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WO2018105926A1 - Composition anti-allergique contenant un extrait d'herbe médicinale composite fermenté par des lactobacilles à titre de principe actif - Google Patents

Composition anti-allergique contenant un extrait d'herbe médicinale composite fermenté par des lactobacilles à titre de principe actif Download PDF

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WO2018105926A1
WO2018105926A1 PCT/KR2017/013239 KR2017013239W WO2018105926A1 WO 2018105926 A1 WO2018105926 A1 WO 2018105926A1 KR 2017013239 W KR2017013239 W KR 2017013239W WO 2018105926 A1 WO2018105926 A1 WO 2018105926A1
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Prior art keywords
extract
lactobacillus
allergic
lactic acid
acid bacteria
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Korean (ko)
Inventor
마진열
유재명
조원경
양주혜
박광일
박은희
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Korea Institute of Oriental Medicine KIOM
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Korea Institute of Oriental Medicine KIOM
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/489—Sophora, e.g. necklacepod or mamani
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A61K36/232—Angelica
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/304—Foods, ingredients or supplements having a functional effect on health having a modulation effect on allergy and risk of allergy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine

Definitions

  • the present invention relates to a composition for the prophylaxis or treatment of allergic diseases, including a fermented extract of lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit, and Polygonum cuspidatum .
  • a composition for the prophylaxis or treatment of allergic diseases including a fermented extract of lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit, and Polygonum cuspidatum .
  • Prophylaxis against atopic dermatitis, asthma, allergic rhinitis or allergic conjunctivitis including lactobacillus fermented extracts of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum
  • the method for preventing or treating an allergic disease comprising administering the composition to a subject.
  • An allergic disease refers to a disease caused by immune hypersensitivity reactions.
  • An allergic reaction is a complex reaction involving various cells and mediators and also causes various diseases. Therefore, it is expected that the treatment of various diseases through the control of allergic reactions is ongoing.
  • Allergic diseases include atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, allergic dermatitis, and the like.
  • Atopic dermatitis is a worldwide disease that can occur at any age, with 70% to 95% occurring in infants under 5 years of age.
  • Korea the social and medical problems caused by atopic dermatitis are already at a serious level.
  • Koreans about 15% of Koreans are atopic dermatitis patients, and 18 to 22 of 100 infants aged 1 to 4 years are known as atopic patients.
  • the number of children with adult atopic dermatitis has also increased, and in severe cases, even social problems such as employment and marriage have become serious social problems. In these cases, severe mental stress can lead to extreme avoidance and depression.
  • atopic dermatitis has become a serious social and medical problem, there is no effective treatment to date.
  • Allergic rhinitis is the most common allergic disease, and its incidence is gradually increasing, affecting the quality of life, and it has become a social and economic problem due to a decrease in production capacity and an increase in the cost of treatment. There is no situation.
  • Currently used drug therapy is a basic treatment for allergic rhinitis, but recurrences are common and serious side effects when the drug is discontinued.
  • Immunotherapeutic may be the only treatment that can restore the only modified immune system, but there are cost problems. .
  • Allergic conjunctivitis is an acute inflammatory disease of the conjunctiva (white egg) caused by certain allergens.Allergens such as pollen in the spring, dust in the air, and dandruff in animals come into contact with the conjunctiva of the eye, causing mast cells, eosinophils or basophils. When an allergic reaction through an immune cell such as a device is induced, various inflammatory substances such as histamine, inflammatory cytokines, and inflammatory lipid metabolites (Prostaglandins & Leukotrienes) are secreted to cause an inflammatory response of the conjunctiva.
  • Such allergic conjunctivitis can be treated with glucocorticoid steroid eye drops.However, these anti-inflammatory drugs can cause sudden systemic weakness, fever, increased intraocular pressure due to acute adrenal insufficiency, etc. Side effects such as glaucoma and cataract may occur, and there is a need for an antiallergic agent without such problems.
  • the present inventors have made intensive efforts to develop a drug that is effective in the treatment of allergic diseases and ensures safety derived from natural products. It was confirmed that the antiallergic effect was remarkably superior to the herbal medicine alone, and thus had an excellent allergic disease prevention or treatment effect. Thus, it was confirmed that the lactic acid bacteria fermentation complex herbal extract can be effectively used for the prevention or treatment of allergic diseases, and completed the present invention.
