KR101803263B1 - Anti-inflammatory, anti-allergic fermented concentrate production and improvement of atopic dermatitis - Google Patents

Abstract

The present invention relates to a fermented product having an anti-inflammatory, anti-allergic and atopic skin improving effect and a process for producing the fermented product, and more particularly to a method for producing fermented product by pulverizing the product from a stem or leaf to obtain a milled powder, By providing intracellular free radical scavenging by inhibiting the activity of lipid peroxidation, lipoxygenase and hyaruronidase by providing a fermented product obtained by inoculation and fermentation, and by providing a cosmetic composition containing the fermented product, The present invention relates to a cosmetic composition containing a seaweed mixed fermentation product having an anti-inflammatory, anti-allergic and atopic skin improving effect which improves inflammation, allergy and atopic skin and prevents side effects on skin irritation.
The present invention also provides a fermented broth having anti-inflammatory, anti-allergic and atopic skin-improving effects, characterized in that the fermentation is carried out by intestinal bacteria or lactic acid bacteria after pulverization, acid hydrolysis and neutralization reaction.
The present invention also relates to a method for preparing a fermented milk, comprising the steps of: (S10) milling millet to obtain milled powder, (S20) obtaining acidic hydrolysis of the milled powder to obtain a milled solution, (S30), obtaining a fermented broth fermented by inoculating the pH-adjusted fermented broth with intestinal bacteria or lactic acid bacteria (S40), and sterilizing, filtering and freeze-drying the fermented fermented broth to obtain a fermentation broth (S50). The method for producing a fermented broth having an anti-inflammatory, anti-allergic and atopic skin improving effect is provided.
In addition, the present invention provides a cosmetic composition comprising a fermented product having an anti-inflammatory, anti-allergic, and atopic skin improving effect.

Description

TECHNICAL FIELD The present invention relates to a fermented fermented product having an anti-inflammatory, anti-allergic and atopic skin improving effect, a process for producing the fermented product, and a cosmetic composition containing the fermented fermented concentrate and an improvement of atopic dermatitis.

The present invention relates to a fermented product having an anti-inflammatory, anti-allergic and atopic skin improving effect and a method for producing the fermented product, and more particularly to a method for producing a fermented product by pulverizing the product from a stem or leaf to obtain a powdery powder, Fermented products having an anti-inflammatory, anti-allergic and atopic skin-improving effect for inhibiting the activity of lipid peroxidation, lipoxygenase and hyaruronidase by intracellular free radical scavenging by providing a fermented product inoculated and fermented, And a method for producing the same.

In addition, the present invention provides a cosmetic composition containing a fermented product of fermentation, thereby improving skin inflammation, allergy and atopic skin, and improving the anti-inflammatory, anti-allergic, and atopic skin- And to a cosmetic composition containing water.

The theory of free radicals was presented by D. Harman in the mid-1950s, and has recently attracted attention from the academic community and industry. There are many causes for the presence of radicals in the human body. These radicals cause cell destruction, breakage of the connective tissue of the dermal layer of the skin, or cross-linking, thus causing wrinkles, skin cancer, cell killing, rheumatoid arthritis, It causes various problems such as acne. The cause of Radical is the phagocytosis of white blood cells, the action of electron transport system, myeloperoxide (MPO), ultraviolet rays, tobacco, normal metabolism, stress, pollutants and bacteria during the process of ATP production in mitochondria.

Of course, there are superoxide dismutase (SOD), catalase, vitamin E, vitamin C, and ubiquinol, which are antioxidants (radical scavengers) in the human body. But gradually, the antioxidant system begins to break down by age, pollution, ultraviolet rays, stress and so on. Therefore, the antioxidant system becomes ineffective and the radicals gradually increase.

The radical destroys collagen, elastin, and hyaluronic acid, which are connective tissues of the dermis, causing settling (wrinkling) on certain parts of the skin and oxidizing the lipid part of the cell membrane, Causing skin diseases, acne, skin cancer and the like.

In addition, this radical participates in the spontaneous oxidation reaction (dopaquinone melanin) during the melanin formation process, and causes the cause such as stain and freckles and wrinkle formation (Korean Journal of Cosmetic Scientists, Vol. 23, No. 1, 75-132).

Inflammation is a physiological response that protects the body from harmful environmental conditions, such as intrusion of external foreign matter such as bacteria and mechanical damage. This inflammation leads to a large number of polymorphonuclear leukocytes (PMNs) and immune substances, and these cells secrete a variety of inflammatory cell products, proteases and cytokines. I will. Enzymes such as elastase, hyalurinidase and lipoxygenase are well known as proteins and lipolytic enzymes produced during inflammation.

On the other hand, these actions sometimes cause deleterious damage to adjacent tissue cells and non-cellular components, so that under proper conditions, inflammation regains normal function after the initial state, but the irritant stimulating agent does not disappear Continued production will result in chronic inflammation resulting in even more severe tissue damage.

As a result, the cells and connective tissues are damaged by the proteolytic enzymes induced by the hyperinflammation, and the damage of the connective tissues reduces the elasticity of the skin, thereby causing wrinkles and also adversely affecting cell regeneration and proliferation Resulting in rapid aging of the skin.

In addition, inflammation of the skin is constituted by local erythema and swelling. When the damage becomes larger, the normal tissue is restored in the dissipation step by the scar tissue, which is the result of fibroblast proliferation and accumulation of connective tissue protein do. However, if the tissue is more severely damaged and the inflammatory response persists, restoring normal tissue structure and function becomes much more difficult.

The production of mediators that promote leukocyte migration along with inflammation is also mediated by arachidonic acid. This pathway consists of cyclooxygenase and lipoxygenase, and leukotriene and prostagladine produced by these enzymes act as inflammation inducers.

