WO2017138788A1 - Anti-inflammatory and antibacterial composition containing, as active ingredients, peptide derived from chungkukjang extract and natural extract or fermentation product, and cosmetic composition for preventing or alleviating immune skin diseases, containing same - Google Patents

Abstract

The present invention relates to an anti-inflammatory, antibacterial, and immunity skin disease prevention or alleviation cosmetic composition containing, as active ingredients, a peptide derived from a chungkukjang (thick soybean paste) extract, and a natural extract or fermentation product. More specifically, the present invention relates to an anti-inflammatory and antibacterial composition containing: an inflammatory cell inhibitory peptide derived from a chungkukjang extract; and an extract or fermentation product of one or more plants selected from the group consisting of Chelidonium majus Linne, Symplocos prunifolia, Thuja orientalis, Nymphaea tetragona, phellodendron barks and Paeonia suffruticosa Andrew roots. It was ascertained that the composition containing, as active ingredients, a peptide derived from a chungkukjang extract, and a natural extract or fermentation product, of the present invention, has excellent anti-inflammatory and antibacterial effects, and the composition of the present invention can be useful as an anti-inflammatory and antibacterial composition and can be useful in a cosmetic composition for preventing and alleviating immune skin diseases.

Description

Anti-inflammatory and antimicrobial composition containing the peptide derived from Cheonggukjang extract and natural extract or fermented product as an active ingredient, and a cosmetic composition for preventing or improving immune skin diseases including the same

The present invention relates to an anti-inflammatory and antimicrobial composition containing the peptide and natural extracts or fermented products derived from the Cheonggukjang extract, more specifically, the cosmetic composition or food composition for anti-inflammatory and antimicrobial containing the composition as an active ingredient, immunogenic It relates to a cosmetic composition for preventing or improving skin diseases, and a pharmaceutical composition for preventing or treating inflammatory and infectious diseases.

Inflammation-related diseases include periodontitis, arthritis, gastritis, enteritis, rhinitis, sinusitis, sore throat, tonsillitis, and pneumonia. Inflammation includes acute and chronic inflammation in preparation for the invasion of pathogens. Inflammatory response is a kind of immune response that protects the human body, but excessive and long-term chronic inflammation is harmful to the human body, such as autoimmunity, allergies, and the like. Chronic inflammation is also associated with cardiovascular disease, cancer, dementia and adult disease. Inducible nitric oxide synthase (iNOS), NO (nitrogen monoxide), NO, transcription factors NF-κB, cytokines TNF-α, and IL-6 are factors that promote cell inflammation.

Drugs such as steroids and aspirin have been developed as an inflammatory agent, but these are known to have side effects such as immunosuppression, gastrointestinal diseases, bleeding, and hepatotoxicity. Accordingly, the development of new or improved therapeutics is essential and urgent. The development of inflammatory drugs is underway in the field of inhibiting the secretion of inflammatory substances, preventing the adhesion of inflammatory cells, and researches actively searching for the active ingredient in natural products are actively conducted in connection with the problem of overcoming the above side effects.

Cheonggukjang is one of Korea's representative fermented foods made from soybean. It is one of the traditional types of soy sauce, soy sauce, soybean paste, and red pepper paste that have been commonly used to this day. The state has played an important role as a food. Cheonggukjang is made by using Bacillus subtilis called Bacillus subtilis attached to rice straw. It gives its unique taste and smell by enzymes produced by Bacillus subtilis during fermentation. In addition, it produces a large amount of viscous substance, which is a mixture of levan form fructan and polyglutamate derived from sugar and protein of raw soybeans. It is a high quality fermented soybean with high fat content.

Recently, many researchers have revealed the active ingredient contained in Cheonggukjang and its role, so isoflavone (isoflavone), phytic acid, saponin, trypsin from soybean origin It is a functional food because it contains a large amount of various bioactive substances such as inhibitors, unsaturated fatty acids, dietary fiber, oligosaccharides, and γ-glutamyltranspeptidase (γ-GTP), antioxidants and thrombolytic enzymes. Its importance is being reexamined.

Among them, isoflavones found in soybean have an anti-inflammatory effect of suppressing the expression of these factors. Soybean fermented food Cheonggukjang is rich in isoflavones such as genistein and daidzein, amino acid polymers and peptides that are proteolytic products.

On the other hand, sebum is an oily secretion secreted from sebaceous gland to the surface of the hair and the surface of the epidermis, and its main chemical component is composed of cholesterol, fatty acids and ester compounds.

Sebum acts as a protective film to help protect the skin, but is mainly produced by the release of testosterone (testosterone), and can be overproduced by age, season, women's menstrual cycle, pregnancy, and stress. When excessive sebum is produced, pores may be widened or bacteria living on the skin may break down and inflame the skin, causing various skin diseases, and acne, which is a representative skin disease, is caused by excessive sebum secretion or the growth of acne bacteria. Seborrheic dermatitis, in particular, is a chronic inflammatory skin disease that occurs in the scalp and face where sebum secretion is active due to increased sebaceous glands, especially the eyebrows, nose, lips, ears, armpits, chest, and inguinal area. It is a skin disease. The best way to treat and prevent seborrheic dermatitis is to control sebum secretion, control inflammatory reactions and eliminate acne.

