KR102270709B1 - Cosmetic composition for skin improvement containing complex ceramide and natural extracts - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin conditions, comprising five types of complex ceramides and natural extracts and, more specifically, to a cosmetic composition for improving skin conditions in which five types of complex ceramides and a leaf extract of Taraxacum officinale have the effect of maintaining the microbiome balance in the skin and strengthening the skin barrier, and to a method for preparing the same.

Description

Cosmetic composition for skin improvement comprising complex ceramide and natural extract {COSMETIC COMPOSITION FOR SKIN IMPROVEMENT CONTAINING COMPLEX CERAMIDE AND NATURAL EXTRACTS}

The present invention relates to a cosmetic composition for improving skin comprising five kinds of complex ceramides and natural extracts, and specifically, five kinds of complex ceramides and dandelion leaf extracts have the effect of maintaining the microbiome balance in the skin and strengthening the skin barrier. It relates to a cosmetic composition for improving skin and a method for manufacturing the same.

Microbiome is a compound word of microbiota, which refers to the microbial community, and genome, which refers to genome. Microbiota is a term that focuses on the microorganism strain itself compared to the microbiome, and refers to microorganisms that inhabit the human body and have a symbiotic relationship. It refers to research and technology based on genomics of interactions between humans. However, recently, microbiota, which means microorganisms, and microflora, and microbiome are commonly used. The term microbiome, better known as the intestinal microbiome, is to suppress harmful bacteria by ingesting lactic acid bacteria to expand the power of intestinal lactic acid bacteria, and both lactic acid bacteria and harmful bacteria correspond to the microbiome.

One of the reasons why the microbiome has been receiving a lot of attention in recent years is that its applicability to human health and disease problems has been greatly expanded. Metabolic diseases such as obesity and diabetes, as well as cancer, infection, immunity, and mental disorders are highly related to the microbiome in the body, and maintaining the balance of the microbiome is known to have a positive effect on the functions of the human body. Interest in research is increasing.

In the case of the microbiome market currently being formed, the functional food or wellness product market centered on probiotics and prebiotics accounts for more than 80% of the total market. As the health functional probiotics market grows rapidly, interest in R&D in cosmetics, medicine, and pharmaceutical fields is also increasing rapidly.

On the other hand, as the relationship between skin acne and staphylococcus aureus and propionibacterium acnes is known, cosmetics have been developed and commercialized for this, so far most cosmetics have an antiseptic effect on harmful bacteria. The development of cosmetics based on the prebiotic function is insufficient.

Korean Patent No. 10-2121006

An object of the present invention is to provide a cosmetic composition for maintaining the microbiome balance in the skin comprising ceramide, dandelion leaf extract and lactobacillus ferment lysate.

One aspect of the present invention for achieving the above object is any selected from the group consisting of ceramide EOP (EOP), ceramide NS (NS), ceramide NP (NP), ceramide ES (AS) and ceramide AP (AP) one or more ceramides; Dandelion leaf extract; And it relates to a cosmetic composition for maintaining the microbiome balance in the skin comprising a lactobacillus ferment lysate.

In the present invention, “microbiome” is a compound word of microbiota and genome, and refers to the entire microbial community and genome coexisting in humans, animals and plants, soil, sea, lake, rock wall, atmosphere, etc. . Among them, the skin microbiome refers to a part of the human microbiome as a group of microorganisms that form a complex interrelationship between microorganisms and microorganisms present in the skin, and between hosts and microorganisms.

The “maintenance of the microbiome balance in the skin” refers to properly maintaining the balance of the microbial community in the skin by suppressing the proliferation and/or activity of harmful bacteria in the skin while enhancing the activity and survival of beneficial bacteria in the skin.

Specifically, maintaining the microbiome balance may be achieved through the promotion of skin beneficial bacteria.

In the present invention, "prebiotics" can be defined as short-chain carbohydrates, that is, oligosaccharides, which are minimally hydrolyzed and absorbed by animals and induce the dominance of beneficial bacteria among intestinal microorganisms. Most of the prebiotics can be selectively used as a substrate by beneficial intestinal bacteria, representatively FOS (fructooligosaccharide), TOS (transgalactooligosaccharide), OF (oligofructose) or inulin (inulin). Probiotics, also called beneficial bacteria, maintain the normal host gastrointestinal microflora and provide beneficial effects on the host, such as increasing resistance to pathogenic bacteria, which support the growth or maintenance of these probiotics. That's what prebiotics are.

