KR101400264B1 - Compositions for enhancing skin barrier comprising mixture extract of Citrus unshiu Markovich and Achyranthes fauriei - Google Patents

Abstract

The present invention relates to a new natural plant mixed extract, and more particularly to a skin barrier strengthening composition containing a mixed extract of dermis and moss.
According to the present invention, the mixed extracts of dermis and ascidian are mixed at a certain ratio, thereby having an excellent synergy effect between the raw materials in strengthening the skin barrier, and a cosmetic composition or skin external preparation containing the extract having such effect as an active ingredient Can be manufactured. The mixed extract of the present invention not only increases the expression of aquaporin-3, auroralin and claudin-1, which are major skin barrier strengthening factors, but also has an effect of inhibiting the expression of TNF-a, an inhibitor of aquaporin-3 expression Respectively. Therefore, it is expected to be used as a cosmetic composition for strengthening skin barrier containing the mixed extract of dermis and ascidus as an active ingredient.

Description

TECHNICAL FIELD The present invention relates to a composition for enhancing skin barrier containing a mixed extract of dermis and shrub,

The present invention relates to a new natural plant mixed extract, and more particularly to a skin barrier strengthening composition containing a mixed extract of dermis and moss.

The skin performs various functions essential for the human body to survive. Barrier function to maintain homeostasis in the human body in response to environmental change, sensory function to recognize external change, and temperature control function belong to the most representative skin functions, and all of them correspond to changes in external environment, And maintains the homeostasis capable of maintaining the same. Among the various functions of the skin, especially the barrier function of the skin, is mainly caused by the stratum corneum which is located at the outermost part of the skin. However, recent studies have shown that the stratum corneum has been reported to affect not only the barrier function but also the function, role, and structure of the inner living cell layer, i.e., the epidermis or dermis, and its importance is continuously increasing . As the importance of the stratum corneum is emphasized, researches on it have been actively carried out. In particular, studies on the structure and function of the keratinocytes in the stratum corneum have been actively carried out.

There are two types of pores associated with the water homeostasis present in the epidermis: Aquaporins (AQP) and Tight Junction (TJ). Transport of water and solvents is carried out through aquaporins (AQPs) through transcellular cells and paracellular through extracellular spaces through tight junctions. Tight junctions and tight junction-proteins act as normal barriers to prevent TEWL in healthy skin and are upregulated when the skin is lost or wounded. So it acts as a rescue system.

Aquaporins (AQP), a skin factor for enhancing skin barrier function and restoring damaged barrier, is an integral membrane protein that acts as a water transporter. There are 13 kinds (0 to 12) in mammals and glycerol can be transported. Among these, AQP1, 3, and 5 are expressed in the skin, and aquaglycerolporin 3 (AQP3) is mainly expressed in the basal layer of the epidermis. In atopic dermatitis and psoriasis, AQP3 expression is more strongly expressed from the basal layer to the anterior layer than normal tissues. As AQP3 increases expression in atopic dermatitis and psoriasis, it plays an important role in moisture transfer and moisturizing, and promotes dry skin skin by promoting skin skin moisture loss.

In addition, TNF-α is produced by multiple cell types in the skin, including keratinocytes, macrophages, and Langerhans cells, and is considered to be a key mediator of skin inflammation. In the skin, TNF-α not only regulates the inflammation and initiation of the response responses, but also has a negative effect on wound healing, so regulation of transient production of TNF-a is essential for skin inflammation.

TNF-α has been shown to decrease AQP3 mRNA expression and promoter activity in AQP3 expression of keratinocytes, suggesting that TNF-α inhibits AQP3 gene transcription and TNF-α Decreased expression of AQP3 expression affects dry skin and skin inflammation.