Therefore, inhibitors of lipoxygenase and cyclooxygenase can inhibit the production of inflammatory substances as well as inhibit cell membrane breakdown and lipid peroxidation by reactive oxygen species produced during inflammation.

In addition, it inhibits proteins and lipolytic enzymes produced by hyperinflammation, thereby protecting cells damaged thereby, thereby inhibiting skin aging.

In addition, atopic skin occurs mainly in early childhood and is called as fever. It is caused by decrease of ceramides content in the stratum corneum. As the moisture retaining function of the stratum corneum is lowered, the skin is dried, skin barrier function is decreased, the stimulating substance enters into the stratum corneum to induce stimulation, and when the antigen protein which is difficult to intrude into the normal skin invades, skin allergy easily occurs and it is prone to pruritus symptoms. Therefore, more than 90% of atopic patients are infected with Staphylococcus aureus, which is caused by scratching the patient because of itching. However, recent reports indicate that the bacteria 's exotoxin stimulates the body' s immune system, It is said that it makes the atopy worse by letting out material. That is, the bacterium itself acts as an allergen.

The cause of such atopic skin has been largely classified into four categories. First, it is caused by a genetic cause. It constitutes the intercellular lipid, which is a binding substance between the phospholipid and the keratinocyte, The lack of gamma-linoleic acid (GLA), which is an essential fatty acid, makes it difficult to make the skin barrier hard, making it easy to lose moisture on the surface of the skin and to produce allergies by external stimuli. Secondly, It is caused by environmental factors. As the life of the apartment becomes more common due to the changes in the living environment of modern people, the living environment is changed to a dry state. As a result, the atopic skin is increasing worldwide. As a result, the water retention is decreased, And allergic reactions occur, and the third is that the bow It is believed that lipid peroxidation is produced by oxygen, which is caused by oxygen, which causes atopic skin. Fourth, it is caused by infection with bacteria, viruses and fungi. Science, Korean Society of Dermatology Publishing Committee, Jae Moon, 1992).

For example, Korean Patent Registration No. 10-1051077 discloses a conventional technique for anti-inflammation, anti-allergy, and atopy, which includes 2-piperazino-4,5-disubstituted-1,3-thiazole derivatives, A preparation method thereof, and a therapeutic agent for inflammation-related diseases caused by SPC receptor activity containing the same as an active ingredient. More particularly, the present invention relates to a 2-piperazino-4,5-disubstituted-1,3-thiazole derivative or optically isolated To an agent for treating inflammation related diseases which is effective for atopic dermatitis induced by SPC receptor activity, inflammation occurring in other diseases, pruritus or skin infections, which contains this substance as an active ingredient.

The above-mentioned prior art is advantageous in that it can be mass-produced and has a low cost.

However, the above-mentioned prior arts have produced a compound through an artificial organic synthesis, so that it is difficult to obtain a substantial effect due to poor stability during synthesis, and side effects are caused by the compound depending on a person.

Therefore, in order to reduce adverse effects on the above-described technology, a composition comprising a natural extract that is less harmful to the human body than an organic synthetic compound is required.

Korean Patent Registration No. 10-1262813 discloses a composition for preventing and treating atopic dermatitis or allergic skin diseases, which comprises extracts of Alaska pollack as an active ingredient. The extracts were concentrated by using a reflux extractor and then lyophilized to obtain an extract of Ganoderma lucidum.

The above-mentioned prior art is a composition for extracting natural extracts, which has fewer side effects than the therapeutic agent extracted by organic synthesis and has an advantage of minimizing destruction of active ingredients.

However, the extraction method using the reflux extractor has the problem that the cost due to the use of high energy is increased and the extraction is performed at a high temperature, so that the effect of atopic dermatitis or allergic skin disease due to the degradation of stability and component modification is reduced.

Therefore, in order to reduce the adverse effect on the above-described technology, there is a need for a method capable of extracting energy loss through fermentation while minimizing the denaturation of the extracted component, rather than heating it to a high temperature.

Korean Patent Registration No. 10-1403523 discloses a conventional method for solving the above problem, which is a method for producing a fermented cosmetic product comprising a fermentation property of germinated grains as a main ingredient. More specifically, And fermenting the fermented cosmetics.

The above-mentioned prior art is to ferment the grains (such as brown rice, mung bean, black sesame, sorghum, yulmu, seolite, and soybean) and fermenting them at room temperature by adding lactobacillus platinum, It is advantageous that there is no loss of the component due to it.

However, the disadvantage is that it takes too long to ferment and there is no anti-inflammatory, anti-allergic or atopic skin effect.

Therefore, a method of shortening the fermentation time of grains is required, and a natural extract having anti-inflammatory, anti-allergic and atopic skin improving effects is required.

Disclosure of the Invention The present invention has been conceived to solve the above-mentioned problems. It is an object of the present invention to provide a process for producing a water- Anti-inflammatory, anti-allergic and atopic skin-improving effect, which reduces the fermentation time and has the anti-inflammatory, anti-allergic and atopic effect, by obtaining the fermented product by fermentation by inoculating Raoultella or Leuconostoc strain And an object of the present invention is to provide a fermented product thereof.

Further, the present invention provides a cosmetic composition containing a fermented product obtained by the process of the present invention, which provides anti-inflammatory, allergic and atopic skin improving effects such as intracellular free radical scavenging effect, lipid peroxidation, inhibition of hyparrhodicase activity, and inhibition of lipoxygenase activity ≪ RTI ID = 0.0 > cosmetic < / RTI >

The present invention provides a fermented broth having an anti-inflammatory, anti-allergic and atopic skin improving effect, characterized in that the fermentation is carried out by intestinal bacteria or lactic acid bacteria after pulverization, acid hydrolysis and neutralization reaction.