However, there are not many reports on the anti-inflammatory and antimicrobial effects using the Cheonggukjang peptides, and in particular, there are no examples of the use of the Cheonggukjang peptides for effective prevention and treatment of skin diseases.

Thus, the present inventors completed the present invention by confirming that the composition containing the cheonggukjang extract-derived peptide and natural extract or fermented product as an active ingredient has excellent anti-inflammatory and antibacterial activity.

The present invention is to solve the above problems, one object of the present invention is to provide an anti-inflammatory and antimicrobial composition comprising a peptide for inhibiting inflammatory cells derived from Cheonggukjang extract, and a plant extract or fermented product.

Still another object of the present invention is to provide an anti-inflammatory and antibacterial cosmetic composition containing the anti-inflammatory and antimicrobial composition as an active ingredient.

Another object of the present invention to provide a cosmetic composition for the prevention or improvement of immune skin diseases containing the anti-inflammatory and antimicrobial composition as an active ingredient.

Another object of the present invention to provide an anti-inflammatory and antimicrobial food composition containing the anti-inflammatory and antimicrobial composition as an active ingredient.

Another object of the present invention to provide a pharmaceutical composition for preventing and treating inflammatory and infectious diseases containing the anti-inflammatory and antimicrobial composition as an active ingredient.

Anti-inflammatory and antimicrobial composition according to a preferred embodiment of the present invention for achieving the object as described above, Peptides for inhibiting inflammatory cells derived from Cheonggukjang extract; And at least one selected from the group consisting of Chelidonium majus Linne, Blackwood tree (Symplocos prunifolia), White matter (Thuja orientalis), Water lily (Nymphaea tetragona), Peach (phellodendron bark) and Peel (Peonia suffruticosa Andrew roots) Plant extracts or fermentations.

Peptides for inhibiting inflammatory cells derived from the extract of Cheonggukjang is for inhibiting inflammatory cells in which the sequence of Alanine (Ala) -phenylalanine (Phenylalanine, Phe) -Proline (Proline, Pro) -Glycine (Gly) is arranged in order. Peptide or Glutamic acid (Glu) -Valine, Val-Tyrosine (Tyr) -Tyrosine (Tyr) -Methionine (Met) -Tyrosine (Tyr) sequence It is preferably a peptide for inhibiting inflammatory cells.

Herein, the mixing ratio of the peptide and the plant extract is preferably 1: 100 to 100: 1 by weight.

Anti-inflammatory and antimicrobial cosmetic composition according to a preferred embodiment of the present invention contains the anti-inflammatory and antimicrobial composition as an active ingredient.

Cosmetic composition for the prevention or improvement of immune skin diseases according to a preferred embodiment of the present invention contains the anti-inflammatory and antimicrobial composition as an active ingredient.

Anti-inflammatory and antimicrobial food composition according to a preferred embodiment of the present invention contains the anti-inflammatory and antimicrobial composition as an active ingredient.

A pharmaceutical composition for preventing and treating inflammatory and infectious diseases according to a preferred embodiment of the present invention contains the anti-inflammatory and antimicrobial composition as an active ingredient.

Cheonggukjang extract derived from the present invention and the composition containing the natural extract or fermented product as an active ingredient was confirmed that the anti-inflammatory and antibacterial effect is excellent.

Therefore, the composition of the present invention can be usefully used as an anti-inflammatory and antimicrobial composition, which can be used for cosmetics and foods. In addition, the composition of the present invention can be usefully used in the cosmetic composition for the prevention and improvement of immune skin diseases, can also be used in pharmaceutical compositions for the prevention and treatment of inflammatory and infectious diseases.

Figure 1 is a photograph of the concentrate after extraction with 70% ethanol and the raw material of two plants (top; bark, bottom; yellowish white) according to an embodiment of the present invention.

2 is an anti-inflammatory effect test results confirming the NO production inhibitory ability for the mixture of the peptide and the plant extract according to an embodiment of the present invention.

Figure 3 is a photograph of the antimicrobial activity against the acne (Propionibacterium Acnes) according to a preferred embodiment of the present invention.

Figure 4 is a photograph of the antimicrobial activity of the acne bacteria (Propionibacterium Acnes) of the Cheonggukjang peptide.

As the invention allows for various changes and numerous embodiments, particular embodiments will be illustrated in the drawings and described in detail in the written description. However, this is not intended to limit the present invention to specific embodiments, it should be understood to include all modifications, equivalents, and substitutes included in the spirit and scope of the present invention.

