KR102553740B1 - Cosmetic composition for modulating skin microbiome - Google Patents

Abstract

The present invention relates to a cosmetic composition for regulating skin microbiome, and more particularly, by including a black garlic extract, beneficial skin bacteria can be promoted and harmful skin bacteria can be suppressed, ultimately suppressing pigmentation, reducing pores, reducing oiliness or wrinkles can show improvement.

Description

Cosmetic composition for controlling skin microbiome {COSMETIC COMPOSITION FOR MODULATING SKIN MICROBIOME}

The present invention relates to a cosmetic composition for regulating skin microbiome.

Black garlic is made by aging raw garlic for a certain period of time. Compared to raw garlic, black garlic has higher sugar content, acidity, and jelly-like properties, so it can be consumed on its own compared to raw garlic. There are advantages to being higher. In addition, black garlic is known to have high antioxidant activity due to browning reaction substances produced during the ripening process, and to have effects related to cancer prevention, cholesterol lowering, arteriosclerosis improvement, heart disease prevention, and oxidative stress.

Microbiome is a compound word of microbiota and genome, and refers to the entire genome and microbial community coexisting in humans, animals and plants, soil, sea, lake, rock wall, and air. In the past, the development of materials to improve health or prevent/treat diseases by controlling the microbial community in the intestines was the main focus, but recently interest in the development of materials to improve skin by regulating the skin microbiome is increasing. . However, until now, the discovery of materials related to skin functional enhancement by directly controlling the skin microbiome has been insufficient. Therefore, it is necessary to develop a material that can improve skin functionality by directly controlling the skin microbiome.

Korean Patent Publication No. 2020-0018879

An object of the present invention is to provide a cosmetic composition for regulating skin microbiome.

1. A cosmetic composition for regulating microbiome in the skin containing black garlic extract.

2. The cosmetic composition according to 1 above, wherein the microbiome control promotes beneficial bacteria in the skin.

3. The cosmetic composition according to 2 above, wherein the beneficial bacteria is at least one selected from the group consisting of Bacteroidetes , Actinobacteria and Fusobacteria phylum.

4. In the above 3, the beneficial bacteria are at least one selected from the group consisting of Bifidobacterium , Alistipes, Barnesiella and Fusobacterium genus Phosphorus, a cosmetic composition.

5. The cosmetic composition according to 1 above, wherein the microbiome control inhibits harmful bacteria in the skin.

6. The cosmetic composition according to 5 above, wherein the harmful bacteria is at least one selected from the group consisting of Proteobacteria and Firmicutes .

7. The cosmetic composition according to 6 above, wherein the harmful bacteria is at least one selected from the group consisting of Escherichia , Shigella , and Listeria .

8. The cosmetic composition according to 1 above, wherein the cosmetic composition has a skin barrier strengthening effect, skin moisturizing effect, skin dry symptom improvement effect, skin pigmentation inhibitory effect, pore shrinking effect, oil reduction effect, wrinkle improvement effect and skin elasticity improvement effect. To exhibit at least one effect selected from the group consisting of, a cosmetic composition.

9. The cosmetic composition according to 1 above, wherein the black garlic extract is contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition.

10. The cosmetic composition according to 1 above, wherein the black garlic extract is prepared by a method comprising immersing black garlic in an alcohol solvent of C1 to C6 for extraction.

11. The cosmetic composition according to 10 above, wherein the method further comprises the step of freeze-drying after the extraction step.

The composition of the present invention can promote beneficial skin bacteria and suppress harmful skin bacteria, and ultimately exhibit effects of suppressing pigmentation, reducing pores, reducing oil content, or improving wrinkles.

Figure 1 shows the result of confirming the increase in the alpha diversity of the skin microbial community by the black garlic extract-containing formulation.
Figure 2 shows the result of confirming the increase in the alpha diversity of the skin microbial community by the black garlic extract-containing formulation.
Figure 3 shows the results of confirming the change in the distribution of skin microorganisms by the black garlic extract-containing formulation.

The present invention provides a cosmetic composition for regulating the microbiome in the skin containing a black garlic extract.