  • composition for the prevention or treatment of allergic diseases including the extract of lactic acid bacteria fermentation of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum To provide.
  • Another object of the present invention comprising administering to a subject a composition comprising a fermented extract of lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum To provide a method for preventing or treating allergic diseases.
  • Another object of the present invention is to ferment lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit , and Polygonum cuspidatum for the manufacture of a medicament for preventing or treating allergic diseases. It is to provide a use of the extract.
  • Another object of the present invention to prepare a health functional food or cosmetics to improve the allergic diseases, of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum It is to provide a use of lactic acid bacteria fermented extract.
  • Still another object of the present invention is to prevent, ameliorate or treat allergic diseases, including extracts of fermented lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum . It is to provide a use.
  • composition comprising the lactobacillus fermentation extracts of Sophora Root, Angelica gigas , Arctium lappa fruit, and Polygonum cuspidatum of the present invention is more effective in producing allergens than conventionally known antiallergic extracts. Since it can be effectively suppressed, it can be usefully used as a composition for preventing or treating allergic diseases.
  • Fig. 2 is a graph comparing the inhibitory effect of ⁇ -hexosaminidase from mast cells on F-PASA extract and KIOM-MA128. (## P ⁇ 0.01 for each concentration group of KIOM-MA128)
  • Figure 3 is a graph plotting the inhibitory effect of ⁇ -hexosaminidase from mast cells of F-PASA extract, woofer fermentation extract, Angelica fermentation extract, and Ginseng fermentation extract. (** P ⁇ 0.01 for DNP-IgE / DNP-HSA group, ## P ⁇ 0.01 for F-PASA group)
  • Fig. 5 is a graph illustrating the comparison of the inhibitory effect of TNF- ⁇ production from mast cells of F-PASA extract, woofer fermentation extract, Angelica fermentation extract, and Ginseng fermentation extract. (** P ⁇ 0.01 for DNP-IgE / DNP-HSA group, ## P ⁇ 0.01 for F-PASA group)
  • FIG. 6 is a graph illustrating the inhibitory effect of IL-4 (Interleukin 4) production of F-PASA extract. (** P ⁇ 0.01 compared to DNP-IgE / DNP-HSA group)
  • FIG. 9 is a graph showing the degree of cytotoxicity induced by F-PASA extract.
  • FIG. 11 is an electrophoretic photograph of the Western blot showing the degree of inhibition of the activity of the F-PASA extract on phosphorylation of the signaling proteins Syk, Fyn, and Lyn of the Fc ⁇ RI receptor signaling system. (** P ⁇ 0.01 compared to DNP-IgE / DNP-HSA group)
  • FIG. 13 is an electrophoretic photograph of Western blot results showing the degree of inhibition of the activation of cytokine production-related mechanisms (phosphorylation of JNK, ERK1 / 2, p38, Akt) of F-PASA extract. (* P ⁇ 0.05 and ** P ⁇ 0.01 compared to DNP-IgE / DNP-HSA group)
  • the present invention includes allergic disease, including lactic acid bacteria fermented extract of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum It provides a composition for the prevention or treatment of.
  • the present invention provides a pharmaceutical composition of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum for the manufacture of a medicament for preventing or treating allergic diseases. It provides the use of lactic acid bacteria fermented extract.
  • the present invention includes the lactic acid bacteria fermented extracts of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum , preventing, improving allergic diseases Or for therapeutic use.
  • Sophora Root is a perennial plant of the dicotyledonous rosewood legume, also called the cane of the thief, the ginseng, and the sperm of the snake. have.
  • angelica gigas is a perennial aromatic herb belonging to the Apiaceae, which is known to have a blood-producing action, an anti-cancer effect, and a blood pressure-lowering effect to produce blood when blood is scarce.
  • Arctium lappa fruit is a dry seed of burdock belonging to the chrysanthemum, the burdock is about 1.5 m in height, the stem is known to grow branches thick and long roots. It is also called a chamber, bookstore, etc., and is known to be effective in urination disorders, eczema, and swelling.
  • Polygonum cuspidatum is a perennial plant of the family Mardiaceae, and it is known that the root is used as a diuretic, a sedative in herbal medicine.
