KR102546410B1 - Cosmetic composition for improving atopic dermatitis, skin moisturizing and skin soothing containing fermented extracts of Gynostemma pentaphyllum, Trifolium pratense and Hedera Helix - Google Patents

Abstract

The present invention relates to a cosmetic composition containing complex fermented extracts of Dole, Red clover, and Pine chinensis leaves as active ingredients. It relates to a cosmetic composition exhibiting skin inflammation relief and skin soothing, atopy improvement, skin moisturizing and skin barrier strengthening activity.

Description

Cosmetic composition for improving atopic dermatitis, skin moisturizing and skin soothing containing fermented extracts of Gynostemma pentaphyllum, Trifolium pratense and Hedera Helix }

The present invention relates to a cosmetic composition containing complex fermented extracts of sphagnum, chinensis chinensis, and pine chinensis as active ingredients. It relates to a cosmetic composition exhibiting inflammation relief and skin soothing, atopic improvement, skin moisturizing and skin barrier strengthening activity.

Skin is a membrane that covers the outside of the body and plays a role in protecting and regulating internal organs and other internal organs by performing various physiological functions that protect the body from various external stimuli, obstacles, and environmental factors such as dryness. do. However, due to biological aging, the secretion of various hormones internally controlling metabolism is reduced, and the function of immune cells and the activity of cells are lowered.

Human skin is an outer layer of the body composed of epidermis, dermis, and subcutaneous fat, and is known to function as a physical barrier that protects the body from various environmental factors, and functions as a tactile sensation and temperature control. When an external stimulus is applied to the skin's defense system, the first sebaceous layer, secondarily the stratum corneum, and in severe cases, the collagen fibers of the dermis act as defenses.

The stratum corneum of the skin is an important factor determining the skin condition as the first barrier between the skin and the external environment. The stratum corneum of healthy skin contains 15 to 20% of water, and when the water content falls below 10%, it becomes dry, loses luster and elasticity, and wrinkles increase. When defects occur due to extrinsic factors such as penetration of irritants or endogenous factors such as aging, hormones, and stress, resistance decreases, resulting in deterioration of the function of the epidermal double lipid film and increased sensitivity to irritants, resulting in sensitive skin and dry skin. .

In addition, as the skin barrier function is weakened by stimulation, irritation-inducing substances penetrate into the stratum corneum and cause irritation to cause skin inflammation, and as the ability to retain moisture in the skin is weakened, the skin becomes easily dry.

Externally, due to the destruction of the ozone layer, the amount of ultraviolet rays in the sun's rays increases, and as environmental pollution becomes more severe, excessive physical and chemical stimulation and stress caused by the increase of free radicals and active free radicals lead to a decrease in the normal function of the skin, It can damage the skin by accelerating the generation of spots and freckles due to melanin deposition and skin aging.

In recent years, cosmetics made of natural materials have been in the limelight by consumers for reasons such as human safety and skin irritation relief, and accordingly, various studies on natural products as cosmetic raw materials are being conducted.

Under this background, the present inventors have studied to develop a cosmetic with atopic skin improvement effect and skin soothing and moisturizing effect, and a complex fermented extract obtained by fermenting and extracting a mixture of stone, red clover, and pine pine leaves, which have been used as conventional cosmetics, in a specific ratio. The present invention was completed by confirming that this synergistic effect not only exhibits excellent atopic skin improvement, anti-inflammatory, antioxidant and soothing effects, but also exhibits excellent skin moisturizing and skin barrier strengthening effects.

(001) Republic of Korea Patent Registration No. 10-1599254 (2016.02.25) (002) Republic of Korea Patent Registration No. 10-0667448 (2007.01.04) (003) Republic of Korea Patent Publication No. 10-2022-0055190 (2022.05.03)

An object of the present invention is to provide a cosmetic composition that contains as active ingredients a complex fermented extract of stone, red clover, and pine pine leaf, and exhibits excellent skin-improving effects.

In order to achieve the above object, according to the present invention, there is provided a cosmetic composition containing as active ingredients a complex fermented extract of stone, trifolium pratense, and western pine leaf.