The term “black garlic” means that raw garlic has turned black by aging under appropriate temperature and humidity conditions for a certain period of time. Compared to raw garlic, black garlic shows effects such as higher antioxidant effect, cancer prevention, cholesterol lowering, arteriosclerosis improvement, and heart disease prevention. In addition, black garlic is aged garlic, and since volatile sulfur compounds that cause odor are reduced, it has a characteristic that there is no unpleasant smell due to odor compared to raw garlic.

The black garlic extract may be included in an amount of 0.0001 to 10% by weight based on the total weight of the cosmetic composition.

For example, the black garlic extract may be included in 0.0001 to 10% by weight, 0.0005 to 5% by weight, 0.001 to 1% by weight, 0.001 to 0.1% by weight, or 0.001 to 0.01% by weight based on the total weight of the cosmetic composition.

The black garlic extract may be prepared by an extraction method comprising the step of immersing black garlic in an alcohol solvent of C1 to C6 and extracting it, and specifically prepared by an extraction method comprising the step of extracting by immersing black garlic in an alcohol solvent of C1 to C3. may have been

The extraction may be performed at 20 °C to 60 °C, 25 °C to 55 °C, 30 °C to 50 °C, or 35 °C to 45 °C.

The extraction method may further include freeze-drying after the extraction.

The term “microbiome” is a compound word of microbiota and genome, and refers to the entire genome and microbial communities that coexist in humans, animals, plants, soil, sea, lake, rock wall, and atmosphere.

Among them, the “microbiome in the skin” refers to a part of the human microbiome as a group of microorganisms that form a complex mutual relationship between microorganisms and hosts and microorganisms existing in the skin. Specifically, there are more than 2 million microorganisms of 1000 types per 2 cm ² at most in the skin.

The term “regulation of the microbiome in the skin” means promoting the activity or survival of beneficial bacteria in the skin and/or inhibiting the proliferation or activity of harmful bacteria in the skin, and through this, the balance of the microbial community in the skin can be properly maintained . On the other hand, maintaining the microbiome balance in the skin is one of the important factors in improving skin condition, but it is easy to break the balance by various lifestyle and environmental factors such as aging, ultraviolet rays, fine dust, hormones, and diet. The need for cosmetics that can properly maintain the balance of the microbial community by controlling it is gradually increasing.

Control of the microbiome in the skin may promote beneficial bacteria in the skin or inhibit harmful bacteria in the skin.

The term "enhancement of beneficial bacteria in the skin" means promoting the promotion of beneficial bacteria in the skin or activating the growth of beneficial bacteria.

The beneficial bacteria may be at least one selected from the group consisting of Bacteroidetes , Actinobacteria and Fusobacteria .

The beneficial bacteria may be at least one selected from the group consisting of Bifidobacterium , Alistipes, Barnesiella and Fusobacterium .

In one embodiment of the present invention, it was confirmed that the cosmetic composition of the present invention promotes the promotion of beneficial skin bacteria, and it was confirmed that the skin condition can be effectively improved through the promotion of beneficial skin bacteria.

The term “inhibition of harmful bacteria on the skin” means inhibiting the growth of harmful bacteria in the skin or inactivating the growth of harmful bacteria.

The harmful bacteria may be at least one selected from the group consisting of Proteobacteria and Firmicutes .

The harmful bacteria may be at least one selected from the group consisting of Escherichia , Shigella and Listeria .

In one embodiment of the present invention, it was confirmed that the cosmetic composition of the present invention inhibits harmful bacteria in the skin, and it was confirmed that it can be effectively used to improve skin conditions through inhibition of harmful bacteria in the skin.

The cosmetic composition of the present invention may exhibit effects of regulating the microbiome in the skin and improving skin conditions.

The skin condition improvement effect may include a skin barrier strengthening effect, a skin moisturizing effect, an effect of improving skin dryness symptoms, an effect of inhibiting skin pigmentation, a pore shrinking effect, an oil reduction effect, a wrinkle improvement effect, and an effect of improving skin elasticity.

The cosmetic composition of the present invention can exhibit excellent skin improvement effects without side effects such as tingling, swelling, and erythema.