  • the ginseng, Angelica, alligator, and knotweed used in the present invention may be used in nature or purchased commercially, but is not limited thereto.
  • red ginseng, Angelica, alligator and Escherichia coli were used, they showed more excellent anti-allergic activity, confirming that they can be used as a composition for preventing or treating allergic diseases (FIGS. 3 and 5).
  • water, C 1 (C 1 ) to C 4 (C 4 ) alcohol preferably methanol, ethanol or butanol, or a mixed solvent thereof may be used, but is not limited thereto.
  • the extraction method may be a solvent extraction, hot water extraction, cold needle extraction, reflux cooling extraction, ultrasonic extraction, or steam extraction, but is not limited thereto.
  • the lactobacillus fermentation is Lactobacillus rhamnosus, Lactobacillus ashdophyllus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus vulgaris, Lactobacillus delbruecchi subspecies lactis, Lactobacillus It may be composed of one or more strains selected from the group consisting of gasseri and Bifidobacterium breve, and more specifically, the lactic acid bacteria fermentation may be made of Lactobacillus rhamnosus .
  • lactic acid bacteria that can be used is not limited to the lactic acid bacteria exemplified above, lactic acid bacteria culture medium is also to be selected from MRS (Man-Rogosa-Sharpe), lactose, M17 and APT medium (Asparagine Enrichment Broth) for each lactic acid bacteria But it is not limited thereto.
  • allergic disease refers to a disease caused by an immune hypersensitivity reaction
  • an allergic reaction is a complex reaction involving various cells and mediators and at the same time causes various diseases.
  • the allergic disease may be atopic dermatitis, asthma, allergic rhinitis or allergic conjunctivitis, but is not limited thereto.
  • Asthma is a chronic disease that causes respiratory distress due to irritability of the airways and intermittent airway contractions due to chronic inflammation of the airways. Asthma is caused by allergens (antigens), such as house dust mites and pollen, present in the external environment, resulting in excessive immune reactions (allergic reactions), resulting in inflammation of airway mucosal tissues. causes asthma symptoms (Lemanske RF Jr. JAMA 1997, 278, 1855-1873). With the recent advances in immunology and molecular genetics, the prevalence of asthma is increasing despite the understanding of the pathophysiology of asthma and the development of new therapeutic agents. In particular, steroids or bronchodilators, which are used as conventional therapeutics, should be used for a lifetime as a medicament for the control and prevention of symptoms rather than the fundamental cure of a disease, which causes a significant economic burden on individuals and society.
  • the present invention comprises the steps of administering to a subject a composition comprising a fermented extract of lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum It includes, it provides a method of preventing or treating allergic diseases.
  • red ginseng, Angelica, alligator, Escherichia coli, fermentation of lactic acid bacteria or allergic disease are as described above.
  • prevention means any action that suppresses or delays the onset of inflammation by the administration of the composition
  • treatment means any action that improves or advantageously changes the symptoms of inflammation by administration of the composition. Means.
  • the term "administration" refers to introducing the pharmaceutical composition of the present invention to an individual in any suitable manner, and the route of administration of the composition is through various routes, oral or parenteral, as long as the target tissue can be reached. It may be administered, and specifically, in a conventional manner via the oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial, transdermal, nasal, inhaled or intradermal routes.
  • fermented extracts of Ginseng, Angelica, alligator, and K. koji rarely induced cytotoxicity (FIG. 9), and Syk, Fyn, which are signaling proteins of the rate-limiting step of the Fc ⁇ RI receptor signal transduction system.
  • Lyn inhibited the phosphorylation of Syk, Lyn (Fig. 11), by inhibiting the activation of Lyn inhibits the degranulation-related mechanism (PLC ⁇ 1 / 2-PKC ⁇ pathway), a downstream signaling system (Fig. 12), cytokine production
  • PLC ⁇ 1 / 2-PKC ⁇ pathway a downstream signaling system
  • Fig. 12 cytokine production
  • JNK, ERK1 / 2, p38, Akt phosphorylation JNK, ERK1 / 2, p38, Akt phosphorylation
  • composition of the present invention may include a pharmaceutically acceptable carrier, excipient or diluent in addition to the above-described active ingredient for administration.