The present invention also relates to a method for preparing a fermented milk, comprising the steps of: (S10) milling millet to obtain milled powder, (S20) obtaining acidic hydrolysis of the milled powder to obtain a milled solution, (S30), obtaining a fermented broth fermented by inoculating the pH-adjusted fermented broth with intestinal bacteria or lactic acid bacteria (S40), and sterilizing, filtering and freeze-drying the fermented fermented broth to obtain a fermentation broth (S50). The method for producing a fermented broth having an anti-inflammatory, anti-allergic and atopic skin improving effect is provided.

In addition, the present invention provides a cosmetic composition comprising a fermented product having an anti-inflammatory, anti-allergic, and atopic skin improving effect.

As described above, the present invention provides a fermented product having anti-inflammatory, anti-allergic and atopic skin-improving effects, and a method for producing the fermented product, which comprises grinding a stem and a leaf, And then fermented by inoculating Raoultella or Leuconostoc strain to obtain a fermented product. By performing intracellular free radical scavenging, lipid peroxidation, Haaruronidase Inhibiting activity, and inhibiting lipoxygenase activity.

In addition, the cosmetic composition containing the fermented product having the anti-inflammatory, anti-allergic and atopic skin improving effect according to the present invention has the effect of improving skin inflammation, allergy and atopic skin and preventing side effects on skin irritation.

FIG. 1 is a flow chart of a method for producing a fermented product having anti-inflammatory, anti-allergic and atopic skin improving effects according to the present invention.

As a result of searching for natural substances which are effective for improving anti-inflammatory, anti-allergic and atopic skin, various natural substances were searched for, and as a result, they exhibited excellent anti-inflammatory, anti-allergic and atopic skin improving effects And solved the safety and safety problems of products by applying this fermented product.

Further, it has been found that the fermented product is blended with a cosmetic composition at a certain concentration or more, and the skin condition is improved even when the result is directly applied to human skin.

Hereinafter, the present invention will be described in detail.

The present invention provides a fermented broth having an anti-inflammatory, anti-allergic and atopic skin-improving effect, characterized in that the fermentation is carried out by intestinal bacteria or lactic acid bacteria after pulverization, acid hydrolysis and neutralization reaction.

Sorghum (Bicolor) is an annual plant of the alopecia plantus. It grows naturally in poor soil and dry soil, and is cultivated as an important food source throughout Asia and Africa. In Korea, rice, barley, wheat and corn It is cultivated as a food resource. Various varieties exist and are classified into grain sorghum, sorgo, and broom-corn depending on the application (Chang et al., 2005). The length of the stem is 20 ~ 30cm, and the fruit is used only as a seed because the shell is thick and the shell does not reach even when threshing. Folk remedies have been used for various symptoms such as improvement of appetite, promotion of digestion, maintenance of body temperature, gastrointestinal protection, and detoxification. Although sorghum is known to contain large amounts of active ingredients such as dietary fiber, phenolic compounds, and tannins, red sorghum does not contain tannin. phenolic compounds are mainly composed of flavonoids, phenolic acids, and antioxidants, and most of them are known as flavonoids (Ryu HS, Kim J, Kim HS 2006. Enhancing effect of sorghum bicolor L. Moench (sorghum, Korean J Food Cookery Sci 22: 363 (2006). The Korean Society for Applied Microbiology and Biotechnology, Seoul, Korea. Effects of waxy and normal sorghum flours on sponge cake properties. Food Eng Prog 9: 199-207. (Kim, KO, Kim, HS, 2006. Effect of Sorghum bicolor L. Moench (sorghum, water-water) Water extracts on mouse immune cell activation. J Korean Diet Assoc 12: 82-88.).

Although the millet can use the whole area of millet, it is preferable to use a stem or a leaf.

The millet is subjected to pulverization, acid hydrolysis, and neutralization to obtain an effective ingredient having anti-inflammatory, anti-allergic and atopic-improving effects, and fermented with intestinal bacteria or lactic acid bacteria to obtain a fermented product.

That is, the fermented fermented product according to the present invention exerts intracellular free radical scavenging, and has an excellent effect of inhibiting the activity of lipid peroxidation, rapeoxygenase, and hyaruronidase.

A method for producing the fermented product using the fermentation broth will be described with reference to the accompanying drawings.

FIG. 1 is a flowchart of a method for producing a fermented product having anti-inflammatory, anti-allergic and atopic skin improving effects according to the present invention.

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. The method for producing fermented broth having anti-inflammatory, anti-allergic and atopic skin improving effects of the present invention preferably includes the following steps Do not.

The present invention relates to a method for producing a fermented milk, comprising the steps of: (S10) milling millet in a pulverizer to obtain a powder; (S20) obtaining an aqueous solution by acid hydrolysis of the powder; (B) adjusting pH by adding a neutralizing agent (S40) of sterilizing, filtering and lyophilizing the fermented broth to obtain a fermented fermented broth (S50); a step (S40) of fermenting the fermented broth by inoculating the pH-adjusted solution with intestinal bacteria or lactic acid bacteria; .

The manufacturing method of the present invention will be described separately for each step.

(S10) of pulverizing millet to obtain a powder is carried out.

It is preferable that the millet is dried in a shade of air and pulverized at a rotation speed of 2,500 to 3,500 rpm in a pulverizer. The particle size of pulverized millet is preferably about 0.1 to 35 mu m.

When the milling speed of the pulverizer is less than 2,500 rpm at the time of milling, the pulverized coal is not sufficiently crushed and pulverized. If the pulverizer has a rotational speed of 3,500 rpm or more, the increase in energy costs, noise generation, The effective component of the silk is destroyed.

Acid hydrolysis of the hydrate powder to obtain an aqueous solution (S20) is carried out.

The transferring powder obtained in the step S10 may be subjected to acid hydrolysis by selecting at least one of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, formic acid and phosphoric acid, more preferably using sulfuric acid, nitric acid or hydrochloric acid, Preferably, sulfuric acid is used for acid hydrolysis.