Cosmetic compositions include solutions, external ointments, creams, foams, nourishing lotions, softening lotions, packs, softening waters, emulsions, makeup bases, essences, soaps, liquid cleansers, bath additives, sunscreen creams, sun oils, suspensions, emulsions, and pastes. , Gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, may be prepared in a formulation selected from the group consisting of patches and sprays, but is not limited thereto.

The cosmetic composition may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and as typical ingredients, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, A colorant, preservative, flavoring agent, etc. may be suitably blended, but is not limited thereto.

Cosmetically acceptable carriers included in the cosmetic composition vary depending on the formulation of the cosmetic composition.

When the formulation of the cosmetic composition is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as a carrier component This 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 cosmetic composition is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene Glycol, 1,3-butyl glycol oil, etc. may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester It 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 cosmetic composition is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, a micro-diluent as a carrier component 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 cosmetic composition may be used as a transdermal administration method such as direct application or spraying to the skin, and the route of administration is not limited as long as it can reach the target tissue.

The amount of the cosmetic composition of the present invention may be appropriately adjusted according to individual differences such as age, severity of lesions, etc., or formulation, and may be applied to the skin once or several times a day and used for one week to several months.

Hereinafter, the configuration and effects of the present invention will be described in more detail by way of examples. However, the following examples are provided only for illustrative purposes to aid understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

Preparation of cosmetic composition containing black garlic extract

1. Example 1

100 g of black garlic obtained from Namhae-gun Black Garlic Co., Ltd. was immersed in 500 ml of 70% ethanol and extracted by stirring at 40 ° C. for 12 hours. After 12 hours, ethane was removed using an evaporator, and black garlic extract powder was obtained through lyophilization.

Purified water, glycerin, butylene glycol, vegetable squalane, hydrogenated lecithin, niacinamide, 1,2-hexanediol, dimethicone, phytantriol, phytosphingosine, ceramide NP, batyl alcohol, 3-O-ethyl A creamy cosmetic composition was prepared by adding black garlic extract powder to ascorbic acid, allantoin, scented geranium oil, gwanggwak scented oil, and ylang ylang flower oil. Black garlic extract powder was added in an amount of 0.005% by weight based on the total weight of the composition.

2. Comparative Example 1

A creamy cosmetic composition was prepared in the same manner as in Example 1, but without adding black garlic extract.

Experimental Example 1: Analysis of skin facial microbial community

Twelve women were treated with Comparative Example 1 (left, L) and Example 1 (right, R) on both cheeks of the face twice a day after washing in the morning and in the evening for a total of 5 days, respectively. Analysis was performed by collecting skin microbiome from both cheeks 6 hours after daily formulation treatment. Specifically, a skin microbiome sample was taken from a 2 x 2 cm area of skin on both cheeks of the subject. Samples were collected on the morning of the first day (day1_control, day1_experimental) and the fifth day (day5_control, day5_experimental) right after facial cleansing during the five days participating in the experiment.

DNA extraction, PCR amplification and sequencing followed the Earth Microbiome Project (EMP) standard protocol for standardized microbiome research (https://www.earthmicrobiome.org/protocols-and-standards/dna-extraction-protocol). . Genomic DNA was extracted with DNeasy Power Soil HTP 96 well kit (Qiagen Inc, CA, USA) according to the instructions provided by 76 manufacturers. (https://www.protocols.io/view/emp-dna-extraction-protocol-nutdewn).

Specifically, the V4 region of 16S rRNA in the skin microbiome sample obtained was amplified using 515F/806R primers. The amplification reaction consisted of 12 ul PCR-grade water (Sigma-Aldrich, MO, USA), 10 ul Gold hot-start Taq PCR master mix (Bioneer, Daejeon, Korea), 1 ul each of 5 uM regular and reverse primers, and 1 ul This was done using template DNA. All samples with a total reaction volume of 25ul were amplified 3-fold. It was denatured at 94 °C for 3 minutes, and amplification cycles of 94 °C for 45 seconds, 50 °C for 60 seconds, and 72 °C for 90 seconds were repeated 35 times. PCR products were quantified using the Quant-iT PicoGreends DNA assay kit (Invitrogen, CA, USA), and fluorescence intensity was measured using a microplate reader (BioTek, Winouski, USA). Then, the product was clean-up using a NucleoSpin PCR clean-up kit (MACHEREY-NAGEL, Duren, Germany). Final amplification products were sequenced using an Illumina Miseq platform (San Diego, CA, USA). Sequencing was performed at Sanigen (Anyang, Korea).