  • the carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • compositions of the present invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, or the like, oral preparations, suppositories, or sterile injectable solutions, respectively, according to conventional methods.
  • oral preparations suppositories, or sterile injectable solutions, respectively, according to conventional methods.
  • it may be prepared using diluents or excipients such as fillers, weighting agents, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used.
  • Solid form preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like.
  • Such solid preparations may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, gelatin and the like in the composition.
  • excipients such as starch, calcium carbonate, sucrose, lactose, gelatin and the like in the composition.
  • lubricants such as magnesium stearate, talc can also be used.
  • It may be prepared by adding various excipients such as humectants, sweeteners, fragrances, preservatives and the like in addition to liquid oral liquids or liquid paraffin for oral use.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and suppositories.
  • non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used.
  • base of the suppository utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • composition of the present invention may be administered orally or parenterally (eg, applied intravenously, subcutaneously, intraperitoneally or topically) according to the desired method, and the dosage is based on the condition and weight of the patient, the degree of disease, Depending on the drug form, route of administration, and time, it may be appropriately selected by those skilled in the art.
  • the present invention provides a cosmetic composition for preventing or ameliorating allergic diseases, including a lactobacillus fermented extract of a mixture of red ginseng, Angelica, allium and ephedra.
  • the present invention is a fermented extract of lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit and Polygonum cuspidatum for preparing cosmetics for improving allergic diseases Serves the purpose of.
  • the formulation of the cosmetic composition of the present invention is not particularly limited, it is preferably a formulation of an external preparation for skin, and one preferred form of the external preparation for skin is a softening cosmetic, nourishing cosmetic, massage cream, nourishing cream, pack, gel or skin
  • Cosmetic compositions having formulations of adhesive cosmetics, and other preferred forms include transdermal dosage forms such as lotions, ointments, gels, creams, patches or sprays.
  • other components than the essential components described above may be appropriately selected and blended by those skilled in the art without difficulty according to the formulation or purpose of use of the other external preparation.
  • the present invention provides a dietary supplement for preventing or ameliorating allergic diseases, including fermented extract of lactic acid bacteria of a mixture of red ginseng, Angelica, alligator and Escherichia coli.
  • the present invention is a lactic acid bacteria of Sophora Root, Angelica gigas , Arctium lappa fruit , and Polygonum cuspidatum Provides the use of fermented extracts.
  • Fermented extract of the present invention may be added to health food for the purpose of improving allergic diseases, it may be added as it is or used with other food or food ingredients, and may be appropriately used according to conventional methods.
  • the mixed amount of the active ingredient can be determined suitably according to the purpose of use (prevention, health or therapeutic treatment).
  • herbal extracts or herbal fermenters are added in amounts of up to 30% by weight, preferably up to 10% by weight relative to the raw materials.
  • the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. .
  • Examples of the food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, dairy products including other noodles, gum, ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.
  • F-PASA extract of the present invention (fermented product of Angelica ginseng, ginseng, allium, e. / w / w / w)) in 10 liters of distilled water and heated to a temperature of 115 °C for about 3 hours.
  • the resulting solution was filtered through a strainer having a diameter of 150-500 ⁇ m and then sterilized by heating for about 5 minutes.
  • Lactobacillus rhamnosus (KFRI 128, FCTC 2182) was inoculated and fermented at 37 ° C for two days.
  • the fermentation was then filtered through a nylon net having a diameter of 60 ⁇ m and allowed to stand for one day.
  • the supernatant was taken and lyophilized to obtain an F-PASA extract (dried).
  • the dry matter was stored at a temperature of -20 ° C and used in all experiments dissolved in 10% DMSO solution.
  • lactic acid bacterium fermentation product of the herbal extract in the same manner as the method described in Korean Patent Laid-Open Publication No. 10-2010-0093901 of the present inventors (KIOM- MA128) was prepared.