In the present invention, sulfuric acid was prepared from 0.1 to 3.0% (v / v) of dilute sulfuric acid, and the acceptor powder obtained in step (S10) was diluted sulfuric acid and heat-treated at 100 to 130 ° C for 15 to 60 minutes. Preferably 0.5 to 2.0% (v / v) of diluted sulfuric acid, and the heat treatment time is 20 to 40 minutes. The obtained result is maintained at room temperature, and then separated from the solid phase and the aqueous solution by centrifugation or membrane filter do.

And adjusting the pH by adding a neutralizing agent to the aqueous solution (S30).

The aqueous solution obtained in the step S20 is selectively neutralized by selecting at least one of calcium carbonate (CaCO ₃ ), sodium hydroxide (NaOH), sodium carbonate (Na ₂ CO ₃ ) or calcium hydroxide (Ca (OH) ₂ ) Calcium carbonate is added to adjust the pH to 5.0 ~ 7.0.

The step (S40) of obtaining a fermented fermented product by inoculating the pH-adjusted aqueous solution with intestinal bacteria or lactic acid bacteria is carried out.

The extract obtained in the above step S30 is filtered using a membrane filter of 0.1-0.5 mu m, and intestinal bacteria or lactic acid bacteria having increased activity through pre-culture are pre-cultured to an _OD570 value of 0.9, and the total weight (V / v).

It is preferable that the intestinal bacteria and the lactic acid bacteria are used alone or in a mixture thereof.

The intestinal bacterium (Raoultella) is preferably composed of one or two or more strains selected from the group consisting of Raoultella ornitholytica, Raoultella planticola, Raoultella terrigena.

The lactic acid (Leuconostoc) are Leuconostoc carnosum, Leuconostoc citreum, Leuconostoc durionis, Leuconostoc fallax, Leuconostoc ficulneum, Leuconostoc fructosum, Leuconostoc garlicum, Leuconostoc gasicomitatum, Leuconostoc gelidum, Leuconostoc inhae, Leuconostoc kimchii, Leuconostoc lactis, Leuconostoc mesenteroides, Leuconostoc pseudoficulneum, Leuconostoc pseudomesenteroides ≪ RTI ID = 0.0 > and / or < / RTI >

After inoculation, a fermentation broth is obtained in a 10 L fermenter under the conditions of a culture temperature of 30 to 37 DEG C, pH of 4.0 to 6.0, stirring speed of 150 to 220 rpm, and culture time of 24 to 48 hours.

The fermented product is sterilized, filtered and lyophilized to obtain a transformant powder (S50).

The fermentation broth is sterilized (121 DEG C, 15 minutes, 1.5 atm) so that the fermentation product obtained in step S40 can not be further fermented. Ethanol was added to the sterilized fermented product so that the fermentation microorganism could not grow any more, and the concentration was adjusted to be a 30% (v / v) ethyl alcohol aqueous solution. In the extractor equipped with a cooling condenser, After 2 to 5 times of hot-water extraction, the mixture was filtered through 100 to 500 mesh filter paper and filtered twice with Edbentek 5 filter paper and Watman GF / C 150 mm filter paper at 30 to 60 ° C, Filtered. Then, the concentrate concentrated at 50 ° C was freeze-dried using a vacuum concentrator to obtain a dry powder.

The fermented product having anti-inflammatory, anti-allergic and atopic skin improving effects according to the present invention improves skin inflammation, allergy and atopic skin, has no skin toxicity, can be applied to sensitive skin, It can be used in various cosmetic compositions.

In addition, the cosmetic composition containing the fermentation broth having anti-inflammatory, anti-allergic and atopic skin improving effects according to the present invention is effective for inhibiting intracellular free radical scavenging effect, lipid peroxidation, hyparrhodicase activity, It is an excellent cosmetic composition which is excellent in efficacy, safety, cheapness and easiness of use because it is excellent in anti-inflammatory and anti-inflammatory effects, safe from side effects in skin irritation test, low in production cost and easy to use.

That is, there is no particular limitation on the formulation of the hydrolyzed cosmetic composition. For example, the hydrogel cosmetic composition may have formulations such as lotion, nutritional lotion, nutritional cream, massage cream, essence, and pack.

The anti-inflammatory, anti-allergic and atopic skin cosmetic composition can be added to the cosmetic in an amount of 0.1% to 30% by weight in terms of the weight of the liquid phase and recovered by evaporating the solvent using a distillation apparatus equipped with a cooling condenser May be added to the cosmetic in an amount of 0.001 to 5 wt%, preferably 0.01 to 5 wt%, in terms of dry weight.

Hereinafter, the structure and effects of the present invention will be described in detail with reference to Examples, Comparative Examples and Experimental Examples of the present invention, but the present invention is not limited thereto.

< Example  1 > The fermented extract of the present invention and the method for producing the powder (solvent-water)

After removing the foreign matter by washing with water flowing through a saw blade or a stem, 1 kg of the pulverized powder obtained by drying in a well-aired shade was pulverized and mixed with 10 L of dilute sulfuric acid prepared so that the concentration of sulfuric acid was 2% (v / v) Followed by heat treatment at 121 占 폚 for 30 minutes. After cooling, the temperature was maintained at room temperature, calcium carbonate (CaCO ₃ ) was added to adjust the pH to 5.5, and the solid-liquid separation was performed using a centrifuge to prepare a pretreatment liquid for fermentation.