In the present invention, it was identified that the dandelion leaf extract and Lactobacillus ferment lysate promote the promotion of beneficial bacteria in the skin, and it was confirmed that the prebiotic function was improved according to the extraction conditions and the origin and type of the ferment lysate.

In the present invention, "Western dandelion leaf ( Taraxacum officinale ) extract" is an extract obtained by extraction treatment of dandelion leaves, a diluted or concentrated solution of the extracts, a dried product obtained by drying the extract, a preparation or purified product of the extract, or It includes extracts of all formulations that can be formed using the extract itself and the extract, such as mixtures thereof.

The extract may be extracted from a natural, hybrid or mutated plant of the dandelion leaf, and may also be extracted from a plant tissue culture.

The method of extracting the extract of the present invention is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, and the like, and these may be performed alone or in combination of two or more methods.

The type of the extraction solvent used in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water (purified water), C ₁ to C ₄ anhydrous or lower alcohol, a mixed solvent of water and lower alcohol, acetone, 1,3-butylene glycol, ethyl acetate, and chloroform. One or more selected from the group may be used alone or in combination of two or more.

The extraction ratio of the active ingredient included in the raw material may be different depending on the polarity of the solvent. In particular, the solvent may preferably be an alcohol having excellent selectivity in extracting the physiologically active substance of the raw material, and preferably, an optimal skin improvement effect may be realized by ethanol extraction.

In particular, since water and ethanol have different polarities, active ingredients extracted according to each polarity may be different, so that the concentration of ethanol may be appropriately controlled so that an optimal skin improvement effect can be realized. However, if the concentration of the ethanol is less than 50%, the active ingredient showing the skin improvement effect may not be sufficiently extracted, and if the concentration is more than 80%, an appropriate yield may not be realized, so the concentration of the ethanol is within the above range can be adjusted appropriately. In one embodiment, the extraction solvent of the dandelion leaf extract may be ethanol, more specifically 70% ethanol.

In the present invention, “fermentation” refers to a process of decomposing organic matter by using the enzyme possessed by the fermenting strain itself. It is known that the effects of raw materials that have undergone the fermentation process through fermentation strains are increased at least 2 times to up to tens of times compared to before fermentation. In particular, fermented metabolites contain antioxidants, including various amino acids and organic acids, which are good for the skin and have an effect of improving the skin.

Specifically, the western dandelion leaf extract may be fermented at a low temperature of 15°C or more and 25°C or less using a Lactobacillus strain.

In one embodiment of the present invention, the fermented lysate of dandelion leaf extract and Lactobacillus strain was mixed to confirm the skin beneficial bacteria growth promoting effect, and in particular, it was confirmed that the western dandelion leaf extract fermented at a low temperature had excellent skin beneficial bacteria promoting effect. .

In the present invention, “lysate” means a solution or suspension in an aqueous medium of cells of a microorganism such as broken Lactobacillus. Cell lysates include, for example, macromolecules such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc. and/or micromolecules such as amino acids, sugars, fatty acids, etc., or fractions thereof. The lysate also contains cell debris, which may be smooth or granular in structure.

As a method for achieving cell lysis of the microorganism, various known methods may be used, and any method capable of achieving cell lysis of the microorganism may be used. For example, cell opening/destruction can be performed enzymatically, chemically or physically. Non-limiting examples of enzymes and enzyme mixtures are proteases such as proteinase K, lipases or glycosidases; Non-limiting examples of chemicals are ion permeable carriers, detergents such as sodium dodecyl sulfate, acids or bases; Non-limiting examples of physical means are high pressure such as French pressing, osmotic pressure, temperature such as heat or cold. In addition, a method using an appropriate combination of an enzyme other than a proteolytic enzyme, an acid, a base, and the like may also be used.

