KR101999374B1 - Composition for Improving Atopic Dermatitis Using Fermented Products of Fruit of Diospyros kaki - Google Patents

Abstract

The present invention discloses a composition for improving atopic dermatitis using a reduced fermented product. Specifically, the present invention relates to the use of Rhodopseudomonas palustris , Lactobacillus < RTI ID = 0.0 > plantarum , and Saccaromyces cerevisiae as an active ingredient. The present invention also provides a composition for improving atopic dermatitis.

Description

FIELD OF THE INVENTION The present invention relates to a composition for improving atopic dermatitis,

The invention persimmon (Diospyros The present invention relates to a composition for improving atopic dermatitis using a fermented product obtained by fermenting a sensate with useful microorganisms such as lactic acid bacteria, photosynthetic bacteria and yeast.

Atopic dermatitis is characterized by severe itching and chronic skin rash. It is the most common form of infantile eczema. It is caused by atopic dermatitis.

The cause of atopic dermatitis has not yet been elucidated yet. However, the cause of atopic dermatitis has not yet been clarified. However, it has been reported that increased IgE production, decreased CD8 + suppression / cytotoxic T lymphocytes, decreased function and dysfunction, monocyte / macrophage infiltration, There have been reported immunological phenomena such as increased numbers of eosinophils, dendritic cells (DCs), epidermal Langerhans cells and CD4 + T lymphocytes, and increased production of chemokines (TSLP, RANTES, CTACK, LARC, MDC, TARC etc.) (Invest. Dermatol., 96: 523-526,1991, J. Invest Dermatol., 97: 389-394,1991, Immunol., 11: 81-88,1999, Curr Drug Targets Inflamm Allergy., 2: International Immunology, 14 (7): 767-773, 2002; Pediatr Allergy Immunol 19: 711-120, 2003; J. Allergy Clin Immunol., 107: 871-877, 2001; Adv. Immunol. 605-613, 2008).

In particular, serum concentrations of macrophage-derived chemokine (MDC) and thymus and activation-regulated chemokine (TARC) have been reported to be significantly increased in patients with atopic dermatitis (Hijnen et al., J. Allergy Clin. Immunol. 2) 334-340), when ciclosporin A or corticosteroids, used as therapeutic agents for atopic dermatitis, were administered to patients with atopic dermatitis, serum concentrations of MDC and TARC were reported to decrease (Y. Shimada et al In vitro studies have shown that MDC and TARC are highly expressed when HaCaT cells are treated with INF-γ or TNF-α, which can inhibit this expression (see, for example, J. Dermatol. Sci., 34, 201-208, 2004) (Horikawa et al., Int. Immunol., 14 (7) 767-773, 2002).

Therefore, a substance that inhibits the expression of MDC or TARC can be used as a remedy for atopic dermatitis.

The present invention discloses a fermented product of useful microorganisms which inhibit the expression of MDC and TARC in HaCaT cells stimulated with IFN-? And TNF- ?, and has an activity of improving atopic dermatitis in actual clinical evaluation.

It is an object of the present invention to provide a composition for improving atopic dermatitis using a fermented microorganism useful as a persimmon.

Other and further objects of the present invention will be described below.

The present inventors treated HaCaT cells treated with hIFN-γ and TNF-α to inhibit the production of MDC and TARC by treating atrophic dermatitis improving activity of a fermented product obtained by fermenting persimmon (fruit) with useful microorganisms such as lactic acid bacteria, photosynthetic bacteria and yeast The results showed that the 70% ethanol extract and the fermented microorganism of the useful microorganism showed higher activity than the 70% ethanol extract of the fermented microorganism. When the fermented microorganism fermented in a cell test was found to have a high activity even after 8 weeks of clinical trials using three nutritional lotions each containing its own fermented product, And showed markedly higher activity of atopic dermatitis improving activity than the fermented product itself.

The present invention is provided based on these experimental results, and the composition for improving atopic dermatitis of the present invention is characterized by containing lactic acid bacteria, photosynthetic bacteria and yeast fermented products as an effective ingredient.

Herein, the meaning of the above-mentioned "active ingredient" means an ingredient that alone exhibits the desired activity or can exhibit activity together with a carrier which is itself inactive.