20 L fermenter was added with 10 L of pre-treated solution, and Raoultella ornitholytica was pre-cultured to an OD ₅₇₀ value of 0.9. The culture was inoculated at 300 ~ 37 ℃, pH 4.0 ~ 6.0, stirring speed 150 ~ 220 rpm , And the incubation time was 48 hours. After the completion of fermentation, the fermentation broth was sterilized (121 ° C, 15 minutes, 1.5 atm) to prevent further fermentation. Water was added as a solvent to the fermented broth sterilized so that the fermentation microorganism could not be further propagated. Extracted from the fermentation broth for 2-3 hours at 70 to 90 ° C in an extractor equipped with a cooling condenser, filtered with 300 mesh filter paper, The mixture was filtered twice with Edbentek No. 5 filter paper and Watman GF / C 150 mm filter paper at a temperature of 50 캜, and finally, only the fermentation extract of 0.1 탆 or less was microfiltered. The concentrate was concentrated to dryness using a vacuum concentrator at 50 ° C to obtain 97.1 g of a dry powder.

< Example  2> Process for producing fermented extract and powder according to the present invention (solvent - 30% (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Example 1 using 30% (v / v) ethanol to obtain 105.1 g of dried powder.

< Example  3> The fermented extract of the present invention and the powder production method (solvent - 50% (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Example 1 using 50% (v / v) ethanol to obtain 103.9 g of dried powder.

< Example  4> The fermented extract of the present invention and the powder preparation method (solvent - 70% (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Example 1 using 70% (v / v) ethanol to obtain 102.7 g of a dry powder.

COMPARATIVE EXAMPLE 1 A method for preparing a water extract and a powder according to the present invention (solvent-water)

After removing the foreign matter by washing with water flowing through a saw blade or a stem, 20 kg of water was added to 1 kg of pulverized powder in a pulverizer which was air-dried, and the pulverized extractor was equipped with a cooling condenser for 5 hours After extraction, the mixture was filtered through 300 mesh to separate the medicinal material and the extract. The mixture was sealed for 3 days at room temperature. The precipitate was filtered twice with Edbentek 5 filter paper and Watman GF / C 293 mm filter paper. The mixture was concentrated at 50 ° C using a vacuum concentrator and lyophilized to obtain 71.2 g of a dry weight.

< Comparative Example  2> Process for producing extract and powder according to the present invention (Solvent-30 % (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Comparative Example 1 using 30% (v / v) ethanol to obtain dry weight of 82.6 g.

< Comparative Example  3> Process for preparing extract and powder according to the present invention (solvent - 50% (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Comparative Example 1 using 50% (v / v) ethanol to obtain 78.3 g of dry weight.

< Comparative Example  4> Process for producing extract and powder according to the present invention (solvent-70 % (v / v) ethanol)

The solvent was extracted, concentrated and lyophilized in the same manner as in Comparative Example 1 using 70% (v / v) ethanol to obtain 77.5 g of dry weight.

< Manufacturing example  1 > The anti-inflammatory, Anti-allergy , Production of lotion containing water-soluble fermented product having atopic skin improving effect

An example of the prescription of lotion (skin lotion) among the cosmetics containing the watering leaves and stem fermented product powder of Example 2 is as follows.

number Raw material Content (% by weight) One The fermentation broth (Example 2) 1.0 2 glycerin 3.0 3 Butylene glycol 2.0 4 Propylene glycol 2.0 5 Polyoxyethylene hardened castor oil 1.0 6 ethanol 10.0 7 Triethanolamine 0.1 8 antiseptic a very small amount 9 Pigment a very small amount 10 Spices a very small amount 11 Purified water Balance

<Manufacturing Method>

2, 3, 4, and 8 are added in the order of No. 11, followed by dissolving by stirring. The mixture is heated by heating at about 60 ° C for 5 times, and then 10 times is added to dissolve. Finally, 1, 6, 7, and 9 are added, agitated sufficiently after stirring.

< Manufacturing example  2 > The anti-inflammatory, anti- Anti-allergy , Production of nutritious lotion containing a fermented product having atopic skin improving effect

Table 2 shows the prescription examples of the nutritional lotion among the cosmetics containing the water-soluble leaves and the stem fermented product powder of Example 2.

number Raw material Content (% by weight) One The fermentation broth (Example 2) 1.0 2  Wax 1.0 3  Polysorbate 60 1.5 4  Sorbitan sesquioleate 0.5 5  Liquid paraffin 10.0 6  Sorbitan stearate 1.0 7  Pro-type glycerin monostearate 0.5 8  Stearic acid 1.5 9 Glyceryl stearate / FG-400 stearate 1.0 10 Propylene glycol 3.0 11 Carboxy polymer 0.1 12 Triethanolamine 0.2 13 antiseptic a very small amount 14 Pigment a very small amount 15 Spices a very small amount 16  Purified water Balance

<Manufacturing Method>

10, 11, 13, and 16 were heated to 80 to 85 ° C while mixing and stirring. Then, the mixture was charged into a manufacturing part, and then an emulsifier was allowed to react. 2, 3, 4, 5, 6, 7, 8, 9, And then emulsified. After the emulsification is completed, the mixture is cooled to 50 ° C with stirring using an agitator, and then the mixture is cooled to 45 ° C, and then the mixture is cooled to 45 ° C.

< Manufacturing example  3> The anti-inflammatory, Anti-allergy , Preparation of nutrition cream containing fermented product having atopic skin improving effect

Table 3 shows the prescription examples of the nutritional cream among the cosmetics containing the water-soluble leaves and the stem fermented product powder of Example 2.

number Raw material Content (% by weight) One The fermentation broth (Example 2) 1.0 2 Stearic acid 2.0 3 Cetyl alcohol 2.0 4 Glyceryl monostearate 2.0 5 Polyoxyethylene sorbitan monostearate 0.5 6 Sorbitan sesquioleate 0.5 7 Glyceryl monostearate / glyceryl stearate /
Polyoxyethylene stearate 1.0 8 Wax 1.0 9 Liquid paraffin 4.0 10 Squalane 4.0 11 Caprylic / capric triglyceride 4.0 12 Carboxyvinyl polymer 0.3 13 Butylene glycol 5.0 14 glycerin 3.0 15 Triethanolamine 0.5 16 Purified water Balance

<Manufacturing Method>

12, 13, 14, and 16 were heated to 80 to 85 ° C while mixing and stirring. Then, the mixture was put into a manufacturing part, The mixture is heated to 85 ° C to dissolve, and the mixture is stirred and charged into the production section and emulsified. After the emulsification is completed, the mixture is cooled to 35 ° C with stirring using an agitator, and the mixture is cooled to 25 ° C by 1 time and aged.

