Anredera cordifolia extract-containing composition, method for producing the same, and cosmetic

Abstract

To provide an Anredera cordifolia extract-containing composition having functionalities particularly useful for cosmetic use.SOLUTION: An Anredera cordifolia extract-containing composition contains an extract component obtained from Anredera cordifolia in an amount of 0.01 wt.% or more as an active ingredient, and has one or more functionalities selected from an anti-aging effect, a barrier function-enhancing effect, a moisturizing effect, and an antioxidant effect.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition containing ingredients contained in Chenopodium album and a method for producing the same. The present invention also relates to a cosmetic product containing a Chenopodium album extract-containing composition. [Background technology]

[0002] Anredera cordifolia is a plant of the genus Anredera in the family Basellaceae, native to South America. Techniques for using extracts of plants from the family Basellaceae in cosmetics are described in, for example, Patent Documents 1 to 3.

[0003] Patent Document 1 discloses a hyaluronic acid production promoter for the purpose of promoting the production of hyaluronic acid, which comprises one or more plants selected from Solena amplexicaulis (Lam.) Gandhi, Laggera alata (D. Don) Sch.-Bip. ex Oliv, Pedilanthus tithymaloides (L.) Poit., Kalimeris indica (L.) Sch.-Bip., Anredera cordifolia (Tenore) Steen, and Bougainvillea glabra Choisy, or an extract thereof.

[0004] Furthermore, Patent Document 2 discloses an external skin preparation containing one or more extracts selected from extracts of Chenopodiaceae plants, Amaranthaceae plants, Nyctaginaceae plants, Phytolaccaeaceae plants, Bulidaceae plants, Portulacaceae plants, Malvataceae plants, or Cactaceae plants, for the purpose of improving rough skin and cutaneous blood circulation. In particular, Patent Document 2 reports that betalains contained in these plants may be involved in effects such as improving rough skin. Note that Patent Document 2 only lists Malva japonica as an example of a plant of the Malvataceae family, and does not mention Chenopodium album.

[0005] Furthermore, Non-Patent Document 1 measures the proliferation effect of NB1RGB skin fibroblasts, which are involved in collagen production, for the purpose of evaluating the physiological activity of 47 species of plants cultivated in Okinawa Prefecture. Non-Patent Document 1 reports that, of these 47 species of plants, no proliferation effect was observed in the methanol extract of the leaves of Anredera cordifolia. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Laid-Open No. 2007-084448 [Patent Document 2] Japanese Patent Application Laid-Open No. 63-192705 [Non-patent literature]

[0007] [Non-Patent Document 1] Takahashi, Makoto, et al. "Screening of medicinal and edible plants in Okinawa, Japan, for enhanced proliferative and collagen synthesis activities in NB1RGB human skin fibroblast cells." Bioscience, Biotechnology, and Biochemistry 76.12, 2317-2320, 2012 Summary of the Invention [Problem to be solved by the invention]

[0008] As mentioned above, Patent Document 1 shows the effect of promoting hyaluronic acid production by Akasakazura, and Patent Document 2 shows the effect of Malvaceae plants (especially Malvaceae) on improving rough skin and skin blood circulation, but does not show any other functions of Malvaceae plants, especially Malvaceae. Furthermore, these documents do not go so far as to guarantee the stability and safety of the composition.

[0009] Therefore, a main object of the present invention is to provide a composition containing Chenopodium album extract that has functionality useful particularly for cosmetic applications. [Means for solving the problem]

[0010] A first aspect of the present invention relates to a composition containing a Chenopodium album extract (hereinafter simply referred to as the "composition"). The composition contains 0.01 wt% or more of an extract obtained from Chenopodium album as an active ingredient and has one or more of the following functionalities: anti-aging activity, barrier function strengthening activity, moisturizing activity, and antioxidant activity. Of these functionalities, the composition preferably has at least an anti-aging activity using an extract obtained from Chenopodium album as an active ingredient. The anti-aging activity specifically refers to the gene expression activity of type I collagen (COL1A1), collagen-degrading enzyme (MMP1), differentiation-related protein (KRT1), vascular endothelial growth factor (VEGF), cortisol-activating enzyme (11β-HSD1), or cortisol-inactivating enzyme (11β-HSD2). Furthermore, the composition preferably has an extract obtained from Chenopodium album as an active ingredient and further has a barrier function strengthening activity or a moisturizing activity in addition to the anti-aging activity. The barrier function strengthening effect is specifically the effect of inducing gene expression of a protein cross-linking enzyme (TGM1) or filaggrin (FLG). Furthermore, the moisturizing effect is specifically the effect of inducing gene expression of water-permeable proteins (AQP1, AQP9). Furthermore, the composition preferably contains an extract obtained from Chenopodium album as an active ingredient and further has antioxidant effect in addition to anti-aging effect. Specifically, the antioxidant effect is SOD (superoxide dismutase)-like effect or radical scavenging activity. The SOD-like effect removes active oxygen generated on the skin or in the body, promoting the improvement of spots and wrinkles. Furthermore, the radical scavenging effect eliminates free radicals with high oxidizing power generated on the skin or in the body. Examples of radicals that can be eliminated include the ABTS radical and the DPPH radical.

[0011] The composition of the present invention preferably contains total saponins, total polyphenols, total free amino acids, and vitexin, or pharmaceutically acceptable salts thereof, as extract components obtained from Chenopodium album. In particular, the composition preferably contains 200 ppm or more of total saponins or pharmaceutically acceptable salts thereof, 80 ppm or more of total polyphenols or pharmaceutically acceptable salts thereof, and 35 ppm or more of total free amino acids or pharmaceutically acceptable salts thereof. By clarifying the required content of the functional components, it is possible to achieve functionality not previously available in Chenopodium album extracts.

