KR20190033908A - Composition comprising the milk-cream fermentation product of lactic acid bacteria for preventing and improving atopic dermatitis - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing, alleviating and improving atopic dermatitis containing a lactobacillus-fermented milk cream liquid as active ingredients, and various applications thereof. The fermented milk cream liquid fermented by a strain of Lactococcus spp., Lactobacillus spp., or Bifidobacterium spp. of the present invention has a skin stabilization or reduction of the amount of transdermal water evaporation through improvements of water content in the skin and reduction of pH value in skin, thereby effectively preventing, alleviating and improving atopic dermatitis. In addition, the present invention improves an amount of water in skin to have an excellent effect in skin moisturization. Moreover, the present invention is used for oral administration or skin application respectively or simultaneously, to alleviate atopic dermatitis, thereby being useful in relevant cosmetics, food, or pharmaceutical industries.

Description

[0001] The present invention relates to a composition for preventing and improving atopic dermatitis, which comprises an oil-in-milk fermentation broth of lactic acid bacteria as an active ingredient,

The present invention relates to a cosmetic composition for preventing, alleviating and improving atopic dermatitis comprising an oil-in-milk fermentation broth of lactic acid bacterium as an active ingredient and its various applications.

Atopic dermatitis is a chronic, recurrent inflammatory skin disease. Skin dryness and pruritus, which can result in eczema and inflammatory skin lesions when scratched, and a vicious cycle in which more severe pruritus and sleep disturbances occur. In addition, if the proper management is not performed at the initial stage, it may progress to disseminated papillary eruption with erythema and scar, and lichenification that thickens and wrinkles the skin. In infancy, eczema usually appears on the bent part of the neck, limbs, and back of the knee. When the eczema is chronic, the eczema of the eczema area progresses, and when atopic dermatitis remains until adulthood, it is easily exposed to the inflammatory reaction There is a tendency to develop eczema or erythema and flushing around the face or limb rather than the body.

Atopic dermatitis is a complex clinical manifestation due to the interaction of hereditary and environmental factors. Therefore, the cause of the disease can not be explained by any one reason. However, the environmental pollution such as soot, increase of use of food additives, And environmental factors such as the increase of allergen causing allergies such as house dust mite caused by the disease, genetic predisposition, immunological reaction, abnormal skin barrier, and family history are considered to be the main cause.

In general, atopic dermatitis patients have abnormalities of skin barrier, decrease of skin moisture content, increase of skin pH value and increase of transdermal water evaporation, as well as colonization of Staphylococcus aureus in atopic dermatitis lesions high.

Currently, the severity of atopic dermatitis is assessed by SCORAD (Scoring of Atopic Dermatitis) and ADSI (Atopic Dermatitis Severity Index), which are clinical scores of patients with atopic dermatitis. Objectively assess skin damage indicators such as skin barrier function and inflammation status by measuring skin surface hydration (SSH), skin pH, and transepidermal water loss (TEWL).

The basic treatment of atopic dermatitis is the use of a moisturizing agent to prevent skin's cleansing and drying, the use of therapeutic agents such as oral antihistamines, steroids, immunosuppressants, antibiotics, antivirals, antifungals, And photochemotherapy. However, when these preparations are used for a long period of time, side effects such as skin atrophy and swelling, enlargement of blood vessels and pores, and skin discoloration are caused, and at the same time, due to the nature of atopic dermatitis, There are also concerns about the side effects of drugs such as conferences and steroids, which may result in poor treatment or a chronic disease that relies on unproven treatments and eventually worsens and improves. Therefore, there is a constant demand for a therapeutic agent for atopic dermatitis derived from natural materials which is free from side effects and safe to human body.

Lactobacillus activates microfibrils in the immune system to instantly detect bacteria or viruses, promote lymphocyte division, increase IgA production in the blood, and promote immunogenicity by promoting gamma interferon production. The anti - allergic effect of some lactic acid bacteria was shown mainly in experimental animals and cultured cells, and its effect was proved especially in the center of atopic dermatitis. In particular, the culture medium of lactic acid bacteria forms an acidic coating thinly, and when applied to the skin together with the moisturizing agent, it gives a luster and a soft touch. Therefore, it is necessary to find new active ingredients for effectively preventing, alleviating and improving atopic dermatitis.

It is an object of the present invention to provide a cosmetic composition for preventing, alleviating and improving atopic dermatitis.

It is also an object of the present invention to provide a cosmetic composition for moisturizing the skin.

It is also an object of the present invention to provide an external preparation for skin for the prevention, alleviation and improvement of atopic dermatitis.

It is also an object of the present invention to provide a food composition for preventing, alleviating and improving atopic dermatitis.

It is also an object of the present invention to provide a pharmaceutical composition for preventing and treating atopic dermatitis.

It is another object of the present invention to provide a method for producing a milk cream fermentation broth for preventing, alleviating and improving atopic dermatitis.

In order to achieve the above object, the present invention provides a milk cream of at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. A cosmetic composition for preventing, alleviating and improving atopic dermatitis comprising a fermentation liquid as an active ingredient.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. And a cosmetic composition for moisturizing the skin.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. Thereby providing an external preparation for skin for preventing, alleviating and improving atopic dermatitis.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. And to provide a food composition for preventing, alleviating and improving atopic dermatitis.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. The present invention provides a pharmaceutical composition for preventing and treating atopic dermatitis.

The present invention also relates to a method for producing a fermentation broth by cultivating at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. The present invention provides a method of producing a milk cream fermentation broth for preventing, alleviating and improving atopic dermatitis.

Lactococcus genus of the present invention (Lactococcus spp.), Lactobacillus genus (Lactobacillus spp.) Or Bifidobacterium (Bifidobacterium spp.) Oil cream fermentation broth of a strain skin with improved retention skin moisture, pH value decreased skin Stabilizing or reducing the amount of transdermal moisture evaporation, thereby effectively preventing, alleviating and improving atopic dermatitis. In addition, it exhibits an excellent effect on moisturizing the skin by improving the moisture retention amount of the skin. In addition, atopic dermatitis can be ameliorated by using them for oral administration or skin application, respectively, or simultaneously treating them, and thus they can be usefully used in the related cosmetics, food or pharmaceutical industries.

FIG. 1 is a graph showing the effect of treating the milk cream fermentation broth of Example 1 of the present invention with atopic dermatitis.

The present invention relates to a method for producing a fermented milk comprising an oil-in-milk fermentation broth of at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. A cosmetic composition for preventing, alleviating and improving atopic dermatitis is provided.

The milk cream fermentation broth of the present invention is a fermentation broth obtained by culturing Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. In a culture medium, a concentrated fermentation broth, Means a dried product, a cultured filtrate, a concentrated culture filtrate, or a dried filtrate, and may be a cultured filtrate containing the strain or the strain after the strain has been removed. The culture is not limited in its formulation, and may be, for example, a liquid, an emulsion, or a solid.

The fermentation broth is prepared by mixing milk-cream, milk, skim milk, Lactobacilli MRS medium, LB (lysogeny broth) medium, R2A medium (MB-R2230) and casein hydrolyzate Containing medium, preferably a milk cream, milk or Lactobacilli MRS medium, or a culture medium for culturing the strain of each of the above-mentioned genus, but the present invention is not limited thereto.

The milk may be at least one selected from the group consisting of crude oil, non-fat milk, low-fat skim milk added with a milk fat content of 1 part by weight or less, skim milk, and the milk is cow, goat, but are not limited to, horses, donkeys, water buffalo, or camel.

The milk cream may be at least one selected from the group consisting of milk fat separated from raw milk and milk, concentrate of the milk fat, processed milk cream, and powdered milk cream, but the milk cream generally contains 30 ~ 40% cream, an extra light cream with a fat content of 3 ~ 10%, a light cream with a milk fat content of 10 ~ 20%, a table cream with a milk fat content of 20 ~ 30% Table cream, sour cream, whipping cream, double cream with a fat content of 48-60% (processed milk of 18% or more of milk fat processed by adding food or food additives) Double cream, or Powdered milk cream (more than 50% milk fat) made by adding food or food additives to milk cream, may be used instead of fresh cream.

The nutrients of the milk cream are affected by the nutrients of the crude oil before the separation of the cream and the water soluble nutrients are reduced while the oil soluble nutrients (vitamins A, D, E, K) are 2 to 3 times, .

The milk cream fermentation broth can treat at least one selected from the group consisting of tyndallization treatment, reduced pressure concentrating treatment and yeast extract treatment, and each of them can be treated alone or after one treatment, Can be processed.

In the present invention, the tyndallized milk cream fermentation broth can be intermittently sterilized by heating the milk cream fermented with the above-mentioned various lactic acid bacteria at a constant pressure of 80 or more for 15 minutes or more. The reduced-pressure concentration treatment may be performed at a temperature of 50-70 minutes or more after the intermittent sterilization treatment, followed by a warming of 50-70 minutes or longer, whereby the reduced pressure can be concentrated more than twice. The yeast extract and the MRS broth treatment may be 0.1% Can be treated on the medium.

The milk cream fermentation broth may be a fermented milk cream fermented with various lactic acid bacteria or a mixed fermentation broth of a crude fermented broth of a lactic acid bacterium or a lactic acid bacterium fermented with various lactic acid bacteria or a mixture of fermented milk cream fermented with various lactic acid bacteria Yeast extract and MRS broth in a milk cream fermentation broth or milk cream fermentation broth.

The Lactobacillus spp. May be selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus gasseri , Lactobacillus sp. A group consisting of Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus reuteri , Lactobacillus salivarius, Lactobacillus fermentum and Lactobacillus acidophilus. , But the present invention is not limited thereto as long as it is a lactobacillus strain strain for preventing, alleviating and improving atopic dermatitis.

Bifidobacterium spp. Is not limited to Bifidobacterium bifidum , but is not limited to, Bifidobacterium spp., For the prevention, alleviation and improvement of atopic dermatitis.

Lactococcus spp. Is not limited to Lactococcus lactis , but Lactococcus sp. Strains for preventing, alleviating and improving atopic dermatitis.

The composition exhibits an effect of increasing skin moisture content, decreasing the pH value of the skin, stabilizing the skin or reducing the amount of transdermal moisture evaporation, but is not limited thereto.

In a specific example of the present invention, the inventors of the present invention confirmed the effect of atopic treatment on the fermented milk cream fermented with various lactic acid bacteria, respectively. As a result, the inventors of the present invention filed a patent application on March 25, 2016 10-2016-0035879) and Lactobacillus lambosus milk cream fermentation broth (B2) of International Application (PCT / KR2017 / 003172) dated March 25, 2017 to increase skin moisture retention, decrease skin pH , It was confirmed that the transdermal water-swellable amount was reduced, and not only the lesion development of atopic dermatitis was suppressed, but also contributed to the alleviation of the lesion and the suppression of lesion recurrence (see Tables 3 to 5).

