KR102454574B1 - Cosmetic composition for promoting skin keratin cycle - Google Patents

Abstract

The cosmetic composition for skin regeneration according to the present invention sufficiently supplies calcium, vitamin D and vitamin E, including almonds, taheebo, and white oyster mushroom, and promotes regeneration of aged skin to remove dead skin cells and improve skin texture has the effect of making

Description

COSMETIC COMPOSITION FOR PROMOTING SKIN KERATIN CYCLE}

The present invention relates to a cosmetic composition for skin regeneration.

The skin is largely divided into three layers: the epidermis, the dermis, and the subcutaneous tissue from the outside, and has the function of protecting each organ in the body from temperature and humidity changes, ultraviolet rays, and other physical and chemical external stimuli.

The skin has a primary function to protect our body from the external environment and a cosmetic function. In addition, cosmetics have an aesthetic function that cleans or beautifully expresses human skin and a defense function that protects the skin from harmful environments.

The role of the skin declines due to various harmful environments such as ultraviolet rays, air pollution, stress, and yellow dust. It not only improves the skin, but also protects the skin and helps the skin to maintain its normal function.

However, as in these days, artificial temperature control of air conditioning and heating according to changes in the environment or life patterns, skin stress caused by various stresses and environmental pollution occurring in social life, frequent washing due to makeup habits, and natural skin due to increasing age Due to various causes such as aging, the skin becomes dry, the surface becomes rough, and the skin becomes brittle, loses moisture and loses vitality. Therefore, the need for skin aging prevention and skin regeneration is increasing.

The skin regeneration process is a phenomenon in which keratinocytes located in the basal layer are keratinized through a differentiation process and replaced with new cells. In aged skin, it is known that the function of keratinocytes decreases and skin regeneration is delayed, and the exfoliation of dead skin cells does not occur properly or is not replaced with new cells, resulting in rough and dull skin.

On the other hand, it is known that calcium (Calcium), vitamin D (Vitamin D), vitamin E (Vitamin E), etc. are related as factors promoting the differentiation of keratinocytes. Calcium contributes to proliferation, regulation of differentiation, and maintenance of homeostasis of keratinocytes, and its absorption rate is low compared to other nutrients, and it is known that consumption of coffee containing caffeine further interferes with calcium absorption. Vitamin D is involved in the absorption of calcium ions, and is mainly synthesized in the body (epidermis) by exposure to sunlight. Modern people rarely get sun exposure due to increased indoor activities, and vitamin D synthesis is reduced even in outdoor activities due to the daily use of sunscreen, causing a vitamin D deficiency problem. Vitamin E is involved in calcium concentration regulation and is known to increase intracellular calcium ion concentration.

Conventionally, various natural extracts have been used to provide moisturizing, skin soothing and anti-aging compositions, but studies on effective cosmetic compositions for skin regeneration by promoting the expression of differentiation factors in keratinocytes have been insufficient.

The present invention provides a composition capable of activating the function of vitamin D in the epidermal layer and promoting the expression of a differentiation factor in keratinocytes to promote skin regeneration.

The cosmetic composition for skin regeneration according to the present invention preferably includes at least one extract selected from an almond extract, a Tabebuia extract, and a Tremella extract.

In addition, the cosmetic composition for skin regeneration according to the present invention comprises: a taheebo extract; It is more preferable to include; almond extract or white wood ear mushroom extract.

In addition, it is more preferable that the cosmetic composition for skin regeneration according to the present invention includes an almond extract, a taheebo extract, and a white wood ear mushroom extract.

In addition, in the cosmetic composition for skin regeneration according to the present invention, the almond extract is preferably included in an amount of 0.1 to 5% by weight of the total composition.

In addition, in the cosmetic composition for skin regeneration according to the present invention, the taheebo extract is preferably included in an amount of 0.1 to 10% by weight of the total composition.

In addition, in the cosmetic composition for skin regeneration according to the present invention, it is preferable that the white wood ear mushroom extract is included in an amount of 0.1 to 20% by weight.