The tight junction (TJ) is located in the intercellular space of the epidermal granule cell layer and is located at the outermost part of the above intercellular junction (ICJ) structures, that is, at the top of the epidermis. It plays a beneficial role in protecting the organ and maintaining the homeostasis of the human body. Such TJ constructs include TJ-associated proteins such as occludin (OC), claudin, and junctional adhesion molecules. They bind to the actin cytoskeleton and cytosolic regulatory proteins via plaque proteins (zona occludens-1 & cingulin) that are present in the cytoplasm, And has a biological barrier function that regulates water-solute and water movement through space. Among the TJs, occludin is a representative protein component of TJ, which is the earliest found, and its expression level is also known to be highly correlated with the biological function of TJ.

The term moisturizers (moisturizers, moisturizers, moisturizers, moisturizers, moisturizers and lotions) often cause a misunderstanding that the ingredients in the formulation directly moisturize the skin. In the past, the concept of moisturizers has been understood as simple replenishment of skin moisture. Recently, however, more attention is focused on restoring the barrier function, which is the most important function of the skin, rather than simple replenishment of water. Or is recognized as a major etiology.

Accordingly, the present inventors have made intensive efforts to overcome the problems of the prior art. As a result of screening for effective substances among various natural substances in order to find a substance having an effect on strengthening skin barrier, the present inventors have found that increase of aquaporin- α, Tight junction (TJ) component of the extracts of the two raw materials mixed at a certain ratio than the dermis / And the present invention was completed.

Accordingly, it is a main object of the present invention to provide a composition for enhancing skin barrier containing a mixed extract of dermis and tsuyu, which is excellent in skin barrier strengthening effect and has no problem in safety and stability.

It is another object of the present invention to provide a cosmetic composition using the composition for enhancing skin barrier containing the mixed extract of dermis and mist.

According to one aspect of the present invention, there is provided a composition for skin barrier enhancement, comprising a mixed extract of dermis and ascidus as an active ingredient.

In the present invention, strengthening the skin barrier means strengthening the barrier function of the stratum corneum at the outermost part of the skin. The stratum corneum, which is the primary barrier of the skin, can be damaged by physical stimulation or wounds. In the case of people with atopic or dry skin, the skin barrier of the stratum corneum is not functioned properly or is damaged. A so-called "moisturizer" is applied to reduce the treatment or damage of atopic or dry skin. A general moisturizer can not solve the fundamental problem simply by replenishing water or covering the skin. Therefore, the mixed extract of the present invention enhances the skin barrier function by increasing or suppressing the expression of factors related to the barrier function of the stratum corneum and solving the fundamental problem.

The dermis (Citrus unshiu Markovich, Citrus reticulata Blanco) is the mature peri of the citrus unshiu Markovich or the relative plants in Korea. In Japan, Citrus unshiu Markovich (Citrus reticulata Blanco: 柑) is used as a dermis and Citrus tachibana (Makino Tanaka: 橘 柑) is used as a citrus peel. Citrus reticulata Blanco : 柑) and its cultivar varieties as dermis. The dermis is said to be better in the long term, and the redder the better, so the name of Hongpie and Dermis is formed. The fruit is called tangerine (Tachibana), the leaf is tachibana (橘 葉), the yellow part where the inside of the fruit bark is removed is called 柑 紅 (橘), the seed is called 橘 核 (橘 核) . This medicine has a peculiar odor, is slightly irritant, has a spicy flavor and is warm in quality. [辛苦 温]

The dermis cleanses the gums and strengthens the function of the spleen, treating only the abdominal cavity, trimming, vomiting, nausea, indigestion, sluggishness, lingering symptoms, and thin stools. Seawater, sputum and diuretic action. It is a medicine that the effect of medicine increases with the reading aloud, mahwang, dandelion, sour oil, Pharmacological action has been reported to cause the digestive tract stimulation, digestion promotion, geomorphic, anti-ulcer, antidiarrheal secretion, hypercapnia, elevated blood pressure, antiallergic, bile secretion promotion, uterine smooth muscle suppression, Its appearance is unevenly shaped shell, outer side is yellowish red, dark tan, and there are many small concave marks due to loss. The inside is white or pale grayish brown, and the vagina is light and easy to crumble.