In addition, a mixed fermentation broth (K11) consisting of 90% by weight of fermented milk of intermittently sterilized milk cream, 90% by weight of Lactobacillus lambosus milk cream fermentation solution and 10% by weight of Lactobacillus laminocus 10-fold concentrate, 80% by weight of Lactobacillus laminosse oil cream fermentation broth, A mixed fermentation broth of 20% by weight of a 10-fold fermented Laminocus concentrate or a mixed fermentation broth of 90% by weight of a lactobacillus lanosus oil cream fermentation broth and 10% by weight of a 10% fermented lactococcus lactis fermentation broth or 90% by weight of a lactobacillus lan- And 10% by weight of Bifidobacterium bifidum 10-fold concentrated fermentation broth, respectively. As a result, the inventors of the present invention filed a patent application (10-2016-0035879) on March 25, 2016, (B2) of Lactobacillus laminosus milk cream (B2) of International Application (PCT / KR2017 / 003172) on 25th of May, the increase of skin moisture content, the decrease of pH value of skin, the transdermal water evaporation The image shows a more superior effect of reduction, etc., as well as to inhibit the expression of the lesions of atopic dermatitis, it was confirmed that contribute to the inhibitory effect of relaxation and the lesion recurrence of lesions (Table 6 to Table 8).

In addition, the effects of atorvastatin treatment of lactobacillus lambsus milk cream yeast fermentation broth, milk cream MRS fermentation broth, lactococcus lactis milk yeast fermentation broth and milk cream MRS fermentation broth were examined. As a result, the present inventors' patent application filed on March 25, 2016 10-2016-0035879) and Lactobacillus rhamnosus milk cream fermentation (B2) of International Application (PCT / KR2017 / 003172) dated March 25, 2017, increase in skin moisture content, decrease in skin pH value, (See Table 10 to Table 12). The results are shown in Table 10 to Table 12, and the results are shown in Table 10 to Table 12.

In addition, a mixture of 25% by weight of the lactobacillus laminosus milk cream fermentation broth, 25% by weight of the lactococcus lactis milk cream fermentation broth, 25% by weight of the lactobacillus seed oil cream fermentation broth, and 25% by weight of the lactobacillus delubruchii cream fermentation broth The inventors of the present invention filed a patent application (10-2016-0035879) on March 25, 2016 and found that the effect of the lactic acid fermentation broth of Lactobacillus fermentation broth or Lactobacillus plantarum milk yeast fermentation broth and milk cream MRS fermentation broth was confirmed. (B2) of Lactobacillus lambosus yream cream (B2) of International Application (PCT / KR2017 / 003172) dated March 25, 2017 to increase skin moisture retention, decrease skin pH value, reduce transdermal water vapor , It was confirmed that not only the expression of lesions of atopic dermatitis was suppressed but also contributed to the relief of lesion and the inhibition of recurrence of lesions (see Tables 6 to 8 and Tables 10 to 12 Reference).

As a result of confirming the effect of atopy therapy after oral mixed Lactobacillus lambatus mixture bacteria, the present inventors' patent application (10-2016-0035879) on March 25, 2016 and international application (PCT / KR2017 / 003172), but it contributes to the suppression of the lesion and the inhibition of lesion recurrence as well as suppressing the lesion development of atopic dermatitis In particular, when taking the lactobacillus mixed bacterial strain of oral Lactobacillus lambatus and applying the creamy fermented milk lotion of the present invention, the present inventors filed a patent application (10-2016-0035879) on March 25, 2016 and the international application (B2) of Lactobacillus laminosus milk cream (PCT / KR2017 / 003172), the increase of skin moisture content of atopic dermatitis lesion, decrease of skin pH value and reduction of transdermal water evaporation The change typically higher, as well as to inhibit the expression of the lesions of atopic dermatitis, it was confirmed that contribute to a higher inhibitory effect of mitigating and lesion recurrence of lesions (see Table 13 to Table 15).

Accordingly, the cosmetic composition of the present invention is useful as a cosmetic composition for relieving or recurring lesions such as dry skin, pruritus, eczema, inflammatory skin lesions and subacute lesions, esophagus, scarring, and chronic lesions, which are mainly found in atopic dermatitis. Inhibitory effect and corresponding alleviation of lesions of allergic dermatitis, enhancement of skin moisture content, reduction of pH value of skin, and decrease of transdermal moisture evaporation amount, so that cosmetic composition for preventing, alleviating and improving atopic dermatitis and external preparation for skin Lt; / RTI >

The composition may be used for oral or skin application and for mixing. For oral use, a live or dead organism strain may be used, and the mixing ratio (w / g) of the live organism or dead organism may be optionally selected in order to achieve the purpose of preventing, alleviating and improving dermatitis.

The cosmetic composition of the present invention can contain, in addition to the above-mentioned effective ingredients, all kinds of ingredients which can be used in ordinary cosmetics such as perfumes, coloring matters, bactericides, antioxidants, preservatives, moisturizers, thickeners, excipients, diluents, inorganic salts, May be further included.

The above-mentioned cosmetic composition may be provided for use in basic cosmetics, makeup cosmetics, body cosmetics, hair cosmetics, scalp cosmetics or cosmetic cosmetics. Examples of the basic cosmetics include a cream, a lotion, a pack, a massage cream, and a milky lotion. The makeup cosmetics include a foundation, a makeup base, a lipstick, an eye shadow, an eyeliner, a mascara, an eyebrow pencil, Examples of the hair cosmetics include shampoo, rinse, hair treatment, hair mousse, hair liquid, pomade, hair color agent, hair blotting agent, soap, liquid cleanser, bathing agent, sunscreen cream, Color scalp, scalp cosmetics include hair tonic and scalp treatment, and shaving cosmetics include aftershave lotion and shaving cream.

In addition, the cosmetic composition of the present invention may further contain other ingredients usually added to cosmetics. More specifically, the other ingredients to be added to the conventional cosmetic composition are selected from the group consisting of a preservative component, a moisturizer, an emollient, a surfactant, an organic pigment and an inorganic pigment, an organic powder, an ultraviolet absorbent, an antiseptic, a bactericide, , Alcohols, coloring matters, fragrances, blood circulation promoters, coolants, restraining agents or purified water.

The above-mentioned sustaining component may specifically be ester-based, hydrocarbon-based, silicon-based, fluorine-based, animal or vegetable oil, and the like.

Examples of the ester-based oil include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isostearic acid But are not limited to, isopropyl myristate, butyl stearate, butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isosilyl myristate, isostearic acid isostearyl, isostearyl palmitate, octyldodecyl myristate, Trimethylolpropane, trimethylolpropane triisostearate, trimethylolpropane triisostearate, tri-2-ethylhexanoate trimethylolpropane, tri-2-ethylhexanoate trimethylolpropane, Ethylhexanoic acid pentaerythritol, ethylhexanoic acid pentaerythritol, caprylic acid cetyl, decyl lauric acid, hexyl laurate, myristyl myristate, myristyl myristate, myristyl acid stearyl stearate, stearyl stearate, Octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, isopropyl stearate, Ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, di (capryl, capric acid) propylene glycol , Propylene glycol dicaprylylate, dicapric acid neopentyl glycol, dioctanoic acid neopentyl glycol, tricarboxylic acid glyceryl, triunsaturated acid glyceryl, triisopalmitic acid glyceryl, triisostearic acid glyceryl, neopentanoic acid octyl Dodecylsuccinic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, octadecanoic acid, Stearyl, octyldecyl isostearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisolealkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate , Triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, sebacic acid di But are not limited to, isopropyl myristate, isopropyl myristate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, Stearyl, 1,2-stearoylhydroxystearic acid, isostearyl, 1,2-stearoylhydroxystearic acid stearate, Or 1,2-stacking egg yl-hydroxy stearic acid and the like cetearyl source.

The hydrocarbon-based oil may be, for example, squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, flowing isoparaffin, polybutene, microcrystalline wax or vaseline.

Examples of the silicone-based oil include polymethyl silicone, methylphenyl silicone, methyl cyclopolysiloxane, octamethyl polysiloxane, decamethyl polysiloxane, dodecamethyl cyclosiloxane, dimethylsiloxane-methyl cetyloxysiloxane copolymer, dimethylsiloxane-methyl stearoxysiloxane copolymer Alkyl-modified silicone oil, amino-modified silicone oil, and the like.

The fluorine-containing oil may be perfluoropolyether or the like.

Also, animal or vegetable oils can be included. Examples of the oil or vegetable oil include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rape seed oil, palm oil, palm oil, Free, sunflower oil, grape seed oil, cottonseed oil, palm oil, cucumber nut oil, wheat germ oil, rice germ oil, shea butter, coltsfoot colostrum, marker daisy nut oil, mead home oil, argan oil, rosehip seed oil, green tea seed oil, , Canned wax, carnauba wax, beeswax, olive wax, liquid lanolin or hardened castor oil and the like can be used.

The surfactant may specifically be an anionic surfactant, a cationic surfactant or a positive surfactant.

Examples of the anionic surfactant include fatty acid soap, alpha-acylsulfonate, alkylsulfonate, alkylarylsulfonate, alkylnaphthalenesulfonate, alkylsulfate, POE alkyl ethersulfate, alkylamide sulfate, alkyl phosphate, POE alkyl An acylated hydrolyzed collagen peptide salt, or a perfluoroalkylsulfate salt such as an alkyl sulfosuccinate salt, an acylated hydrolyzed collagen peptide salt, or a perfluoroalkyl Phosphate esters and the like.

The cationic surfactant includes, for example, alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenylbromide Trimethylammonium chloride, benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate or lanolin derivative quaternary ammonium salt, and the like.

Examples of the amphoteric surfactant include carboxybetaine, amidebetaine, sulfobetaine, hydroxysulfobetaine, amidosulfobetaine, phosphobetaine, aminocarboxylate, imidazoline derivative or amide Amine-type amphoteric surfactant, and the like.

The pigment may be an organic pigment, an inorganic pigment, a composite pigment thereof, or the like.

Examples of the inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, Bengala, clay, bentonite, titanium mica, titanium oxide, bismuth oxychloride, zirconium oxide, magnesium oxide, Aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, chromamine and combinations thereof.

Examples of the organic pigments include polyamides, polyesters, polypropylenes, polystyrenes, polyurethanes, vinyl resins, urea resins, phenol resins, fluororesins, silicon resins, acrylic resins, melamine resins, epoxy resins, polycarbonate resins, Benzene-styrene copolymer, silk powder, cellulose, CI Pigment Yellow, CI Pigment Orange, and a complex thereof. Examples of the organic powder include metallic soap such as calcium stearate; Metal salts of alkyl phosphates such as sodium zinc cetylate, zinc laurylate and calcium lauryl laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc and N-lauroylglycine calcium; Amidosulfonic acid multivalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-acyls such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylglycine, N-alpha-palmitoyllium N-alpha-lauroyl arginine and N- Basic amino acids; N-acylpolypeptides such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-aminocaprylic acid and alpha-aminolauric acid; Polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, ethylene tetrafluoride, and the like.

The cosmetic composition may further comprise a known sunscreen agent.

The ultraviolet screening agent may be an organic ultraviolet screening agent and / or an inorganic ultraviolet screening agent. The cosmetic composition may be prepared in any form conventionally produced in the art.

The term " cultivation " in the present invention means cultivation of microorganisms under moderately artificially controlled environmental conditions.

The culture medium may contain nutrients required for culturing, that is, a microorganism to be cultured, in order to cultivate a specific microorganism, and may be a mixture in which a substance for a special purpose is further added and mixed. The medium is also referred to as an incubator or a culture medium, and is a concept including both natural medium, synthetic medium and selective medium. Bacillus subtilis strain GCB-13-001 can be cultured according to a conventional culture method.