In addition, the cosmetic composition for skin regeneration according to the present invention contains 2 of Loricrin, Involucrin, Cytokeratin10 and Desmoglein2, which are skin cell differentiation factors related to skin regeneration. It is preferred to express more than one species.

In addition, the cosmetic composition for skin regeneration according to the present invention preferably expresses at least one of vitamin D and vitamin D receptor, which are factors related to skin cell differentiation.

In addition, the cosmetic composition for skin regeneration according to the present invention preferably has β-glucosidase enzyme activity.

The cosmetic composition for skin regeneration according to the present invention sufficiently supplies calcium, vitamin D and vitamin E, including almonds, taheebo, and white oyster mushroom, and promotes regeneration of aged skin to remove dead skin cells and improve skin texture has the effect of making

1 schematically shows the experimental results for the identification of cellular aging markers.
2 schematically shows the experimental results for confirming the skin regeneration cycle.
Figure 3 shows the results of testing the promoting effect of the skin cell differentiation factor of Examples 1 to 3 of the present invention.
4 shows the results of testing the promoting effect of the skin cell differentiation factor of Example 4 of the present invention.
5 schematically shows the results of testing the effect of increasing vitamin D receptor expression of Example 4 of the present invention.
6 and 7 schematically show the results of the skin keratin improvement test in Example 4 of the present invention in images and diagrams.
8 schematically shows a photograph of a skin texture improvement test result of Example 4 of the present invention.
9 schematically shows an exterior photograph of Example 4 of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. The embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art. Accordingly, the embodiment of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below.

Throughout the specification of the present invention, when a part "includes" a certain component, it does not exclude other components unless otherwise stated, but further includes other components.

Throughout the specification of the present invention, when a step is located “on” or “before” another step, this means not only a case in which a step is in a direct time-series relationship with another step, but also a step of mixing after each step and Likewise, the order of two stages may include the same rights as in the case of an indirect time series relationship in which the time series order may change.

The terms "about", "substantially", etc. to the extent used throughout the specification of the present invention are used in or close to the numerical value when manufacturing and material tolerances inherent in the stated meaning are presented, and the present invention It is used to prevent an unconscionable infringer from using the disclosure in which exact or absolute figures are mentioned to help understand. As used throughout this specification, the term “step of (to)” or “step of” does not mean “step for”.

The epidermis of the skin consists of a basal layer, a spinous layer, a granular layer, and a cornified layer. Keratinocytes generated from the basal layer, the lowest layer of the epidermis, pass through the stratum corneum and granular layer for 28 days to form the stratum corneum. When the final differentiation proceeds, structural proteins such as Involucrin and Loricrin are bound to the inside of the cell membrane to form a cornified cell envelope, a hard structure that does not dissolve in water.

Desmoglein2 is a glycoprotein present in desmosomes and is expressed in the spinous layer of the epidermis. In the process of differentiation of keratinocytes, it is involved in cell-cell junctions. Cyrokeratin10 is directly involved in the regulation of the cell cycle in the early stage of differentiation and is known to be mainly distributed between the stratum corneum and the stratum corneum of the epidermis. Loricrin and Involucrin are markers expressed in the cornified layer of the epidermis. Loricrin is involved in regulating cell differentiation, proliferation, and migration, and involucrin is a protein that connects lipids between keratinocytes and keratinocytes. As the final differentiation progresses, the expression level decreases. will increase

Although there may be various methods for promoting skin regeneration, using a cosmetic composition is advantageous in terms of beauty and convenience.

Natural extracts that can be used in these cosmetic compositions for skin regeneration contain various nutrients and compounds, so their actions are complex.

Among various natural extracts, for skin regeneration, skin cell differentiation factors related to skin regeneration of human keratinocytes are Loricrin, Involucrin, Cytokeratin10 and Desmoglein2. It is preferable to be able to promote the expression of.