Preferably, the composite fermented extract is characterized in that it is prepared by fermenting and extracting a mixture of stone, red clover, and pine pine leaf mixture with lactic acid bacteria.

The lactic acid bacteria used for fermentation are Lactobacillus plantarum, Lactococcus lactis, Lactobacillus bulgaricus, Lactobacillus rhamnosus, Pediococcus cerevisiae (Pediococcus cereviciae), Lactobacillus breve, Leuconostoc mesenteroides, Bifidobacterium bifidum, Lactobacillus sakei and Lactobacillus crispatus ( Lactobacillus crispatus) is at least one selected from the group consisting of.

Preferably, the composite fermented extract is extracted by mixing and extracting stone, trifolium pratense, and western pine leaves in a weight ratio of 1 to 2: 1 to 2: 1 to 2, respectively.

The complex fermented extract as an active ingredient is contained in an amount of 0.01 to 15% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is characterized in that it is for antioxidant, skin inflammation relief, skin soothing, atopic skin improvement, skin moisturizing or skin barrier strengthening.

The complex fermented extracts of doloeia, red clover, and pine pine leaf of the present invention exhibit excellent antioxidant, inflammation and irritation relief, moisturizing and skin barrier strengthening effects due to their synergistic effects, so they are for antioxidant, skin inflammation relief, skin soothing, It can be usefully used as a cosmetic for improving atopic skin, moisturizing the skin, and strengthening the skin barrier.

Figure 1 is a graph showing the effect of increasing the expression of β-defensin-2 of the combined fermented extracts of P. chinensis, P. chinensis, and Pine chinensis leaf according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail.

According to the present invention, there is provided a cosmetic composition containing as active ingredients a composite fermented extract of dolwei, red clover, and western pine leaves.

Gynostemma pentaphyllum is a perennial vine of the dicotyledonous gourd family. It grows in forests of mountains and fields. The rhizome stretches to the side, has white hairs at the nodes, grows tangled, but climbs up with tendrils. Leaves are alternate, compound leaves with 5 small leaves, narrow egg-shaped elliptical or narrow egg-shaped. The small leaf at the end is 4-8cm long and 2-3cm wide with a small petiole, has a sharp tip, has fine hairs on the front leaf vein, and has sawtooth on the edge. It is a raw material of tea and is effective for obesity, neuralgia, and high blood pressure as a health herb. In the present invention, leaves, stems or outposts other than stone are used.

Red clover (Trifolium pratense) is a dicotyledonous perennial plant belonging to the rose order legume family. It is also called black tea construction, red clover flower, red ginseng leaf (紅三葉), and gold flower leaf (金花菜) and is native to Europe. grows in grass It grows up to 30~60cm in height and has hairs all over. Leaves are alternate and compound leaves divided into 3 parts. The small leaf is a long oval shape with a round end or slightly hollowed out, fine sawtooth on the edge, and a white pattern of the number 8 in the center of the surface. The stipule is attached to the lower part of the petiole, and the end is pointed and the edge is flat. Flowers bloom in June-July, are reddish purple, and hang round in the axils. The calyx is tubular, the tip split into 5, pointed, and has many hairs. The lifespan of the plant is short, but the roots go deep. It is planted as fodder, but the spread is growing wild. It is similar to Shamrock, but has almost no peduncle, no bracts, and has scattered hairs on the main stem. It is widely cultivated in Europe. In the present invention, the leaves, stems or outposts of red clover are used.

Western songak (Hedera Helix) is an evergreen vine plant of the dicotyledonous umbel Araliaceae. Western songak is widely distributed in Europe and North Africa, and was introduced as a houseplant in Korea and spread under the name Helix. In the original habitat, it grows up to 30 m in height. Leaves are alternate, glossy dark green, 10 cm long, split into 3 to 5 branches, and the leaves of old branches are egg-shaped without splitting. On young branches and pedicels, there are grayish-white stray hairs (star-shaped hairs split into several branches). Flowers are bisexual and run in an umbel. There are 5 petals, 5 stamens and 5 styles each, and 5 ovaries. There are several types of horticultural varieties, and the representative species, the tricolor species, has large leaves with light green and milky patterns on a green background, irregularly, and milky at low temperatures. The part is stained with a light red color. In the present invention, the leaves of Western Songak are used.