[0012] The composition of the present invention preferably does not cause skin sensitization. Specifically, the composition preferably has a negative skin sensitization test result. This ensures the safety of the composition.

[0013] The composition of the present invention preferably does not exhibit any precipitation or sedimentation of its components when stored in a dark place at room temperature for at least three months. Room temperature means 15 to 25°C, and a dark place means a place not exposed to direct sunlight. This ensures the stability of the composition.

[0014] The composition according to the present invention may contain ethanol or butylene glycol as a solvent, which can more reliably ensure the safety and stability of the Chenopodium album extract.

[0015] A second aspect of the present invention is a cosmetic product containing the Chenopodium album extract-containing composition according to the first aspect described above, thereby providing a cosmetic product that can claim the functionality of Chenopodium album extract.

[0016] A third aspect of the present invention is a promoter for CAMSAP3 (Calmodulin-regulated spectrin-associated protein 3) expression in epidermal keratinocytes (NHEK: Normal Human Epidermal Keratinocytes). The CAMSAP3 expression promoter of the present invention contains an extract obtained from Chenopodium album L. as an active ingredient. The content of the extract obtained from Chenopodium album L. may be, for example, 0.0001 wt% or more, 0.00015 wt% or more, or 0.001 wt% or more, or 0.0015 wt% or more. The present inventors have confirmed that a composition containing Chenopodium album L. extract promotes CAMSAP3 gene expression in epidermal keratinocytes. CAMSAP3 is a protein belonging to the CAMSAP family and contributes to the formation of non-centrosomal microtubules. Microtubules are known to be located near the cell membrane in epithelial cells and confer polarity. In particular, in the skin, depending on epidermal turnover, they convert from centrosome-dependent to non-centrosomal microtubules, contributing to tissue function. Compositions containing Akasakazura extract promote the expression of CAMSAP3 in epidermal keratinocytes, thereby promoting the formation of non-centrosomal microtubules and regulating epidermal turnover, contributing to the maintenance of skin homeostasis.

[0017] A third aspect of the present invention relates to a method for producing a composition containing a Chenopodium album extract. Specifically, the production method according to the third aspect relates to a method for producing a composition containing a Chenopodium album extract according to the first aspect described above. This production method includes a drying step, an extraction step, and a separation and purification step. The drying step is a step of drying the above-ground parts of Chenopodium album. Note that the structure of a plant is broadly classified into the above-ground parts (shoot system) and the underground parts (root system), and the above-ground parts of Chenopodium album include stems, leaves, buds, flowers, and fruits. The extraction step is a step of extracting the dried above-ground parts of Chenopodium album with a solvent to obtain an extract component. The separation and purification step is a step of separating and purifying the extract component obtained here. This results in a composition containing a Chenopodium album extract containing the extract component obtained from Chenopodium album as an active ingredient. The composition obtained here preferably has an extract of Chenopodium album as an active ingredient and has one or more of the following functions: anti-aging effect, barrier function strengthening effect, moisturizing effect, and antioxidant effect.

[0018] In the production method according to the present invention, the separation and purification step includes filtering the extracted component at least once. Before filtration, the extracted component may be left in a cool, dark place to precipitate insoluble components. A cool, dark place refers to a place at 1 to 15°C that is not exposed to direct sunlight. When the extracted component is left in a cool, dark place, the period of time may be at least one day, but may also be at least seven days or at least 14 days.

[0019] In the production method according to the present invention, the solvent is preferably a 30 to 100 wt % aqueous ethanol solution or an aqueous butylene glycol solution. [Effects of the Invention]

[0020] According to the present invention, it is possible to provide various functionalities of Chenopodium album that are particularly useful for cosmetic applications. [Brief explanation of the drawings]

[0021] [Figure 1] FIG. 1 shows an example of a manufacturing process for a composition containing Chenopodium album extract. [Figure 2] FIG. 2 shows the results of component analysis of a composition containing Chenopodium album extract. [Figure 3] FIG. 3 shows the results of measuring the total saponin concentration, total polyphenol concentration, and total free amino acid concentration of the composition containing Chenopodium album extract. [Figure 4] FIG. 4 shows the results of measuring the SOD-like activity (IC50) of a composition containing Chenopodium album extract. [Figure 5] FIG. 5 shows the measurement results (inhibition rate) of the SOD-like activity of a composition containing Chenopodium album extract. [Figure 6] FIG. 6 shows the measurement results (IC50) of the ABTS radical scavenging activity of compositions containing Chenopodium album extract. [Figure 7] FIG. 7 shows the measurement results (inhibition rate) of the ABTS radical scavenging activity of a composition containing Chenopodium album extract. [Figure 8] FIG. 8 shows the results of measuring the gene expression levels (keratinocytes) of a composition containing Chenopodium album extract. [Figure 9] FIG. 9 shows the results of measuring gene expression levels (fibroblasts) in a composition containing Chenopodium album extract. [Figure 10] FIG. 10 shows the results of measuring the CAMSAP3 expression promoting effect (epidermal keratinocytes) of a composition containing Chenopodium album extract. [Figure 11] FIG. 11 shows the results of a stability test of a composition containing Chenopodium album extract. [Figure 12] FIG. 12 shows, as a reference example, the measurement results of the CAMSAP3 expression promoting effect (epidermal keratinocytes) of Kabuchi Citrus Fruit Peel extract. DETAILED DESCRIPTION OF THE INVENTION

[0022] Hereinafter, embodiments of the present invention will be described with reference to the drawings. The present invention is not limited to the embodiments described below, and includes appropriate modifications of the embodiments described below within the scope obvious to those skilled in the art.

[0023] In the present specification, the expression "A to B" representing a numerical range means "A or more and B or less."