The present invention has an effect of promoting the expression of differentiation factors in stages by participating in the overall process of differentiation of keratinocytes in the epidermis, including almond extract, taheebo extract, and white wood ear mushroom extract among various natural products.

First, before confirming the effect of the cosmetic composition according to the present invention, cellular aging factors that can distinguish young skin from aging skin were identified as follows, and the skin regeneration cycle of young skin and aged skin was confirmed.

(Experimental Example 1)

Path changes (p4~p9) in Human Epidermal Keratinocyte neonatal (HEKn) were performed by western-blot to determine whether it could be classified into models of young skin and aged skin according to the cell pathway (p4~p9). ) according to the expression level of p16INK4a (a marker of cellular aging) was compared.

Specifically, human keratinocytes of P4~p9 were inoculated into a 60 mm plate at a concentration of 100 x 10 ⁴ cells, cultured for 24 hours, and then replaced with a medium not containing growth factors and cultured for 24 hours. After incubation, the protein was recovered by treatment with RIPA Buffer, and a sample was prepared so that the amount of protein was 30 μg through BCA quantification. Proteins were separated from the prepared sample using sodiumdodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a PVDF membrane using electrotransfer, and intracellular expression levels were measured using immunoblotting. Confirmed.

As a result, as shown in FIG. 1 , the expression of the cell aging marker p16INK4a increased as the cellular pathway increased. It was confirmed that the early passage (Early passage: p4~p5) and the late passage (P8~p9) can be used as models for young skin and aging skin, respectively.

(Experimental Example 2)

Loricrin and involucrin, which are skin cell differentiation factors related to skin regeneration of human keratinocytes in the early pathway (p4) and late pathway (p8), to check whether the skin regeneration cycle is delayed in aging skin than in young skin The expression levels of (Involucrin), Cytokeratin10 (Cytokeratin10) and Desmoglein2 (Desmoglein2) were compared by Western-blot.

Specifically, human keratinocytes of the early pathway (p4) and late pathway (p8) were inoculated into a 60 mm plate at a concentration of 100 x 10 ⁴ cells, cultured for 24 hours, and then replaced with a growth factor-free medium. Then incubated for 24 hours. After incubation, the protein was recovered by treatment with RIPA Buffer, and a sample was prepared so that the amount of protein was 30 μg through BCA quantification. Proteins were separated from the prepared sample using SDS-PAGE (Sodiumdodecylsulfate-polyacrylamide gel electrophoresis), transferred to a PVDF membrane using Electrotransfer, and intracellular expression levels were measured by immunoblotting. Confirmed.

As a result, as shown in FIG. 2 , the expression timing of Loricrin, Involucrin, Cytokeratin10 and Desmoglein2 was longer in the late pathway than in the early pathway. confirmed late. That is, it was confirmed that skin regeneration was delayed in aging skin.

The present invention provides a cosmetic composition that promotes skin regeneration by sufficiently supplying calcium, vitamin D and vitamin E.

Specifically, it is preferable that the cosmetic composition for skin regeneration according to the present invention includes at least one extract selected from an almond extract, a taheebo extract, and a white wood ear mushroom extract.

In addition, the cosmetic composition for skin regeneration according to the present invention comprises: a taheebo extract; It is more preferable to include; almond extract or white wood ear mushroom extract.

In addition, it is more preferable that the cosmetic composition for skin regeneration according to the present invention includes an almond extract, a taheebo extract, and a white wood ear mushroom extract.

The almond extract is preferably included in an amount of 0.1 to 5% by weight of the total composition.

When almond extract is included in an amount of less than 0.1% by weight, it is not possible to significantly increase the expression level of skin cell differentiation factors related to skin regeneration. When included in excess of 5% by weight, the increase in the effect according to the content increase is not clear, and due to the high concentration, it may cause cytotoxicity and irritation to the skin. It can also cause turbidity in the formulation and cause precipitation to make the formulation unstable.

The taheebo extract is preferably included in an amount of 0.1 to 10% by weight of the total composition.