For example, cells of the microorganisms of the present invention are lysed by freezing and thawing, more preferably freezing at a temperature below -70°C and thawing at a temperature above 30°C, in particular freezing at a temperature below -75°C is preferred. It is preferred and preferred to thaw at temperatures above 35°C, most preferred are freezing temperatures below -80°C and thawing temperatures above 37°C. The freezing/thawing may be repeated one or more times, more preferably two or more times, more preferably three or more times, particularly preferably four or more times, and most preferably five or more times.

Specifically, the fermentation lysate may be a Lactobacillus fermentation lysate, and more specifically, may be a Lactobacillus fermentation lysate separated from kimchi.

As a result of measuring the skin microbiome balance maintenance efficacy using fermented lysates of microbial strains isolated from various foods such as breast milk, cheese, and kimchi in an embodiment of the present invention, all of them were effective in promoting skin beneficial bacteria, but It was confirmed that the Lactobacillus ferment lysate not only showed a remarkable effect of promoting beneficial bacteria, but also of reducing the activity of harmful bacteria on the skin.

The cosmetic composition may further include the use of strengthening the skin barrier and alleviating skin redness.

By adding ceramide to a cosmetic composition comprising the dandelion leaf extract and lactobacillus ferment lysate, it is possible to restore the damaged skin barrier as the microbiome balance in the skin is broken.

In the present invention, “ceramide” is a kind of sphingolipid-based compound having a structure in which fatty acids are linked to sphingosine or phytosphingosine. Specifically, it may be represented by the following Chemical Formula 1, wherein R represents an alkyl moiety of a fatty acid.

[Formula 1]

The ceramide is a major lipid component constituting the stratum corneum, and plays an important role in the formation of moisturizing ability of the skin and the occurrence of skin barrier diseases. Ceramide accounts for about 40 to 50% or more of the lipid between the keratinocytes constituting the stratum corneum of the skin, and is an essential component that shows the structure or function of the stratum corneum. It acts as a lipid barrier to inhibit water evaporation in the stratum corneum and maintains the orderly structure of the stratum corneum. function to make

The ceramides are sphingosine, pytosphingosine, and 6-OH-sphingosine, which are basic structures, respectively, and are given skeletal names as S, P, and H, respectively, depending on the type of binding of fatty acids. It can be divided into EOS(ceramide1), EOP(ceramide9), EOH(ceramide4), NS(ceramide2), NP(ceramide3), NH(ceramide8), AS(ceramide5), AP(ceramide6), AH(ceramide7), etc. can

The ceramides each have different polarities, and may be classified into ceramide EOP, ceramide NS, ceramide NP, ceramide AS, and ceramide AP according to the degree of polarity.

In the case of ceramide EOP, it is known that dermatitis such as atopic dermatitis occurs because the keratinocytes of the skin do not make skin lipids well when the production capacity is low, and therefore it is mainly used to improve the skin condition that feels dry and itchy.

In addition, ceramide NP is a complex active ingredient, and as the most important substance among oils and fats contained in keratin of the skin, it has a particularly excellent function in moisturizing the skin. It is known to mainly play a key role in giving shine, luster and elasticity to hair, strengthening the lipid barrier of the skin, and forming a skin protective film against the external environment.

Specifically, the ceramide may include one or more selected from the group consisting of ceramide EOP, ceramide NS, ceramide NP, ceramide AS and ceramide AP.

More specifically, the ceramide may be composed of ceramide EOP, ceramide NS, ceramide NP, ceramide AS and ceramide AP in a ratio of 1-1.5: 1-1.5: 1-3: 1-1.5: 1-1.5.

In an embodiment of the present invention, a composition containing the five kinds of complex ceramide, dandelion leaf extract and lactobacillus ferment lysate was prepared in liquid, cream, and cream mist formulations to conduct human application experiments. As a result, skin barrier function was strengthened. And it was confirmed that the redness improvement effect was excellent.

The cosmetic composition of the present invention is a solution, external ointment, cream, liquid, foam, nourishing lotion, softening lotion, pack, softening water, body wash, emulsion, makeup base, essence, soap, liquid detergent, bath agent, sunscreen cream, sunscreen Oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, foam cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays can be prepared in a formulation selected from the group consisting of, but limited thereto. it is not going to be

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers to be formulated in general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, and thickener. , a chelating agent, a colorant, a preservative, a fragrance, and the like may be appropriately blended, but the present invention is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohard propellants such as, but not limited to, locarbon, propane/butane or dimethyl ether. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butylglycol oil and the like can be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, fatty acid esters of polyethylene glycol or sorbitan may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, as a carrier component a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used, but is not limited thereto. These may be used alone or in combination of two or more.