Terms including ordinal numbers such as first and second may be used to describe various components, but the components are not limited by the terms. These terms are only used to distinguish one component from another.

In the following description, the detailed description can be omitted for techniques well known to those skilled in the art. In addition, in describing the present invention, when it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the subject matter of the present invention, the detailed description may be omitted. Also, the terminology used herein is a term used to properly express a preferred embodiment of the present invention, which may vary depending on a user, an operator's intention, or customs in the field to which the present invention belongs.

Therefore, the definitions of the terms should be made based on the contents throughout the specification. Throughout the specification, when a part is said to "include" a certain component, it means that it can further include other components, without excluding other components unless specifically stated otherwise.

Hereinafter, the present invention will be described in detail.

In one embodiment of the present invention, the present invention provides a peptide for inhibiting inflammatory cells derived from the extract of Cheonggukjang; And at least one selected from the group consisting of Chelidonium majus Linne, Blackwood tree (Symplocos prunifolia), White matter (Thuja orientalis), Water lily (Nymphaea tetragona), Peach (phellodendron bark) and Peel (Peonia suffruticosa Andrew roots) It provides an anti-inflammatory and antimicrobial composition comprising a natural extract or fermentation.

Peptides for inhibiting inflammatory cells derived from the extract of Cheonggukjang is for inhibiting inflammatory cells in which the sequence of Alanine (Ala) -phenylalanine (Phenylalanine, Phe) -Proline (Proline, Pro) -Glycine (Gly) is arranged in order. Peptide or Glutamic acid (Glu) -Valine, Val-Tyrosine (Tyr) -Tyrosine (Tyr) -Methionine (Met) -Tyrosine (Tyr) sequence It is preferably a peptide for inhibiting inflammatory cells.

Here, the mixing ratio of the peptide and the plant extract and the fermentation product is preferably 1: 100 to 100: 1 by weight.

Cheonggukjang extract according to the present invention can be obtained as follows.

First, the soybeans are washed, immersed in water, and then boiled using a pressurized steamer. The soybeans are inoculated with Bacillus licheniformis, preferably Bacillus licheniformis B1, and fermented at 30-50 ° C. for 2-4 days, preferably 40-3 days; Lyophilizing the fermented fermented product to powder it; Dissolving the powdered powder in water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water, ethanol, butanol or a mixed solvent thereof, more preferably ethanol; Distilling the powder dissolved in the ethanol under reduced pressure; And lyophilizing the vacuum distilled powder.

The soybean includes foreign soybeans such as Korean, Chinese, and Japanese.

In general, Bacillus subtilis (Bacillus subtilis) bacteria are used in the preparation of Cheonggukjang using soybeans, but the present invention specifically Bacillus licheniformis, more preferably Bacillus rickeniformis B1 bacteria. It is characterized by using.

In the present invention, in order to obtain an extract from soybean fermented products, extraction methods such as hot water extraction, cold sediment extraction, reflux cooling extraction, or ultrasonic extraction may be used. Particularly, as a solvent, water, ethanol, butanol or a mixed solvent thereof, Preferably, by using ethanol, more preferably 80% ethanol, it was possible to obtain an extract effective for inflammation.

In order to isolate the peptide, the inventors separated the peptide by performing HPLC from the Cheonggukjang extract as described in Example 2 below.

The peptide may be isolated from the extract of Cheonggukjang as described above, and may also be easily prepared by known peptide synthesis methods.

The peptide may be a peptide for inhibiting inflammatory cells. In this case, the inflammatory cell may be a macrophage (macrophage) activated by lipopolysaccharide (LPS). In addition, the present invention is an expression of at least one selected from the group consisting of inducible nitric oxide synthase (iNOS), nitric oxide (NO), transcription factors NF-κB, cytokine TNF-α, and IL-6 Or it may be a peptide for inhibiting inflammatory cells, characterized in that to reduce the activity.

In order to confirm the anti-inflammatory activity of the peptide, the present inventors treated LPS (lipopolysaccharide), which causes inflammation, to Raw264.7 cells, which are immune cells, to increase the concentration of intracellular nitric oxide (NO), and then peptides and extracts. The concentration of NO reduced by inhibiting the inflammatory response was measured using a Griess reagent. As a result, it was confirmed that the peptide of the present invention has excellent anti-inflammatory activity (see FIG. 2).

Treatment of macrophage with LPS (Lipopolysaccharide) activates macrophages, increases protein and NO production of iNOS, inflammatory markers, iNOS mRNA expression, and greatly increases NF-κB activity, TNF-α and The expression of IL-6 is also greatly increased. However, as shown in the embodiment of the present invention, the reaction of the chungkukjang extract extracted with ethanol to LPS-treated macrophages reduced the protein, NO production, iNOS mRNA expression of iNOS, NF-κB depending on the concentration of the cheonggukjang extract Was significantly reduced, and the expression of TNF-α and IL-6 was also markedly decreased. In addition, even when using the peptide represented by SEQ ID NO: 1 and the peptide represented by SEQ ID NO: 2 according to the present invention isolated from the extract of the Chungkukjang was confirmed that the expression level of iNOS protein, iNOS and TNF-α mRNA also decreased . Accordingly, the present invention can be used for the treatment, amelioration and / or prevention of inflammatory diseases including the proliferation or metastasis of inflammation related cells or markers.