[0024] [1. Composition containing Chenopodium album extract] The present invention relates to a composition containing a red vine extract. This composition contains an extract (red vine extract) obtained from red vine at 0.01 wt% or more as an active ingredient. There is no upper limit to the amount of the extract, but it may be, for example, 1 wt% or less. Red vine specifically refers to the above-ground parts of red vine, and the red vine extract mainly contains the following components: (A) Total saponin (or a pharmaceutically acceptable salt thereof) (B) Total polyphenols (or pharmaceutically acceptable salts thereof) (C) Total free amino acids (or pharmaceutically acceptable salts thereof) (D) Vitexin (or a pharmaceutically acceptable salt thereof) The composition of the present invention may contain the above components (A) to (D) and their pharmaceutically acceptable salts in an amount of 0.01 to 1 wt %. Other components can be added to this composition as long as their functionality (anti-aging effect, barrier function strengthening effect, moisturizing effect, antioxidant effect) is not lost.

[0025] The above-mentioned components (A) to (D) can be extracted or isolated from the above-ground parts of Chenopodium japonica. When Chenopodium japonica is extracted using ethanol or butylene glycol, which can be used in cosmetics or quasi-drugs, as an extraction solvent, the contents of each component in the extract are generally in the following order: 1. Total saponins 2. Total polyphenols 3. Total free amino acids 4. Vitexin

[0026] More specifically, when the total amount of the above components (A) to (D) is taken as 100%, the ratio (weight ratio) of each component is as follows: Total saponins: 50-75% Total polyphenols: 15-30% Total free amino acids: 5-20% Vitexin: 0.1~1.5%

[0027] The preferred contents of the components (A) to (D) in the composition (mass concentration) are as follows: Total saponins: 200~1000ppm Total polyphenols: 80~400ppm Total free amino acids: 35-175 ppm Vitexin: 1-20 ppm

[0028] In addition to the above components (A) to (D), the composition may contain pharmaceutically acceptable salts thereof. Examples of pharmaceutically acceptable salts include sodium salts, potassium salts, magnesium salts, calcium salts, barium salts, ammonium salts, monoethanolamine salts, diethanolamine salts, triethanolamine salts, monoisopropanolamine salts, triisopropanolamine salts, hydrochlorides, and sulfates.

[0029] The composition may also contain a solvent in addition to the Chenopodium album extract. Examples of the solvent include water, alcohol, and aqueous alcohol solutions. Preferred examples of the alcohol are ethanol and butylene glycol. As the butylene glycol, it is particularly preferred to use 1,3-butylene glycol. In addition, the solvent may contain a polyhydric alcohol used as a moisturizer in cosmetics, etc. Examples of polyhydric alcohols include glycerin, diglycerin, polyglycerin-3, polyglycerin-10, 1,3-butylene glycol, propylene glycol, 3-methyl-1,3-butanediol, 1,3-propanediol, 2-methyl-1,3-propanediol, trimethylolpropane, pentaerythritol, hexylene glycol, ethylene glycol, diethylene glycol, triethylene glycol, polyethylene glycol, dipropylene glycol, 1,2-butanediol, 1,2-pentanediol, 1,2-hexanediol, 1,2-octanediol, 1,2-decanediol, polypropylene glycol, sorbitol, maltitol, and trehalose. The solvent may be used alone or in combination of two or more thereof. The content of the solvent in the Chenopodium album extract-containing composition can be adjusted as appropriate, for example, to 10 to 99%, 30 to 99%, or 50 to 99%. Alternatively, the Chenopodium album extract-containing composition may consist only of Chenopodium album extract and solvent, with the remainder of the Chenopodium album extract being the solvent.

[0030] As shown in the examples below, the composition of the present invention has the functionalities of anti-aging activity, barrier function strengthening activity, moisturizing activity, and antioxidant activity. Therefore, the composition of the present invention can be used as an anti-aging agent, barrier function strengthening agent, moisturizing agent, and antioxidant agent. The anti-aging activity specifically refers to the gene expression activity of type I collagen (COL1A1), collagen-degrading enzyme (MMP1), differentiation-related protein (KRT1), vascular endothelial growth factor (VEGF), cortisol-activating enzyme (11β-HSD1), or cortisol-inactivating enzyme (11β-HSD2). The barrier function strengthening activity specifically refers to the gene expression activity of protein cross-linking enzyme (TGM1) or filaggrin (FLG). The moisturizing activity specifically refers to the gene expression activity of water-permeable proteins (AQP1, AQP9). The antioxidant activity specifically refers to the SOD-like activity or radical scavenging activity. The SOD-like action removes active oxygen generated in the skin or body, promoting the improvement of spots and wrinkles. The radical scavenging action also eliminates highly oxidizing free radicals generated in the skin or body. Examples of radicals that can be eliminated include the ABTS radical and the DPPH radical.

[0031] As demonstrated in the Examples below, the composition of the present invention has the effect of promoting CAMSAP3 expression in epidermal keratinocytes. Therefore, the composition of the present invention can be used as an agent for promoting CAMSAP3 expression in epidermal keratinocytes. CAMSAP3 is a protein involved in the formation and stabilization of microtubules. Microtubules are important protein polymers that constitute the cytoskeleton and are responsible for functions such as maintaining cell morphology, cell division, and intracellular transport. CAMSAP3 specifically binds to the minus ends of non-centrosomal microtubules and promotes their stabilization. It is known that epidermal keratinocytes switch from centrosome-dependent to non-centrosomal microtubule formation during cell differentiation, and CAMSAP3 plays an important role in this process. Non-centrosomal microtubule formation is deeply involved in the formation of epidermal cell polarity and epidermal turnover. Thus, CAMSAP3 is an important factor that controls epidermal cell differentiation and epidermal turnover through microtubule formation and stabilization. By promoting the expression of CAMSAP3, the composition of the present invention promotes the formation of non-centrosomal microtubules, regulating epidermal turnover and contributing to the maintenance of skin homeostasis. Specifically, the effect of promoting CAMSAP3 expression in epidermal keratinocytes is shown to be 1.7 to 2.5 times higher than that of the control when a composition containing Akasakazura extract is added at, for example, 0.00015 to 0.0015 wt%. The Akasakazura extract content may be, for example, 0.0001 wt% or more, 0.00015 wt% or more, or 0.001 wt% or more, or 0.0015 wt% or more. There is no particular upper limit to the Akasakazura extract content, but it may be 1 wt% or less, 0.1 wt% or less, or 0.01 wt% or less.