The Achyranthes fauriei is called a whistle because the shape of the root is similar to that of a cow's knee. Appearance is a long, long, circumferential circular root, or a circular root with a side root. The root head is attached or removed with a little rhizome. The roots are usually rod-shaped or slightly curved, and the outer surface is yellow or yellowish brown with many vertical wrinkles and sparse ridges. If you use raw whiskey, you can eliminate the ejaculation and boils, and if you grind it, you will see the liver and the gods and strengthen the muscles and skeleton. It removes eosinophilia, and it is used for physiological impoverishment, postpartum abdominal pain. It replenishes bone marrow, it is used for arthritis with good sound, and it treats the mouth and tongue rash caused by the diarrhea. The pharmacological effects of uterine stimulation, cholesterol lowering, diuretic action, hypoglycemic action, and liver function improvement have been reported.

In the present invention, the mixed extract may be extracted by any extraction method known in the art, and may be extracted with water, anhydrous or hydrous ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, benzene, chloroform, glycerin, , Propylene glycol, or a mixed solvent thereof, and is preferably ethanol. The composition of the present invention is characterized by being a composition for skin barrier enhancement.

According to the present invention, there is provided a composition for enhancing skin barrier characterized in that the mixed extract is extracted at a mixed weight ratio of 3 to 7: 1 to 5: dermis: ascorbic acid.

In the present invention, the mixed extract is contained in an amount of 0.01 to 10.0 (W / V)% by dry weight.

In the present invention, the mixed extract is extracted by immersing the dermis and the extract at 5 to 37 DEG C for 1 to 15 days in a solvent, or extracting by heating under a solvent at 50 to 100 DEG C for 3 to 20 hours A composition for strengthening skin barrier is provided.

Specifically, after 5 to 20 times volume of at least one solvent selected from the group consisting of water, anhydrous or hydrous ethanol, methanol, glycerin, and butylene glycol as an extraction solvent is added to the dried dermis and the wax at a weight, Is extracted by heating at 50 to 95 ° C for 3 to 20 hours in a state where the solvent is prevented from being evaporated or the active ingredient is extracted by immersing at 5 to 37 ° C for 1 to 15 days. The extracted extract is concentrated using a distillation apparatus equipped with a cooling condenser. The solvent is evaporated, and the concentrate is concentrated under reduced pressure. The extract is freeze-dried or spray-dried to obtain a dried extract. It is preferable to appropriately select in a wide range from 0.01 to 10.0% by weight in terms of solid content of the mixed extract thereafter.

In accordance with the present invention, there is provided a composition for skin barrier enhancement, wherein the mixed extract has an effect of increasing the expression of aquaporin-3, acrodine and claudin-1 in keratinocytes.

As shown in the examples of the present invention, the mixed extract according to the present invention showed excellent synergy of enhancing skin barrier between raw materials when raw materials were mixed and extracted from each single extract. Specifically, when the mixed extract is treated with human epidermal cells, the expression of aquaporin-3, which plays an important role in moisture transfer and moisturization, is increased. Thus, it is possible to prevent dry skin induction by suppressing skin moisture loss This means enhancing the skin barrier function. In addition, it increases the expression of occludin (OC) and claudin, a structure of TJ (tight junction), which protects the internal organ from the external environment and maintains the homeostasis of the human body. Indicating that it is possible to more effectively control the migration of water-containing solutes and moisture through the peri-cellular space, which means enhancing the skin barrier function.

The present invention provides a composition for skin barrier enhancement, which has an effect of inhibiting tumor necrosis factor-alpha (TNF-α) expression of keratinocytes.

As shown in the examples of the present invention, the mixed extract according to the present invention showed excellent synergy of enhancing skin barrier between raw materials when raw materials were mixed and extracted from each single extract. Specifically, treatment of mixed extracts with human epidermal cells inhibited the expression of TNF-a, which affects the expression of aquaporin-3 due to stimulant mediators, which results in the production of aquaporin caused by transient production of TNF- -3 reduction, thereby preventing skin dryness and irritation of the skin, which means enhancing the skin barrier function.

In the present invention, the skin barrier strengthening agent reduces the moisture loss of the skin and prevents skin dryness and skin inflammation.