As used herein, the term "inflammation" disease is a general term for a disease that has inflammation as a main lesion, but is not limited thereto, and preferably, atopy, psoriasis, dermatitis, allergy, arthritis, rhinitis, otitis media, sore throat, tonsillitis and cystitis , Nephritis, pelvic inflammatory disease, Crohn's disease, ulcerative colitis, ankylosing spondylitis, systemic lupus erythematodes (SLE), atherosclerosis, asthma, arteriosclerosis, edema, rheumatoid arthritis, delayed allergy (type IV allergy), Graft rejection, graft-versus-host disease, autoimmune encephalomyelitis, multiple sclerosis, inflammatory bowel disease, arthritis, cystic fibrosis, diabetic retinopathy, rhinitis, ischemic-reperfusion injury, vascular restenosis, glomerulonephritis or gastrointestinal allergy, more Preferably edema, rheumatoid arthritis, systemic lupus erythematosus, atherosclerosis, pelvic inflammatory disease, asthma, atopic dermatitis, psoriasis, inflammatory bowel disease, ankylosing chuck Salt, and it may be multiple sclerosis, graft-versus-host disease.

In one embodiment of the present invention, the natural extract or fermented product may be a plant-derived extract or fermented product.

The plant extract or fermented product consists of Chelidonium majus Linne, Black ash tree (Symplocos prunifolia), White liquor (Thuja orientalis), Water lily (Nymphaea tetragona), Peach (phellodendron bark) and Peel (Paeonia suffruticosa Andrew roots) It is preferable to select at least 1 type from the group.

When the plant extract of the present invention is obtained by treating the plant with an extraction solvent, various extraction solvents may be used. According to an embodiment of the present invention, a polar solvent or a nonpolar solvent may be used. Suitable as polar solvents include water, alcohols, preferably methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol Suitable as nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- Pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro Methane, 1,2-dichloroethane, anneal, diethylamine, ether, carbon tetrachloride and THF.

Here, the extraction of the plant is preferably hot water extraction and ultrasonic extraction using water and ethanol, wherein the concentration of ethanol is suitable 30 ~ 90%. The ratio of plant and solvent is preferably 10: 100 to 10: 800 by weight, and the extraction time is suitably between 1 hour and 48 hours.

The plant extract of the present invention may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the plant fermentation may be carried out by inoculating the plant with a microorganism capable of fermenting the plant, wherein the microorganism used may be used without limitation as long as it can ferment the plant. Bacillus bacteria used in general plant fermentation may be used, and in general, Bacillus subtilis bacteria are used, but the present invention is particularly Lactobacillus acidophilus, more preferably Lactobacillus Lactobacillus acidophilus S1-76 can be used.

In the present invention, the term "antibacterial" is specifically preferably having antimicrobial activity against skin disease related bacteria, in particular acne (Propionibacterium Acnes), but is not limited thereto.

The plant extract of the present invention extracted with 70% ethanol or 1: 1 mixture of plant fermentation using Lactobacillus axidophyllus S1-76 was diluted to 1 mg / ml, 0.5 mg / ml, 0.25 mg / ml The antibacterial activity against acne bacteria was tested (see Experimental Example 1 below). As a result, in the case of a mixture of 70% ethanol extract of Paeonia suffruticosa Andrew roots, Nymphaea tetragona, phellodendron bark, and fermented product, it was positive control against the acne causative propionibacterium acne. Excellent antimicrobial activity was confirmed.

In addition, the experiment of the mixture including the plant extract and fermented product and one of the Cheonggukjang peptide (alanine-phenylalanine-proline-glycine) was conducted in the same manner. In the case of peptides, the antimicrobial activity of the peptide alone was not confirmed (see FIG. 4), but the mixture of the peptide and the plant extract showed better antimicrobial activity than that of the simple plant extract or the fermented product. It was confirmed that the induction of synergistic effect to improve the antibacterial activity of the plant extract or fermentation.

As an embodiment of the present invention, the present invention provides an anti-inflammatory and antimicrobial cosmetic composition containing the anti-inflammatory and antimicrobial composition as an active ingredient.

The cosmetic composition of the present invention includes components commonly used in cosmetic compositions in addition to the anti-inflammatory and antimicrobial compositions, and includes conventional auxiliaries such as sulfates, stabilizers, solubilizers, vitamins, pigments and flavors, and carriers. .

In the cosmetic composition of the present invention, the peptide and the plant extract of the present invention may be added in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight to the cosmetic composition usually contained.

Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanta, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of spray, additionally chloro fluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Soluble cellulose, aluminum metahydroxy, bentonite, agar or tragacanta and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

As an embodiment of the present invention, the present invention provides a cosmetic composition for the prevention or improvement of immune skin diseases comprising the anti-inflammatory and antimicrobial composition as an active ingredient.

As used herein, the term "prevention" refers to any action that inhibits or delays the development of an inflammatory disease by administration of the composition, and "treatment" refers to any action that improves or benefits the symptoms of the inflammatory disease by administration of the composition. it means.

In the present invention, the term "immune skin disease" refers to a skin disease related to immunity, and specifically, atopy, seborrheic dermatitis, psoriasis, and the like, but are not limited thereto.

As an embodiment of the present invention, the present invention provides an anti-inflammatory and antimicrobial food composition comprising the anti-inflammatory and antimicrobial composition as an active ingredient.

Food composition according to the present invention can be prepared in the form of a mixture by mixing with known active ingredients known to be effective in the prevention and treatment of inflammatory diseases, functional food (nutritional supplement), health food ( It includes all forms, such as health food and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

In the food composition of the present invention, a mineral-containing compound such as vitamin B, C, E or beta-carotene, Ca, Mg, Zn or phospholipids such as lecithin or maltose, oligosaccharide, amino acid as auxiliary ingredients together with the peptide and plant extract or fermented product. Compounds such as these may be used. Among them, the use of a mixture of two or three components among vitamins C, E, beta carotene, maltol, and the like is more preferable because it can enhance the biological activity effect.

In addition to the above ingredients, in order to enhance the taste as a known additive, natural flavors such as plum, lemon, pineapple or herb, or natural fruit juice, chlorophyllin (Chlorophyllin), in addition to the above ingredients in order to enhance the taste Natural flavors such as plum, lemon, pineapple or herb, or natural juices, natural pigments and sweeteners such as chlorophyllin and flavonoids, such as fructose, honey, sugar alcohol, sugar and citric acid and sodium citrate Acidic agents can be mixed and used.

A preferred content of the peptide, the plant extract and the fermented product in the food composition of the present invention may include about 0.01 to 10% by weight based on the total weight of the food.

The present invention provides a functional food in the form selected from the group consisting of tablets, granules, and liquids comprising the food composition. Functional food refers to foods that are designed and processed to fully exhibit biological control functions such as biological defense, biological rhythm control, and disease prevention and recovery.

The functional food production is not particularly limited other than those included in the food composition of the present invention, it may further contain the necessary ingredients for each food as a conventional food acceptable food supplement.

Tablets are easy-to-take and accurate dosages of powdered or crystalline materials that are aggregated and pressed into rounded discs or cones. Functional food tablets of the present invention may include, but are not limited to, excipients such as lactose, starch, white sugar, mannitol, sorbitol, inorganic salts, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, May contain binders such as sodium carboxymethylcellulose and / or polyvinylpyrrolidine, optionally disintegrating or boiling mixtures such as starch, agar, alginic acid or its sodium salt and / or absorbents, colorants, flavors, And sweeteners. The formulations may be prepared by conventional mixing, granulating or coating methods.

Granules refer to a round, granulated formulation, which can always take a certain proportion and are easy to take for mixed formulations. If necessary, colorants, fragrances, etc. may be added to the granules of the present invention, and may be coated with a suitable coating agent.

A liquid formulation refers to a liquid formulation, and stabilizers, copulating agents, preservatives, etc. may be added as necessary for the present invention. In addition, the functional beverage composition of the present invention may further include fruit extract for flavor improvement. Fruit extracts have the advantage of being able to use various active ingredients with fruit with flavor improvement. The fruit used in the fruit extract is not limited thereto, but may include at least one selected from pineapple, lemon, citrus, orange, apple, pear, grapefruit, apricot, strawberry, peach, melon, guava, lemon, and plum. Can be. Other functional beverage compositions of the present invention may include known additives. As these known additives, nicotinic acid amide, vitamins A, B, C, D, E, folic acid, etc. may be used as vitamins, and L-glycine, L-lysine, DL-methionine, and L- as amino acids. Valine, biotin, L-cysteine, L-cystine, L-tryptophan, L-threonine, L-phenylalanine, etc. may be used, and as herbal preparations, wolfberry, ogapi, baekrye, earl, green, red ginseng, ginseng, ginseng, sulphurine Can be used, such as yellow, mixed flavors, apple flavors, yogurt flavors, drink flavors and the like can be used, sweeteners are white sugar, glucose, fructose, fructooligosaccharides, isomaltoligosaccharides, malto oligosaccharides, Chitosan oligosaccharide, chito oligosaccharide, soy oligosaccharide, xylo oligosaccharide, isosaccharide (high fructose), invert sugar, aspartame, stevioside, sorbitol, mannitol, etc. can be used, and citric acid, DL-apple acid, succinic acid, etc. Can be, as a stabilizer Among the sorbates, polysorbate 20, polysorbate 40, polysorbate 80 and the like may be used, and as preservatives, benzoic acid and its derivatives, dehydroacetic acid and its salts, sorbic acid and its salts and the like may be used. .