[0032] The composition may contain Chenopodium album extract as is, or may contain ingredients used in cosmetics or quasi-drugs, as appropriate, within the scope that does not impair the effects of Chenopodium album extract. Examples of such ingredients include oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents, chelating agents, excipients, and film-forming agents.

[0033] The composition of the present invention can be incorporated into cosmetics or quasi-drugs, primarily for use as external skin preparations. Examples of the dosage forms of cosmetics and quasi-drugs include lotions, creams, emulsions, gels, aerosols, essences, packs, cleansers, bath products, foundations, dusting powders, lipsticks, ointments, and patches. The composition of the present invention can also be incorporated into soaps, body washes, facial cleansers, shampoos, rinses, treatments, and toothpastes, for example.

[0034] [2. Method for producing a composition containing Chenopodium album extract] Next, a preferred embodiment of a method for producing a composition containing Chenopodium album extract will be described with reference to Figure 1. As shown in Figure 1, the production method according to this embodiment includes, in this order, a washing and drying step (step S1), an extraction step (step S2), a vacuum concentration step (step S3), a solvent substitution step (step S4), a sedimentation step (step S5), a concentration adjustment step (step S6), and a microfiltration step (step S7). Each step will be described below.

[0035] The washing and drying process (step S1) involves washing the raw material, the aboveground parts of the Japanese ivy, and then drying them. While any washing and drying method is acceptable, it is preferable to use a method that does not degrade the components of the Japanese ivy, such as a method that does not use ultrasound or heat. For example, the aboveground parts of the Japanese ivy can be washed with water or hypochlorous acid water, and then dried at a low temperature under reduced pressure at 30 to 50°C (preferably 40°C) using a vacuum dryer to obtain a dried product. Alternatively, the aboveground parts of the Japanese ivy can be washed with water or hypochlorous acid water, and then placed on a colander or drying net to dry in the sun for 2 to 3 days. The dried aboveground parts of the Japanese ivy can then be crushed.

[0036] The extraction step (step S2) is a step of obtaining an extract (Chinese vine extract) from the dried aerial parts of Chenopodium capsicum using an extraction solvent. The extraction solvent is preferably ethanol, 1,3-butylene glycol, or an aqueous solution thereof. The ethanol aqueous solution preferably has an ethanol concentration of 30 to 70 wt%, and more preferably about 50 wt% (±5 wt%). The 1,3-butylene glycol aqueous solution preferably has a 1,3-butylene glycol concentration of 30 to 70 wt%. For example, the extraction conditions may be such that the dried material is immersed in the extraction solvent, and the temperature of the extraction solvent is set to room temperature (15 to 25°C) or 15 to 50°C, and extraction is performed under atmospheric pressure (about 1013 hPa) for at least 5 hours and up to one week (168 hours). The extracted solution is filtered to separate the extraction residue, yielding an extract containing the extraction solvent and Chenopodium capsicum extract. The first filtration after the extraction step may be carried out using, for example, filter paper or a filtration filter with a mesh size of 1 to 10 μm. If necessary, an auxiliary filter material such as diatomaceous earth may be used for the filtration.

[0037] The vacuum concentration step (step S3) is a step in which the extract from which the residue has been separated is distilled under reduced pressure to remove the extraction solvent and concentrate the components constituting the Chenopodium album extract to obtain a solid content. The vacuum distillation can be carried out, for example, using an evaporator at 50°C or below.

[0038] The solvent substitution step (step S4) is a step in which the solid fraction obtained in the reduced pressure concentration step is substituted with another solvent. The solvent used here is preferably ethanol, 1,3-butylene glycol, or an aqueous solution thereof, similar to the extraction solvent described above. It is particularly preferable to use a 30 to 100 vol% aqueous ethanol solution or a 30 to 100 vol% aqueous 1,3-butylene glycol solution. In this way, the solid fraction obtained by the reduced pressure concentration is redissolved in a solvent to obtain a solvent-substituted solution.

[0039] The filtration step (step S5) is a step in which the solvent replacement solution is filtered to obtain a filtrate from which insoluble components have been removed. The precipitating process can be carried out using a method that promotes precipitation, such as a centrifuge, or a membrane filter. A step of removing the insoluble components (sediment) from the solvent replacement solution in a cool, dark place before filtration can also be added. Specifically, the solvent replacement solution is first allowed to stand in a cool, dark place for at least two weeks to precipitate the insoluble components. The cool, dark place conditions may be an environment at 1 to 10°C, preferably 1 to 5°C, and away from direct sunlight. The solvent replacement solution may be allowed to stand in a cool, dark place for at least one day, but may also be for at least seven or 14 days.