According to another aspect of the present invention, there is provided a skin external application composition for skin barrier enhancement, which comprises a mixed extract of dermis and moss.

According to another aspect of the present invention, there is provided a cosmetic composition for skin barrier enhancement, which comprises the dermal mixture of dumbbell and mist as an active ingredient.

As shown in Table 8, which is the result of Experiment 1, TEWL and moisture content of mixed extract-containing creams were measured according to the presence or absence of mixed extracts. As a result, the moisture loss of the mixed extract- This means that the skin barrier is strengthened.

In the present invention, the composition may be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutritional lotion, a massage cream, a nutritional cream, a moisturizer cream, a hand cream, A formulation of a composition selected from the group consisting of an essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser formulation. Such formulations can be prepared using methods known in the art.

The ingredients contained in the cosmetic composition of the present invention may contain, in addition to the above-mentioned extracts, the components conventionally used in cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments and fragrances, And a carrier.

INDUSTRIAL APPLICABILITY As described above, according to the present invention, the mixed extract of dandruff and whey has a synergistic effect of strengthening skin barrier between each ingredient, and a skin barrier strengthening cosmetic composition or skin external preparation containing an extract having such effect as an active ingredient Can be manufactured.

FIG. 1 shows the results of measurement of cytotoxicity by concentration of mixed extract.
FIG. 2 shows the results of measurement of the increase in expression of aquaporin-3 by the concentration of the mixed extract.
FIG. 3 shows the results of measuring the inhibitory effect of TNF-α on the concentration of the mixed extract.
FIG. 4 is a graph showing the results of measurement of the increase in expression of acridine and claudin-1, which are constitutive factors of tight junctions according to the concentration of the mixed extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example  1. Dermis and Wuss  Preparation of mixed extracts

700 g of completely dried dermis and 300 g of astaxanthin were placed in 5 kg of 70% ethanol, and the mixture was heated for 5 hours using an extractor equipped with a cooling condenser, followed by filtration through a 400-mesh filter cloth. After cooling to room temperature, the mixture was allowed to stand at 5 to 15 ° C for 7 days and then aged. Then, the mixture was filtered through filter paper No. 2 of Wattsman. The filtered extract was filtered through a 0.45 μm filter, and then concentrated under reduced pressure at 60 ° C. in a distillation apparatus equipped with a cooling condenser to obtain 18.5 g (dry weight) of an ethanol mixed extract.

Experimental Example 1. Cytotoxicity of each raw material constituting the mixed extract Measure

Each raw material was extracted with 70% ethanol, filtered, and concentrated under reduced pressure to obtain an extract of each of dermis and mist.

The mixed extract was obtained from the mixed extract of Example 1, which was extracted with 70% ethanol at a ratio of 70: 30.

In order to observe the cytotoxicity and cell viability of the mixed extracts and the respective raw materials, succinate dehydrogenase or mitochondrial dehydrogenase present in the living cell mitochondria was used to measure the water solubility and yellowing of MTT (3- [4,5-dimethylthiazole -2-yl] -2,5-diphenyl tetrazolium bromide) is reduced to a blue formazan derivative which is water insoluble. The MTT assay is an experimental method widely used for cell proliferation and toxicity by measuring the number of living cells. Formazan derivatives are prepared by dissolving a dissolving agent (usually dimethylsulfoxide) and measuring the absorbance. The above method will be described in detail as follows.

First, cells of Human keratinocyte (CRL-2310, ATCC) were used. The cells were inoculated on a 24-well culture plate with a cell number of 8 x 10 < ⁵ > After 24 hours of inoculation, the medium was replaced with serum-free medium, the test sample was added by concentration, and then cultured in a 5% CO ₂ incubator at 37 ° C for 1 day. After the incubation, the supernatant was removed, and further washed with 200 μl of 5% PBS (phosphate buffered saline). The MTT solution was added to each well in an amount of 1.0 ml per well. After 4 hours, MTT was removed and 1.0 ml of DMSO After that, the absorbance was measured at 570 nm. The control group of this experiment was a group cultured only with the extraction solvent from which the extract of the present invention was excluded, and the cell viability was calculated according to the following equation (1).

