Stratum corneum peeling improver

Abstract

To provide a desquamation improver with an excellent effect of inhibiting or improving the phenomenon that the degradation by a degrading enzyme is inhibited caused by the aggregation of horny layer cell adhesion factors, formation of polymeric products, and changes in protein structure.SOLUTION: The present invention has solved the above problems by using one or more selected from the whole plant extract of Artemisia dracunculus and the petal extract of Lilium rubellum.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a stratum corneum peeling improving agent. More specifically, the present invention relates to various agents and uses that suppress or improve the phenomenon in which structural changes in stratum corneum cell adhesion factors cause inhibition of degradation of stratum corneum cell adhesion factors by degrading enzymes. [Background technology]

[0002] The stratum corneum, located at the outermost layer of the skin, functions as a physical and chemical barrier between the body and the external environment, as well as regulating the texture and appearance of the skin. Maintaining a healthy stratum corneum is essential for maintaining a healthy body and skin. Maintaining a healthy stratum corneum requires the proper progression of epidermal turnover, i.e., the proliferation and keratinization of epidermal cells in the basal layer of the epidermis, and the process of stratum corneum desquamation (also known as desquamation). In particular, stratum corneum desquamation is a complex process in which the degradation of cell adhesion factors is regulated by various enzymes, and the amount and function of these molecules must be precisely controlled.

[0003] In normal skin, the cell adhesion factors that connect stratum corneum cells to each other are degraded smoothly, weakening the adhesion between stratum corneum cells in the outermost layer of the stratum corneum and allowing natural desquamation. On the other hand, abnormal proliferation of epidermal cells, decreased enzyme activity, and abnormalities in stratum corneum desquamation enzyme inhibitors disrupt the precise control of the cell adhesion factor degradation process, resulting in rough skin texture and appearance and skin diseases. Furthermore, the local occurrence of these abnormalities clearly distinguishes the affected area from the surrounding unaffected skin, making abnormalities in stratum corneum desquamation particularly undesirable from a cosmetic standpoint. Therefore, maintaining a normal stratum corneum desquamation process is extremely important for maintaining a healthy body and skin.

[0004] Corneocytes contain corneodesmosomes, a type of cell adhesion molecule. Corneodesmosomes are desmosomes that undergo structural changes when keratinocytes transform from granular cells into keratinocytes. Corneodesmosomes are composed of desmoglein 1 (DSG1) and desmocollin 1 (DSC1), the main components of desmosomes in granular cells, as well as corneodesmosin (CDSN), supplied from intracellular lamellar granules, in the extracellular portion.

[0005] Kallikrein-related peptidases (KLKs) are known to be enzymes involved in the degradation of corneodesmosomes. KLKs are a family of 15 serine proteases, consisting of KLK1 to KLK15. In the stratum corneum, the trypsin-like serine protease KLK5 degrades DSG1 and DSC1, while the chymotrypsin-like serine protease KLK7 degrades DSC1. Both are also known to degrade CDSN. The action of KLKs on corneodesmosomes contributes significantly to stratum corneum desquamation, and inhibition of corneodesmosome degradation by KLKs also inhibits stratum corneum desquamation.

[0006] <Inhibitory factors for stratum corneum peeling and issues to be resolved / what is unknown> It is believed that if the components of corneodesmosomes undergo some kind of change, they will not be degraded and stratum corneum desquamation will be inhibited. Regarding the inhibition of corneodesmosome desquamation related to the inhibition of corneodesmosome degradation, it has been reported that the formation of aggregated and polymerized products of corneosinocyte adhesion factors or changes in protein structure inhibit the sufficient function of degrading enzymes on corneosinocyte adhesion factors, thereby inhibiting smooth stratum corneum desquamation (Patent Document 1). It has also been suggested that protein modifications such as glycation, carbonylation, and nitration in the stratum corneum are involved in the formation of aggregated and polymerized products of corneosinocyte adhesion factors and changes in protein structure. Based on these findings, it is believed that for stratum corneum desquamation, it is important to maintain corneodesmosomes intact, or, if changes do occur, to resolve those changes and make them degradable by degrading enzymes. In other words, if the phenomenon of stratum corneum desquamation being inhibited due to the insufficient function of degrading enzymes on corneosinocyte adhesion factors could be inhibited or improved, it would be possible to improve the delay in stratum corneum desquamation. However, no active ingredients capable of inhibiting or improving this phenomenon have been discovered, and the development of new active ingredients has been desired.