As used herein, "atopic dermatitis" is defined as including any disease classified as atopic dermatitis in the art, regardless of the cause, either directly or indirectly, of its occurrence. In general, atopic dermatitis is classified into infantile atopic dermatitis, pediatric atopic dermatitis, adult atopic dermatitis, and maternal atopic dermatitis according to the onset period or the object of the invention. In the present specification, Of atopic dermatitis.

In the present specification, the term "fermented product" means a culture obtained by adding a carbon source to a sensory fruit to be fermented and obtained by inoculating and culturing lactic acid bacteria, photosynthetic bacteria and yeast. The culture may be used as it is, or may be used in liquid or powder form by concentration under reduced pressure and / or lyophilization, or may be used in the form of a liquid or a powder, or an alcohol having 1 to 4 carbon atoms such as methanol and ethanol, acetone, ethyl acetate, chloroform, methylene chloride, It can be used by extracting with a mixed solvent. It may also be extracted with an alcohol having 1 to 4 carbon atoms such as methanol and ethanol, acetone, ethyl acetate, chloroform, methylene chloride, water or a mixed solvent thereof. Such an extract may be in liquid or powder form in which part or all of the extraction solvent has been removed.

The carbon source of the microorganism inoculated in the above is well known in the art, and thus any of those known in the art can be used. Usually, oligosaccharides, lactose, glucose, fructose, sugar, molasses, dextrose, will be. These carbon sources can be added in an arbitrary range in consideration of the intended fermentation time, degree of fermentation, and the like. It is usually added in the range of 1 to 70 parts by weight based on 100 parts by weight of the fermentation target.

In addition, the cultivation after inoculation of the microorganisms can be carried out at a temperature condition of 5 ° C to 45 ° C, particularly at a temperature of 25 ° C to 40 ° C. If the incubation temperature is lower than the above range, the fermentation rate may be slowed and unwanted fermentation products may be produced. Even if the incubation temperature is higher than the above range, the fermentation rate may be similarly slowed down due to the death of the fermentation microorganisms, . In the case of the incubation time, it is preferable to continue until the growth of the inoculated microorganism reaches a maximum. The time at which the inoculated microorganism reaches its maximum growth will depend on the incubation temperature, the amount of fermentation material, the amount of microorganisms inoculated, and so on.

The composition for improving atopic dermatitis of the present invention may contain any amount (effective amount) of the active ingredient decaf fermented product, depending on the purpose of use, formulation, compounding purpose, and the like, which can exhibit atopic dermatitis improving activity. An effective amount will be determined within the range of from 0.001 wt% to 99.900 wt% based on the total weight of the composition. The term "effective amount" as used herein refers to an amount of an effective ingredient capable of preventing, ameliorating, treating, or delaying the onset of symptoms of atopic dermatitis. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

The composition for improving atopic dermatitis of the present invention can be identified as a food composition in a specific embodiment.

When the composition of the present invention is identified as a food composition, the form of the food is not particularly limited as far as it is a beverage such as tea, juice, carbonated beverage, ionic drink, processed oil such as milk and request route, gum, rice cake, Food preparations such as foods, powders, tablets, capsules and the like.

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

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

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

Examples of the physiologically active substance include catechins such as catechin, epicatechin, gallocatechin and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine and riboflavin.

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

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

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

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

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

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

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

The composition for improving atopic dermatitis of the present invention can be identified as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention may be formulated into a wide variety of dosage forms, including pharmaceutically acceptable carriers, excipients and the like, in addition to the active ingredient, and may be formulated into a wide variety of dosage forms such as a localized preparation such as cream, lotion, ointment (semi-solid external drug), micro loo emulsion, (TTS) (e.g., patches, bandages, etc.), and the like.

The term "pharmaceutically acceptable" as used herein means that the application (prescribing) subject does not have the above-mentioned toxicity that is adaptable without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (e.g., corn starch, potato starch and the like), cellulose, derivatives thereof (e.g. sodium carboxymethylcellulose, ethylcellulose, etc.), malt, gelatin, talc, (E.g., peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin and the like), alginic acid, and the like. Depending on the formulation of the pharmaceutical composition of the present invention, one or more suitable ones may be selected and used. Suitable pharmaceutically acceptable carriers and formulations are described in Remington's Pharmaceutical Sciences (19th ed., 1995). The pharmaceutical composition of the present invention may further comprise an emulsifier (e.g., TWEENS), a wetting agent (e.g., sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, a preservative, water, saline, a phosphate buffer solution and the like.