< Manufacturing example  4> The anti-inflammatory, anti- Anti-allergy , An atopic skin improving effect The fermented product  Manufacture of essences containing

Table 4 shows prescription examples of essences in cosmetics containing the water-insoluble leaves and stem fermented product powders of Example 2.

number Raw material Content (% by weight) One  The fermentation broth (Example 2) 1.0 2 Sitosterol 1.7 3  Polyglyceryl 2-oleate 1.5 4  Ceramide 0.7 5 Steareth-4 1.2 6 cholesterol 1.5 7 Dicetyl phosphate 0.4 8 Concentrated glycerin 5.0 9 Macadamia oil 15.0 10 Carboxyvinyl polymer 0.2 11  Xanthan gum 0.2 12  antiseptic a very small amount 13  Spices a very small amount 14  Purified water Balance

<Manufacturing Method>

2, 3, 4, 5 and 6 are homogenized at a constant temperature to form nonionic amphipathic lipids. The non-ionic amphipathic lipids were mixed with 1, 7, 8 and 14, homogenized at a constant temperature, passed through a microfluidizer and then gradually added at a constant temperature to homogenize the microfluidizer again. And 10, 11, 12 and 13 are added to stabilize and mature.

< Experimental Example  1> Assessment of cell non-toxicity and cell proliferation effect

The cell proliferation effects of the fermented dry powders prepared in Examples 1 to 4 and the extract powders prepared in Comparative Examples 1 to 4 were measured using cell culture.

1) Experimental method

The cytotoxicity and proliferation experiments were carried out in the following manner. The cells were cultured in a 96-well microplate for 5,000 cells / well and incubated for 30 minutes in a thermostatic chamber. The cells were treated with 25, 50, 100, 200 (μg / ml) And cultured for 72 hours. Thiazoline blue was added and the cells were further cultured for 4 hours. The culture medium was discarded, and the reaction suspension was added to each well of the microplate. After stirring for 5 minutes, the absorbance at 570 nm was measured. The control group was co-cultured with 10% Fetal Bovine Serum (FBS) medium as an optimal condition for cell growth, and the cell proliferation rate of the test sample was calculated with the cell proliferation of the control group as 100% .

2) Experimental results

The cell proliferation effect was calculated by Equation 1, and the results are shown in Table 5 below.

[ Equation 1 ]

density
(μg / ml) Cell proliferation effect (%) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Control group 100 100 100 100 100 100 100 100 25 101.8 105.5 102.4 103.4 100.7 103.5 101.3 101.2 50 105.1 118.7 106.8 110.4 101.5 104.6 102.1 100.9 100 110.3 123.6 112.7 117.2 104.2 105.8 105.2 104.3 200 116.1 131.6 120.4 121.6. 104.7 110.8 107.8 105.9

This experiment was conducted to confirm the cytotoxicity and proliferation of the fermentation broth (Examples 1 to 4) and the hydrolyzed powder (Comparative Examples 1 to 4). As shown in Table 5, when the cell proliferation at the optimal cell growth conditions was taken as 100%, the proliferation effect of the cellulase-extracted powders (Comparative Examples 1 to 4) was negligible, but the fermentation broth 1 ~ 4) cell proliferation was very good.

< Experimental Example  2> free radicals ( Free radical ) Evaluation of erase effect

Free radical scavenging effects of the fermented dry powders prepared in Examples 1 to 4 and the extract powders prepared in Comparative Examples 1 to 4 were measured.

1) Experimental method

Experiments were carried out using the DPPH (1,1-diphenyl-2-picryl-hydrazyl) method (Blois, MSNature 181, 1190, 1958). DPPH and quercetin were purchased from Sigma . Absorbance was measured at 517 nm in Examples 1 to 4 and Comparative Examples 1 to 4 at various concentrations (5, 10, 20, 50 and 100 μg / ml) in 1 ml of a 0.2 mM DPPH methanol solution. Instead, purified water was used.

Scavenging activity (SC ₅₀ ) represents the concentration required to reduce the free radical concentration by 50%. In the SC ₅₀ The lower the value, the better the free radical scavenging activity.

The free radical scavenging effect was calculated using the following equation 2, and the results are shown in Table 6 below.

[ Equation 2 ]

2) Experimental results

density
(μg / ml) Free radical scavenging effect (%) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 5 9.1 19.5 10.5 13.5 2.2 3.7 3.3 1.1 10 30.4 51.6 45.4 43.1 11.8 15.9 12.5 9.2 20 57.1 83.6 73.7 67.6 38.6 38.2 30.6 27.6 50 79.1 98.2 90.4 85.4 72.5 83.5 76.6 68.1 100 96.4 98.9 97.3 92.7 87.7 86.5 86.1 89.5 SC ₅₀ 34.45 18.45 27.21 32.28 44.70 42.29 45.33 50.24

As shown in Table 6, the free radical scavenging concentrations (SC ₅₀ ) of the fermentation broths of Examples 1 to 4 were 34.45, 18.45, 27.21 and 32.28 μg / ml, while the extractive powders of Comparative Examples 1 to 4 the free radical 50% scavenging concentration (SC ₅₀₎ are 44.70, 42.29, 45.33, example 1-4 in that the free radical scavenging effect of cane fermentation powder superior to the Comparative examples 1-4 cane extract powder to 50.24 μg / ml I was able to confirm that it was excellent.