[0040] The concentration adjustment step (step S6) is a step of adjusting the solids concentration of the filtered solution to a predetermined amount. Specifically, the solids concentration of the filtered solution is measured, and the same solvent as used in the solvent substitution step (step S4) is added to adjust the solids concentration to a predetermined amount. For example, if a 30 vol% 1,3-butylene glycol aqueous solution is used in the solvent substitution step, the solids concentration of this solution can be adjusted using the same aqueous solution. The solids referred to here are those derived from Chenopodium album and specifically include total saponins, total polyphenols, total free amino acids, and vitexin. In this way, a Chenopodium album extract-containing composition is obtained. In this concentration adjustment step, for example, the concentration can be adjusted by adding the desired amounts of water and 1,3-butylene glycol so that the solids concentration of the Chenopodium album extract-containing composition becomes 0.5 wt%. However, when adjusting the solid content concentration of the Chenopodium album extract-containing composition, it is not necessary to use the same solvent as in the solvent substitution process; a different solvent can also be used to adjust the Chenopodium album extract-containing composition. The predetermined solid content concentration is not particularly limited, but should be set to a concentration that adequately exhibits the various functionalities of Chenopodium album and ensures stability and safety. Specifically, the solid content concentration of the Chenopodium album extract-containing composition is preferably 0.01 wt% or more, and particularly preferably 0.03 wt% or more or 0.05 wt% or more. The upper limit of the solid content concentration of the Chenopodium album extract-containing composition is preferably 2 wt% or less, and particularly preferably 1 wt% or less.

[0041] The microfiltration step (step S7) is a step in which the concentration-adjusted composition containing Chenopodium album extract is precisely filtered using a membrane filter or the like. For example, the composition containing Chenopodium album extract may be filtered using a membrane filter with a mesh size of 0.45 μm or less. In this step, the composition containing Chenopodium album extract may be sterilized by filtration and filled into a sterilized container. This microfiltration step is carried out in a clean environment, such as a clean room or clean bench. [Example]

[0042] Next, the Chenopodium album extract-containing composition and the method for producing the same according to the present invention will be described in more detail with reference to examples.

[0043] [Example 1] In Example 1, the above-ground parts of Chenopodium japonica were dried, and then a 50 wt% aqueous ethanol solution was added. Extraction was carried out for 168 hours (7 days) at room temperature (extraction temperature 15°C). The extracted solution was then filtered to separate the extract from the extraction residue. Next, the extract was concentrated under reduced pressure using an evaporator, and water and 1,3-butylene glycol were added to achieve a 30 wt% 1,3-butylene glycol concentration, followed by filtration. The filtrate was left to stand at 4°C for 336 hours (14 days) to precipitate and separate the insoluble matter, which was then separated by filtration (precipitation treatment). The final solid concentration was adjusted to 0.5 wt% with a 30 wt% aqueous 1,3-butylene glycol solution, and the filtrate filtered through a 0.45 μm filter was filled into sterilized containers to obtain a Chenopodium japonica extract-containing composition.

[0044] [Example 2] In Example 2, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction time was changed to 5 hours.

[0045] [Example 3] In Example 3, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction time was changed to 24 hours.

[0046] [Example 4] In Example 4, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction time was changed to 72 hours.

[0047] [Example 5] In Example 5, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction time was changed to 120 hours.

[0048] [Example 6] In Example 6, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction temperature was changed to 50°C and the extraction time was changed to 2 hours.

[0049] [Example 7] In Example 7, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction temperature was changed to 50°C and the extraction time was changed to 5 hours.

[0050] [Example 8] In Example 8, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction solvent was changed to a 30 wt % aqueous solution of 1,3-butylene glycol.

[0051] [Example 9] In Example 9, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction solvent was changed to a 50 wt % aqueous solution of 1,3-butylene glycol.

[0052] [Example 10] In Example 10, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction solvent was changed to a 70 wt % aqueous solution of 1,3-butylene glycol.

[0053] [Example 11] In Example 11, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the period of the sedimentation treatment was shortened to one day.

[0054] [Example 12] In Example 12, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the period of the sedimentation treatment was shortened to 3 days.

[0055] [Example 13] In Example 13, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the period of the sedimentation treatment was shortened to 7 days.

[0056] [Example 14] In Example 14, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the period of the sedimentation treatment was shortened to 10 days.

[0057] [Example 15] In Example 15, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the period of the sedimentation treatment was shortened to one night and the number of filtrations was increased to two.

[0058] [Example 16] In Example 16, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction solvent was changed to water and the extraction time was changed to 5 hours.

[0059] [Example 17] In Example 17, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction temperature was changed to 50°C and the extraction time was changed to 5 hours.

[0060] [Example 18] In Example 18, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the extraction solvent was changed to water, the extraction temperature was changed to 90°C, and the extraction time was changed to 5 hours.

[0061] [Example 19] In Example 19, a composition containing Chenopodium album extract was produced in the same manner as in Example 1, except that the sedimentation treatment was omitted.

[0062] [Test method] The above-described Examples and Comparative Examples were subjected to component analysis, functionality testing, stability testing, and safety testing by the methods described below.

[0063] 1.Component analysis (total saponins) The total saponin concentration of each composition in the examples was simply measured using the phenol-sulfuric acid method (Dubois et al., 1956) after removing water-soluble sugars using a Sep-Pak C18 cartridge (Waters). Saponin is a general term for glycosides composed of polycyclic compounds and sugars, and this simple quantification method utilizes the color development of the sugar moiety. Here, a calibration curve was created using soybean saponin as a standard substance, and the soybean saponin equivalent saponin was calculated.

[0064] (total polyphenols) The total polyphenol concentration of each composition in the examples was measured using the Folin-Ciocalteu method (ISO 14502-1:2005), an official ISO method. The Folin-Ciocalteu method measures absorbance by changing the color of a reagent due to the reduction of phenolic hydroxyl groups. A calibration curve was created using gallic acid monohydrate as a standard substance, and the polyphenols equivalent to gallic acid were calculated.