The excipient may be selected according to the formulation of the pharmaceutical composition of the present invention. For example, suspending agents such as sodium carboxymethyl cellulose, methyl cellulose, hydropropyl methyl cellulose, sodium alginate, polyvinyl pyrrolidone, .

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

The composition for improving atopic dermatitis of the present invention can be identified as a cosmetic composition in another specific embodiment.

The cosmetic composition of the present invention may contain, in addition to the active ingredient which exhibits the atopic dermatitis-improving activity, an active ingredient such as a conventionally used ingredient in the cosmetic composition such as a stabilizer, a solubilizing agent, a surfactant, a vitamin, Adjuvants, and carriers.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

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

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention can be produced according to a method for producing a cosmetic composition which is usually carried out in the art, except that it contains a sensitive fermentation product exhibiting atopic dermatitis improving activity.

As described above, the present invention can provide a composition for improving atopic dermatitis using a reduced fermented product.

The composition of the present invention can be used as a cosmetic product, medicine, food or the like.

Figures 1 and 2 show the results of showing inhibitory activities of TARC and MDC on the reduced fermented product.
Fig. 3 shows the cytotoxicity test result of the reduced fermented product.

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

< Example > Traditional  By useful microorganisms Fermented  Produce

100 parts by weight of crushed product obtained by purchasing, washed and washed by purchasing Diospyros kaki (fruit) cultivated in Jeju, 50 parts by weight of molasses on the basis of 100 parts by weight of the crushed product, and 300 parts by weight of crushed product , And the photosynthetic bacterium prepared in the LB medium for this mixture was also used as Rhodopseudomonas 2% (v / v) of the culture medium of Lactobacillus palustris , Lactobacillus plantarum prepared in the MRS liquid medium plantarum) broth 2% (v / v) and the yeast Saccharomyces prepared in the PDB Seth Serenity badge My kids Vichy (Saccaromyces cerevisiae culture medium 2% (v / v) were inoculated and cultured at a temperature range of 30 to 40 캜 to obtain a decidual fermented product. The fermented product was sterilized by irradiating ultraviolet rays, concentrated under reduced pressure, and lyophilized to be used in the experiment.

< Comparative Example > Sensory extracts and useful microorganisms themselves Fermented  Produce

&Lt; Comparative Example 1 > Preparation of persimmon extract

Five percent by weight of 70% ethanol was added to 100 parts by weight of the pulverized product obtained by purchasing a persimmon grown in Jeju, washed and ground, and the mixture was extracted at room temperature for 24 hours, filtered, concentrated under reduced pressure and lyophilized Sensitive extract.

&Lt; Comparative Example 2 > Preparation of self- fermented microorganism

A fermented product of useful microorganisms was prepared in the same manner as in the above example except that 100 parts by weight of molasses and 600 parts by weight of water were mixed except for the pulverized product, and the photosynthetic bacteria, lactic acid bacteria and yeast were inoculated therein, . The fermented product was sterilized by irradiating ultraviolet rays, concentrated under reduced pressure, and lyophilized to be used in the experiment.

< Experimental Example > Anti-atopic  Active experiment

<Experimental Example 1> Evaluation of inhibitory activity of TARC and MDC , a causative agent of atopic dermatitis

HaCaT cells (3.0 × 10 ⁵ cells / mL) were inoculated into a 24-well plate and DMEM culture medium. Cells were cultured in a 5% CO ₂ incubator for 18 hours. After the medium was removed, HaCaT cells were treated with 50 μl of sample and 450 μl of TNF-α (10 ng / ml) + IFN-γ (10 ng / ml) Respectively. After 24 hours, the culture medium was centrifuged (12,000 rpm, 3 minutes), and the amount of atopy inducing factor produced in the supernatant was measured. Determination of TARC and MDC was quantified using the mouse enzyme-linked immnunosorbent assay (ELISA ) kit (R & D Systems Inc., Minneapolis, MN, USA) r 2 value of the standard curve for the standard was at least 0.99.