The present invention relates to a technique for producing a complex fermented extract by fermenting and extracting the above-mentioned stone, trifolium pratense, and pine chinensis leaves, and using it as a cosmetic.

Preferably, the composite fermented extract is characterized in that it is prepared by fermenting and extracting a mixture of stone, red clover, and pine pine leaf mixture with lactic acid bacteria.

The lactic acid bacteria used for fermentation include Lactobacillus plantarum, Lactobacillus lactis, Lactobacillus bulgaricus, Lactobacillus rhamnosus, Pediococcus cerevisiae Pediococcus cereviciae, Lactobacillus breve, Leuconostoc mesenteroides, Bifidobacterium bifidum, Lactobacillus sakei and Lactobacillus crispatus (Lactobacillus crispatus) may be used, and preferably Lactobacillus plantarum is used.

According to one embodiment of the present invention, the complex fermented extracts of the stone, red clover, and western pine leaves are prepared by the following method.

First, after preparing a mixture consisting of stone, trifolium chinensis, and pine chinensis, it is suspended in purified water, and lactic acid bacteria are inoculated and cultured in the suspension to obtain a culture. At this time, Lactobacillus plantarum can be used as the lactic acid bacteria, and the culture is performed at a culture temperature of 25 to 35 ° C for 1 to 3 days, preferably at a culture temperature of 30 ° C for about 3 days. . The resulting culture is centrifuged to obtain a supernatant, which is filtered to prepare a complex fermentation extract.

Preferably, the mixture is made by mixing stones, trifolium pratense, and cypress leaves in a weight ratio of 1 to 2: 1 to 2: 1 to 2, respectively, more preferably in a weight ratio of 2: 1: 1 It is done.

According to another embodiment of the present invention, the complex fermented extract may be prepared by preparing fermented extracts of stone, trifolium pratense and western pine leaves, respectively, and then mixing them.

The composite fermented extracts of the present invention prepared in this way have excellent antioxidant effects (Test Example 1), skin inflammation and irritation relief, and atopic skin improvement effects (Test Examples 2 to 4) by their synergistic action. ), skin moisturizing effect (Test Example 5) and skin barrier strengthening effect (Test Example 6) were confirmed.

In addition to the active ingredients, the complex fermented extract of chinensis, red clover, and pine pine leaf may be added in an amount of 0.01 to 15% by weight based on the total weight of the cosmetic composition. When added at less than 0.01% by weight, the effect is weak, and when it exceeds 15% by weight, the increase in the effect is reduced compared to the increase in content, which is rather inefficient.

The cosmetic composition of the present invention can be prepared in the form of a general emulsified formulation and a solubilized formulation. Emulsified cosmetics include nutrition lotion, cream, essence, etc., and solubilized cosmetics include softening lotion. In addition, by containing a dermatologically acceptable medium and base other than cosmetics, it can be prepared in the form of an adjuvant that can be applied topically and systemically, which is commonly used in the field of dermatology.

In addition, the formulation of the cosmetic composition in the present invention is not limited, but skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, essence, It may be applied in any one formulation selected from the group consisting of pack, soap, shampoo, treatment, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, and eye shadow.

[Example]

Hereinafter, the present invention will be described in more detail based on the following examples and test examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited by the following examples.

Examples 1 to 6: Preparation of composite fermented extract

After drying, pulverized stones, red clover, and pine pine leaves were mixed in the ratio shown in Table 1 below, suspended in purified water corresponding to 10 times the dry weight, and then sterilized at 121 ° C. for 15 minutes. Lactobacillus plantarum was inoculated into the sterilized medium and fermented for 3 days at a temperature of 30°C.