Miseq sequencing data were analyzed using the Quantitative Insights Into Microbial Ecology (Qiime2, version 2020.2) pipeline. Reads were demultiplexed according to unique barcode sequences using the QIIME2 demux plugin. Demultiplexing sequences were clustered using QIIME2 DADA2 to generate a table of amplicon sequence variants (ASVs). Representative sequences of each amplicon sequence variant were aligned using the QIIME2 alignment plugin. Next, the FastTree program was used by applying the QIIME2 phylogenetic plugin to build a rooted phylogenetic tree. Taxonomic analysis was performed with the QIIME2 feature classifier plugin using 99% identity to the Silva database (Quast et al. 2012). Alpha diversity and beta diversity analysis were performed using QIIME2 plugins alignment, phylogeny, diversity, and emperor. Phylogenetic diversity was used to plot the alpha rarefaction and beta diversity of the ASVs table. The phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 (PICRUSt2) v 2.3.0 bioinformatics software package was used to predict the functional metagenome of the 16S rRNA gene. Using the ASVs table and representative sequences, Kyoto Encyclopedia of Genes and Genomes (KEGG) orthology (KO) metagenome abundance was predicted. Then, KEGG pathway abundance was inferred using the predicted functional profile. A linear discriminant analysis effect size (LEfSe) algorithm was used to determine significant differences between groups (LDA score > 2.0-fold).

Significant differences in alpha diversity between groups were measured using the non-parametric Kruskal-Wallis test. Statistical significance between groups was measured in PCoA plots using permutational multivariate analysis of variance (PERMANOVA), and distances between groups were evaluated using non-parametric t-tests. Linear discriminant analysis (LDA) effect size (LEfSe) was used to confirm significant differences in the relative abundance of bacterial composition between groups. The α value of the factorial Kruskal-Wallis test was 0.05, and the critical value of the LDA score was 3.0. Statistical comparison of frequencies of different ASVs between groups was confirmed in QIIME by Kruskal-Wallis tests. Spearman's correlation coefficient was calculated through QIIME to estimate the correlation between biological parameters of skin and bacterial abundance.

As a result of confirming the difference in phylogenetic diversity (PD) between the microbial community obtained from the left cheek of the face (Comparative Example 1 treatment group) and the microbial community obtained from the right cheek (Example 1 treatment group), a significant value was obtained from the right face It was confirmed that the phylogenetic diversity values of the microbial community were high (DAY_1_R vs DAY_1_L and DAY_5_R vs DAY_5_L) (see Fig. 1). In addition, as a result of confirming the difference in phylogenetic diversity between the microbial community secured on the 1st day of treatment and the microbial community secured on the 5th day of treatment, it was confirmed that the phylogenetic diversity value of the microbial community secured on the 5th day of treatment was high (DAY_1_L vs. DAY_5_L and DAY_1_R vs. DAY_5_R ) (see Fig. 2). That is, it was confirmed that the face treated with Example 1 containing the black garlic extract exhibited a high level of phylogenetic diversity value. Based on the results of previous studies showing higher alpha diversity in the skin of women in their 20s and 30s than in the skin microbiome of women in their 60s and 70s ( Sci Rep . 2019 Nov 14;9(1):16748) , It was confirmed that the subject's skin microbiome could be controlled similarly to that of young skin by treatment with a black garlic-containing formulation.

In addition, as a result of confirming changes in the skin microbiome at the phylum level and genus level when the cosmetic composition containing the black garlic extract was treated, compared to the left cheek treated with Comparative Example 1 (LEFT in FIG. 3) In the right cheek (RIGHT in FIG. 3) treated with Example 1 containing black garlic extract, beneficial bacteria such as Bifidobacterium , Alistipes, Barnesiella and Fusobacterium ) increased, and it was confirmed that harmful bacteria Escherichia , Shigella , and Listeria were suppressed (see FIG. 3).