[0065] (total free amino acids) The total free amino acid concentration of each composition in the examples was measured according to the automated pre-column derivatization amino acid analysis method using a Nexera X2 (Shimadzu Corporation) under the following measurement conditions. The pre-column derivatization method involves reacting amino acids with a derivatization reagent, separating the reaction products by reverse-phase chromatography, and measuring them using a fluorescence detector. The free amino acids measured here were aspartic acid, glutamic acid, serine, histidine, glycine, threonine, arginine, alanine, tyrosine, valine, methionine, cystine, phenylalanine, isoleucine, leucine, lysine, proline, and tryptophan, and the total concentration of these was defined as the "total free amino acid concentration." <Measurement conditions> Derivatization reagent: orthophthalaldehyde and 9-fluorenylmethyl chloroformate Column: Shim-pack Velox C18 (2.7 μm, 100 × 3.0 mm)

[0066] (Vitexin) The vitexin concentration of the composition according to Example 1 was measured by high performance liquid chromatography (HPLC) under the following measurement conditions. <Measurement conditions> HPLC equipment: Nexera X2 (Shimadzu Corporation) Column: Shim-pack VP-ODS (Φ4.6×150mm, 5μm) Column temperature: 40℃ Elution: 0.1 vol% formic acid:acetonitrile = 80:20 ·Flow rate: 1.0mL / min ·Injection volume: 5μL Detection: 350nm

[0067] 2. Functionality test (SOD-like activity) The SOD inhibitory activity of the composition containing Chenopodium album extract was measured using the SOD Assay Kit - WST (Dojindo Laboratories). The sample and reagent were reacted (37°C, 20 min), and the absorbance (450 nm) of the solution was measured using a plate reader to determine the IC50 (50% inhibitory concentration, 50% inhibitory activity).

[0068] (ABTS radical scavenging activity) The ABTS radical scavenging activity of compositions containing Chenopodium album extract was measured using the ABTS radical scavenging test. The sample was reacted with the ABTS radical solution (37°C, 4 min), and the absorbance (734 nm) of the solution was measured using a plate reader to determine the IC50 (50% inhibitory concentration, 50% inhibitory activity).

[0069] (Gene expression test) The gene expression activity of the composition containing Chenopodium album extract was measured as follows: Normal human epidermal keratinocytes and human neonatal skin fibroblasts were placed in a 60 mm dish at 5 × 10 4Cells were seeded at 1000 cells / dish. After 24 hours, samples were added to a final concentration of 0.03%. Medium without sample was used as a control. After 24 hours of culture, RNA was extracted from the cells using TRI reagent (Merck KGaA, Darmstadt, Germany). Using this RNA as a template, cDNA was synthesized by reverse transcription with an oligo dT primer using a Primescript RT reagent kit (Takara Bio, Shiga, Japan). mRNA levels were quantified by PCR using primers for various genes on a LightCycler 96 (Roche, Basel, Switzerland) with Luna Universal qPCR Master Mix (New England Biolabs, MA, USA). Cq values ​​were calculated using the delta Ct method, and the expression levels of each gene were expressed as relative values ​​based on the Cq value of GAPDH. To confirm the anti-aging effect, we measured the gene expression of type I collagen (COL1A1), collagen-degrading enzyme (MMP1), differentiation-related protein (KRT1), vascular endothelial growth factor (VEGF), cortisol-activating enzyme (11β-HSD1), and cortisol-inactivating enzyme (11β-HSD2). To confirm the barrier function strengthening effect, we measured the gene expression of protein cross-linking enzyme (TGM1) and filaggrin (FLG). To confirm the moisturizing effect, we measured the gene expression of water-permeable proteins (AQP1, AQP9).

[0070] 3. Stability testing The red ivy extract composition was placed in a 30 mL glass container and left to stand in a room (20°C) or in a thermostatic bath (5°C, 40°C, 50°C, cycles of -10°C to 20°C, fluorescent light irradiation at 30°C), where the temperature was kept constant, and the changes over time were visually observed over a period of three months.

[0071] 4.Safety Testing The safety of the Chenopodium album extract composition to the skin was determined using an in vitro skin sensitization test.

[0072] [Results and Discussion] The results of component analysis of the Chenopodium album extract-containing compositions according to Examples 1 to 19 are shown in FIG.

[0073] 1.Component analysis (Total saponins, total polyphenols, total free amino acids) The quantitative results of total saponins, total polyphenols, and total free amino acids of the Chenopodium album extract composition are shown in Figure 3. The total saponin concentration was higher in the 50 wt% ethanol extract (Examples 1 to 7) and the 1,3-butylene glycol aqueous solution extract (Examples 8 to 10) than in the water extract (Examples 16 to 18), indicating that an alcohol-containing solvent is suitable as an extraction solvent. Furthermore, the total polyphenol concentration was higher in the 50 wt% ethanol extract (Examples 1 to 7), but no significant difference was observed in the total free amino acid concentration among the Examples. Examples 1 and 7 showed similar values, providing the best results.

[0074] (Vitexin) The vitexin concentration was measured for Example 1, which showed good results for the concentrations of total saponins, total polyphenols, and total free amino acids. The vitexin concentration in Example 1 was 5.0 mg / L or higher, confirming that the sample contained polyphenol components characteristic of Japanese ivy.

[0075] 2. Functionality test For Examples 1 and 7, which gave good results in the component analysis, a functionality test was carried out.

[0076] (SOD-like activity) The results of the SOD inhibitory activity of Example 1 are shown in Figure 4. The 50% inhibitory concentration (IC50) of ascorbic acid, the positive control (pc), was 0.169 mg / mL, while that of Example 1 was 0.527 mg / mL, confirming relatively strong activity. Furthermore, the results of the solids concentration and inhibition rate in the Chenopodium album extract-containing composition for Example 1 are shown in Figure 5. 17.7% inhibition was confirmed at a solids concentration of 0.01%.