As an embodiment of the present invention, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory and infectious diseases comprising the anti-inflammatory and antimicrobial composition as an active ingredient.

The pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier, excipient or diluent. As used herein, the term "pharmaceutically acceptable" means a drug that does not cause a side effect and a sufficient amount to produce a therapeutic effect, the type of disease, the age, weight, health, sex, sensitivity of the patient to the drug It can be easily determined by those skilled in the art according to well-known factors in the medical field, such as the route of administration, the method of administration, the frequency of administration, the duration of treatment, the combination or the drug used simultaneously.

Carriers usable in the present invention include macromolecules that are slowly metabolized, such as liposomes, polysaccharides, polylactic acid, polyglycolic acid, polymeric amino acids, and amino acid copolymers. Salts of inorganic acids such as, for example, hydrochloride, hydrobromide, phosphate and sulfate; Pharmaceutically acceptable salts such as salts of organic acids such as acetates, propionates, malonates and benzoates; Liquids such as water, saline, glycerol and ethanol, and auxiliary substances such as wetting agents, emulsifiers or pH buffering materials can be used. Pharmaceutically acceptable carriers are described in Remingtion's Pharmaceutical Sciences, Mack Publishing Company, 1991. Compositions comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. When formulated, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used. Solid preparations for oral administration may include tablet pills, powders, granules, capsules, and the like, and such solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose or lactose ( lactose), gelatin and the like can be mixed. In addition to the simple excipients, lubricants such as magnesium stearate, talc and the like can also be used. Liquid preparations for oral administration include suspensions, solution solutions, emulsions, and syrups, and various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, 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.

In addition, the pharmaceutical composition of the present invention is a group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories It can have any one formulation selected from.

In addition, the composition may be formulated in a unit dosage form suitable for intrabody administration of a patient according to a conventional method in the pharmaceutical field, preferably in the form of a formulation useful for the administration of a peptide medicine, and is commonly used in the art. Oral or intradermal, intravenous, intramedullary, intramedullary, intraventricular, lung, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal tract, topical, sublingual, intravaginal or rectal Administration may be by parenteral administration routes including, but not limited to, routes.

The total effective amount of the composition of the present invention may be administered to a patient in a single dose and may be administered by a fractionated treatment protocol that is administered in multiple doses for long periods of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the extent of the disease. The pharmaceutical composition according to the present invention is not particularly limited to its formulation, route of administration and method of administration as long as the effect of the present invention is shown.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

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

Example 1 Preparation of Cheonggukjang Extract

Boiled soybeans were inoculated with Bacillus licheniformis B1 bacteria, fermented at 40 ° C. for 3 days, and vacuum-dried to powder. Then, the powder was dissolved in 80% ethanol, distilled under reduced pressure, and vacuum-freeze-dried again to obtain the Chunggukjang extract.

Example 2 Isolation of Peptides from Cheonggukjang Extract

In order to obtain the peptide present in the ethanol extract of Cheonggukjang obtained in Example 1, ODS (manufactured by Waters, 5C18-AR-300 (4.6mm)) column, and CH3CN containing 20% TFA (20-50% ) As a solvent, HPLC was performed under conditions of a flow rate of 0.5 ml / min.

As a result, four kinds of peptides were obtained as follows.

1st peptide: amino group terminal-Ala-Phe-Pro-Gly-carboxyl group terminal

-2nd peptide: amino terminal-Glu-Val-Tyr-Tyr-Met-Tyr-carboxyl terminal

Third peptide: amino group terminal-Lys-Pro-carboxyl group terminal

4th peptide: amino terminal-Gln-Lys-carboxyl terminal

Example 3 Plant Extract Preparation

As shown in FIG. 1, well-dried forage roots (Rubia cordifolia var. Pratensis), sturgeon (Styrax japonica), zombie ivy (Hosta minor), celticus (Chelidonium majus Linne), black ash tree (Symplocos prunifolia), Extract 70% ethanol in 1:10 ratio of the raw materials of Thuja orientalis, Nymphaea tetragona, phellodendron bark, and Peel (Paeonia suffruticosa Andrew roots). Filtered and filtered with 0.45 μm membrane filter. The filtrate was filtered under reduced pressure at 50 ° C. in a distillation apparatus equipped with a cooling condenser and then evaporated under reduced pressure to obtain a concentrate. The obtained concentrate was lyophilized to give a dried product.