The medium used for cultivation should meet the requirements of a specific strain in a suitable manner while controlling temperature, pH, etc. in a conventional medium containing a suitable carbon source, nitrogen source, amino acid, vitamin, and the like. The carbon sources that can be used include glucose and xylose mixed sugar as main carbon sources, and sugar and carbohydrates such as sucrose, lactose, fructose, maltose, starch and cellulose, soybean oil, sunflower oil, castor oil, Oils and fats such as oils and the like, fatty acids such as palmitic acid, stearic acid and linoleic acid, alcohols such as glycerol and ethanol, and organic acids such as acetic acid. These materials may be used individually or as a mixture. Nitrogen sources that may be used include inorganic sources such as ammonia, ammonium sulfate, ammonium chloride, ammonium acetate, ammonium phosphate, ammonium carbonate, and ammonium nitrate; Amino acids such as glutamic acid, methionine and glutamine, and organic nitrogen sources such as peptone, NZ-amine, meat extract, yeast extract, malt extract, corn steep liquor, casein hydrolyzate, fish or their decomposition products, defatted soybean cake or decomposition products thereof . These nitrogen sources may be used alone or in combination. The medium may include potassium phosphate, potassium phosphate and the corresponding sodium-containing salts as a source. Potassium which may be used include potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding sodium-containing salts. As the inorganic compound, sodium chloride, calcium chloride, iron chloride, magnesium sulfate, iron sulfate, manganese sulfate and calcium carbonate may be used. Finally, in addition to these materials, essential growth materials such as amino acids and vitamins can be used.

In addition, suitable precursors may be used in the culture medium. The above-mentioned raw materials can be added to the culture in the culture process in a batch manner, in an oil-feeding manner or in a continuous manner by an appropriate method, but it is not particularly limited thereto. Basic compounds such as sodium hydroxide, potassium hydroxide, ammonia, or acid compounds such as phosphoric acid or sulfuric acid can be used in a suitable manner to adjust the pH of the culture.

The present invention also relates to a method for producing a fermented milk fermented milk, which comprises using at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. Thereby providing an external preparation for skin for preventing, alleviating and improving atopic dermatitis.

In the present invention, the external preparation generally refers to a concept covering the entire composition used for external use. For example, it is widely used as a cosmetic composition containing various cosmetics such as basic cosmetics, makeup cosmetics, and hair cosmetics, various medicines such as ointment, , Preferably a cosmetic composition, more preferably a functional cosmetic composition or a functional cosmetic composition.

The external preparation may contain, in addition to the active ingredient, a component that is usually formulated in an external preparation according to the intended use and the properties of the composition for external use, such as a moisturizer, an ultraviolet absorber, a vitamin, an animal extract, an exfoliant, a whitening agent, a vasodilator, And the like may be further blended.

The formulation of the external preparation may take any appropriate form depending on the intended use and the nature of the composition for external application. For example, an aqueous solution, a solubilization system, an emulsion system, an emulsion system, a gel system, a paste system, an ointment system, Water-oil-powder three-layer system, and the formulations and the form of the external preparation of the present invention are not limited by the formulations.

In addition, the external preparation may further include a mechanism necessary for infiltrating or transferring the active ingredient into the dermal tissue.

The present invention relates to a fermented milk-containing fermented milk of at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. A cosmetic composition for moisturizing the skin is provided.

Since the cosmetic composition includes a cosmetic preparation containing the above-mentioned active ingredient, the overlapping contents of the composition of the present invention described above are omitted in order to avoid the excessive complexity of the present invention due to the overlapping description of the present invention.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. And to provide a food composition for preventing, alleviating and improving atopic dermatitis.

The active ingredient may be added to a health supplement such as food, beverage or the like.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including dairy products, meat, sausage, bread, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen noodles, other noodles, gums, ice cream, Beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, all of which include health functional foods in a conventional sense.

The milk cream fermentation broth of the present invention can be added as it is to the food or can be used together with other food or food ingredients, and can be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement). Generally, the amount of the compound in the health food may be 0.1 to 90 parts by weight of the total food. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional beverage composition of the present invention is not particularly limited to the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 of the composition of the present invention.

In addition to the above, the milk cream fermentation broth of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and heavy stabilizers (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the fermentation broth of the present invention may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages.

These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the extract of the present invention or fractions thereof.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. The present invention provides a pharmaceutical composition for preventing and treating atopic dermatitis.

The pharmaceutical composition of the present invention may further comprise an adjuvant in addition to a milk cream fermentation solution of Lactococcus spp., Lactobacillus spp. Or Bifidobacterium spp. have. Such adjuvants may be used without limitation as long as they are known in the art, but may include, for example, Freund's complete or incomplete adjuvant to increase its immunity.

The pharmaceutical composition according to the present invention can be prepared by incorporating the active ingredient into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers for use in the pharmaceutical compositions of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition of the present invention can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or the like, oral preparations, suppositories or sterilized injection solutions according to a conventional method .

In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin And the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used diluents such as water and liquid paraffin, various excipients such as wetting agents, sweetening agents, fragrances and preservatives . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Propellants, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of suppositories, witepsol, tween 61, cacao paper, laurin, and glycerogelatin can be used.

The pharmaceutical composition according to the present invention can be administered to the individual by various routes. All modes of administration may be expected, for example, by oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the age, weight, sex, physical condition, etc. of the individual. A serine protease inhibiting peptide contained in the pharmaceutical composition; It is obvious that the concentration of the expression vector may be variously selected depending on the subject. Preferably, the concentration of the expression vector is 0.01 to 5,000 / / ml in the pharmaceutical composition. If the concentration is less than 0.01 μg / ml, the pharmaceutical activity may not be exhibited. If the concentration is more than 5,000 μg / ml, it may be toxic to the human body.

The present invention also relates to a method for producing a fermentation broth by cultivating at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. The present invention provides a method of producing a milk cream fermentation broth for preventing, alleviating and improving atopic dermatitis.

Milk-cream, milk, skim-milk, Lactobacilli MRS medium, LB (lysogeny broth) medium, R2A medium (MB-R2230) and casein hydrolyzate acid-containing medium, but the present invention is not limited thereto.

The method may include treating at least one selected from the group consisting of tyndallization treatment, reduced pressure concentration treatment, and yeast extract treatment to the milk cream fermentation broth, but the present invention is not limited thereto.

And encapsulating the milk cream fermentation broth to make it for oral use, but it is a step included in the production for oral use, but is not limited thereto.

The milk cream fermentation broth may be a fermented milk cream fermented with various lactic acid bacteria or a mixed fermentation broth of a crude fermented broth of a lactic acid bacterium or a lactic acid bacterium fermented with various lactic acid bacteria or a mixture of fermented milk cream fermented with various lactic acid bacteria Yeast extract and MRS broth treated with fermentation broth of milk cream, fermented milk cream or milk cream, and fermentation broth of milk cream which is obtained by treating yeast extract and MRS broth. In order to prevent, alleviate and improve atopic dermatitis, Is not limited.

The present invention also provides a fermented milk cream composition comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. A quasi-drug composition for preventing, alleviating and improving atopic dermatitis, comprising

In the present invention, the term " quasi-drug " means a fiber, a rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a human or an animal, a weak action on the human body, Or similar to those not of machinery, for the purpose of diagnosing, treating, alleviating, treating or preventing diseases of humans or animals, including those intended for sterilization, insecticide and similar uses for the prevention of infection Means an article, except machinery, machinery or equipment, used for the purpose of giving pharmacological effects to the structure and function of humans or animals, It also includes sanitary goods.

When the composition of the present invention is contained in a quasi-drug for the purpose of preventing, alleviating and improving atopic dermatitis, the composition may be used as it is, or may be used together with other quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The quasi-drug of the present invention is not particularly limited, but may be used in the form of, for example, a cream, a lotion, an aerosol, a shampoo, an gel or a pack.

In the case of a cream, an ointment, a shampoo, a gel, or a pack, it is preferable to use white petrolatum, yellow petrolatum, lanolin, bleached wax, cetanol, stearyl alcohol, stearic acid, hydrogenated oil, Base of; Solvents and dissolution aids such as oleic acid, myristic acid isopropyl, triisooctanoic acid glycerin, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acid, fatty acid ester, aliphatic alcohol and vegetable oil; Antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, and butylhydroxyanisole; Preservatives such as parahydroxybenzoic acid ester; Humectants such as glycerin, propylene glycol and sodium hyaluronate; Surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; Carboxyvinyl polymer, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salt, hydroxypropylcellulose, hydroxypropylmethylcellulose and the like.

In the case of the aerosol preparation, the white petrolatum, the yellow petrolatum, the lanolin, the bleached wax, the cetanol, the stearyl alcohol, the stearic acid, the stearic acid, Base oils such as hydrogenated oils, gelled hydrocarbons, polyethylene glycols, liquid paraffin, squalane and the like; Oleic acid, myristic acid isopropyl, diisopropyl adipate, isobutyl sebacate, glycerin triisooctanoate, crotamiton, diethyl sebacate, hexyl laurate, fatty acid esters, aliphatic alcohols and vegetable oils Solvents and dissolution aids; Antioxidants such as tocopherol derivatives, L-ascorbic acid, dibutylhydroxytoluene, and butylhydroxyanisole; Preservatives such as parahydroxybenzoic acid ester; Humectants such as glycerin, propylene glycol and sodium hyaluronate; Surfactants such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, and lecithin; Thickening agents such as carboxyvinyl polymer, xanthan gum, carboxymethyl cellulose, carboxymethyl cellulose sodium salts, hydroxypropyl cellulose, hydroxypropylmethyl cellulose and the like; In addition, various stabilizers, buffers, mating agents, suspending agents, emulsifiers, fragrances, preservatives, solubilizers, and other suitable additives may be added.

In addition, a stabilizer, a preservative, an absorption promoter, a pH adjuster, and other suitable additives may be added as necessary.

In addition, the composition of the present invention may be used in combination with other atopic dermatitis agents known to date in order to further enhance the activity of preventing, alleviating and improving atopic dermatitis.

The present invention will be described in more detail with reference to the following examples. However, the following examples are only for the purpose of illustrating the present invention, and thus the present invention is not limited thereto.

Example 1 Preparation of Fermented Milk Cream Fermented with Various Lactic Acid Bacterium Strains, Intermittently Sterilized Milk Cream Fermented Liquid, Preparation of Fermented Concentrate and Its Treatment for Atopic Dermatitis

<Example 1-1> Preparation of a milk cream fermentation broth fermented with various lactic acid bacteria strains

Lactococcus lactis subsp. Lactis KCTC 3115 (K1), Lactobacillus plantarum ATCC 8014 (K2), and Lactobacillus parasitol were preincubated in 1 liter of milk containing 35% or more milk fat. Lactobacillus paracasei subsp. Paracasei KCTC 3510 (K3), Lactobacillus gasseri KCTC 3162 (K4), Lactobacillus delbrueckii subsp. Bulgaricus KCTC 3635 (K5), Lactobacillus lutei (Lactobacillus reuteri KCTC 3594) (K6 ), Lactobacillus salivarius (Lactobacillus salivarius subsp. salicinius KCTC 3600 ) (K7), Lactobacillus Planta Room (Lactobacillus fermentum KCTC 3112) (K8 ), Lactobacillus ash also filler's (Lactobacillus acidophilus KCTC 3140) (K9) at a concentration of 1 × 10 ⁶ cfu / ㎖ or more, and cultured in a 37 ° C incubator for more than 24 hours.