As a result of the measurement, it can be confirmed that no cytotoxicity is observed at all the concentrations treated with the mixed extract of the present invention, as shown in Table 1 and Fig.

&Quot; (1) "

Cell survival rate (%) = (sample OD570) / (control OD570) 100

sample cell Survival rate  (%) Control group 100 Dermis extract  0.1 μg / ml 100.93  1 μg / ml 102.55  5 μg / ml 115.16  10 μg / ml 109.28  25 μg / ml 106.06 50 μg / ml 105.15 100 μg / ml 113.78 Raspberry extract  0.1 μg / ml 100.3  1 μg / ml 103.84  5 μg / ml 114.79  10 μg / ml 104.14  25 μg / ml 104.30 50 μg / ml 98.43 100 μg / ml 111.27 The mixed extract of Example 1 0.1 μg / ml 98.46  1 μg / ml 102.29  5 μg / ml 111.64  10 μg / ml 107.63  25 μg / ml 108.20 50 μg / ml 106.88 100 μg / ml 95.74

Experimental Example 2. Measurement of the amount of aquaporin-3 expressed in mixed extracts, dermis, and ascidian

Real-time PCR (Real-time polymerase chain reaction) was performed to increase the expression of Aquaporin-3 gene. Human keratinocyte (CRL-2310, ATCC) cells were used. Samples were added for each concentration and incubated for 24 hours. One mL of trizol (Trizol, Invitrogen, USA) was added to the cells and separated according to the RNA isolation method of Invitrogen. RNA was quantitated at 260 nm using an ultraviolet detector, and then cDNA (complementary DNA) was synthesized and subjected to real-time PCR. For cDNA synthesis, cDNA synthesis kit (High Capacity RNA-to-cDNA Kit, Applied Biosystems, USA) was used. Real-time PCR was performed using the Real-time PCR Kit (Power SYBR Green PCR Master Mix, Applied Biosystems, CA, USA) and was performed using the kit method. Applied Biosystems 7500 FAST Real-Time PCR System , CA, USA), and amplified products were quantitatively analyzed. The conditions are as follows. (positive control; Retinoic acid 1 [mu] M)

As a result of the measurement, as shown in the following Table 2 and FIG. 2, the expression of aquaporin-3 was increased in a concentration-dependent manner at all concentrations of the mixed extract of the present invention, It showed good synergy effect. The expression level of the extract was about twice as high as that of the retinoic acid, a positive control, indicating that the mixed extract had an excellent synergistic effect on the increase of the expression of aquaporin-3.

sample Aquaporin -3 expression level (%) Control group 100 Retinoic acid 1 μM 121.34 Dermis extract  10 μg / ml 131.12 50 μg / ml 182.41 100 μg / ml 184.67 Raspberry extract  10 μg / ml 114.1 50 μg / ml 122.2 100 μg / ml 110.15 The mixed extract of Example 1  10 μg / ml 152.7 50 μg / ml 196.4 100 μg / ml 206.5

EXPERIMENTAL EXAMPLE 3 Inhibitory Effect of TNF-α Expression in Mixed Extracts, Dermis, and Wheat

Real-time PCR (Real-time polymerase chain reaction) was performed to observe TNF-α gene expression inhibition. Human keratinocyte (CRL-2310, ATCC) cells were used. Samples were added at different concentrations and treated with PMA to induce TNF-α gene expression. The cells were cultured for 24 hours, and 1 mL of Trizol (Invitrogen, USA) was added to the cells and the cells were separated according to the RNA isolation method of Invitrogen. RNA was quantitated at 260 nm using an ultraviolet detector, and then cDNA (complementary DNA) was synthesized and subjected to real-time PCR. For cDNA synthesis, cDNA synthesis kit (High Capacity RNA-to-cDNA Kit, Applied Biosystems, USA) was used. Real-time PCR was performed using the Real-time PCR Kit (Power SYBR Green PCR Master Mix, Applied Biosystems, CA, USA) and was performed using the kit method. Applied Biosystems 7500 FAST Real-Time PCR System , CA, USA), and amplified products were quantitatively analyzed. The conditions are as follows.