The composition of the present invention may be used as a transdermal administration method such as directly applied to the skin or sprayed, and the administration route of the composition of the present invention may be administered through any general route as long as it can reach the target tissue.

The usage amount of the composition of the present invention may be appropriately adjusted according to individual differences or formulations such as age, degree of lesion, etc., and may be used for one week to several months by applying drops to the skin once to several times a day.

The cosmetic composition of the present invention can maintain the microbiome balance in the skin through the excellent prebiotic function of dandelion leaf extract and Lactobacillus ferment lysate.

In addition, the skin barrier function damaged due to the breakdown of the balance of beneficial and harmful bacteria in the skin can be restored through 5 types of complex ceramides, and furthermore, it can relieve skin redness and have a soothing effect on the skin. It can be utilized as a cosmetic composition for cosmetic use.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 shows the results of confirming the effect of soothing and alleviating damaged skin for the product using Example 1 (A: liquid formulation, B: cream formulation, C: cream mist formulation).
2 shows the results of confirming the redness relief effect of the product using Example 1 (A: liquid formulation, B: cream formulation, C: cream mist formulation).
3 shows the results of confirming the skin barrier strengthening function for the product using Example 1 (A: liquid formulation, B: cream formulation, C: cream mist formulation).
4 shows the skin improvement efficacy and quality questionnaire evaluation results of the product using Example 1 (A: liquid formulation, B: cream formulation, C: cream mist formulation).

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples are merely illustrative of the present invention, and the present invention is not limited by the following examples.

Preparation Example 1. Sample preparation of dandelion leaf extract

1-1. Preparation of dandelion leaf extract

After washing the dandelion leaf with water, it was completely dried at room temperature and pulverized to obtain 100 g of a pulverized product. 100 g of the pulverized product was placed in a flask, and extracted by cooling at 25° C. for 3 days with 1000 g of distilled water (extraction solvent). The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a dandelion leaf extract.

1-2. Preparation of low-temperature fermented dandelion leaves using low-temperature (15 - 25℃) fermentation process

The Lactobacillus strain isolated from kimchi or cheese was inoculated into the dandelion leaf extract prepared in Preparation Example 1-1. Thereafter, fermented dandelion leaves were prepared by fermentation at 250 rpm for 72 to 96 hours at a temperature of 15 - 25°C.

1-3. Preparation of high temperature fermented dandelion leaf using high temperature (35 - 40℃) fermentation process

The Lactobacillus strain isolated from kimchi or cheese was inoculated into the dandelion leaf extract prepared in Preparation Example 1-1. Thereafter, fermented dandelion leaves were prepared by fermentation at 250 rpm for 72 to 96 hours at a temperature of 35-40°C.

Preparation Example 2. Sample preparation of lactic acid bacteria fermentation lysate

2-1. Fermentation lysate sample preparation for each strain

After crushing and homogenizing the kimchi, 1.0 g of this was taken, suspended in 9.0 ml of sterile physiological saline (0.85% NaCl), diluted stepwise, and plated on the MRS growth medium shown in Table 1 below and each corresponding selective medium, respectively. After culturing at 37° C. for 24 to 48 hours, strains having different colonies were selected and purified through several pure culture processes. The purely isolated strain was spread on MRS solid medium and cultured at 37°C for 24 hours, and the purely isolated colonies were cultured in MRS liquid medium and mixed with 20% (v/v) glycerol in a 1:1 ratio. It was stored in a -80°C cryogenic freezer. Thereafter, the strain was cultured in a culture medium and fermented, sterilized with a sterilizer, and impurities were filtered through a filter having a size of 0.1 to 0.3 μm.

strain optional badge growth medium Lactobacillus spp. LBS
(Lactobacillus selection) MRS Lactococcus spp. M17 MRS Pediococcus spp. PSM MRS Leuconostoc spp. PES-30 MRS