Or more than 1 x 10 &lt; ⁶ &gt; cfu / ml of Bifidobacterium bifidum KCTC 3202 (K10) preincubated in 1 liter of milk cream containing 35% or more of milk fat in the same manner as above, 37 fermentation broth for 24 hours or more to remove the oxygen in the fermentation broth by anaerobic jar and carbon dioxide gas pack (Anaerogen 2.5L, Thermo scientific, USA) .

As a control for the milk cream fermentation broth of the various lactic acid bacteria strains, Lactobacillus rhamnosus KCTC 5033 milk cream fermentation broth (B2) was used according to the method described in the prior art (10-2016-0035879). The Lactobacillus lambosus was inoculated with 1 x 10 ⁶ cfu / ml of Lactobacillus lambosus strain pre-cultured in 1 l of a milk cream containing 35% or more of milk fat, and then cultured in a 37 ° C incubator for 72 hours. &Lt; / RTI &gt;

<Example 1-2> Preparation of Lotion Formulation Containing Various Lactic Acid Bacterial Cream Fermented Liquid

Lotion formulations containing 75% by weight of the total weight of the formulation of the various lactic acid bacterial oil-containing fermentation broths of Example 1-1 were prepared according to the following Table 1. As a control, the lotion (B2) containing the fermentation broth of Lactobacillus laminosus milk cream was prepared according to the method described in the prior art (10-2016-0035879, PCT / KR2017 / 003172), 75 units of fermented sterilized milk cream (g), olive - derived wax 5 unit (g), napre 1 unit (g) as an oil base, and 3 drops of lavender essential oil.

Ingredient Unit (g) Lotion W Fermented sterilized milk-cream (K1, K2, K3, K4, K5, K6, K7, K8, K9, K10) 75 O Grape seed oil 19 Olive derived wax 5 A Napre One Total weight 100 K3, fermented milk-cream with KCTC 3510; K4, fermented milk-cream with KCTC 3115; K5, fermented milk-cream with ATCC 8014; K3, fermented milk- fermented milk-cream with KCTC 3635; K6, fermented milk-cream with KCTC 3594; K7, fermented milk-cream with KCTC 3600; K8, fermented milk-cream with KCTC 3112; milk-cream with KCTC 3202); O, oil base; A, additives. This biological resources used in this research were distributed from KCTC.

<Example 1-3> Preparation of Lactobacillus lambosus milk cream fermentation broth

In the same manner as in < Example 1-1 &gt;, 1 x 10 ⁶ cfu / ml or more of Lactobacillus lambus oocyte precultured in milk cream containing 35% or more of milk fat was inoculated, And cultured to obtain a milk cream fermentation broth.

<Example 1-4> Preparation of Tyndallized Lactobacillus Lambsus milk cream fermentation broth

The lactobacillus laminosse oil cream fermentation broth obtained in the same manner as in <Example 1-3> was heated to 80 or higher at a constant atmospheric pressure for 15 minutes or longer to obtain an intermittently sterilized milk cream fermentation broth.

Example 1-5 Preparation of Various Lactic Acid Fermentation Concentrate

Lactobacilli MRS or skim milk was inoculated with 1 × 10 ⁶ cfu / ml or more of lactobacillus lambus and Lactococcus lactis pre-cultured in the same manner as in Example 1, And cultured for more than 24 hours to obtain various lactic acid fermentation broths. Thereafter, various lactic acid fermentation broths were subjected to intermittent sterilization by heating at a constant pressure of 80 or more for 15 minutes or the like, followed by heating at 60 or more for 60 minutes or longer. .

&Lt; Example 1-6 > Preparation of lotion formulation containing variously prepared milk cream fermentation broth

Tyndallized Lactobacillus laminosse oil cream fermentation broth (K11) obtained in the above <Example 1-4>, Lactobacillus laminosse oil cream fermentation broth obtained in the above <Example 1-3> and Example (K12, K13, K14, and K15) obtained by blending various lactic acid-enriched fermented broths in the proportions shown in Table 2 below, the lactobacillus lanxosus strains obtained in the above <Example 1-3> 25% by weight of the fermentation broth of lactocococcus lactis oil cream obtained in the above <Example 1-1>, 25% by weight of the fermentation broth of lactobacillus gasseri oil cream, (K16) mixed with 25% by weight of a milk cream fermentation broth was prepared at a ratio of 75% by weight of the total weight of the formulation (Table 2).

The lotion containing the fermentation broth of Lactobacillus laminosus milk cream was the same as that of the Bacillus rhamnosus oil cream formulation (B2) of Example 1, and was used as a comparative group of the various lactic acid bacteria oil cream lotions.

Ingredient Unit (g) Lotion W Fermented sterilized milk cream (K11, K12, K13, K14, K15, K16) 75 O Grape seed oil 19 Olive derived wax 5 A Napre One Total weight 100 K13, mixture, added 10% 10-fold concentrated fermented broth with KCTC 5033 to fermented milk-cream with KCTC 5033; K13, mixture, added 10-fold concentrated fermented broth with KCTC 5033 to fermented milk-cream with KCTC 5033; K14, mixture, added 10% 10-fold concentrated fermented broth with KCTC 5033 to fermented milk-cream with KCTC 5033; Fermented milk-cream with KCTC 3133, fermented milk-cream with KCTC 5033; fermented milk-cream with KCTC 3162, fermented milk-cream with KCTC 3162 and fermented milk-cream with KCTC 3635); O, oil base; A, additives.

<Experimental Example 1-1> Selection of subjects to be tested for atopic dermatitis

Among the participants who had atopic dermatitis lesions recruited through the clinical recruitment announcement, those who showed intermittent or continuous symptoms of atopic dermatitis for 6 months or more were selected first, and within 6 months of them, local and systemic steroids, Immunomodulators and inhibitors, antihistamines, and those who have received similar treatment, except those who wish to participate in the final test. The duration of the test was 6 weeks (42 days).

&Lt; Experimental Example 1-2 >

In order to measure the exact state of the skin lesion, the MPA580 (Multi Probe Adapter system, C + K Electronic GmbH, Germany) was used in the skin measurement room where the room temperature and humidity were set at 20 to 25 at room temperature and 40 to 60% Cologne, Germany). The test subjects were prohibited to apply the lotion for 12 hours before entering the skin measurement room for accurate evaluation of the lesion area, and they were measured after taking the stabilization for about 30 minutes after admission.

In addition, skin measurements were made twice before and after the clinical trial, and two subjects were measured in the same way for more objective results. Skin measurements were performed by specifying skin surface hydration (SSH), skin acidity (pH), and transepidermal water loss (TEWL) as the severity of the lesions of the test subjects , And the objective lesion site was measured after the clinical trial.

In addition, lesion sites of participating subjects were photographed twice before and after the test using a digital camera (SAMSUNG, VLUU PL150). To obtain objective results, one identical photographer took the same lesion site as the mechanical measurement site at the designated location.

<Experimental Example 1-3> Improvement of atopic dermatitis lesion after application of lotion containing fermented milk cream fermented with various lactic acid bacteria

<1-3-1> Confirmation of skin moisture content (SSH)

The lotion containing the fermented milk cream fermented with various lactic acid bacteria prepared in the same manner as in <Example 1-2> was applied to the atopic dermatitis lesion site using the fermentation broth obtained in <Example 1-1> Changes in the moisture content of the skin before and after was confirmed.

As a result, as shown in the following Table 3, the Lactobacillus laminosse oil cream fermentation broth (B2) was found to be about 22% significant as in Experimental Example 1-3 of the present invention's patent application (10-2016-0035879) Lactobacillus lactis (K1) was significantly increased by about 21%, Lactobacillus plantarum (K2) was increased by about 12%, Lactobacillus paracasei (K3) was increased by about 18% Lactobacillus delbrueckii (K5) was increased by about 24%, Lactobacillus lutea (K6) was increased by about 16%, Lactobacillus delbrueckii (K5) Salivary (K7) was significantly increased by about 23%, Lactobacillus fermentum (K8) was increased by about 19%, Lactobacillus acidophilus (K9) was increased by about 17%, Bifidobacterium Bifidum (K10) increased skin moisture content by about 22% The lotions containing the fermented milk cream fermented with a lactic acid bacterium were found to increase the skin moisture content of the atopic dermatitis lesion area similarly to the case of applying the fermentation broth of lactobacillus rhamnosus oil cream (B2) (Table 3).

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) SSH B2 21 22.12 ± 7.98 27.05 7.99 -4.92 + 4.53 -4.981 (.000 ^*** ) K1 3 20.04 ± 3.14 24.31 ± 2.17 -4.27 ± 1.05 -7.056 (.020 ^* ) K2 3 19.77 ± 2.63 22.10 ± 2.81 -2.33 + - 0.61 -6.614 (.022 ^* ) K3 3 23.85 ± 3.20 28.13 + - 2.69 -4.28 ± 0.52 -14.288 (.005 ^** ) K4 3 20.33 + - 3.99 24.64. + -. 3.73 -4.31 ± 0.31 -23.834 (.002 ^** ) K5 3 21.41 + - 4.25 26.48 ± 2.01 -5.07 ± 2.67 -3,292 (.081) K6 3 26.59 ± 2.74 30.91 + 2.80 -4.32 ± 0.31 -24.289 (.002 ^** ) K7 3 19.50 ± 1.04 24.07 ± 1.59 -4.56 ± 1.76 -4.484 (.046 ^* ) K8 3 21.94 ± 3.12 26.07 ± 3.25 -4.12 ± 0.24 -30.279 (.001 ^** ) K9 3 23.52 + 1.32 27.51 + - 1.15 -3.99 ± 0.28 -24.267 (.002 ^** ) K10 3 20.72 ± 2.64 25.27 + - 3.44 -4.55 + 0.88 -8.927 (.012 ^* ) ^* p <.05, ^** p <.01, ^*** p <.001
Abbreviation: B2, testing products made from fermented milk-cream with KCTC 5033; K1, testing products made from fermented milk-cream with KCTC 3115; K2, testing products made from fermented milk-cream with ATCC 8014; K3, testing products made from fermented milk-cream with KCTC 3510; K4, testing products made from fermented milk-cream with KCTC 3162; K5, testing products made from fermented milk-cream with KCTC 3635; K6, testing products made from fermented milk-cream with KCTC 3594; K7, testing products made from fermented milk-cream with KCTC 3600; K8, testing products made from fermented milk-cream with KCTC 3112; K9, testing products made from fermented milk-cream with KCTC 3140; K10, testing products made from fermented milk-cream with KCTC 3202; N, the number of people in group.

<1-3-2> Determination of acidity (pH) of skin surface

The change in acidity of the skin surface before and after the test was confirmed in the same manner as in <1-3-1> above.