  • the herbal extract is a herbal preparation prepared by extracting red ginseng, licorice, gilt, angui, venom, mokyang, windproof, sanjoin, eoseongcho, yeonkyo, woobangja, yinyanggak, ginseng, japonica, fat milk, cheongung, hyeonsam and jangjanggeun , 160 g of red ginseng, 80 g of licorice, 80 g of gold and silver coins, donkey 80 g, poisonous g 80 g, tree root 80 g, windproof 80 g, Sanjoin 80 g, Eoseongcho 160 g, kyokyo 80 g, alligator 160 g, Yin Yang 80 g, ginseng A herbal mixture was prepared at a ratio of 160 g, 80 g root, 80 g fat, 80 g cheongung, 160 g of Hyunsam and 80 g of rye root
  • Lactobacillus ashdophyllus (KFRI # 128), Lactobacillus casei (KFRI # 127), Lactobacillus plantarum (KFRI # 144, KFRI # 402), Lactobacillus fermentum KFRI # 164), Lactobacillus bulgaricus (KFRI # 344), Lactobacillus delbrueki subspecies Lactis (KFRI # 442), Lactobacillus gasseri (KFRI # 658, KCTC 3163) and Bifidobacterium breve (KFRI # 744), and the specific fermentation process is as described in Korean Patent Publication No. 10-2010-0093901.
  • fermented alligator extract F-allergen
  • fermented Angelica extract F-Dangri
  • fermented red ginseng extract F-Ginseng
  • a method for preparing a specific fermented extract was prepared according to Example 1, and the strain for each fermentation was disclosed in Korean Patent Publication No. 10-2010-0054272; Seo, Min-Jun et al. Journal of the Korean Herb Society, vol. 28, no. 39-44, 2013; And the strain described in Korean Patent Publication No. 10-2014-0008618.
  • RBL-2H3 cells were dispensed in 24-well plates at a concentration of 1 ⁇ 10 5 cells / well, 100 units / mL penicillin, 100 ⁇ g / mL streptomycin and 5% (v / v) fetal bovine serum. Incubated overnight at 37 ° C., 5% CO 2 , humidified conditions in MEM- ⁇ medium. After incubation, cells were washed with 1 ⁇ PBS buffer and incubated with DNP-IgE 50 ng / mL for 24 hours with Sigma, Co., Ltd to sensitize the cells.
  • DNP-HSA Dinitrophenyl-human serum albumin
  • Inhibition of secretion of allergens from mast cells was determined by measuring the activity of ⁇ -hexosaminidase, a marker of degranulation.
  • the F-PASA extract of the present invention can inhibit the secretion of ⁇ -hexosaminidase from mast cells in a concentration-dependent manner, and the IC 50 value was 510.9 ⁇ g / mL.
  • F-PASA extract was able to inhibit more than 75% of allergen secretion at a concentration of 2 mg / mL (Fig. 1).
  • TNF- ⁇ tumor necrosis factor- ⁇
  • the concentration of TNF- ⁇ in the supernatant of the allergic reaction-induced mast cells of Experimental Example 1-1 was measured according to the manufacturer's manual using an ELISA kit (e-Bioscience, Inc.) and the results are shown. 4 and 5, respectively.
  • the F-PASA extract of the present invention exhibited a superior ability to inhibit TNF- ⁇ production of mast cells compared to the extract of Comparative Example 2 regarding a fermentation product of a single substance (FIG. 5).
  • IL-4 Interleukin 4
  • PGD 2 Prostaglandin D 2
  • LTC 4 Leukotriene C 4
  • the F-PASA extract of the present invention significantly inhibits the production of IL-4 of FIG. 6, PGD 2 of FIG. 7, and LTC 4 of FIG. 8.
  • the F-PASA extract of the present invention inhibited the production of IL-4 close to 100% at a concentration of 0.2 mg / mL, and confirmed that about 98% of PGD 2 production at a concentration of 0.5 mg / mL.
  • RBL-2H3 cells were dispensed in 96-well plates at a concentration of 1 ⁇ 10 4 cells / well, followed by 37 ° C. in MEM- ⁇ medium containing 5% (v / v) fetal bovine serum. Incubate overnight at temperature. The cells that were cultured the next day were washed with 1 ⁇ PBS buffer and incubated with 50 ng / mL of DNP-IgE (Dinitrophenyl-immunoglobulin E; Sigma, Co., Ltd.) for 24 hours to sensitize the cells.