The supernatant was separated from the fermented culture using continuous centrifugation at 7,000 rpm. Lactobacillus plantarum was removed from the supernatant using a membrane filter equipped with a 0.45 μm filter to obtain composite fermented extracts of Dole, chinensis chinensis, and Pine chinensis.

weight ratio Example One 2 3 4 5 6 out of stone One One One One 2 One red clover - One - One One 2 western pine leaves - - One One One One

Comparative Examples 1 to 3: Preparation of mixed extracts

After drying, pulverized stone, red clover, and pine pine leaves were mixed in the ratio shown in Table 2 below, and then purified water corresponding to 10 times the dry weight was added and extracted at 121 ° C. for 2 hours. After this, it was filtered using a 2G3 Glass-filter (PYREX 32940 FNL, JAPAN) and then a hot water extract was obtained.

weight ratio comparative example Comparative Example 1 Comparative Example 2 Comparative Example 3 out of stone One One 2 red clover - One One western pine leaves - One One

Test Example 1: Antioxidant effect

In comparison with Examples 1 to 6, the antioxidant effect was measured by measuring the free radical scavenging ability for the samples 1 to 3.

The electron donating ability of the sample was measured as follows. 100 μl of 5 × 10 ^-4 M DPPH (1-1-diphenyl-2-picryl-hydrazyl) was added to 100 μl of the solution for each concentration of each sample, stirred for 1 minute, and reacted for 30 minutes in a 25 ° C incubator. , and absorbance was measured at 517 nm using a microplate counter. In order to compare the actual results, the experiment was conducted using vitamin C, which is known to have excellent free radical scavenging activity, as a control group. The results are shown in Table 3 below.

sample Concentration (ug/ml) Free radical scavenging activity (%) Example 1 250 32.2 1,000 57.5 Example 2 250 32.8 1,000 59.4 Example 3 250 33.0 1,000 59.1 Example 4 250 38.2 1,000 65.6 Example 5 250 42.7 1,000 76.4 Example 6 250 40.5 1,000 71.9 Comparative Example 1 250 30.6 1,000 51.6 Comparative Example 2 250 33.2 1,000 59.7 Comparative Example 3 250 32.8 1,000 58.9 control group
(Vitamin C) 250 48.4 1,000 84.5

As can be seen from Table 3, both the examples and the comparative samples showed free radical scavenging ability. It was confirmed that the effect of inhibiting cell damage and skin aging due to was excellent.

Test Example 2: COX-2 and PGE-2 expression reduction effect by stimulation

In order to examine the effect on cytokine production of Human Epidermal Keratinocyte (HaCaT), 100 ul of 5×10 ⁵ cells/ml cells were put into a 96 well plate, cultured in a CO ₂ incubator for 48 hours, and then the medium was discarded. After washing the surface with a PBS solution, the medium was replaced with a medium without FBS, followed by starvation for 18 hours and cultured for 24 hours. After culturing, stimulation factors UV-B (100mJ/cm ² ), LPS (10ng/ml), and SLS (10ppm) were treated to stimulate the cells, and then each of Examples 1 to 6 and Comparative Examples 1 to 3 Samples were treated with 0.1% each, and after culturing for 24 hours, the amount of mRNA expression was examined using RT-PCR.

Using the mRNA expression level increased by each of the stimulating factors as a control, the expression decrease rate was calculated by the following formula, and the results are shown in Table 4.

mRNA expression reduction rate (%) = [1-(sample mRNA expression amount / control group mRNA expression amount)] × 100

division COX-2 expression reduction rate Decreased rate of PGE2 expression UV-B LPS SLS UV-B LPS SLS Example 1 17.0 12.2 23.6 18.4 11.2 10.8 Example 2 19.7 16.8 20.0 16.6 11.8 11.2 Example 3 19.4 16.4 20.4 16.7 12.4 11.1 Example 4 26.3 20.1 27.5 22.9 18.7 16.5 Example 5 29.6 23.2 29.2 26.7 20.9 18.0 Example 6 25.9 18.8 26.8 22.3 18.0 16.9 Comparative Example 1 9.2 7.7 10.3 11.1 8.4 5.3 Comparative Example 2 9.6 8.2 10.7 13.4 9.8 8.5 Comparative Example 3 10.5 9.6 12.1 15.1 10.0 9.8 Dexamethasone 27.4 20.5 28.9 23.2 19.1 16.7

As confirmed in Table 4, the complex fermented extracts of P. chinensis, P. chinensis, and Pine pine leaf of Examples 4 to 6 of the present invention were compared to the samples of other Examples and Comparative Examples, and COX-2, an inflammatory factor induced by stimulation, and The expression of PGE-2 was further reduced, and the most excellent effect was shown in Example 5.