As a result of the measurement, as shown in the following Table 3 and FIG. 3, TNF-α expression was induced by PMA and TNF-α expression inhibition effect of each sample was examined. As a result, TNF- But the mixed extracts showed better inhibitory effects than the respective extracts. In particular, at a concentration of 100 μg / ml, the inhibitory effect was similar to that of the control group.

sample TNF-α expression level (%) Control group 3.44 PMA 100 nM 100 PMA + dermis 1 μg / ml 43.26 10 μg / ml 29.08 100 μg / ml 29.09 PMA + 1 μg / ml 56.3 10 μg / ml 46.1 100 μg / ml 38.2 PMA + Example 1 1 μg / ml 32.1 10 μg / ml 18.3 100 μg / ml 11.5

Experimental Example  4. Mixed extract and dermis, Wuss Oak Runin , Claudin -1 expression level

Real-time PCR (Real-time polymerase chain reaction) was performed to observe the increase of gene expression of the tight junction components, aclidine and claudin-1. Human keratinocyte (CRL-2310, ATCC) cells were used. Samples were added for each concentration and incubated for 24 hours. One mL of trizol (Trizol, Invitrogen, USA) was added to the cells and separated according to the RNA isolation method of Invitrogen. RNA was quantitated at 260 nm using an ultraviolet detector, and then cDNA (complementary DNA) was synthesized and subjected to real-time PCR. For cDNA synthesis, cDNA synthesis kit (High Capacity RNA-to-cDNA Kit, Applied Biosystems, USA) was used. Real-time PCR was performed using the Real-time PCR Kit (Power SYBR Green PCR Master Mix, Applied Biosystems, CA, USA) and was performed using the kit method. Applied Biosystems 7500 FAST Real-Time PCR System , CA, USA), and amplified products were quantitatively analyzed. The conditions are as follows. (positive control; CaCl ₂ 1.2 mM + A23187 0.1 쨉 g / ml)

The results of measurement of the expression levels of acrodin and claudin-1, constituents of tight junctions, are shown in Table 4 and FIG. As a result, both of acridine and claudin-1 showed synergistic effect of increasing the expression level in the mixed extracts. Particularly, in the case of acordin, the effect of the expression was further enhanced. Thus, it has been proved that the expression of the factors enhancing the skin barrier is increased to show a good effect for strengthening the skin barrier.

sample Oak Runin  Expression level (%) Claudin -1 expression level (%) Control group 100 100 CaCl ₂ 1.2 mM 108.1 104.13 CaCl ₂ 1.2 mM + A23187 0.1 μg / ml 128.37 122.89 Raspberry extract  1 μg / ml 124.32 121.92 50 μg / ml 130.18 129.60 100 μg / ml 141.47 130.93 Dermis extract  1 μg / ml 113.6 107.52 50 μg / ml 127.97 110.52 100 μg / ml 145.32 114.7 Mixed extract
Example 1  1 μg / ml 134.5 129.4 50 μg / ml 147.4 132.5 100 μg / ml 162.6 139.8

Prescription example  1. Lotion (skin lotion)

Table 5 shows the prescription examples of lotion (skin lotion) among the cosmetic compositions containing the mixed extract of Example 1.

number Raw material Content (% by weight) One  The mixed extract (Example 1) 3.0 2  glycerin 3.0 3  Butylene glycol 2.0 4  Propylene glycol 2.0 5  Polyoxyethylene hardened castor oil 1.0 6  ethanol 10.0 7  Triethanolamine 0.1 8  antiseptic a very small amount 9  Pigment a very small amount 10  Spices a very small amount 11  Purified water Balance

<Manufacturing Method>

2, 3, 4, and 8 are sequentially added to and mixed with 11, followed by dissolving by stirring. The mixture is heated at 60 ° C to dissolve 5 times, and then 10 times are added thereto. Finally, 1, 6, 7, and 9 are added, agitated sufficiently after stirring.