2-2. Lactobacillus fermentation lysate sample preparation by origin

After crushing and homogenizing breast milk, cheese, and kimchi, respectively, take 1.0 g of them, suspend them in 9.0 ml of sterilized physiological saline (0.85% NaCl), and dilute them stepwise, and add them to the MRS growth medium of Table 1 and each corresponding selective medium, respectively. smeared. After culturing at 37° C. for 24 to 48 hours, strains having different colonies were selected and purified through several pure culture processes. The purely isolated strain was spread on MRS solid medium and cultured at 37°C for 24 hours, and the purely isolated colonies were cultured in MRS liquid medium and mixed with 20% (v/v) glycerol in a 1:1 ratio. It was stored in a cryogenic freezer at -80°C. The Lactobacillus was cultured in a culture medium, fermented, and then sterilized with a sterilizer to filter out impurities with a filter having a size of 0.1 to 0.3 μm.

Examples and Comparative Examples

Example ingredient
[Content (parts by weight)] Kimchi-derived Lactobacillus Ferment Lysate Dandelion Leaf Extract Western Dandelion Leaf
high temperature fermented extract Western Dandelion Leaf
Low-Temperature Fermented Extract 5 types of complex ceramides One 2 One - - One 2 2 - One - One 3 2 - - One One

comparative example ingredient
[Content (parts by weight)] Kimchi-derived Lactobacillus Ferment Lysate Cheese-derived Lactobacillus Ferment Lysate Lactobacillus ferment lysate derived from breast milk Kimchi-derived Lactococcus Ferment Lysate Dandelion Leaf Extract 5 types of complex ceramides One 2 - - - - One 2 2 - - - One - 3 - - - 2 One One 4 - 2 - - One One 5 - - 2 - One One

Experimental Example 1. Confirmation of skin barrier strengthening effect of 5 types of complex ceramides

In the present invention, an experiment was performed to confirm the effect on the skin according to the combination of five types of ceramides.

HaCaT cells were seeded in a 24-well plate at 1.0 × 10 ⁵ cells/mL and cultured at 37° C., 5% CO ₂ conditions for 18 hours. The samples were exchanged for serum-free DMEM and incubated for 24 hours. After that, each group was treated with PBS containing no drugs that could affect protein quantification after removing the culture medium and washing with PBS. The protein was collected after lysing the cells by repeating low-temperature and room-temperature incubation. Quantification was carried out by using the Filaggrin-ELISA kit (Elabscience Biotechnology Co., Ltd) and the method provided by the manufacturer.

Example ingredient
[Content (parts by weight)] ceramide Kimchi-derived Lactobacillus Ferment Lysate Dandelion Leaf Extract EOP NS NP AS AP 1-1 - - One - - 2 One 1-2 0.5 - 0.5 - - 2 One 1-3 0.2 0.2 0.2 0.2 0.2 2 One 1-4 0.17 0.17 0.32 0.17 0.17 2 One Comparative Example 2 - - - - - 2 One

Increase in filaggrin (FLG) production (%) - density % control - 100 Example 1-1 50 130.5 Example 1-2 50 130.2 Examples 1-3 50 145.7 Examples 1-4 50 148.1 Comparative Example 2 50 107.5

As a result, in the case of Examples 1-4 containing the five kinds of ceramide complex extracts, the expression level of filaggrin was increased by 29.87%, and especially when the content ratio of ceramide NP (NP) was high, it was confirmed that the skin barrier protection effect was excellent. did.

Experimental Example 2. Confirmation of skin beneficial bacteria enhancement effect

An experiment was performed to confirm the effect of Examples 1 to 3 on the promotion of beneficial skin bacteria.

2-1. Leukonostok mecenteroides ( Leuconostoc mesenteorides ) Confirm the effect of promoting bacterial growth

Leuconostoc mesenteorides belonging to lactic acid bacteria as beneficial bacteria, it was confirmed whether there is a proliferation promoting effect of the ( Leuconostoc mesenteorides ) strain.