[0077] (Gene expression using keratinocytes) The results of the gene expression effect using the keratinocytes of Example 7 are shown in Figure 8. The results obtained suggest that an increase in differentiation-related protein (KRT1) will have an anti-aging effect (turnover effect), an increase in protein cross-linking enzyme (TGM1) and the main protein in keratinocytes, filaggrin (FLG), will have a barrier function strengthening effect, and an increase in water molecule permeable proteins (AQP1 and AQP9) will have a moisturizing effect. The structure of the human epidermis is, from the outside in, the stratum corneum, stratum granulosum, stratum spinosum, and stratum basale, with most of these layers being made up of keratinocytes. In the stratum basale, keratinocyte proliferation, keratinization, and stratum corneum peeling occur continuously, and this process of regeneration is called turnover. As we age, mother cells become deficient in energy, and the turnover cycle in which new cells are born gradually slows down, so promoting this turnover leads to anti-aging. Keratin, a fibrous protein, is the most abundant component of epidermal cells, and KRT1 (keratin 1) and other proteins are used as indicators of increased turnover.

[0078] (Gene expression using fibroblast cells) The results of the gene expression effect using the fibroblasts of Example 7 are shown in Figure 9. The results obtained suggest that an increase in type I collagen (COL1A1) will promote collagen production, a decrease in collagen-degrading enzyme (MMP1) will inhibit collagen degradation, an increase in vascular endothelial growth factor (VEGF) will promote angiogenesis, and a decrease in cortisol-activating enzyme (11β-HSD1) and an increase in cortisol-inactivating enzyme (11β-HSD2) will have an anti-inflammatory effect.

[0079] (CAMSAP3 expression effect) In Example 1, the expression of CAMSAP3 in epidermal keratinocytes (NHEK) was measured. Specifically, 1 × 10 NHEK cells were cultured. 5The cells were dispersed in a medium at a density of 1.5 × 10 cells / dish and seeded on a 60 mm dish. After the NHEK cells adhered, the medium was replaced, and the next day, a medium containing 0.00015 to 0.0015 wt% of a composition containing Chenopodium album extract was added, followed by culturing for 24 hours. Specifically, the concentration (wt%) of the composition containing Chenopodium album extract was 1.5 × 10 -4 wt%, 5 × 10 -4 wt%, 1.5 × 10 -3 The concentration was adjusted to 100% by weight. RNA was then recovered using TRI reagent. As a control, a medium containing no Chenopodium album extract-containing composition was added, and RNA was recovered in the same manner. After purifying the obtained RNA, cDNA was synthesized and the gene expression level was evaluated by PCR. As a result, it was confirmed that the expression level of CAMSAP3 was increased 1.7 to 2.5 times compared to the control. CAMSAP3, a member of the CAMSAP family, has been reported to contribute to the formation of non-centrosomal microtubules, which are known to be located near the cell membrane in epithelial cells and confer polarity. It has also been reported that in skin, microtubules change from centrosome-dependent to non-centrosomal microtubules depending on epidermal turnover, contributing to tissue function. This suggests that the composition containing Akasakazura extract according to the present invention promotes the formation of non-centrosomal microtubules via CAMSAP3, thereby regulating epidermal turnover and contributing to the maintenance of skin homeostasis.

[0080] 3. Stability testing The stability test results for Examples 1, 11, 12, 13, 14, 15, and 19 are shown in Figure 10. Examples 1, 11, 12, 13, and 14, in which the decantation process was carried out over 1 to 14 days and followed by one filtration (through a 0.45 μm or smaller membrane filter); Example 15, in which the decantation process was carried out overnight and followed by two filtrations (through a 0.45 μm or smaller membrane filter); and Example 19, in which the decantation process was not carried out and the filtration (through a 0.45 μm or larger membrane filter) was carried out immediately after the decantation process, all showed almost no precipitation when stored at 5°C for more than three months, and were therefore rated as "Good" for stability. Therefore, the stability of the Chenopodium album extract can be reliably guaranteed by filtering Chenopodium album extract components through a 0.45 μm or smaller membrane filter at least once as a means of separating and purifying the extract.

[0081] 4.Safety Testing An in vitro skin sensitization test was carried out on the Chenopodium album extract-containing compositions of Examples 1, 7, and 8. All results were negative, confirming that the Chenopodium album extract-containing compositions were safe.

[0082] In the above, the present specification has described the embodiments and examples of the present invention in order to express the contents of the present invention. However, the present invention is not limited to the above embodiments and examples, and includes modifications and improvements that are obvious to those skilled in the art based on the matters described in the present specification.

[0083] [Reference example] As a reference example, we will explain Kabuchi Citrus Peel Extract and its CAMSAP3 expression activity.

[0084] [Reference example 1] In Reference Example 1, the juice of kabuchi fruit was extracted, and the peel separated from the juice residue was used as the raw material. The kabuchi peel was washed with hypochlorous acid water and then dried at low temperature under reduced pressure at 40°C using a vacuum dryer to obtain dried kabuchi peel (citrus fruit peel). The dried kabuchi peel was crushed using a crusher and sieved to a particle size of 1.0 mm or less. To prevent deterioration of the raw material, the crushing was performed immediately before extraction. 100 mL of 50 vol% ethanol aqueous solution was added to 10 g of dried kabuchi peel powder, and extraction was performed for 24 hours at room temperature (20°C) (liquid temperature: 15°C) while stirring. The extract was then suction filtered using 7 μm filter paper to separate the extract from the extraction residue. The extract was then evaporated to dryness at 40°C using an evaporator. The extract was then dried under reduced pressure overnight (12 hours) using a vacuum dryer to completely evaporate the ethanol aqueous solution, obtaining the components of the kabuchi citrus fruit peel extract. The components were redissolved in a 30 wt% 1,3-butylene glycol aqueous solution, adjusting the component concentration to 1.6 wt% by weight. The solution was then left to stand overnight (12 hours) at 4°C to precipitate and precipitate the insoluble matter, which was then separated by suction filtration through a 0.45 μm membrane filter. The resulting filtrate was then left to stand overnight (12 hours) at 4°C, followed by suction filtration through a 0.22 μm membrane filter, repeating this process twice. The final solids concentration was adjusted to 1.0 wt% with a 30 wt% 1,3-butylene glycol aqueous solution, followed by sterilization filtration in a clean bench and filling into sterilized containers. This is how Kabuchi Citrus Peel Extract was produced.