Example 4 Plant Fermentation

Well-dried forage root (Rubia cordifolia var. Pratensis), sturgeon (Styrax japonica), zombie leek (Hosta minor), white buckwheat (Chelidonium majus Linne), black ash tree (Symplocos prunifolia), cypress (Thuja orientalis), water lily (Nymphaea tetragona), phellodendron bark and Paeonia suffruticosa Andrew roots (1:10 ratio) of water, inoculated with Lactobacillus axidobilius S1-76. It fermented for 3 days at 37 degrees. It was then filtered through 380 mesh and filtered through a 0.2 μm membrane filter. The filtrate was filtered under reduced pressure at 40 ° C. in a distillation apparatus equipped with a cooling condenser and then evaporated under reduced pressure to obtain a concentrate. The obtained concentrate was lyophilized to give a dried product.

Example 5 Preparation of a Mixture of Peptides, Plant Extracts and Fermentations and Confirmation of Anti-inflammatory Effects

LPS (lipopolysaccharide), which causes inflammation, was treated to Raw264.7 cells, which are immune cells, to increase the concentration of NO in the cells. It measured using (refer FIG. 2). Detailed experimental method was carried out as described below.

1. After thawing raw cells, incubate HDFn in DMEM-FBS10% to prepare cells. (1.5X106cells / 100mm dish)

2. Seed the cultured Raw264.7. (1.5X105 cells / 48well)

3. Incubate for 24h in DMEM-FBS10% medium after 0.5ml / 48well using DMEM-FBS10% medium. DMEM is prepared by CO2-37C-incubation for drug treatment and washing. (O / N)

4. Washing: Remove DMEM-FBS10% completely, replace with DMEM and place in incubator.

5. Drug Treatment (DMSO 0.2% in DMEM): 0.3ml / 48well

 1) 1ug / ml LPS treatment: 10ug / ml LPS in DMEM-FBS10% added 30ul / 48well (0.3ml)

 2) Prepare a test sample by diluting one of the Chunggukjang peptides (alanine-phenylalanine-proline-glycine), a mixture of the peptide, extract and fermented product to an appropriate concentration range (at least three concentration ranges).

3) Take out 48well seeded with Raw264.7, remove DMEM, and exchange with DMEM to which test sample prepared in 3) is added. (0.5ml / 48well)

 4) Incubate for 24h.

6. NO measurement

1) After mixing 50ul of Griess reagent R1 with 100ul of the recovered culture supernatant, react for 5 minutes at room temperature.

2) After 5 minutes, 50ul R2 solution is mixed and reacted at room temperature for 10 minutes.

3) Measure the absorbance at 540 nm.

Experimental Example 1 Paper Disc Antimicrobial Experiment

The plant extract extracted with 70% ethanol of Example 3 was diluted to 1mg / ml, 0.5mg / ml, 0.25mg / ml to test the antimicrobial activity against acne bacteria by concentration. In this case, phenoxy ethanol was used as a positive control in order to compare and contrast the antimicrobial activity of the extracts. The results are shown in Table 1 and FIG. 3.

 Antimicrobial Activity of 70% Ethanol Plant Extracts against Acne Bacteria Clear Zone Size (cm)

division		Concentration (mg / ml)
		One	0.5	0.25
Plant extracts	Filial piety	1.5	1.2	0
	Neck skin	1.9	1.4	0.9
	White	1.6	1.2	0.7
	Black ash wood	1.7	One	0
	training	1.9	1.1	0
	Yellow white	2.2	1.6	1.1
	Phenoxy ethanol	1.8	1.3	0.8
Plant fermentation	Filial piety	1.4	1.3	0.1
	Neck skin	2.1	1.6	1.0
	White	1.7	1.4	0.9
	Black ash wood	1.3	1.1	0.2
	training	2.0	1.3	0
	Yellow white	2.1	1.7	1.0
Plant Extracts and Fermentations + Peptides	White Oyster Extract and Fermentation + Peptide	1.5	1.2	0
	Peel Extract and Fermentation + peptide	2.3	1.7	1.2
	White matter extract and fermentation + peptide	1.6	1.2	0.7
	Black Ash Tree Extract and Fermentation + peptide	1.7	One	0
	Water lily extract and fermented product + peptide	1.9	1.1	0
	Sulfur Extract and Fermentation + peptide	2.6	1.8	1.3

The specific experimental method was carried out as described below.

1. Spread the cultured acne (Propionibacterium Acnes; P acne) on a plate medium with a certain number of bacteria.

2. Drop the paper disc on the plated flat media and apply the test solution to the paper disc.

3. After 72 hours of incubation, determine the antimicrobial activity by measuring the diameter of the clear zone (cm).

From the results of Table 1 above, 70% ethanol extract of Paeonia suffruticosa Andrew roots, water lily (Nymphaea tetragona), phellodendron bark, and fermented product, Propionibacterium acne (Propionibacterium) Acne) showed excellent antimicrobial activity compared to the positive control.