The results are shown in Fig. 1 and Fig. 2. (Sample 1, 100, 200 and 400 / / ml treatment) or the useful microorganism self-fermentation product (Sample 3, 100, 200 and 400 / / ml treatment) with reference to FIG. 1 and FIG. Showed no specific MDC and TARC production inhibitory activity, but fermented microorganism fermented products (Sample 2, 100, 200 and 400 ㎍ / ㎖ treated) showed distinct MDC and TARC production inhibitory activity.

EXPERIMENTAL EXAMPLE 2 Evaluation of cytotoxicity ( LDH assay )

RAW 264.7 cells (1.5 × 10 ⁵ cells / ml) were co-treated with DMEM medium and LPS (1 μg / ml) in DMEM medium for 24 hours, and then cultured for 3 minutes at 3,000 rpm for 5 minutes. The LDH (lactate dehydrogenase) assay was performed using a non-radioactive cytotoxicity assay kit (Promega). 50 μl of the culture medium obtained by centrifugation on a 96-well plate and 50 μl of the reconstituted substrate mix were added and reacted at room temperature for 30 minutes After adding 50 μl of stop solution, absorbance was measured at 490 nm using a microplate reader (Bio-TEK Instruments Inc., Vermont, WI, USA). The average absorbance values for each sample group were determined and compared with the absorbance values of the control (LDH control, 1: 5000) to evaluate cytotoxicity.

The results are shown in Fig. As shown in FIG. 3, the fermented microorganism fermented product showed slight cytotoxicity at a concentration of 400 μg / ml, but all of the samples showed no specific cytotoxicity. This confirms that the MDC and TARC production inhibitory activity of the useful fermented microorganism is not the result of cytotoxicity.

<Experimental Example 3> Clinical trial for improving atopic dermatitis

Nutritional lotions were prepared with the ingredients and contents shown in the following Table 1 for the clinical test for the improvement of atopic dermatitis.

Ingredients and quantities of nutrition lotion ingredient content
(weight%) The samples of Examples or Comparative Examples 2.0 Stearic acid 2.0 Cetyl alcohol 2.0 Glyceryl monostearate 2.0 Polyoxyethylene sorbitan monostearate 0.5 Sorbitan sesquioleate 0.5 Glyceryl monostearate / glyceryl stearate / polyoxyethylene stearate 1.0 Wax 1.0 Liquid paraffin 4.0 Squalane 4.0 Caprylic / capric triglyceride 4.0 Carboxyvinyl polymer 0.3 Butylene glycol 5.0 glycerin 3.0 Triethanolamine 0.5 Purified water Balance

The activity of improving atopic dermatitis was divided into three groups of 60 men and women having symptoms of atopic dermatitis over 5 years old and each nutrient lotion containing the extract of the above example or comparative example was administered twice or three times a day For 8 weeks.

Before and after the start of the study, the SCORAD Index was developed by the European Task Force on Atopic Dermatitis, and the severity of atopic symptoms was high.

The results are shown in Table 2 below as mean ± standard deviation.

SCORAD Index division 0 parking 8 parking Useful micro-organisms fermented in the sense of the example Nourishing lotion 36.4 ± 6.8 22.6 ± 6.2 * The nutritious lotion containing the 70% ethanol extract of Comparative Example 1 34.8 ± 7.2 32.5 ± 5.7 The nutrient lotion containing the useful microorganism fermented product of Comparative Example 2 37.5 8.3 35.6 ± 6.2 * p < 0.05

In clinical trials, the SCORAD index was significantly reduced in the nutrient lotion containing fermented microorganism fermented products, but the other nutrient lotions showed no significant effect.

Claims (4)

Diospyros kaki fruit fermented product as an active ingredient,
The persimmon fermented product is obtained by fermenting Rhodopseudomonas palustris , Lactobacillus plantarum and Saccaromyces cerevisiae after adding a carbon source to persimmon fruit, And,
The persimmon fermented product has an activity of inhibiting the formation of MDC (macrophage-derived chemokine) and TARC (thymus and activation-regulated chemokine) which are atopic dermatitis inducing factors
Wherein the atopic dermatitis is at least one kind selected from the group consisting of:
The method according to claim 1,
The composition for improving atopic dermatitis is a food composition.
The method according to claim 1,
The composition for improving atopic dermatitis is a pharmaceutical composition.
The method according to claim 1,
The composition for improving atopic dermatitis is a cosmetic composition.

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