[0085] [Test method] The above-mentioned Reference Example 1 was subjected to component analysis and functionality testing by the following methods.

[0086] 1.Extraction rate The weight of the solid content in each extract of Reference Example 1 was measured using the following procedure. 2 g of the extract was placed in an evaporating dish and dried on a hot plate at 105°C. After drying under reduced pressure at 105°C using a vacuum dryer, the weight was measured using a precision balance. Measurements were repeated until there was no change in weight, and the solid content was determined. The extraction rate was calculated from this solid content using the following formula. TIFF2025161747000002.tif15166

[0087] 2.Component analysis method (Quantitative analysis of polymethoxyflavonoids) The total amount of polymethoxyflavonoids in each extract according to Reference Example 1 was measured by high performance liquid chromatography (HPLC) under the following measurement conditions. The polymethoxyflavonoids that were quantified were sinensetin, isosinensetin, nobiletin, heptamethoxyflavone, natudaidine, and tangeretin. <Measurement conditions> HPLC equipment: NexeraX2 (Shimadzu Corporation) Column: YMC-Pack ODS-AM (Φ4.6 × 150 mm, 5 μm) Column temperature: 30℃ Gradient conditions: 0-10min;MeOH:water=50:50 10-35min;MeOH:water=50-98:50-2 35-40 min; MeOH:water = 98:2 40-50min;MeOH:water=50:50 ·Flow rate: 1.0mL / min ·Injection volume: 10μL Detection: 330 nm (other than tangeretin), 367 nm (tangeretin)

[0088] 3. Functionality test method (CAMSAP3 expression effect) In Reference Example 1, the expression of CAMSAP3 in epidermal keratinocytes (NHEK) was measured. 5The cells were dispersed in a medium at a density of 100 cells / dish and seeded on a 60 mm dish. After the NHEK cells adhered, the medium was replaced, and the next day, a medium containing 0.00015-0.0015 wt% Kabuchi Citrus Peel extract was added and cultured for 24 hours. Specifically, the concentration (wt%) of Kabuchi Citrus Peel extract was 3 × 10 -4 wt%, 1×10 -3 wt%, 3 × 10 -3 The concentration of the extract was adjusted to 100% by weight. RNA was then extracted using TRI Reagent. As a control, RNA was extracted in the same manner using a medium containing no Kabuchi Citrus Peel Extract. The RNA was purified, cDNA was synthesized, and gene expression levels were evaluated using PCR. The results confirmed that CAMSAP3 expression levels increased 1.4-9.5-fold compared to controls. CAMSAP3, a member of the CAMSAP family, has been reported to contribute to the formation of non-centrosomal microtubules, which are known to align near the plasma membrane in epithelial cells and confer polarity. It has also been reported that in skin, microtubules change from centrosome-dependent to non-centrosomal microtubules depending on epidermal turnover, contributing to tissue function. These findings suggest that Kabuchi Citrus Peel Extract promotes the formation of non-centrosomal microtubules via CAMSAP3, thereby regulating epidermal turnover and contributing to the maintenance of skin homeostasis.

Claims

1. Contains an extract obtained from red vine (Anredera cordifolia) at 0.01 wt% or more as an active ingredient, It has one or more of the following functions: anti-aging, barrier function strengthening, moisturizing, and antioxidant effects. A composition containing red vine extract.

2. The composition contains, as an extract component obtained from the Chenopodium album, Total saponin or a pharmaceutically acceptable salt thereof is 200 ppm or more, Total polyphenols or pharmaceutically acceptable salts thereof are 80 ppm or more, Total free amino acids or pharmaceutically acceptable salts thereof are 35 ppm or more, and Contains vitexin or a pharmaceutically acceptable salt thereof A composition containing the Chenopodium album extract according to claim 1.

3. The composition containing Chenopodium album extract does not cause skin sensitization A composition containing the Akasakazura extract according to claim 1 or claim 2.

4. The composition containing the Chenopodium extract does not separate or precipitate components for more than three months in a dark place at room temperature. A composition containing the Akasakazura extract according to claim 1 or claim 2.

5. The composition containing Chenopodium album extract contains ethanol or butylene glycol as a solvent. A composition containing the Akasakazura extract according to claim 1 or claim 2.

6. A cosmetic comprising the Chenopodium album extract-containing composition according to claim 1 or 2.

7. Contains an extract obtained from Japanese ivy as an active ingredient. An agent for promoting CAMSAP3 expression in epidermal keratinocytes.

8. A cosmetic comprising an agent for promoting CAMSAP3 expression in epidermal keratinocytes.

9. The CAMSAP3 expression promoter contains an extract obtained from Chenopodium album. The cosmetic product according to claim 8.

10. A method for producing a composition containing Chenopodium album extract, comprising: A drying process for drying the above-ground parts of the Japanese ivy; An extraction step of extracting the dried aerial parts of Chenopodium album with a solvent to obtain an extract; A separation and purification step of separating and purifying the extracted components is included. A method for producing a composition containing Chenopodium album extract.

11. The separation and purification step includes filtering the extracted components at least once. A method for producing a composition containing the Akasakazura extract according to claim 10.

12. The solvent is a 30 to 100 wt % aqueous solution of ethanol or butylene glycol. A method for producing a composition containing the Akasakazura extract according to claim 10.

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