Test Example 3: Effect of reducing NF-kB and TNF-α expression by stimulation

In order to examine the effect on cytokine production of Human Epidermal Keratinocyte (HaCaT), 100 ul of 5×10 ⁵ cells/ml cells were put into a 96 well plate, cultured in a CO ₂ incubator for 48 hours, and then the medium was discarded. After washing the surface with a PBS solution, the medium was replaced with a medium without FBS, followed by starvation for 18 hours and cultured for 24 hours. After culturing, the cells were stimulated by treatment with stimulation factors UV-B (100mJ/cm ² ), LPS (10ng/ml), and SLS (10ppm), and then treated with 0.1% of each sample in Examples and Comparative Examples, 24 After culturing for a period of time, the cells were examined for mRNA expression using RT-PCR.

Using the mRNA expression level increased by each of the stimulating factors as a control, the expression decrease rate was calculated by the following formula, and the results are shown in Table 5.

mRNA expression reduction rate (%) = [1-(sample mRNA expression amount / control group mRNA expression amount)] × 100

division NF-kB expression reduction rate Decreased rate of TNF-α expression UV-B LPS SLS UV-B LPS SLS Example 1 19.6 11.1 14.4 15.1 13.2 12.0 Example 2 22.8 15.7 18.3 18.2 21.8 16.5 Example 3 23.0 19.4 18.9 19.9 23.6 17.3 Example 4 24.3 21.0 21.9 24.6 27.7 21.5 Example 5 30.4 23.2 24.6 27.2 30.9 24.2 Example 6 24.3 21.7 23.1 25.0 28.4 22.9 Comparative Example 1 12.0 6.0 11.2 10.8 6.7 7.2 Comparative Example 2 15.3 8.6 12.7 13.7 9.6 9.0 Comparative Example 3 16.7 9.4 13.0 14.5 10.3 9.8 Dexamethasone 27.8 20.3 23.9 25.2 31.4 21.7

As confirmed in Table 5, the composite fermented extracts of P. chinensis, P. chinensis, and Pine pine leaf of Examples 4 to 6 of the present invention were compared to the samples of other Examples and Comparative Examples 1 to 3, inflammatory factor NF induced by stimulation -kB and TNF-α expression It can be seen that further reduction

Test Example 4: β-defensin secretion promoting effect test

For the expression of β-defensin-2 (hBD-2, Human -defensin-2), a human-derived antibacterial peptide having the extracts of the above Examples and Comparative Examples to increase the skin immune barrier function and improve atopic skin, as samples The degree of influence was measured.

HaCaT cells were cultured by dispensing at a concentration of 1×10 ⁶ cells/well in a 6-well plate, and then replaced with FBS-free medium, starvated for 18 hours, and cultured for 24 hours. HaCaT cells were treated with 1.0% of each sample, and after culturing for 24 hours, the cells were examined for Human-defensin-2 (hBD-2) mRNA expression using RT-PCR. The results are shown in Figure 1.

As confirmed in Figure 1, when treated at the same concentration, the combined fermented extracts of P. chinensis, P. chinensis, and Pine leaf of Examples 4 to 6 showed a superior effect of increasing β-defensin-2 expression than the extract samples of other Examples or Comparative Examples. was

Among them, the complex fermented extract of Example 5 showed a more excellent expression increasing effect, and compared to the control pimecrolimus, it showed a similar effect, confirming that the atopy improving activity was excellent.

