA). 620nm) and the absorbance was adjusted and uniformly mixed in a culture dish in which the medium was dispensed according to the pour-plate method and then hardened at room temperature. After sterilizing the paper disc on this medium to the number of samples and adhered to each sample was absorbed at the concentration of [Table 2] below. After culturing at 37 ° C for 24 hours, the size of the clear zone (mm) produced around the disc was measured to compare the antimicrobial activity.

The results are shown in [Table 2].

Antimicrobial activity

The results of Table 2 show that the red sea ginseng extract showed antibacterial activity on Staphylococcus aureus and Staphylococcus epidermidis in a concentration-dependent manner.

Claims (8)

Antihistamine composition comprising red sea ginseng extract as an active ingredient.
The method of claim 1,
The red sea ginseng extract is antihistamine composition, characterized in that obtained by extracting red sea ginseng with water, ethanol, acetone or a mixed solvent thereof.
The method according to claim 1 or 2,
The composition is an antihistamine composition, characterized in that the pharmaceutical composition.
delete As an antimicrobial composition comprising red sea ginseng extract as an active ingredient,
The antimicrobial activity is characterized in that the antimicrobial activity against Staphylococcus aureus or Staphylococcus epidermis.
The method of claim 1,
The red sea ginseng extract is antimicrobial composition, characterized in that obtained by extracting red sea ginseng with water, ethanol, acetone or a mixed solvent thereof.
delete The method according to claim 5 or 6,
The composition is an antimicrobial composition, characterized in that the pharmaceutical composition.