Prescription Example 2. Nutrition Lotion

Table 5 shows the prescription examples of the nutritional lotion among the cosmetic compositions containing the mixed extract of Example 1.

number Raw material Content (% by weight) One  The mixed extract (Example 1) 3.0 2  Wax 1.0 3  Polysorbate 60 1.5 4  Sorbitan sesquioleate 0.5 5  Liquid paraffin 10.0 6  Sorbitan stearate 1.0 7  Pro-type glycerin monostearate 0.5 8  Stearic acid 1.5 9  Glyceryl stearate / FG-400 stearate 1.0 10  Propylene glycol 3.0 11  Carboxy polymer 0.1 12  Triethanolamine 0.2 13  antiseptic a very small amount 14  Pigment a very small amount 15  Spices a very small amount 16  Purified water Balance

<Manufacturing Method>

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

Prescription example  3. Nourishing Cream

Table 7 shows the prescription examples of the nutritional cream among the cosmetics containing the mixed extract of Example 1.

number Raw material Prescription Example 3
Content (% by weight) Comparative Example 3 One The mixed extract (Example 1) 5.0 - 2 Stearic acid 2.0 2.0 3 Cetyl alcohol 2.0 2.0 4 Glyceryl monostearate 2.0 2.0 5 Polyoxyethylene sorbitan monostearate 0.5 0.5 6 Sorbitan sesquioleate 0.5 0.5 7 Glyceryl monostearate /
Glyceryl stearate /
Polyoxyethylene stearate 1.0 1.0 8 Wax 1.0 1.0 9 Liquid paraffin 4.0 4.0 10 Squalane 4.0 4.0 11 Caprylic / capric triglyceride 4.0 4.0 12 Carboxyvinyl polymer 0.3 0.3 13 Butylene glycol 5.0 5.0 14 glycerin 1.0 3.0 15 Triethanolamine 0.5 0.5 16 Purified water Balance Balance 17 water - 10.0

<Manufacturing Method>

Prescription Example 3: 12, 13, 14, and 16 were heated to 80 to 85 ° C with mixing and stirring, 11 is heated to between 80 and 85 ° C and then charged to 15 to emulsify. After the emulsification is completed, the mixture is cooled to 35 ° C with stirring using an agitator, and the mixture is cooled to 25 ° C by 1 time and aged.

Comparative Example 3: 12, 13, 14, and 16 were heated to 80 to 85 ° C while mixing and stirring. 11 is heated to between 80 and 85 ° C and then charged to 15 to emulsify. After the emulsification is completed, the mixture is cooled to 35 ° C with stirring using an agitator, and 17 times is added thereto, followed by cooling to 25 ° C and aging.

apply Experimental Example  1. Skin barrier enhancement measurement

Measurement of transepidermal water loss (TEWL), which measures water loss through the stratum corneum from the dermis, is commonly used to evaluate the stratum corneum status and to indirectly evaluate skin barrier function. The moisture content was measured by measuring the capacitance according to the moisture content, and the relative size of the moisture content was measured.

Twenty healthy adults with no previous history of skin disease were included in the study. The age ranged from 23 to 27 years with an average age of 24.9 years.

The total duration of the experiment was 8 weeks. Subjects were asked to wash their forearms with soap, wipe them with physiological saline, dry them naturally, square each 4 x 4 cm square symmetrically on the forearm, then apply TEWL, The moisture content was measured. 0.5 ml of Prescription Example 3 (mixed extract-containing cream) was applied uniformly to the marked area of the left forearm, and the right foreleg was applied to the control group of Comparative Example 3 (cream containing no mixed extract). Each cream prescription was applied 3 times a day for 8 weeks and TEWL and moisture content of skin surface were measured at intervals of one week during application.

 TEWL was measured with an evaporimeter Tewameter TM 210 (Courage + Khazaka, Cologne, Germany) after exposing the measurement site in a constant temperature and humidity room with an average temperature of 25.0 ° C and an average humidity of 50% and Khazaka, Germany).

The measurement results for 8 weeks are shown in Table 8.