Experimental Example 3 Evaluation of Lipid peroxydation Inhibitory Effect

The lipid peroxydation inhibitory effect of the fermented dry powders prepared in Examples 1 to 4 and the extract powders prepared in Comparative Examples 1 to 4 was measured, and a green tea extract was used as a comparative substance.

1) Experimental method

After dissolving linolenic acid to 0.1% in 8% sodium lauryl sulfate solution, 3.9 ml was taken and 0.1 ml of sample was added to each concentration (100, 300, 500, 700, 1000, 1500 μg / ml) For one hour. 1 ml of 1.5% TBA aqueous solution and 1.5 ml of 20% acetic acid (pH 3.5) were added to the flask. After cooling for 1 hour with stirring, 1 ml of purified water and 5 ml of n-BuOH: pyridine (15: After shaking and centrifugation, the n-BuOH layer was taken and the absorbance at 532 nm was measured.

The inhibitory concentration (IC ₅₀ ) represents the concentration required to reduce the activity of lipid peroxidation by 50%. The IC ₅₀ The lower the value, the better the scavenging activity of lipid peroxidation.

The lipid peroxidation effect was determined using the following equation 3 and the results are shown in Table 7 below.

[ Equation 3 ]

2) Experimental results

density
(μg / ml) Inhibitory effect of lipid peroxidation (%) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Green tea powder 100 5.6 12.7 9.7 8.2 3.4 7.2 6 4 2.3 300 15.3 29.3 22.3 19.7 7.9 13.1 15.7 10.6 9.1 500 30.2 54.6 48.9 35.4 19.3 28.6 34.1 19.7 15.2 700 59.6 78.1 70.4 60.7 45.7 50.5 51.9 37.8 23.6 1000 83.6 97.6 87.6 70.4 81.5 83.6 81.8 65.9 59.4 1500 91.2 98.9 95.9 94.6 82.8 88.7 87.6 85.4 96.1 IC ₅₀ 495.47 279.79 383.84 420.36 636.45 581.86 567.32 715.68 737.33

As shown in Table 7, the fermented water powders of Examples 1 to 4 had a lipid peroxidation inhibitory concentration (IC ₅₀ ) of 495.74, 278.79, 383.84, and 420.36 μg / ml, respectively, and those of Comparative Examples 1 to 4 and green tea extract powder Lipid peroxidation effect was superior to those of 636.45, 581.86, 567.32, 715.68 and 737.33 μg / ml of the lipid peroxidation inhibitory concentration (IC ₅₀ ).

Experimental Example 4 Evaluation of Inhibition of Lipoxygenase Activity

The inhibitory effects of Lpoxygenase activity on the fermented dry powders prepared in Examples 1 to 4 and the extract powders prepared in Comparative Examples 1 to 4 were measured.

1) Experimental method

Experiments were performed using TBAS method. Linoleic acid, Lipoxygenase, Thiobabitulic acid and Quercetin used in the experiment were used by SIGMA. To 1 ml of 1 mM linoleic acid, 0.05 ml of quercetin was added to each of the concentrations (10, 30, 50, and 100 μg / ml) of Examples 1 to 4 and Comparative Examples 1 to 4 as comparative substances, and 0.95 ml of Lipoxygenase After the administration, the mixture is stirred well and reacted at 25 ° C for 10 minutes. Then, 0.5 ml of trichloroacetic acid is added, and 1 ml of thiobarbituric acid is added. After heating for 10 minutes, the reaction is terminated by cooling in ice water for 2 to 3 minutes. To the reaction mixture, 2 ml of butanol was added, and the mixture was centrifuged at 4,000 × g for 5 minutes. Absorbance was measured at 535 nm, and purified water was used instead of each sample.

The inhibitory concentration (IC ₅₀ ) represents the concentration required to reduce the activity of the lipoxygenase by 50%. The IC ₅₀ The lower the value, the better the activity of the lipoxygenase elimination.

The inhibitory effect of the lipoxygenase activity was determined using the following equation 4, and the results are shown in Table 8 below.

[ Equation 4 ]

2) Experimental results

density
(μg / ml) Lipoxygenase activity inhibitory effect (%) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 10 6.6 13.9 10.1 5.1 3.4 7.5 6.7 2.2 30 32.4 45.1 37.5 29.4 19.7 25.7 22.5 16.2 50 55.7 72.5 67.2 59.7 43.7 55.6 49.3 44.7 100 83.1 96.7 88.6 80.6 85.2 82.3 81.1 79.5 IC ₅₀ 55.17 40.97 59.14 61.07 68.79 65.47 67.74 68.19

As shown in Table 8, the fermentation powders of Examples 1 to 4 were 55.17, 40.97, 59.14, and 61.07 μg / ml in the inhibitory concentration (IC ₅₀ ) of the lipoxygenase 50% (IC ₅₀ ) of 68.78, 65.47, 67.74, and 68.19 g / ml, respectively.

&Lt; Experimental Example 5 > Evaluation of inhibitory effect on hyparonidase activity

The ferulic dry powders prepared in Examples 1 to 4 and the extract powders prepared in Comparative Examples 1 to 4 were measured for their inhibitory activity against hyparrhonidase activity.

1) Experimental method

Morgan-Elson method is applied. Inhibition of the inactivated hyaluronidase in the activation step of the hyaluronidase by 400 NF unit / ㎖, final concentration of HA of 0.4 mg / ml and addition of Compound 48/80 buffer solution (0.1 mg / ml) Hyaluronidase activity was measured centrally. The sample was dissolved in buffer to prepare a sample solution. As a control, a buffer was used instead of the sample solution. For each blank, a buffer was used instead of the enzyme solution, and a green tea extract was used as a control.