In addition, the paper disk test method was carried out by diluting the concentration of one species (alanine-phenylalanine-proline-glycine) in the Cheonggukjang peptide. And the experiment of the mixture containing 1 mg / ml of plant extracts and fermented products and one type of (Cheonggukjang peptides (alanine-phenylalanine-proline-glycine)) was carried out in the same manner.

In the case of peptides, the antimicrobial activity of the peptide alone was not confirmed, but the mixture of the peptide, plant extract and fermented product showed better antimicrobial activity than that of the simple plant extract and fermented product. Especially in case of Hwangbaek and Mokdanpi, Cheonggukjang peptides are known to induce synergistic effects to improve the antibacterial activity of plant extracts and fermented products.

Figure 3 is a photograph of the antimicrobial activity of the plant extract and fermented product of the present invention, Propionibacterium acne, a cause of acne of the plant extract and fermented product and peptide mixture, the plant extract and fermentation concentration It can be seen that the antibacterial effect is implemented in proportion.

Experimental Example 2 Measurement of Minimum Inhibitory Concentration (MIC)

In order to further demonstrate the antimicrobial activity against acne causative agents by the extract of the pine needles, MIC (minimal inhibitory concentration) measurement experiment on acne causative organisms was performed.

Extracts and fermentation of Chelidonium majus Linne, Black ash tree (Symplocos prunifolia), Thuja orientalis, Nymphaea tetragona, Phellodendron bark, Paekonia suffruticosa Andrew roots prepared in Example 1 The mixture of water and the peptide was added by disinfection filtration to a concentration suitable for RCM (reinforced clostridial broth), inoculated 1% of each of the activated acne strain culture in the experimental zone, and then put the sample in each concentration. After anaerobic culture for 72 hours under 37 ° C. and CO ₂ conditions, 100 μl aliquots were injected into RCA (reinforced clostridial agar), followed by anaerobic culture. In order to confirm the result obtained, the above experiment was repeated three times. The results are shown in Table 2 below.

Minimal inhibitory concentration (MIC) measurement experiment on acne bacteria of 70% ethanol plant extract

division		Lowest inhibitory concentration (%)
Plant extracts	White oyster	0.05
	Neck skin	0.03
	White	0.05
	Black ash wood	0.08
	training	0.06
	Yellow white	0.02
	Phenoxy ethanol	1.8
Plant fermentation	White oyster	0.03
	Neck skin	0.02
	White	0.05
	Black ash wood	0.07
	training	0.04
	Yellow white	0.02
Plant extracts and fermented products + peptide	White Oyster Extract and Fermentation + peptide	0.04
	Peel Extract and Fermentation + peptide	0.02
	White matter extract and fermentation + peptide	0.05
	Black Ash Tree Extract and Fermentation + peptide	0.08
	Water lily extract and fermented product + peptide	0.05
	Sulfur Extract and Fermentation + peptide	0.01

It was confirmed through the minimum inhibitory concentration experiment that most of the plant extracts have excellent acne antimicrobial effect, in particular, baekbaek, it was superior to the bark skin. And it was confirmed that a mixture of 0.1 mg / ml of a plant extract, fermented product, and one species (alanine-phenylalanine-proline-glycine) among the peptides has a synergistic effect even at the minimum inhibitory concentration experiment.

Claims (7)

1. Peptides for inhibiting inflammatory cells derived from extracts of Cheonggukjang; And

   At least one member selected from the group consisting of Chelidonium majus Linne, Black ash tree (Symplocos prunifolia), White matter (Thuja orientalis), Water lily (Nymphaea tetragona), Peach (phellodendron bark) and Peel (Peonia suffruticosa Andrew roots) Plant extracts or fermented products;

   Anti-inflammatory and antimicrobial composition comprising a.
2. The method of claim 1,

   The peptide is an alanine (Ala) -phenylalanine (Phenylalanine, Phe)-proline (Proline, Pro)-glycine (Glycine, Gly) sequence of peptides for inhibiting inflammatory cells or glutamic acid (Glutamic acid, Glu) -Valine (Va), Tyrosine (Tyrosine, Tyr)-Tyrosine (Tyrosine, Tyr)-Methionine (Methionine, Met) Tyrosine (Tyrosine) characterized in that the peptides for inhibiting inflammatory cells arranged in sequence Anti-inflammatory and antimicrobial composition.
3. The method of claim 1,

   The mixing ratio of the peptide and the plant extract or fermentation is an anti-inflammatory and antimicrobial composition, characterized in that 1: 1: 100 ~ 100: 1 by weight.
4. An anti-inflammatory and antibacterial cosmetic composition containing the composition of claim 1 as an active ingredient.
5. Cosmetic composition for the prevention or improvement of immune skin diseases containing the composition of claim 1 as an active ingredient.
6. Food composition for anti-inflammatory and antimicrobial containing the composition of claim 1 as an active ingredient.
7. A pharmaceutical composition for preventing or treating inflammatory and infectious diseases, comprising the composition of claim 1 as an active ingredient.

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