Experiment 0 weeks 8 weeks Improvement effect TEWL Comparative Example 3 16.32 + - 11.2 15.48 ± 9.62 -0.84 + -8.53 Prescription Example 3 17.22 + - 5.60 10.85 ± 10.11 -6.37 ± 5.67 Moisture content Comparative Example 3 46.48 ± 10.25 50.04 + - 10.82  3.56 ± 7.59 Prescription Example 3 45.39 + - 9.30 67.83 + - 12.28 22.44 + - 9.77

In the case of TEWL, the skin TEWL was 16.32 ± 11.27 at the first measurement and 15.48 ± 9.62 at the 8th week in Comparative Example 3 in which the mixed extract was not contained. In the case of the Prescription Example 3 containing the mixed extract, 17.22 ± 5.60, and 10.85 ± 10.11 after 8 weeks. The average change in TEWL before and after use of the cream was observed to be -0.84 ± 8.53 in the control product and -6.37 ± 5.67 in the test product and decreased after 8 weeks of using the test product. This resulted in reduced water loss after 8 weeks of use, which is a result of enhanced skin barrier due to reduced water loss by using the cream of Formulation Example 3 containing mixed extracts.

In the case of the water content, the skin moisture content was 46.48 ± 10.25 at the first measurement and 50.04 ± 10.82 at the 8th week in Comparative Example 3 in which the mixed extract was not contained. In the case of the Prescription Example 3 containing the mixed extract, 45.39 ± 9.30 , And 67.83 ± 12.28 after 8 weeks. The average change in skin moisture content before and after use of the cream was observed to be 3.56 +/- 7.59 in Comparative Example 3, 22.44 9.77 in Formulation Example 3, which is the test product, and 8 weeks after using Formulation Example 3. This is a result of increased skin moisture content after 8 weeks of use and increased skin barrier due to increased moisture content by using cream containing mixed extracts.

As described above, according to the present invention, the mixed extract increases the expression of aquaporin-3 and not only effectively inhibits TNF-a but also inhibits the formation of tight junction (TJ) constituents occludin, (claudin-1) expression in the brain. Therefore, it is expected to be used as a skin barrier strengthening cosmetic composition containing the dandelion and moss mixed extract as an active ingredient.

Claims (11)

A composition for enhancing skin barrier comprising an extract of Citrus unshiu Markovich and Achyranthes fauriei as an active ingredient.
The method according to claim 1, wherein the mixed extract is selected from the group consisting of water, anhydrous or hydrous ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, benzene, chloroform, glycerin, butylene glycol, propylene glycol, Wherein the composition is extracted with at least one solvent selected from the group consisting of:
[Claim 2] The composition for enhancing skin barrier according to claim 1, wherein the mixed extract is extracted at a mixed weight ratio of 3 to 7: 1 to 5.
The skin barrier strengthening composition according to claim 1, wherein the mixed extract is contained in an amount of 0.01 to 10.0% by weight in terms of dry weight.
[Claim 3] The method according to claim 2, wherein the mixed extract is extracted by immersing dermis and booster in a solvent at 5 to 37 DEG C for 1 to 15 days, or extracting by heating at 50 to 100 DEG C for 3 to 20 hours under a solvent. Composition for skin barrier enhancement.
The composition for enhancing skin barrier according to claim 1, wherein the mixed extract has an effect of increasing keratinocyte aquaporin 3, acrodine, and cloodin-1 expression.
[Claim 2] The composition for enhancing skin barrier according to claim 1, wherein the mixed extract has an effect of inhibiting tumor necrosis factor-alpha (TNF-a) expression of keratinocytes.
The skin barrier strengthening composition according to claim 1, wherein the skin barrier strengthening reduces moisture loss of the skin and prevents skin dryness and skin inflammation.
A dermis, and a mixed extract of moss and dermis.
A dermis, and a mixed extract of moss and moss.
The composition according to claim 10, wherein the composition is at least one selected from the group consisting of a lotion (skin lotion), a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, Wherein the formulation has a formulation selected from the group consisting of a nutritional essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser formulation.

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