As a result, as shown in the following Table 4, as in Experiment 1-3 of the present invention's patent application (10-2016-0035879), the lactobacillus laminosse oil cream fermentation broth (B2) (K2) was decreased by about 2%, Lactobacillus paracase (K3) was decreased by about 5%, Lactobacillus lactis (K1) was decreased by about 6% (K4) decreased by about 5%, Lactobacillus delbrueckii (K5) decreased by about 7%, Lactobacillus lutea (K6) decreased by about 4%, Lactobacillus salivarius (K7) Lactobacillus acidophilus (K9) decreased by about 4%, Lactobacillus acidophilus (K9) decreased by about 3%, Bifidobacterium bifidum (K10) decreased by about 6%, Skin surface pH , And lotions containing a fermented milk cream fermented with various lactic acid bacteria were also found to contain Lactobacillus lambs It was confirmed that similarly to reduce the pH of the skin lesions of atopic dermatitis and, if the application of the cream's organic broth (B2) (Table 4).

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) pH B2 21 5.56 + - 0.48 5.21 + - 0.38 0.34 0.39 3.963 (.001 ^** ) K1 3 5.62 ± 0.18 5.27 ± 0.37 0.35 + 0.22 2.714 (.113) K2 3 5.45 ± 0.41 5.31 0.32 0.13 + - 0.12 2.753 (.111) K3 3 5.56 ± 0.08 5.30 ± 0.15 0.26 ± 0.11 4.274 (.051) K4 3 5.73 ± 0.32 5.47 ± 0.37 0.26 ± 0.06 7.403 (.018 ^* ) K5 3 5.55 0.21 5.16 + 0.06 0.39 ± 0.18 1.429 (.289) K6 3 5.50 0.25 5.26 ± 0.38 0.23 ± 0.20 2.056 (.176) K7 3 5.47 ± 0.24 5.15 ± 0.17 0.31 ± 0.09 5.742 (.029 ^* ) K8 3 5.55 + - 0.10 5.33 ± 0.17 0.22 0.08 4.801 (.041 ^* ) K9 3 5.51 + - 0.15 5.33 + - 0.14 0.17 ± 0.10 3.159 (.087) K10 3 5.56 + - 0.42 5.15 + 0.26 0.31 ± 0.01 4.093 (.055) ^* p <.05, ^** p <.01
Abbreviations were the same as Table 3.

<1-3-3> Determination of Percutaneous Moisture Evaporation Rate (TEWL)

The change in the amount of transdermal water evaporation before and after the test was confirmed in the same manner as in <1-3-1> above.

As a result, as shown in the following Table 5, the lactobacillus lambsos milk cream fermentation broth (B2) of about 32% was obtained as in Experimental Example 1-3 of the present invention's patent application (10-2016-0035879) Lactobacillus plantarum (K2) was significantly reduced by about 20%, and Lactobacillus paracasei (K3) was decreased by about 23%, while that of Lactococcus lactis (K1) was decreased by about 32% (K4) was significantly reduced by about 29%, Lactobacillus delbrueckii (K5) was decreased by about 29%, Lactobacillus lutein (K6) was decreased by about 24%, Lactobacillus saliva Lactobacillus fermentum (K8) was significantly decreased by 26%, Lactobacillus acidophilus (K9) was decreased by about 27%, and Bifidobacterium bifidus The supplement (K10) showed a significant 31% decrease in skin TEWL, The lotions containing the fermented milk cream fermentation solution also confirmed that the TEWL of the atopic dermatitis lesion area was reduced similarly to the case of applying the fermentation broth (B2) of Lactobacillus laminosus milk cream (Table 5).

Therefore, the fermented milk cream fermentation broth mainly fermented by various other lactic acid bacteria mainly increases the skin moisture content, decreases the pH value of the skin, decreases the transdermal water evaporation amount, and increases the lactose content of the conventional lactose (10-2016-0035879, PCT / KR2017 / 003172) It was confirmed that it not only suppresses the expression of lesions of atopic dermatitis but also alleviates the lesion and inhibits the recurrence of lesions similarly to the case of applying the fermentation broth of B2 cream of Bacillus laminosus milk.

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) TEWL B2 21 38.94 + 21.58 26.62 ± 14.36 12.32 ± 13.59 4.154 (.000 ^*** ) K1 3 33.68 ± 3.10 22.93 + 1.57 10.75 ± 2.57 7.229 (.019 ^* ) K2 3 32.65 ± 1.15 26.24 + 1.49 6.41 ± 1.48 7.479 (.017 ^* ) K3 3 34.59 ± 3.34 26.78 ± 3.19 7.81 ± 2.72 4.967 (.038 ^* ) K4 3 31.60 ± 1.97 22.48 ± 0.63 9.12 + 1.36 11.658 (.007 ^** ) K5 3 35.35 + - 4.17 24.96 ± 6.32 10.39 + - 2.41 7.461 (.017 ^* ) K6 3 29.54 + - 7.23 22.38 + - 4.76 7.17 ± 3.27 3.802 (.063) K7 3 35.09 + - 4.58 23.97 + - 4.73 11.13 ± 1.19 16.257 (.004 ^** ) K8 3 29.32 ± 3.29 21.68 ± 2.86 7.64 ± 0.71 18.544 (.003 ^** ) K9 3 36.23 + - 6.23 26.30 ± 2.67 9.94 + - 5.41 10.640 (.009 ^** ) K10 3 34.89 + - 4.33 24.06 + - 3.02 10.83 ± 2.04 9.195 (.012 ^* ) ^* p <.05, ^** p <.01, ^*** p <.001
Abbreviations were the same as Table 3.

<Experimental Example 1-4> Confirmation of improvement effect of atopic dermatitis lesions after application of variously prepared lotions containing Lactobacillus rhamnosus milk cream fermentation broth

<1-4-1> Confirmation of skin moisture content (SSH)

The lotion containing the milk cream fermented liquid prepared variously by the method of <Example 1-5> was applied to the atopic dermatitis lesion site, and then the change of the skin moisture content before and after the test was confirmed.

As a result, as shown in the following Table 6, the Lactobacillus laminosse oil cream fermentation broth (B2) was found to be about 22% significant as in Experimental Example 1-3 of the present invention's patent application (10-2016-0035879) (K11) increased by about 29%, the mixed milk cream fermentation solution of 90% by weight of the lactobacillus laminosse oil cream fermentation broth and 10% by weight of the lactobacillus laminocus 10-fold concentrated solution, while the tyndallized milk cream fermented broth (K11) (K12) was significantly increased by about 32%, and the mixed milk cream fermented liquid (K13) of 80% by weight of the lactobacillus laminocyte cream fermentation broth and 20% by weight of the lactobacillus lan- nomus 10-fold concentrate was significantly increased by about 31% The mixed milk cream fermentation broth (K14) containing 90% by weight of the fermentation broth of Bacillus lambosus milk cream and 10% by weight of the lactococcus lactis 10-fold concentrate significantly increased by about 29%, and 90% by weight of the fermentation broth of Lactobacillus laminosus milk cream gambling The mixed fermentation broth (K15) containing 10% fermented liquor by 10% fermented concentrate showed a significant increase in skin moisture content by about 30%, showing that the skin of atopic dermatitis lesion area was significantly higher than that of Lactobacillus laminosse oil cream fermentation liquid (B2) (Table 6). In addition, it was found that the water content was higher than that of the control.

On the other hand, a mixture of 25% by weight of a fermentation broth of Lactobacillus laminosus milk cream, 25% by weight of a lactococcus lactis milk cream fermentation broth, 25% by weight of a lactobacillus seed oil cream fermentation broth, and 25% The milk cream fermentation broth (K16) showed a significant increase in moisture content of the skin by about 22%, and it was confirmed that the skin moisture content of the atopic dermatitis lesion area was increased similarly to the case of applying the fermentation broth (B2) of Lactobacillus laminosus milk cream (Table 6).

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) SSH B2 21 22.12 ± 7.98 27.05 7.99 -4.92 + 4.53 -4.981 (.000 ^*** ) K11 3 21.30 ± 3.29 27.47 ± 1.22 -6.17 + - 4.38 -2.439 (.135) K12 3 22.57 + - 4.42 29.80 + - 5.77 -7.23 + 1.94 -6.458 (.023 ^* ) K13 3 18.30 ± 2.86 24.03 + - 3.92 -5.73 ± 1.10 -9.015 (.012 ^* ) K14 3 19.00 ± 1.92 24.46 + - 2.50 -5.46 ± 1.10 -8.568 (.013 ^* ) K15 3 23.55 + 1.82 30.59 ± 1.78 -7.03 ± 1.20 -10.124 (.009 ^** ) K16 3 21.30 ± 2.81 26.02 ± 3.69 -4.72 + 0.88 -9.261 (.012 ^* ) ^* p <.05, ^*** p <.001
Abbreviation: B2, testing products made from fermented milk cream with KCTC 5033; K11, testing products made from tyndalized fermented milk-cream broth with KCTC 5033; K12, testing products made from mixture, ad 10% 10-fold concentrated fermented skim milk broth with KCTC 5033 to fermented milk cream with KCTC 5033; K13, fermented milk cream with KCTC 5033; KCTC 5033 to fermented milk cream with 10-fold concentrated fermented skim milk broth with KCTC 5033; K14, testing products made from mixture, ad 10% 10-fold concentrated fermented broth with KCTC 3115 to fermented milk cream with KCTC 5033; K15, 10-fold concentrated fermented broth with KCTC 3202 to fermented milk cream with KCTC 5033; K16, testing products made from fermented milk-cream mixture, mixing fermented milk-cream KCTC 5033, KCTC 3115, KCTC 3162 and KCTC 3635; N, the number of people in group.

<1-4-2> Determination of acidity (pH) of skin surface

The change in acidity of the skin surface before and after the test was confirmed in the same manner as in <1-4-1> above.

As a result, as shown in the following Table 7, the lactobacillus lambsos milk cream fermentation broth (B2) of about 6% was obtained as in Experimental Example 1-3 of the present invention's patent application (10-2016-0035879) , While the tyndallized milk cream fermentation broth (K11) was reduced by about 7%, a mixed milk cream fermentation liquid of 90% by weight of the lactobacillus lanosus milk cream fermentation broth and 10% by weight of the lactobacillus lanxus 10-fold concentrated solution (K12) was significantly decreased by about 9%, and the mixed milk cream fermented liquid (K13) of 80% by weight of the lactobacillus laminocyte cream fermentation broth and 20% by weight of the lactobacillus laminocus 10-fold concentrated concentrate was significantly decreased by about 8% The mixed milk cream fermentation broth (K14) of 90% by weight of the fermentation broth of Bacillus lambosus milk cream and 10% by weight of the lactococcus lactis 10-fold concentrate was significantly reduced by about 8%, and 90% by weight of the fermentation broth of Lactobacillus laminosus milk cream, Gambling terry The mixed fermentation broth (K15) containing 10% by weight of the 10-fold fermentation concentrate of Bifidum 10% decreased the pH of the skin surface by about 8% and the skin pH of the atopic dermatitis lesion site was lower than that of the fermentation broth of Lactobacillus laminosis milk cream (Table 7).

 On the other hand, 25% by weight of lactobacillus lanosus milk cream fermentation broth was mixed with 25% by weight of fermentation broth of lactococcus lactis milk cream, 25% by weight of fermentation broth of lactobacillus gasseri oil cream, and 25% by weight of fermentation broth of lactobacillus delbrueckii cream It was confirmed that the cream fermentation broth (K16) decreased the pH of the skin surface by about 6%, thereby decreasing the skin pH of the atopic dermatitis lesion similarly to the case of applying the fermentation broth (B2) of Lactobacillus lymphosus milk cream (Table 7) .