  • DNP-IgE Dinitrophenyl-immunoglobulin E
  • F-PASA (0 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL) prepared in Example 1 was added to the sensitized cells and further 1 hour After incubation, 10 ⁇ L of WST-1 reagent (Daeil Lab Service. Co., Ltd.) and 0.1 ⁇ g / mL of DNP-HSA (Dinitrophenyl-human serum albumin) were added at the same time. Incubated for hours. In order to confirm cell viability, absorbance at 450 nm was measured using a micro-plate reader (Emax, Molecular Devices Inc. Sunnyvale, California, USA), and the results are shown in FIG. 9. As a result, it was confirmed that the F-PASA extract of the present invention did not cause any cytotoxicity up to a concentration of 2 mg / mL (Fig. 9).
  • Allergic reaction-induced cells prepared in Experimental Example 1-1 were lysed to extract total protein, and then proteins of the Fc ⁇ RI receptor signaling system were quantified by Western blot.
  • Antibodies used in Western blot were Cell signaling Technology, Inc. It was purchased and used.
  • Electrophoresis pictures of the Western blot results are shown in FIGS. 10 to 13.
  • ⁇ -actin was used as a loading control.
  • the F-PASA extract of the present invention is a cPLA 2 (Cytosolic phospholipase A 2 ) and 5-LO (5- (5), which are related enzymes of Arachdonate cascade, a subsignal of Syk (Spleen Tyrosine Kinase). Lipoxygenase) phosphorylation and COX-2 (Cyclooxygenase-2) can be confirmed to inhibit the expression.
  • F-PASA extract also inhibited the expression of PGDS (Prostaglandin-D synthase) and LTC 4 S (Leukotriene C 4 synthase) (FIG. 10).
  • cPLA 2 is an enzyme involved in the rate step of arachidonic acid chain reaction
  • 5-LO is an enzyme involved in the rate step of leukotriene production
  • COX-2 is an enzyme involved in the rate step of prostaglandin production.
  • PGDS is a PGD 2 production related enzyme
  • LTC 4 S is an LTC 4 production related enzyme.
  • the F-PASA extract of the present invention inhibited the phosphorylation of Syk, Lyn among Syk, Fyn, and Lyn, the signaling proteins of the rate-limiting step of the Fc ⁇ RI receptor signal transduction system (FIG. 11). It was confirmed to inhibit the mechanism of degranulation related to the delivery system (PLC ⁇ 1 / 2-PKC ⁇ pathway) (FIG. 12). Furthermore, it was confirmed that the F-PASA extract of the present invention can inhibit the activation of cytokine production-related mechanisms (JNK, ERK1 / 2, p38, Akt phosphorylation) (Fig. 13), usefully used as an antiallergic substance It can be confirmed that.
  • DNP-IgE 100 ng was injected intradermal into the ears of ICR mice. After 1 day, 0.5% Evans blue aqueous solution containing 0, 12.5, 25, 50 and 100 mg / kg of F-PASA was administered orally and 100 ⁇ g of DNP-HSA after 1 hour. Was injected into the vein.
  • dexamethasone which is commonly used as an antiallergic drug, was orally administered. After 30 minutes, the mice were euthanized, their ears were cut out, placed in 1 mL of formamide, and the extracts were extracted for 2 hours at a temperature of 80 ° C. The extract was centrifuged and then the supernatant was taken and measured for absorbance at 620 nm, as shown in FIG. 14.

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Abstract

La présente invention concerne une composition pour prévenir ou traiter des maladies allergiques, la composition contenant des extraits fermentés par des lactobacilles de racines de Sophora, Angelica gigas, fruit d'Arctium lappa, et Polygonum cuspidata, et en particulier, une composition contenant des extraits fermentés par des lactobacilles de racine de Sophora, Angelica gigas, fruit d'Arctium lappa, et Polygonum cuspidata, pour prévenir ou traiter la dermatite atopique, l'asthme, la rhinite allergique, ou la conjonctivite allergique, et une méthode destinée à prévenir ou à traiter les maladies allergiques comprenant une étape d'administration de la composition à un individu.
PCT/KR2017/013239 2016-12-09 2017-11-21 Composition anti-allergique contenant un extrait d'herbe médicinale composite fermenté par des lactobacilles à titre de principe actif Ceased WO2018105926A1 (fr)

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