When the taheebo extract is included in less than 0.1% by weight, it is not possible to significantly increase the expression level of skin cell differentiation factors related to skin regeneration. When included in excess of 10% by weight, the increase in the effect according to the content increase is not clear, and due to the high concentration, it may cause cytotoxicity and irritation to the skin. It can also cause turbidity in the formulation and cause precipitation to make the formulation unstable.

The white wood ear mushroom extract is preferably included in an amount of 0.1 to 20% by weight of the total composition.

When the white wood ear mushroom extract is included in an amount of less than 0.1% by weight, it is not possible to significantly increase the expression level of skin cell differentiation factors related to skin regeneration. When included in excess of 20% by weight, the increase in the effect according to the content increase is not clear, and due to the high concentration, it may cause cytotoxicity and irritation to the skin. It can also cause turbidity in the formulation and cause precipitation to make the formulation unstable.

Hereinafter, the present invention will be described in more detail through specific examples and experimental examples of the cosmetic composition for skin regeneration according to the present invention.

(Examples 1-3) Preparation of a single extract

Grind each of 10-50 g of dried almonds, 10-50 g of taheebo, and 10-50 g of white oyster mushroom, add purified water and 1-5 kg of butylene glycol as extraction solvents, and adjust the pH to 5 using 1M hydrochloric acid solution and 1M sodium hydroxide solution. It was adjusted to ~6. After stirring and extracting for 1 hour at 600 rpm at 25-37° C., it was filtered with 1 μm filter paper. After mixing the filtrate with purified water and butylene glycol, it was prepared by final filtration through a 0.2 μm PES membrane filter. The results are shown in Table 1 below.

Example 1 Example 2 Example 3 Butylene Glycol (% by weight) 45 45 45 Almond Extract (wt%) 5 Taheebo extract (wt%) 5 White wood ear mushroom extract (% by weight) 5 Purified water (wt%) 50 50 50 pH 5-6 5-6 5-6

(Experimental Example 3) Evaluation of enzymatic activity of a single extract

In order to set the optimal extraction conditions, β-glucosidase enzyme activity was confirmed for the single extracts obtained from Examples 1-3. A single extract and a substrate solution (1 mM p-nitrophenyl-D-glucopyranoside, p-NPG) were mixed in a 1:1 ratio and stabilized at room temperature for 2 hours. After reacting in a water bath at 50° C. for 30 minutes, 2 ml of a reaction stop solution (0.25 M sodium carbonate) was added to terminate the reaction, and absorbance was measured at 420 nm. β-glucosidase enzyme activity was calculated compared with the standard solution (P-nitrophenol solution). The results are shown in Table 2 below.

Example 1 Example 2 Example 3 β-glucosidase activity (U/ml) 0.57 4.26 12.81

(Experimental Example 4)

In order to confirm that the single extract of Examples 1 to 3 of the present invention prepared above can promote the delayed skin regeneration cycle in aging skin, after treating Examples 1 to 3 on human keratinocytes, skin regeneration and The expression levels of related skin cell differentiation factors, Loricrin, Involucrin, Cytokeratin10, and Desmoglein2, in the late pathway were confirmed by Western-blot.

Specifically, the human keratinocytes of the late pathway (p9) were inoculated into a 60 mm plate at a concentration of 100 x 10 ⁴ cells, cultured for 24 hours, and then in a medium containing no growth factors in Examples 1, 2, Example 3 was added and cultured for 24 hours. After incubation, the protein was recovered by treatment with RIPA Buffer, and a sample was prepared so that the amount of protein was 30 μg through BCA quantification. Proteins were separated from the prepared sample using SDS-PAGE (Sodiumdodecylsulfate-polyacrylamide gel electrophoresis), transferred to a PVDF membrane using Electrotransfer, and intracellular expression levels were measured by immunoblotting. Confirmed.