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) pH B2 21 5.56 + - 0.48 5.21 + - 0.38 0.34 0.39 3.963 (.001 ^** ) K11 3 5.59 + - 0.12 5.20 0.30 0.39 0.37 1.814 (.211) K12 3 5.64 ± 0.28 5.15 ± 0.20 0.49 + 0.11 7.621 (.017 ^* ) K13 3 5.66 ± 0.23 5.21 ± 0.15 0.45 + 0.17 4.525 (.046 ^* ) K14 3 5.38 ± 0.25 4.97 ± 0.23 0.41 + 0.04 7.581 (.017 ^* ) K15 3 5.57 ± 0.27 5.11 ± 0.11 0.46 0.28 2.859 (.104) K16 3 5.68 ± 0.28 5.35 + 0.14 0.32 ± 0.29 1.951 (.190) ^* p <.05, ^** p <.01
Abbreviations were the same as Table 6.

<1-4-3> Determination of Percutaneous Moisture Evaporation Rate (TEWL)

The change in the amount of percutaneous water evaporation before and after the test was confirmed in the same manner as in <1-4-1> above.

As a result, as shown in the following Table 8, the Lactobacillus laminosse oil cream fermentation broth (B2) was about 32% significant as in Experimental Example 1-3 of the present invention's patent application (10-2016-0035879) (K11) was significantly reduced by about 36%, and that of 90% by weight of the fermentation broth of Lactobacillus laminosus milk cream and 10% by weight of the Lactobacillus laminocus 10-fold concentrated fermentation broth The mixed fermentation broth (K12) was significantly reduced by about 41%, the mixed fermentation broth (K13) containing 80% by weight of the fermentation broth of the lactobacillus rhamnosus milk cream and 20% by weight of the 10-fold concentrated fermentation broth of the lactobacillus lambsosus was significantly reduced by about 40% The mixed fermentation broth (K14) containing 90% by weight of the fermentation broth of Bacillus laminosseus cream and 10% by weight of the 10-fold concentrated fermentation broth of Lactococcus lactis significantly decreased by about 38%, and 90% by weight of the fermentation broth of Lactobacillus laminosse milk cream, Lium The K15 fermentation broth (K15) with a 10-fold concentration of 10-fold fermented soybean milk significantly decreased the amount of transdermal water evaporation by about 39%, and decreased the TEWL of the atopic dermatitis lesion site compared with the fermentation broth of lactobacillus lan- nomus milk cream (Table 8).

On the other hand, a mixture of 25% by weight of a fermentation broth of Lactobacillus laminosus milk cream, 25% by weight of a lactococcus lactis milk cream fermentation broth, 25% by weight of a lactobacillus seed oil cream fermentation broth, and 25% The milk cream fermentation broth (K16) showed a significant decrease in the transdermal water evaporation amount by about 31%, and it was confirmed that the TEWL of the atopic dermatitis lesion area was reduced similarly to the case of applying the fermented broth of lactobacillus laminosis oil cream (B2) 8).

Therefore, a fermentation broth of a lactobacillus laminosus milk cream fermented with intermittent sterilization (tyndallization) or a mixed fermentation broth of 90% by weight of a fermentation broth of lactobacillus laminosseus cream with 10% by weight of a 10-fold concentrated fermentation broth of Lactobacillus lambosus, Or a mixed fermentation broth of 20% by weight of lactobacillus lan- myus 10-fold concentrated fermentation broth or a mixed fermentation broth of 90% by weight of a fermentation broth of lactobacillus rhamnosus milk cream and 10% by weight of a 10-fold concentrated fermentation broth of Lactobacillus lactis, The mixed fermentation broth containing 90% by weight of the fermented cream liquid and 10% by weight of the Bifidobacterium bifidum 10-fold concentrate all exhibited an increase in the skin moisture content, a decrease in the pH value of the skin and a decrease in the transdermal water evaporation amount, (10-2016-0035879) and the International Application (PCT / KR2017 / 003172) on March 25, 2017, the Lactobacillus rhamnosus oilcream (B2), it was confirmed that not only the expression of lesions of atopic dermatitis was suppressed but also the lesion was alleviated and the lesion recurrence was greatly suppressed.

In addition, 25% by weight of the lacto-koccus lactis milk cream fermentation liquid arbitrarily selected for 25% by weight of the lactobacillus laminosus milk cream fermentation liquid, 25% by weight of the lactobacillus gasseri oil cream fermentation broth selected from the lotions containing the fermented milk cream fermented with various lactic acid bacteria %, Lactobacillus delbrueckii fermentation broth of 25% by weight of the fermentation broth of lactic acid bacteria was found to be effective in increasing the skin moisture content of atopic dermatitis lesions, decreasing the skin pH value and reducing the transdermal water evaporation amount, (B2) of Lactobacillus rhamnosus milk cream (B2) of International Patent Application (PCT / KR2017 / 003172) on March 25, 2017, and the expression of lesions of atopic dermatitis But also contributes to suppression of lesion and inhibition of lesion recurrence.

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) TEWL B2 21 38.94 + 21.58 26.62 ± 14.36 12.32 ± 13.59 4.154 (.000 ^*** ) K11 3 30.23 + - 6.62 19.20 ± 4.54 11.03 + - 2.87 6.660 (.022 ^* ) K12 3 29.34 ± 5.99 17.27 + - 4.62 12.08 ± 1.57 13.316 (.006 ^** ) K13 3 39.83 + - 2.30 23.77 + - 2.56 16.06 + - 4.18 6.657 (.022 ^* ) K14 3 37.50 + - 7.15 23.11 + - 6.47 14.39 ± 2.63 9.481 (.011 ^* ) K15 3 34.91 + 2.94 21.24 + - 4.58 13.67 ± 1.85 12.808 (.006 ^** ) K16 3 34.17 ± 7.11 23.47 ± 5.67 10.70 ± 1.71 10.818 (.008 ^** ) ^* p <.05, ^** p <.01, ^*** p <.001
Abbreviations were the same as Table 6.

Example 2 Preparation of Various Oil-Cream Fermented Liquids Containing Yeast Extract in Various Lactic Acid Bacterium Strains and Treatment of Atopic Dermatitis

&Lt; Example 2-1 > Preparation of various milk cream media fermentation broth

Lactobacillus lambatus KCTC 5033 was added to 1 liter of a milk cream yeast medium containing 99% by weight of milk cream containing 35% or more of milk fat and 1% by weight of yeast extract (Yeast extract, Becton, Dickinson & Co, Calif., USA) × 10 ⁶ cfu / ml or more, and cultured in a 37 ° C. incubator for more than 24 hours to obtain a lactobacillus lambsus milk cream yeast fermentation solution (M1).

Lactobacillus lambosus KCTC 5033 was added to 1 liter of milk cream MRS medium containing 5% by weight of Lactobacilli MRS broth (Becton, Dickinson & Co, Calif., USA) in 95% by weight of milk cream containing 35% × 10 ⁶ cfu / ml or more and then cultured in a 37 ° C incubator for 24 hours or more to obtain a lactobacillus lambsos milk cream MRS fermentation broth (M2).

Lactococcus lactis cream yeast fermentation broth (M3) and lactococcus lactis milk cream MRS fermentation broth (M4) were each inoculated into milk cream yeast medium and milk cream MRS medium in the same manner as above, .

In the same manner as above, Lactobacillus plantarum, which had the lowest change in the fermented milk cream fermented with various lactic acid bacteria, was inoculated into milk cream yeast medium and milk cream MRS medium respectively to obtain Lactobacillus plantarum milk yeast fermentation broth M5), and Lactobacillus plantarum milk cream MRS fermentation broth (M6), respectively.

&Lt; Example 2-2 > Preparation of Lotion Formulation Containing Various Fermented Milk Creams

A lotion formulation containing 75 wt% of the total weight of the formulation was prepared as follows (Table 13). The milk cream yeast fermentation broth and milk cream MRS fermentation broth obtained in Example 2-1 were prepared as follows.

In addition, a lotion containing a fermented liquid of Lactobacillus lansosus milk cream was prepared according to the method described in the prior art (10-2016-0035879, PCT / KR2017 / 003172), 75 units (g) of fermented sterilized milk cream as a water base, The lotion was made up of 19 unit (g) of grape seed oil, 5 unit (g) of olive-derived wax, 1 unit of napre (g) as an additive and 3 drops of lavender essential oil.

The lotion containing the lactococcus lactis milk cream fermentation broth and the lactobacillus plantarum milk cream fermentation liquid was the same as the oil cream lotion formulation prepared in the above Example 1-6, Respectively.

Ingredient Unit (g) Lotion W Fermented sterilized milk cream (M1, M2, M3, M4, M5, M6) 75 O Grape seed oil 19 Olive derived wax 5 A Napre One Total weight 100 Fermented milk-cream & yeast broth with KCTC 5033; M2, fermented milk-cream & MRS broth with KCTC 5033; M3, fermented milk-cream & yeast broth with KCTC 3115; M4, fermented milk -cream & MRS broth with KCTC 3115; M5, fermented milk-cream & yeast broth with ATCC 8014; M6, fermented milk-cream & MRS broth with ATCC 8014; Milk-cream & yeast broth, mixture of 99% and 1% of yeast extract; Milk-cream & MRS broth, mixture of 95% of milk-cream and 5% of MRS broth); O, oil base; A, additives.

<Experimental Example 2-3> Confirmation of improvement effect of atopic dermatitis lesions after applying lotion containing various milk cream liquid fermentation broth

<2-3-1> Confirmation of skin moisture content (SSH)

The lotion was applied to the atopic dermatitis lesion site in the <Example 2-2> with the fermentation liquid obtained in the <Example 2-1>, and then the change in the skin moisture content before and after the test was confirmed.

As a result, as shown in Table 10 below, the lactobacillus laminosse oil cream fermentation broth (B2) was significantly increased by about 22%, whereas the lactoferricylcholactus (K1) 21%, while Lactobacillus plantarum (K2) was significantly increased by about 12%, which was slightly lower than that of other lactic acid bacterium creamy fermentation broths.

Lactobacillus lactis milk cream yeast fermented milk (M1) was significantly increased by about 24%, Lactobacillus lambsus milk cream MRS fermented milk (M2) was increased by about 23% M3) was significantly increased by about 24%, and the lactoferricosis lactic acid milk cream MRS fermentation liquid (M4) was significantly increased by about 23%, which was slightly higher than that of the milk cream single culture fermentation liquid. On the other hand, Yeast fermentation broth (M5) was increased by about 18%, and Lactobacillus plantarum milk cream MRS fermentation broth (M6) was increased by about 20%, but it was lower than that of Lactobacillus laminosus milk cream fermentation broth.

As a result, the milk cream fermented milk, milk yeast fermented milk and milk cream fermented milk fermented by Lactobacillus plantarum showed a slightly lower increase than the lactobacillus fermented milk (B2), but the atopic dermatitis lesion Lactobacillus lambsus milk cream yeast fermented milk, Lactobacillus lambosus milk cream MRS fermentation solution, Lactococcus lactis milk yeast fermentation solution, Lactococcus lactis milk cream MRS fermentation broth It was confirmed that it is better to increase the skin moisture content of atopic dermatitis lesion area than that of Lactobacillus laminocus milk cream fermentation (B2) (Table 10).