First, Leukonostok mecenteroides was isolated from kefir grains using a selective medium. Anaerobic culture was performed at 30° C. for 72 hours using 0.002% BPB-MRS medium. Thereafter, lactic acid bacteria colonies were selected and identified by 16S rRNA sequencing. The turbidity of the bacteria that had been woken up two days ago was adjusted to 2.0, and then injected into a tube containing 3 mL of MRS broth ^{, diluted to 10 1} CFU/mL, and the initial number of bacteria was measured. Then, 5 mg of each of the compositions of Examples 1 to 3 was added to 3 mL MRS broth, and after ultrasonic treatment for 5 minutes (50 seconds operation, 10 seconds interval, repeat 5 times), 0.1% DMSO was injected. In the same manner, the compositions of Comparative Examples 1 to 5 were added to 3 mL MRS broth, and the number of bacteria (CFU/mL) was measured after incubation. The results are shown in Table 6 below.

division Number of bacteria (CFU/mL) Example 1 3.16 x 10 ⁷ Example 2 3.17 x 10 ⁷ Example 3 3.51 x 10 ⁷ Comparative Example 1 7.38 x 10 ⁶ Comparative Example 2 3.15 x 10 ⁷ Comparative Example 3 2.85 x 10 ⁷ Comparative Example 4 3.05 x 10 ⁷ Comparative Example 5 3.02 x 10 ⁷

As a result, when the Leukonostok mecenteroides strain was cultured in Examples 1 to 3, the growth was significantly increased compared to that in Comparative Example 1 which did not contain the dandelion leaf extract. Furthermore, it was confirmed that the beneficial bacteria enhancing effect was significantly increased compared to the Lactococcus fermented product (Comparative Example 3) and the Lactobacillus fermented product of different origin (Comparative Example 4, Comparative Example 5).

2-2. Confirmation of reduction of harmful bacteria in facial skin

For 10 female subjects aged 25 to 45 years (average age 33.8 years), the cosmetic composition of the liquid formulation prepared according to Table 7 below was applied to the facial skin twice a day for 2 weeks and 4 weeks Changes in harmful bacteria were confirmed in the posterior facial skin.

ingredient content (wt%) Examples 1-3 or
Comparative Examples 1 to 5 2.0 Butylene Glycol 5.0 1-2-hexanediol 3.0 Ethylhexylglycerin 0.05 Purified water remaining amount synthesis 100

First, the microbes on the skin surface of both cheeks of the face were swabbed in a state where temperature and humidity were constant (temperature 23±2℃, humidity 50±5%). Swabs were performed using Quick Swab (3M Microbiology, USA). Afterwards, cut the swapped head part with sterile scissors, put it in a 50 mL conical tube, add 2X beads of FastDNA SPIN Kit (Goma Biotech, Korea), sodium phosphate, MT buffer, and vortex for 20 minutes. After the genome was extracted, PCR was performed using Bac9F/Bac541R primers (see Meisel et al., 2016; Kong, 2016). After purifying the PCR product, the purified DNA sample was ^{quantified using the Quant-iTTM PicoGreen ®} dsRNA assay kit (Invitrogen, USA) to prepare each sample to contain the same amount of DNA. Thereafter, pyrosequencing was performed using a 454 GS FLX titanium sequencing system (Roche, Germany). The pyro-sequencing was performed using GS FLX titanium reagent according to the manufacturer's instructions. Microbiome using bioinformatics software of phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) based on OTUs from 16S rDNA gene. of meta-genetic analysis was performed. Harmful bacteria were Propionibacterium acnes , Corynebacterium minutissimum and Staphylococcus aureus . The harmful bacteria inhibition results of the cosmetic composition of the present invention are shown in the table below. As shown in 8.