As a result, as shown in Figure 3, Example 1 promoted the expression of involucrin, cytokeratin 10 (Cytokeratin10) and desmoglein2 (Desmoglein2), Example 2 was loricrin (Loricrin) ), involucrin, cytokeratin 10 and desmoglein 2 were promoted. Example 3 was confirmed to have an effect of promoting the expression of Loricrin, Involucrin, and Desmoglein2.

As shown in FIG. 3 , in Examples 1 to 3, factors having the greatest effect are different from each other. Example 1 has the greatest effect of promoting the expression of Cytokeratin10, and Example 2 has the greatest effect of promoting the expression of Loricrin and Involucrin. Example 3 has a large effect of promoting the expression of desmoglein2 (Desmoglein2).

When only Examples 1 and 2 are combined, the effect of promoting the expression of desmoglein 2 may be low, and when only Examples 2 and 3 are combined, the expression of cytokeratin 10 is promoted may be less effective. In addition, when only Examples 1 and 3 are combined, the effect of promoting the expression of Loricrin and Involucrin may be low.

In particular, Example 1 has the effect of accelerating the differentiation of the lower stage by promoting the expression of cytokeratin 10 (Cytokeratin) involved in the earliest stage of differentiation.

Therefore, as in Example 4 below, Examples including all of Examples 1 to 3 were tested.

(Example 4) Almond, taheebo, white wood ear mushroom mixed extract preparation

10 g ~ 50 g of dried almonds were ground, purified water and 1 ~ 5 kg of butylene glycol were added as extraction solvents, and the pH was adjusted to 5 ~ 6 using 1 M hydrochloric acid solution and 1 M sodium hydroxide solution. After stirring and extracting for 1 hour at 600 rpm at 25 ~ 37 ℃, filtered through a 1 ㎛ filter paper to prepare an almond extract. 15 g of taheebo and 25 g of white wood ear mushroom were mixed and pulverized in almond extract, and extracted with stirring at 600 rpm at room temperature for 1 hour, followed by filtration with 1 μm filter paper. After mixing the filtrate with purified water and butylene glycol, it was prepared by final filtration through a 0.2 μm PES membrane filter. The results are shown in Table 3 below.

Example 4 Butylene Glycol (% by weight) 45 Almond Extract (wt%) One Taheebo extract (wt%) 1.5 White wood ear mushroom extract (% by weight) 2.5 Purified water (wt%) 50

In order to observe the appearance properties of the composition prepared above, a photograph was taken and shown in FIG. 9 .

(Experimental Example 5)

To determine whether the mixed extract of the present invention can promote the delayed skin regeneration cycle in aging skin. After the human keratinocytes were treated with Example 4, skin cell differentiation factors related to skin regeneration, such as Loricrin, Involucrin, Cytokeratin10 and Desmoglein2, were The degree of expression in the late pathway was confirmed by Western-blot.

Specifically, human keratinocytes of the late pathway (p7) were each inoculated into a 60 mm plate at a concentration of 100 x 10 ⁴ cells and cultured for 24 hours, followed by the addition of Example 4 to a medium not containing growth factors for 24 hours. cultured. After incubation, the protein was recovered by treatment with RIPA Buffer, and a sample was prepared so that the amount of protein was 30 μg through BCA quantification. Proteins were separated from the prepared sample using SDS-PAGE (Sodiumdodecylsulfate-polyacrylamide gel electrophoresis), transferred to a PVDF membrane using Electrotransfer, and intracellular expression levels were measured by immunoblotting. Confirmed.

As a result, as shown in FIG. 4 , it was confirmed that Example 4 had the effect of promoting the expression of all skin cell differentiation factors.

That is, Example 4, in which Examples 1, 2 and 3 are mixed in an appropriate ratio, has a high effect at a lower concentration compared to the effect each of Examples 1, 2 and 3 has alone looks like In addition, in Example 4, by mixing Examples 1, 2, and 3 in an appropriate ratio, the keratinocytes formed in the basal layer of the skin pass through the stratum corneum and granular layer to finally reach the stratum corneum. It has the effect of accelerating the skin regeneration cycle as a result.