The inhibitory concentration (IC ₅₀ ) represents the concentration required to reduce the activity of the hyaruronidase by 50%. The IC ₅₀ The lower the value, the better the activity of scavenging the hyaruronidase.

The inhibitory effect of the hyparrhoidase activity was determined using the following equation 5, and the results are shown in Table 9 below.

[ Equation 5 ]

2) Experimental results

density
(μg / ml) Inhibitory effect of hyaluronidase activity (%) Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 green tea
Extract powder 75 8.4 13.5 10.8 9.4 4.1 7.9 5.6 3.3 6.5 150 14.5 28.7 23.7 19.5 12.5 18.1 13.7 9.6 10.9 300 49.8 63.5 57.6 51.4 30.7 32.6 33.6 23.5 25.1 600 81.7 91.4 87.6 80.6 80.3 86.23 85.2 78.2 75.1 1200 93.4 99.1 98.1 96.6 83.1 88.9 87.6 84.5 99.3 IC ₅₀ 460.53 293.29 504.60 512.67 527.72 503.57 626.87 559.39 544.34

As shown in Table 9, the inhibitory concentration (IC ₅₀ ) of hyaruronidase in the fermented broth of Examples 1 to 4 was 460.53, 293.29, 504.60, and 512.67 μg / ml, respectively, (IC ₅₀ ) 527.72, 503.57, 626.87, 559.39 μg / ml, and 544.34 μg / ml of the green tea extract powder as a comparative substance.

<Experimental Example 6> Evaluation of improvement effect of atopic skin

The clinical evaluation of the atopic skin improvement effect on the cosmetic preparation of Production Example 3 was measured.

Production Example 3 In order to evaluate the improvement effect of the atopic skin of the cosmetic, the following experiment was carried out on 15 adult patients who were admitted to a clinic or a hospital and had treatment with atopic skin. Test samples were divided into right and left sides by double-blinded test, respectively, and applied to the whole body, but the use of other moisturizing agents which could affect the test sample's effect as much as possible was prohibited. The effects of SCORAD (SCORAD; Scoring Atopic Dermatitis) were evaluated after 1, 2, 3, and 4 weeks after the test sample was applied, and the results are shown in Table 10 below.

1) Acceptance criteria

① Extent criteria: area = damaged area / 100

② Intensity criteria

   - Erythema (1/2/3)

   - Edema (1/2/3)

   - Excretion (1/2/3)

   - Skin exfoliation (1/2/3)

   - Degree of firing (1/2/3)

   - Degree of drying (1/2/3)

③ Subjective criteria

   - Itching (1-10)

   - Insomnia (1-10)

* Scored calculation = (precision standard / 5) + (strength standard / 7) + subjective standard

Classification 0 weeks 1 week 2 weeks 3 weeks 4 weeks Production Example 3 65.7 35.2 22.3 12.7 9.8 0.00% 46.42% 66.06% 80.67% 85.08% Comparative Example 5 69.3 51.3 42.8 38.2 21.5 0.00% 25.97% 38.24% 44.88% 68.98%

In Comparative Example 5, a nutritive cream was prepared according to the formulation of Preparation Example 3, except that the fermented water powder was removed, and purified water was added to prepare a nutritive cream.

As shown in Table 10, it was confirmed that the nutritional cream containing the hydrolyzed powder according to Production Example 3 (Example 2) was superior to the nutritional cream of Comparative Example 5 in improving the atopic skin.

In particular, the results after 4 weeks show that the nutritional cream of Preparation Example 3 has an improvement of atopic skin of about 85%, which is superior to the nutritional cream of Comparative Example 5.

The present invention is not limited to the above embodiments, but may be applied to various fermented products having various anti-inflammatory, anti-allergic and atopic skin improving effects, And the present invention is not limited to the above-described embodiments, and various changes and modifications may be made without departing from the scope of the present invention. But only by the scope of the claims.

Claims (6)

The fermentation is carried out by intragastric bacterium which is a Raoultella strain after acid hydrolysis and neutralization by selecting at least one of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, formic acid and phosphoric acid,
The intestinal bacteria is Raoultella ornitholytica,
The fermented product according to any one of claims 1 to 3, wherein the sorghum is a stem or a leaf. The fermentation is carried out by intragastric bacterium which is a Raoultella strain after acid hydrolysis and neutralization by selecting at least one of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, formic acid and phosphoric acid,
The acid hydrolysis was carried out by adding 10 L of dilute sulfuric acid having a concentration of 2% (v / v) sulfuric acid and heat-treating at 121 캜 for 30 minutes,
The fermentation is carried out by inoculating Raoultella ornitholytica,
The fermented product according to any one of claims 1 to 3, wherein the sorghum is a stem or a leaf. 3. The method according to claim 1 or 2,
Wherein the neutralization reaction is selected by neutralizing at least one of calcium carbonate (CaCO ₃ ), sodium hydroxide (NaOH), sodium carbonate (Na ₂ CO ₃ ) or calcium hydroxide (Ca (OH) ₂ ) A fermented product having a skin improving effect. The method according to claim 1,
Wherein said acid hydrolysis is carried out by acid hydrolysis by adding diluted sulfuric acid after crushing millet, thereby obtaining a fermented product having anti-inflammatory, anti-allergic and atopic skin-improving effects. A cosmetic composition comprising anhydrous fermented product according to any one of claims 1, 2 and 4, and a fermented product having an anti-inflammatory, anti-allergic and atopic skin improving effect. 6. The method of claim 5,
Wherein the cosmetic composition is formulated into a formulation of a lotion, a nutrient lotion, a nutritional cream, a massage cream, an essence or a pack, wherein the cosmetic composition comprises a fermented product having an anti-inflammatory, anti-allergic and atopic skin improving effect.

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