Experimental Example 2: Confirmation of skin condition improvement effect

The cosmetic composition containing the black garlic extract was confirmed to have an effect of improving skin conditions. Specifically, 12 women were treated with Comparative Example 1 (left, L) and Example 1 (right, R) on both cheeks of the face after cleansing every morning for a total of 5 days, respectively. The skin condition improvement effect of 12 women was confirmed on the 1st and 5th day of formulation treatment. The skin condition improvement effect was measured using Bomtech Electronics' A-One simple device, and the skin condition was measured immediately after sampling for microbiome analysis every day during the test period. In order to minimize the effect of changes in external humidity, the humidity level was maintained at 50% and the skin condition improvement effect was measured. Table 1 below shows the results of measuring the wrinkle improvement effect and the U-zone oil reduction effect on the 1st and 5th days after applying the cosmetic composition containing the black garlic extract of the present invention to the right cheek as average values. Each value in Table 1 is the index of the measuring device, and means the value compared with the best skin condition calculated as 0 and based on it.

DAY 1 DAY 5 p-value wrinkle improvement 3.28 2.83 0.15 Reduces U-zone oil 231 151 0.25

Experimental Example 3: Evaluation of side effects, efficacy and feeling of use

After treating 12 women with Example 1 containing the black garlic extract, the ratio (%) of side effects was confirmed based on the number of people with side effects among 12 people (see Table 2). As a result of the side effect evaluation, it was confirmed that more than 90% of the 12 women showed no side effects.

erythema edema scale ache stiffness tingling doesn't exist 83 92 100 92 92 92 Weakness 0 0 0 8 0 8 medium 8 0 0 0 0 0 severity 8 8 0 0 8 0 Value = (Number of people with a corresponding degree of side effects / Number of people tested) * 100, Unit: %

In addition, after treating 12 women with Example 1 containing the black garlic extract, the improvement rate of surface moisture and inner skin moisture was evaluated and the average value was calculated. As a result, it was evaluated that the feeling of moisture and the feeling of moisture in the skin improved by about 20% (see Table 3).

surface moisture moisture in the skin skin improvement rate 21 21 Value = Moisture improvement rate of each test subject (%) / number of test subjects, unit: %

In addition, after treating 12 women with Example 1 containing the black garlic extract, the feeling of use of the cosmetic containing the black garlic extract was evaluated (see Table 4). As a result of sensory evaluation, about 50% or more of 12 women showed satisfaction.

Moisture spreadability absorptivity oily feeling satisfaction very unsatisfied 0 0 0 0 0 dissatisfaction 0 0 25 17 25 commonly 33 17 50 50 25 Satisfaction 58 67 25 25 50 very good 8 17 0 8 0 Value = (Number of people with that level of satisfaction / Number of people tested) * 100, Unit: %

Claims (11)

A cosmetic composition for increasing species diversity of microbiome in facial skin, comprising an extract of black garlic.
The cosmetic composition according to claim 1, wherein beneficial bacteria of the microbiome are promoted.
The cosmetic composition according to claim 2, wherein the beneficial bacteria is at least one selected from the group consisting of Bacteroidetes , Actinobacteria and Fusobacteria .
The cosmetic composition according to claim 3, wherein the beneficial bacteria is at least one selected from the group consisting of Bifidobacterium , Alistipes , Barnesiella and Fusobacterium .
The cosmetic composition according to claim 1, wherein harmful bacteria of the microbiome are inhibited.
The cosmetic composition according to claim 5, wherein the harmful bacteria is at least one selected from the group consisting of Proteobacteria and Firmicutes .
The cosmetic composition according to claim 6, wherein the harmful bacteria is at least one selected from the group consisting of Escherichia , Shigella , and Listeria .
The cosmetic composition according to claim 1, wherein the cosmetic composition exhibits at least one effect selected from the group consisting of a skin moisturizing effect, skin dryness improvement effect, oil reduction effect, wrinkle improvement effect, and skin elasticity improvement effect.
The cosmetic composition according to claim 1, wherein the black garlic extract is contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition.
The cosmetic composition according to claim 1, wherein the black garlic extract is prepared by a method comprising extracting black garlic by immersing it in a C1 to C6 alcohol solvent.
The method according to claim 10, wherein the method further comprises the step of freeze-drying after the extraction, the cosmetic composition.

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