Test Example 5: Measurement of hyaluronic acid production

Hyaluronic acid production was measured for the samples of Examples and Comparative Examples. HaCaT cells were cultured by dispensing in a 96 well plate at a concentration of 1×10 ⁵ cells/well, then replaced with FBS-free medium, starvated for 18 hours, and cultured for 24 hours. HaCaT cells were treated with 0.1% of each sample, cultured for 24 hours, and the collected culture medium was measured for hyaluronic acid (HA) production using an ELSIA kit. The results are shown in Table 6.

division HA production (ug/ml) Example 1 31.5 Example 2 34.8 Example 3 34.2 Example 4 41.6 Example 5 43.5 Example 6 42.0 Comparative Example 1 21.3 Comparative Example 2 25.8 Comparative Example 3 27.9 hyaluronic acid 43.4

As confirmed in Table 6, the composite fermented extracts of P. chinensis, P. chinensis, and Pine pine leaf of Examples 4 to 6 of the present invention showed higher production of hyaluronic acid.

Example 7: Preparation of Serum Containing Complex Fermented Extracts of Stone, Red Clover, and Western Pine Leaf

A serum containing the complex fermented extracts of dolweed, trifolium pratense, and pine pine leaf of Example 5 was prepared according to a conventional method with the composition and content shown in Table 7 below.

ingredient weight% Example 7 Comparative Example 4 Complex fermentation extract (Example 5) 1.0 - carbomer 0.1 0.1 Methylgluses-20 2.0 2.0 glycereth-26 2.5 2.5 jojoba oil 2.0 2.0 MCT Oil 1.0 1.0 dimethicone 2.0 2.0 Citric Acid 0.3 0.3 propylene glycol 5.0 5.0 Butylene Glycol 5.0 5.0 1,2-Hexanediol 2.0 2.0 allantoin 0.2 0.2 betaine 0.2 0.2 Spices a very small amount a very small amount Purified water TO 100 TO 100

Test Example 6: Confirmation of skin barrier function strengthening effect

In order to confirm the effect of enhancing the skin barrier function, after damaging the skin barrier using the taping stripping method on the upper arm of a person, the serums prepared in Example 7 and Comparative Example 4 were applied to a 5 cm ² area After application, the degree of enhancement of the barrier function was evaluated by evaluating the degree of reduction by measuring the amount of transdermal water loss (TEWL) after 3 hours, 6 hours, 12 hours, and 24 hours. The amount of epidermal water loss was measured by DermaLab Combo from Cortex Technology (Denmark). The results are shown in Table 8 below after comparing the change over time based on the amount of epidermal water loss in the initial damaged barrier state (100%).

division 0 hours 3 hours 6 hours 12 hours 24 hours Comparative Example 4 100 47 63 85 92 Example 7 100 30 41 67 71

As confirmed in Table 8, as a result of the skin barrier function enhancement test, the serum composition of Example 7 containing the complex fermented extracts of P. chinensis, Red clover, and Pine chinensis of the present invention was superior to the composition of Comparative Example 4 without them. It showed the effect of strengthening the skin barrier function.

Claims (8)

A complex fermented extract of Dole, trifolium pratense and chinensis chinensis, which is prepared by fermenting and extracting a mixture obtained by mixing 1~2:1~2:1~2 in a weight ratio, respectively, with lactic acid bacteria, is effective. A cosmetic composition containing as an ingredient. delete The method of claim 1, wherein the lactic acid bacteria are Lactobacillus plantarum, Lactococcus lactis, Lactobacillus bulgaricus, Lactobacillus rhamnosus, Pediococcus serei Pediococcus cereviciae, Lactobacillus breve, Leuconostoc mesenteroides, Bifidobacterium bifidum, Lactobacillus sakei and Lactobacillus crispa Cosmetic composition, characterized in that at least one selected from the group consisting of tooth (Lactobacillus crispatus). delete The cosmetic composition according to claim 1, wherein the complex fermented extract as an active ingredient is contained in an amount of 0.01 to 15% by weight based on the total weight of the cosmetic composition. The cosmetic composition according to claim 1, wherein the cosmetic composition is for antioxidation. The cosmetic composition according to claim 1, wherein the cosmetic composition is for relieving skin inflammation, soothing skin, or improving atopic skin. According to claim 1, wherein the cosmetic composition is a cosmetic composition characterized in that for skin moisturizing or for strengthening the skin barrier.

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