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) SSH B2 21 22.12 ± 7.98 27.05 7.99 -4.92 + 4.53 -4.981 (.000 ^*** ) K1 3 20.04 ± 3.14 24.31 ± 2.17 -4.27 ± 1.05 -7.056 (.020 ^* ) K2 3 19.77 ± 2.63 22.10 ± 2.81 -2.33 + - 0.61 -6.614 (.022 ^* ) M1 3 20.66 + 4.07 25.54 + - 4.86 -4.88 ± 1.29 -6.557 (.023 ^* ) M2 3 20.82 ± 3.54 25.67 ± 2.37 -4.84 ± 1.32 -6.362 (.024 ^* ) M3 3 22.74 + - 2.45 28.17 + - 2.69 -5.42 ± 1.13 -8.350 (.014 ^* ) M4 3 22.46 ± 1.66 27.65 + - 0.70 -5.19 ± 1.60 -5.635 (.030 ^* ) M5 3 21.19 + - 3.52 25.17 + - 2.63 -3.98 + 1.31 -5.275 (.034 ^* ) M6 3 23.04 + - 2.55 27.67 ± 2.01 -4.62 ± 1.97 -4.066 (.056) ^* p <.05, ^** p <.01, ^*** p <.001
Abbreviation: B2, testing products made from fermented milk-cream with KCTC 5033; K1, testing products made from fermented milk-cream with KCTC 3115; K2, testing products made from fermented milk-cream with ATCC 8014; M1, testing products made from fermented milk-cream & yeast broth with KCTC 5033; M2, testing products made from fermented milk-cream & MRS broth with KCTC 5033; M3, testing products made from fermented milk-cream & yeast broth with KCTC 3115; M4, testing products made from fermented milk-cream & MRS broth with KCTC 3115; M5, testing products made from fermented milk-cream & yeast broth with ATCC 8014; M6, testing products made from fermented milk-cream & MRS broth with ATCC 8014; N, the number of people in group.

<2-3-2> Determination of acidity (pH) of skin surface

The change in acidity of the skin surface before and after the test was confirmed in the same manner as in <2-3-1> above.

As a result, as shown in the following Table 11, the lactobacillus lambsus milk cream fermentation broth (B2) was significantly reduced by about 6%, whereas the lactococcus lactis (K1) 6%, while that of Lactobacillus frantarium (K2) decreased by about 2%, which was somewhat lower than that of other lactic acid bacterium creamy fermentation broths.

Lactobacillus lactis milk cream yeast fermented milk (M3) decreased by about 7%, lactobacillus laminosus milk cream MRS fermented milk (M2) decreased by about 6% (M4) decreased by about 6%, while lactoferricar milk cream yeast fermentation broth (M5) decreased slightly compared to the milk cream single culture fermentation solution, while Lactococcus lactis milk cream decreased by about 6% (M6) was decreased by about 5%, but the decrease of MRS fermented milk (M6) was about 5% lower than that of Lactobacillus lymphosus milk cream fermentation.

As a result, the decrease in milk fermented milk fermented with lactobacillus plantarum, milk yeast fermented milk and milk cream fermented milk was lower than that of lactobacillus fermented milk (B2) Lactobacillus lambsus milk cream yeast fermented milk, Lactobacillus lambosus milk cream MRS fermentation solution, Lactococcus lactis milk cream yeast fermentation solution, Lactococcus lactis milk cream. In the case of MRS fermentation broth, lactose It was confirmed that the skin pH of the atopic dermatitis lesion was better than that of the Bacillus laminocus milk cream fermentation (B2) (Table 11).

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) pH B2 21 5.56 + - 0.48 5.21 + - 0.38 0.34 0.39 3.963 (.001 ^** ) K1 3 5.62 ± 0.18 5.27 ± 0.37 0.35 + 0.22 2.714 (.113) K2 3 5.45 ± 0.41 5.31 0.32 0.13 + - 0.12 2.753 (.111) M1 3 5.45 ± 0.38 5.09 0.26 0.36 + 0.37 1.704 (.230) M2 3 5.33 ± 0.25 5.00 + 0.08 0.34 ± 0.19 3.002 (.095) M3 3 5.39 ± 0.21 5.03 + - 0.14 0.36 + 0.08 9.827 (.010 ^* ) M4 3 5.50 + - 0.42 5.16 ± 0.28 0.34 0.21 2.767 (.110) M5 3 5.37 ± 0.05 5.10 ± 0.23 0.27 ± 0.20 2.304 (.148) M6 3 5.57 ± 0.25 5.26 ± 0.22 0.30 ± 0.07 7.128 (.019 ^* ) ^* p <.05, ^** p <.01
Abbreviations were the same as Table 10.

<2-3-3> Determination of Percutaneous Moisture Evaporation Rate (TEWL)

The change in the amount of transdermal water evaporation before and after the test was confirmed in the same manner as in <2-3-1> above.

As a result, as shown in Table 16 below, the lactobacillus lambsus milk cream fermentation broth (B2) showed a significant decrease in TEWL by about 32%, whereas Lactococcus lactis (K1) significantly decreased in fermentation broth of Lactobacillus lambosus milk cream (K2) was significantly decreased by about 20%, while the decrease of lactobacillus frantarium (K2) was somewhat lower than that of other lactic acid bacterium creamy fermentation broth.

Lactobacillus lactis milk cream yeast fermentation broth (M1) was significantly decreased by about 35%, Lactobacillus laminosis milk cream MRS fermentation broth (M2) was decreased by about 34% M3) was significantly decreased by about 36%, and the lacto-coccus lactis milk cream MRS fermentation liquid (M4) was significantly decreased by about 34%, which was slightly smaller than that of the milk cream single culture fermentation liquid. On the other hand, Yeast fermentation broth (M5) was significantly decreased by about 28%, while Lactobacillus plantarum milk cream MRS fermentation broth (M6) was decreased by about 30%, but the decrease was smaller than that of Lactobacillus laminosse oil cream fermented broth 12).

As a result, the decrease in milk fermented milk fermented with lactobacillus plantarum, milk yeast fermented milk and milk cream fermented milk was lower than that of lactobacillus fermented milk (B2) Lactobacillus lambsus milk cream yeast fermentation broth, Lactobacillus laminosus milk cream MRS fermentation broth, Lactococcus lactis milk cream yeast fermentation solution, Lactococcus lactis milk cream. In the case of the MRS fermentation broth, It was confirmed that TEWL of the atopic dermatitis lesion site was better than that of the case of application of the fermentation broth (B2) of rhamnosseus milk (Table 12).

Therefore, even in the case of milk cream yeast fermentation broth fermented with Lactobacillus plantarum and milk cream fermented with milk, which showed the lowest change in milk fermentation broth fermented with various lactic acid bacteria, inhibition of lesion expression of atopic dermatitis, inhibition of lesion and inhibition of lesion recurrence , And various milk cream fermentation broths increased the skin moisture content, lowered the pH value of the skin, decreased the transdermal water evaporation amount, and applied to the present inventors' patent application filed on Mar. 25, 2016 (10-2016-0035879 (B2) of Lactobacillus rhamnosus oil cream (B2) of International Application (PCT / KR2017 / 003172) dated March 25, 2017, as well as suppressing the lesion development of atopic dermatitis And to suppress the recurrence of the lesion.

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ t (p) TEWL B2 21 38.94 + 21.58 26.62 ± 14.36 12.32 ± 13.59 4.154 (.000 ^*** ) K1 3 33.68 ± 3.10 22.93 + 1.57 10.75 ± 2.57 7.229 (.019 ^* ) K2 3 32.65 ± 1.15 26.24 + 1.49 6.41 ± 1.48 7.479 (.017 ^* ) M1 3 39.18 + - 2.61 25.35 + 3.08 13.83 + - 2.25 10.638 (.009 ^** ) M2 3 38.73 + - 2.74 25.53 + - 2.68 13.20 ± 2.36 9.696 (.011 ^* ) M3 3 37.02 + - 4.14 23.82 + - 5.61 13.20 ± 2.01 11.379 (.008 ^** ) M4 3 38.50 ± 1.25 25.32 + 5.07 13.18 + - 4.30 5.316 (.034 ^* ) M5 3 37.98 + - 2.75 27.22 ± 1.13 10.76 ± 3.00 6.224 (.025 ^* ) M6 3 39.83 + - 2.30 27.87 + - 4.40 11.97 ± 2.68 7.744 (.016 ^* ) ^* p <.05, ^** p <.01, ^*** p <.001
Abbreviations were the same as Table 10.

Example 3 Confirmation of atopy treatment efficacy after oral lactobacillus rhamnosus mixed bacterium or oral lactobacillus rhamnosus mixed bacterium application and oil cream fermentation liquid application

&Lt; Example 3-1 > Preparation of capsules for oral lactobacillus lambusus mixed bacteria

Lactobacillus lambsosus (KCTC 5033), which had been preincubated with Lactobacilli MRS or skim milk, was inoculated at 1 × 10 ⁶ cfu / ml or more in the same manner as in <Example 1-5> Hour to obtain a Lactobacillus lanxus fermentation broth.

Then, the Lactobacillus laminocus fermentation broth was heated at 60 or less for 1 hour or more, concentrated under reduced pressure, and lyophilized to obtain Lactobacillus laminosse viable bacteria.

In order to kill the live bacteria of the Lactobacillus lanxus fermentation broth, it was subjected to intermittent sterilization (heating for 15 minutes or more at a constant atmospheric pressure of 80 or more), followed by heating at 60 or less for 1 hour or longer, And the resultant powder was used as the Lactobacillus lanamsus microbial cells.

For the purpose of the present experiment, an oral Lactobacillus lambusus mixed bacterium was prepared by mixing 50% by weight of a live cell and 50% by weight of a lactobacillus laminocyte cell, and filling the capsule into a capsule. The dose was 500 mg / capsule, Was 5.0 × 10 ¹⁰ CFU or more, and the number of bacteria was 5.0 × 10 ¹⁰ CFU or more and the total weight was 10.0 × 10 ¹⁰ CFU or more.

As a control, the oral Lactobacillus laminosus live cells were packed in capsules containing 100% by weight of Lactobacillus laminocyte cells, the dose was 500 mg / capsule, the viable cell count per capsule was 10.0 × 10 ¹⁰ CFU or more, Lactobacillus rhamnosus strains were packed in capsules containing 100% by weight of Lactobacillus laminocyte cells, each having a dose of 500 mg / capsule and a number of bacteria per capsule of 10.0 × 10 ¹⁰ CFU or more.

<Experimental Example 3> Confirmation of atopy treatment effect after oral lactobacillus lambusus mixed bacterium or oral lactobacillus lambosus mixed bacterium and simultaneous application of milk cream fermentation broth

<3-1> Confirmation of skin moisture content (SSH)

The atopic dermatitis test (H1), the fungus capsule (H2), and the mixed microbial capsule (H3) obtained in Example 3 were obtained by the method of Experimental Example 1-1 The subjects were given one capsule of water and once a day for 30 days, respectively, and then the change in skin moisture content before and after the test was confirmed. In addition, the inventors of the present invention have applied the oral lactobacillus lambosus mixed microbial capsules prepared in the above Example 3 to the present inventors' application filed on March 25, 2016 (10-2016-0035879) and on March 25, 2017 Lactobacillus laminosus oil cream of the application (PCT / KR2017 / 003172) Lactobacillus laminosse oil cream lotion prepared in the same manner as the lotion containing the fermentation liquid was applied to the atopic dermatitis lesion site twice a day (H4) Changes in the moisture content of the skin before and after was confirmed.