(Experimental Example 6)

To determine whether the mixed extract of the present invention affects the function of vitamin D in aging skin. After the human keratinocytes were treated with the mixed extract of Example 4, the expression level of the vitamin D receptor involved in normal vitamin D function in the late pathway was confirmed by Western-blot.

Specifically, the human keratinocytes of the late pathway (p7) were each inoculated into a 60 mm plate at a concentration of 100 x 10 ⁴ cells and cultured for 24 hours, and then the mixed extract of Example 4 was added to the growth factor-free medium. and cultured for 24 hours. After incubation, the protein was recovered by treatment with RIPA Buffer, and a sample was prepared so that the amount of protein was 30 μg through BCA quantification. Proteins were separated from the prepared sample using SDS-PAGE (Sodiumdodecylsulfate-polyacrylamide gel electrophoresis), transferred to a PVDF membrane using Electrotransfer, and intracellular expression levels were measured by immunoblotting. Confirmed.

As a result, as shown in FIG. 5 , it was confirmed that the mixed extract of Example 4 has the effect of promoting the expression of vitamin D receptor.

That is, the mixed extract of Example 4 increases the expression of vitamin D receptors in skin cells so that vitamin D synthesized in the skin functions normally, thereby promoting the expression of differentiation factors in keratinocytes, and consequently promoting the skin regeneration cycle. show the effect.

(Experimental Example 7) Skin keratin improvement confirmation test

In order to confirm whether the mixed extract of Example 4 of the present invention helps to improve skin keratin, 5 males aged 40 to 60 years were tested.

The mixed extract of Example 4 was applied to the right face once in the morning and in the evening for a total of 4 weeks, and changes in skin keratin were measured every 2 weeks using a SOFT OLUS (Keratin probe) device.

As a result, as shown in FIGS. 6 and 7 , it was confirmed that the amount of dead skin cells decreased over time in the right face to which the mixed extract of Example 4 was applied compared to the left face to which nothing was applied (untreated).

(Experimental Example 8) Skin texture improvement confirmation test

In order to confirm whether the mixed extract of Example 4 of the present invention helps improve skin texture, it was tested on 5 males aged 40 to 60 years.

The mixed extract of Example 4 was applied to the right face once in the morning and in the evening for a total of 4 weeks, and skin texture changes were measured every 2 weeks using OBSERVE equipment.

As a result, as shown in FIG. 8 , it was confirmed that the skin texture of the right face to which the mixed extract of Example 4 was applied was improved compared to the left face to which nothing was applied.

Although the embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and variations are possible within the scope without departing from the technical spirit of the present invention described in the claims. It will be apparent to those of ordinary skill in the art.

Claims (9)

A cosmetic composition for improving the skin keratin cycle that expresses at least one of vitamin D and vitamin D receptor, which are factors related to skin cell differentiation, including almond extract, taheebo extract, and white wood ear mushroom extract. delete delete The cosmetic composition for improving the keratin cycle of the skin according to claim 1, wherein the almond extract is contained in an amount of 0.1 to 5% by weight of the total composition. The cosmetic composition for improving the keratin cycle of the skin according to claim 1, wherein the taheebo extract is contained in an amount of 0.1 to 10% by weight of the total composition. [Claim 2] The cosmetic composition for improving the keratin cycle of the skin according to claim 1, wherein the white wood ear mushroom extract is contained in an amount of 0.1 to 20% by weight. According to claim 1, wherein the cosmetic composition is a skin cell differentiation factor related to skin regeneration Loricrin (Loricrin), involucrin (Involucrin), cytokeratin 10 (Cytokeratin10) and desmoglein 2 (Desmoglein2) at least two types A cosmetic composition for improving skin keratin cycle, characterized in that it expresses delete The cosmetic composition for improving the keratin cycle of the skin according to claim 1, wherein the cosmetic composition has β-glucosidase enzyme activity.

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