division pathogenic strains Change over time (%) 0 weeks 1 week 2 weeks Example 1 Propionibacterium acnes 70.04 60.00 37.99 Corynebacterium minuticium 0.025 0.020 0.009 Staphylococcus aureus 0.061 0.010 0 Example 2 Propionibacterium acnes 70.04 61.09 38.11 Corynebacterium minuticium 0.025 0.019 0.010 Staphylococcus aureus 0.055 0.014 0 Example 3 Propionibacterium acnes 70.04 55.18 30.18 Corynebacterium minuticium 0.025 0.005 0.001 Staphylococcus aureus 0.055 0.003 0 Comparative Example 1 Propionibacterium acnes 70.04 67.01 45.13 Corynebacterium minuticium 0.025 0.023 0.014 Staphylococcus aureus 0.055 0.009 0.003 Comparative Example 2 Propionibacterium acnes 70.04 65.71 44.21 Corynebacterium minuticium 0.025 0.022 0.012 Staphylococcus aureus 0.055 0.011 0.002 Comparative Example 3 Propionibacterium acnes 70.04 65.15 42.05 Corynebacterium minuticium 0.025 0.025 0.012 Staphylococcus aureus 0.055 0.017 0 Comparative Example 4 Propionibacterium acnes 70.04 64.08 40.05 Corynebacterium minuticium 0.025 0.024 0.010 Staphylococcus aureus 0.055 0.014 0 Comparative Example 5 Propionibacterium acnes 70.04 64.42 41.99 Corynebacterium minuticium 0.025 0.025 0.011 Staphylococcus aureus 0.055 0.015 0

As a result, when the products of Examples 1 to 3 are applied to the skin for a certain period of time, all of the activities of Propionibacterium acnes, Corynebacterium minuticium and Staphylococcus aureus, known as skin harmful bacteria, are reduced. Confirmed.

Experimental Example 3. Human application test results to confirm the skin soothing and skin barrier function strengthening effects of 5 types of complex ceramides and natural extracts

To confirm the skin improvement efficacy of the liquid, cream, and cream mist formulation of Example 1 containing 5 types of complex ceramides of the present invention, natural extract and lactobacillus ferment lysate, human application to Human Skin Clinical Test Center The test was requested, and the test was conducted according to the Cosmetics Guidelines of the Ministry of Food and Drug Safety, the Declaration of Helsinki, and the Human Skin Clinical Test Center Standard Operating Instructions (SOP).

The number of test subjects was 22 healthy Korean male and female subjects between the ages of 19 and 60, and there were no dropouts. The average age of the subjects was 48.7 years, the highest age was 60 years, and the lowest age was 28 years. All measurements and evaluations were carried out in a space where there is no movement of air or direct sunlight, and constant temperature and humidity conditions (22±2℃, 50±5%) were maintained, with the subject taking skin stability, and evaluation was performed before and after using the test product. carried out.

3-1. Check skin soothing effect

To evaluate the skin soothing effect, 2% SLS aqueous solution was occluded on the forearm area for 2 hours to induce chemical irritation, and then the forearm was photographed using Antera 3D (Miravex Ltd., Ireland) before and 1 week after using the test product. Skin color lightness (a*) was analyzed.

As a result, skin-colored redness significantly increased in both the application site of the test product of Example 1 and the non-applied control site after stimulation induction compared to before induction of chemical stimulation. However, after 1 week of use of the test product, it was confirmed that the amount of redness reduction was significantly higher in the area where the test product was applied than in the uncoated control area ( FIGS. 1 and 2 ).

3-2. Confirmation of skin barrier function strengthening effect

In order to evaluate the skin barrier enhancement effect, 2% SLS aqueous solution was occluded on the forearm for 2 hours to induce chemical irritation, and then, before and 1 week after using the test product, a Vapometer (Delfin Technologies Ltd., Finland) was used to of transdermal water loss (g/m ² h) was measured. The Vapometer is a device that evaluates Transepidermal Water Loss (TEWL), an indicator of water loss through the epidermis. If the barrier function of the skin is damaged, water loss increases and the measured value increases.

^{As a result, the amount of transdermal moisture loss (g/m 2} h) was significantly reduced in both the application site of the test product of Example 1 and the control area without application after stimulation induction compared to before the induction of chemical stimulation. It was confirmed that the amount of decrease in the amount of transdermal water loss was significantly higher than that of the site (FIG. 3).

3-3. Results of questionnaire evaluation on efficacy and quality after use

A questionnaire on efficacy and quality was conducted to 22 test subjects after 1 week of use. The response scale is as follows. 1: Strongly disagree, 2: Disagree, 3: Neutral, 4: Agree, 5: Strongly agree.

Questionnaire on the efficacy of the test product 1, the redness seems to have decreased. 2. The skin seems to be moist. 3. Skin dryness seems to have decreased. Questionnaire on the quality of test products 1. It is well absorbed into the skin. 2. Not sticky. 3. It smells good. 4. Doesn't seem to irritate the skin. 5. Overall I am satisfied. 6. I am willing to recommend to others.