As a result, as shown in the following Table 13, the lactobacillus lambsos milk cream fermentation broth (B2) was found to be about 22% significant as in Experiment Example 3 of the present invention's patent application (10-2016-0035879) (H1) and the case of taking the fungus capsules (H2), respectively, showed an increase of about 7% and 14%, respectively, compared with those of the oral Lactobacillus rhamnosus live bacterial capsules (H3) showed a tendency to increase by about 16%. On the other hand, when the lotion containing an oral lactobacillus laminocus was administered and the lotion containing the creamy fermentation broth was applied (H4), it was increased by about 29% (Table 13).

Therefore, it was found that the use of dead bacteria in the case of taking Lactobacillus rhamnosus live oral bacteria for oral use was more effective in increasing the skin moisture content of atopic dermatitis lesions, and in particular, the oral Lactobacillus lambosus It was confirmed that when the mixed bacteria was used and the lotion containing the milk cream fermented liquid was applied, the skin moisture content of the atopic dermatitis lesion was higher than that of the Lactobacillus laminosus milk cream fermentation liquid (B2).

Variable Group N Measurement (Mean ± SD) Analysis of variance Before After t ₁ -t ₂ (M + SD) t (p) SSH B2 21 22.12 ± 7.98 27.05 7.99 -4.92 + 4.53 -4.981 (.000 ^*** ) H1 3 25.58 ± 7.86 27.26 + - 5.85 -1.68 ± 9.25 -0.406 (0.706) H2 3 18.53 + - 2.35 21.20 ± 9.75 -2.66 ± 7.46 -.619 (.599) H3 3 19.11 + - 2.52 22.10 ± 3.36 -2.99 ± 1.59 -3.250 (.083) H4 3 16.30 ± 2.80 20.96 ± 3.84 -4.66 ± 1.07 -7.494 (.017 ^* ) ^* p <.05, ^*** p <.001
Abbreviation: B2, lotion made from fermented milk cream with L. rhamnosus ; H1, ate oral L. rhamnosus, live cells; H2, ate oral L. rhamnosus , dead cells; H3, ate oral L. rhamnosus, mixed cells; H4, ate oral L. rhamnosus , mixed cells and applied lotion made from fermented milk cream with on skin.

<3-2> Determination of acidity (pH) of skin surface

The change in acidity of the skin before and after the test was measured in the same manner as in <3-1> above.

As a result, as shown in the following Table 14, the fermentation broth of lactobacillus laminosus milk cream (B2) was significantly reduced by about 6%, whereas when the oral lactobacillus lambsosus live cell capsule (H1) was taken, about 0.5% (H2) showed a tendency to decrease by about 2% and the mixed bacterial capsules (H3) showed a tendency to decrease by about 3%. On the other hand, oral lactobacillus laminosis mixed bacteria (H4) showed a significant decrease of about 7% when the fermented liquid of Lactobacillus laminosus milk was applied (Table 14).

Therefore, it is more effective to take the bacteria than the oral Lactobacillus rhamnosus live bacteria, and to take the mixed bacteria more than the oral lactobacillus rhamnosus live bacteria to reduce the skin pH of the atopic dermatitis lesions. In particular, the oral Lactobacillus lambosus mixture (H4) was found to be more effective in inducing skin pH stabilization of atopic dermatitis lesions than when lactobacillus rhamnosus oil cream fermented liquid (B2) was applied in the case of applying the bacterium and applying the lotion containing the milk cream fermented liquid (H4) Respectively.

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ (M + SD) t (p) pH B2 21 5.56 + - 0.48 5.21 + - 0.38 0.34 0.39 3.963 (.001 ^** ) H1 3 5.50 + - 0.69 5.47 ± 0.81 0.03 ± 0.72 .072 (.950) H2 3 5.61 ± 0.88 5.49 + - 0.61 0.12 + - 0.37 .592 (.614) H3 3 5.27 ± 0.16 5.12 ± 0.10 0.16 ± 0.08 3.584 (.070) H4 3 5.57 + - 0.13 5.16 ± 0.08 0.41 + 0.15 4.759 (.041 ^* ) ^* p <.05, ^** p <.01
Abbreviations were the same as Table 13.

<3-3> Determination of Percutaneous Moisture Evaporation Rate (TEWL)

The change in the amount of percutaneous water evaporation was measured in the same manner as in <3-1> above.

As a result, as shown in Table 15 below, the TEWL was significantly reduced by about 32% in the lactobacillus lanosus milk cream fermentation broth (B2), whereas in the case of taking the oral Lactobacillus laminosis broth capsule (H1) (H3) showed a decrease of about 15%, while oral lactobacillus lambs had a tendency to increase by 0.2% (H4) showed a significant decrease of about 36% (Table 15) when the lotion containing milk cream fermented liquid was applied at the same time.

Therefore, it is more effective to take the bacteria than the oral Lactobacillus rhamnosus live bacteria, and to take the mixed bacteria more effectively than the oral Lactobacillus rhamnosus live bacteria to reduce the TEWL of atopic dermatitis lesions. In particular, (H4) was more effective in reducing TEWL of atopic dermatitis lesions than in the case of Lactobacillus rhamnosus milk cream application (B2), while at the same time it was applied lotion containing Lactobacillus laminosus milk cream fermentation solution (H4) (Table 15)

(10-2016-0035879) of March 25, 2016 and International Application (PCT / KR2017 / 003172) of March 25, 2017, when the oral lactobacillus Lamb- Of the Lactobacillus rhamnosus oil cream lotion of the present invention is slightly lower than that of (B2), but it is confirmed that it not only inhibits the expression of lesions of atopic dermatitis but also contributes to the suppression of lesion and the inhibition of lesion recurrence (10-2016-0035879) of the present inventors and the present application filed on March 25, 2016 (10-2016-0035879) and 2017 (HLA) of the present inventors in the case of taking the lactobacillus laminocyte mixture with oral lactobacillus lambosus, (B2) of Lactobacillus rhamnosus oil cream lotion of International Application (PCT / KR2017 / 003172) on March 25, 2001, compared with the case of B2, the skin moisture content of atopic dermatitis lesion increased, By further changing significantly higher reduction in minutes, etc. All of evaporation, as well as to inhibit the expression of the lesions of atopic dermatitis, it was confirmed that contribute to a higher inhibitory effect of mitigating and lesion recurrence of lesions.

Variable Group N Measurement
(Mean ± SD) Analysis of variance Before After t ₁ -t ₂ (M + SD) t (p) TEWL B2 21 38.94 + 21.58 26.62 ± 14.36 12.32 ± 13.59 4.154 (.000 ^*** ) H1 3 25.08 + - 1.67 25.13 + - 0.75 -0.05 1.58 -.095 (.926) H2 3 30.61 + - 9.60 26.96 ± 12.73 3.65 ± 5.60 1.596 (.171) H3 3 42.07 ± 2.68 35.90 + - 4.33 6.17 ± 3.15 3.387 (.077) H4 3 41.78 ± 3.46 26.74 + - 4.85 15.04 + - 2.61 9.969 (.009 ^** ) ^** p <.01, ^*** p <.001
Abbreviations were the same as Table 13.

Claims (19)

A milk-cream fermentation broth of at least one strain selected from the group consisting of Lactococcus spp. , Lactobacillus spp. And Bifidobacterium spp. A cosmetic composition for preventing, alleviating and improving atopic dermatitis. The fermented milk cream according to claim 1, wherein the milk cream is milk-cream, milk, skim milk, Lactobacilli MRS medium, LB (lysogeny broth) medium, R2A medium (MB- And a culture medium containing casein hydrolyzate (casamino acid). The cosmetic composition for prevention, alleviation and improvement of atopic dermatitis according to claim 1, [3] The milk according to claim 2, wherein the milk is at least one selected from the group consisting of crude oil, non-fat milk, low-fat fat added with a fat content of 1 part by weight or less and skim milk. A cosmetic composition for improvement. The method according to claim 1, wherein the milk cream is at least one selected from the group consisting of milk fat separated from raw milk and milk, a concentrate of a fat pad, a processed milk cream and a powdered milk cream. And a cosmetic composition for improvement. The method according to claim 1, wherein the milk cream fermentation liquid is treated with at least one selected from the group consisting of tyndallization treatment, reduced pressure concentration treatment and yeast extract treatment. A cosmetic composition for improvement. The method according to claim 1, wherein the Lactobacillus spp. Is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus cassia Lactobacillus gasseri , Lactobacillus delbrueckii subsp. Bulgaricus , Lactobacillus reuteri , Lactobacillus salivarius, Lactobacillus fermentum and Lactobacillus acidophilus (Lactobacillus sp. Lactobacillus acidophilus) . The cosmetic composition for preventing, alleviating and improving atopic dermatitis according to claim 1, wherein the at least one cosmetic composition is at least one selected from the group consisting of Lactobacillus acidophilus . The cosmetic composition for preventing, alleviating and improving atopic dermatitis according to claim 1, wherein Bifidobacterium spp. Is Bifidobacterium bifidum . The cosmetic composition for preventing, alleviating and improving atopic dermatitis according to claim 1, wherein the Lactococcus spp. Is Lactococcus lactis . The cosmetic composition for preventing, alleviating and improving atopic dermatitis according to claim 1, wherein the composition exhibits an effect of improving the skin moisture content, decreasing the pH value of the skin, or stabilizing the skin or reducing the amount of evaporation of transdermal moisture. The cosmetic composition for preventing, alleviating and improving atopic dermatitis according to claim 1, wherein the composition is for oral or skin application. 11. The cosmetic composition for preventing, alleviating or improving atopic dermatitis according to claim 10, wherein the oral use is a live or dead organism strain. The method according to claim 1, wherein the fermented milk cream of at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. Wherein the cosmetic composition is a cosmetic composition for skin moisturizing. Prevention of atopic dermatitis comprising as an active ingredient an oil-in-milk fermentation broth of at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. , An external skin preparation for alleviation and improvement. A method for producing atopic dermatitis comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. atopic dermatitis). A method for producing atopic dermatitis comprising at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. atopic dermatitis). Culturing at least one strain selected from the group consisting of Lactococcus spp., Lactobacillus spp. And Bifidobacterium spp. In an oil cream medium to obtain a fermentation broth; Wherein the atopic dermatitis prevention, alleviation, and improvement are carried out in the same manner as in the above. The method according to claim 16, wherein the milk cream fermentation broth is milk-cream, milk, skim milk, Lactobacilli MRS medium, LB (lysogeny broth) medium, R2A medium (MB- And a culture medium containing a casein hydrolyzate (casamino acid). The method for producing a milk cream fermentation broth for preventing, alleviating and improving atopic dermatitis. 17. The method of claim 16, wherein the step of treating at least one selected from the group consisting of tyndallization treatment, reduced pressure concentrating treatment and yeast extract treatment is applied to the milk cream fermentation broth. (Method for manufacturing fermented milk cream for relief and improvement). The method of claim 16, further comprising encapsulating the milk cream fermentation broth to be used for oral use.

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