As a result, satisfaction with the effect of reducing redness and maintaining skin moisture was excellent in relation to skin efficacy, and in relation to product quality, the product received particularly high ratings for its spreadability and hypoallergenicity (FIG. 4).

Experimental Example 4. Confirmation of human skin hypoallergenic test results of 5 types of complex ceramides and natural extracts

To confirm the skin improvement efficacy of the liquid, cream, and cream mist formulation of Example 1 containing 5 types of complex ceramides of the present invention, natural extract and lactobacillus ferment lysate, human application to Human Skin Clinical Test Center The test was requested, and the test was conducted according to the Cosmetics Guidelines of the Ministry of Food and Drug Safety, the Declaration of Helsinki, and the Human Skin Clinical Test Center Standard Operating Instructions (SOP).

The number of subjects was 31 healthy Korean male and female subjects between the ages of 19 and 60, and there were no dropouts. The subjects had an average age of 47.39 years, the highest age was 60 years, and the lowest age was 23 years.

All measurements and evaluations were conducted in a space where there is no movement of air or direct sunlight and constant temperature and humidity conditions (indoor temperature 20±2℃, relative humidity 50±5%) were maintained. 20 μl of the test product was taken from a site without skin damage or skin damage, and occluded patch was applied for 24 hours. The skin reaction was observed 30 minutes and 24 hours after the patch was removed, and the presence and extent of skin irritation was visually evaluated by a dermatologist according to the evaluation criteria.

The skin irritation level of the test product was evaluated based on the skin irritation level classified by applying the Draize method after calculating the skin irritation index based on Frosh & Kligman (1979) and CTFA guideline (2007). Skin irritation index criteria and criteria are shown in Tables 10 and 11 below.

sign score Explanation - 0 no response + One Weak but distinct erythema reaction ++ 2 Erythema reaction accompanied by papules or swelling +++ 3 Strong erythematous reaction with edema and papules ++++ 4 Strong erythematous reaction with edema and blisters

skin irritation index Determination of skin irritation 0.00~0.25 Non (non-irritating) 0.26 to 1.00 mild irritant 1.01~2.50 Moderate irritation 2.51~4.00 strong irritant

division 30 min 24 hrs Average
skin reaction Judgment result One 2 3 4 One 2 3 4 Example 1
(Liquid) - - - - - - - - 0.00 non-irritating Example 2 (cream) - - - - - - - - 0.00 non-irritating Example 3 (Cream Mist) - - - - - - - - 0.00 non-irritating

As a result, as shown in Table 12, the test product of Example 1 was judged to be non-irritating regardless of the formulation, and it was confirmed that there was no adverse reaction observed during the test period. Those of ordinary skill in the art to which the invention pertains will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and likewise components described as distributed may be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.

Claims (6)

Ceramide EOP (EOP), Ceramide NS (NS), Ceramide NP (NP), Ceramide ACE (AS) and Ceramide AP (AP) 1~1.5 : 1~1.5 : 1-3 : 1~1.5 : 1~1.5 Ceramide consisting of a ratio of;
Dandelion leaf extract; and
Lactobacillus fermentation lysate; cosmetic composition for maintaining the microbiome balance in the skin comprising a. delete According to claim 1,
The Lactobacillus fermentation lysate is a cosmetic composition comprising a lysate of a Lactobacillus strain isolated from kimchi. According to claim 1,
The solvent of the dandelion leaf extract is 70% ethanol, and the cosmetic composition is fermented at a low temperature at 15°C or more and 25°C or less. According to claim 1,
The cosmetic composition further comprises the use of strengthening the skin barrier and alleviating skin redness. According to claim 1,
The cosmetic composition includes a solution, an external ointment, a cream, a liquid, a foam, a nourishing lotion, a flexible lotion, a pack, a soft water, a body wash, an emulsion, a makeup base, an essence, a soap, a liquid detergent, a bath agent, a sunscreen cream, a sun oil, Suspension, emulsion, paste, gel, lotion, powder, soap, foam cleansing, oil, powder foundation, emulsion foundation, wax foundation, a cosmetic composition in any one formulation of a patch and spray.

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