[0007] Tarragon (Artemisia dracunculus) is a perennial plant of the Asteraceae family, originally native to southern Europe and Siberia, and is distributed throughout southern Russia and Central Asia. It grows to a height of 60-100 cm, with erect, well-branched stems and opposite, slender, pointed, glossy, and deep yellow-green leaves. Tarragon is known to inhibit the production of lipid peroxides (Patent Document 2) and to scavenge active oxygen species (Patent Document 3) when an oxidizing agent is added to plasma obtained from mice orally administered tarragon. However, its ability to inhibit and improve protein modification and to improve stratum corneum peeling was unknown.

[0008] The maiden lily (Lilium rubellum) is a member of the Liliaceae family and is also known as the Hime Sayuri (Princess Early Lily or Princess Small Lily). It grows to a height of approximately 30-50 cm and features pale pink flowers without spots. While a similar lily is the Sasayuri (Sasa Lily), the maiden lily is distinguished by the yellow tips of its stamens. The roots of certain species of lilies, primarily those in the Lilium genus (scientific name: Lilium), in the Liliaceae family, such as the Tiger Lily and the Hakata Lily, are known for their tonic, diuretic, and cough suppressant properties and have long been used as herbal medicines. Furthermore, numerous studies have been published on the effects of Madonna lily root extract on promoting elastin production (Patent Document 4), and on the effects of ceramide production promotion (Patent Document 5) and antioxidant effects (Patent Document 6) on the bulbs of the Liliaceae family. Although there are few reports on parts other than the root, it has been reported that plants of the genus Lilium in the Liliaceae family have the effect of promoting glycosaminoglycan production (Patent Document 7), and that the leaves of Longiflorum lilies and the petals of Asiatic lilies have the effect of inhibiting hyaluronidase activity (Patent Document 8). It has also been reported that extracts of the buds of plants belonging to the genus Lilium in the Liliaceae family have antioxidant and protein glycation inhibitory effects (Patent Document 9). However, these effects are not observed in extracts of flowers, stems, or leaves, and have been shown to be unique to bud extracts. Although numerous studies have been conducted on lilies for a long time, their ability to inhibit and improve protein modification, and to improve stratum corneum peeling, was unknown. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Patent Publication No. 2021-148492 [Patent Document 2] Japanese Patent Application Laid-Open No. 2011-236149 [Patent Document 3] Japanese Patent Application Laid-Open No. 2001-122765 [Patent Document 4] Japanese Patent Application Laid-Open No. 2012-056933 [Patent Document 5] Japanese Patent Application Laid-Open No. 2002-370998 [Patent Document 6] Japanese Patent Application Laid-Open No. 2011-016760 [Patent Document 7] Japanese Patent Application Laid-Open No. 2010-018594 [Patent Document 8] Japanese Patent Application Laid-Open No. 2003-012489 [Patent Document 9] Japanese Patent Application Laid-Open No. 2011-225564 Summary of the Invention [Problem to be solved by the invention]

[0010] The purpose of this invention is to provide a stratum corneum peeling improving agent that is highly effective in suppressing the phenomenon in which the degradation of stratum corneum cell adhesion factors by degrading enzymes is inhibited due to the formation of aggregates and polymerized products of stratum corneum cell adhesion factors and changes in protein structure, or in improving the state in which the degradation of stratum corneum cell adhesion factors by degrading enzymes is inhibited. [Means for solving the problem]

[0011] In light of the phenomenon in which aggregation and formation of polymerized products of stratum corneum adhesion factors and changes in protein structure occur, which in turn inhibit the degradation of stratum corneum adhesion factors by degradative enzymes, the present inventors have conducted extensive research to suppress or ameliorate this phenomenon. As a result, they have found that an extract of the whole plant of tarragon (Artemisia dracunculus) and an extract of the petals of maidenhair lily (Lilium rubellum) not only suppress but also ameliorate the phenomenon in which the degradation of stratum corneum adhesion factors by degradative enzymes is inhibited, leading to the completion of the present invention.

[0012] That is, the present invention solves the above problems by using one or more extracts selected from the whole plant extract of tarragon and the petal extract of maidenhair lily. [Effects of the Invention]

[0013] According to the present invention, by suppressing or improving the structural changes of stratum corneum cell adhesion factors, the stratum corneum cell adhesion factors are decomposed by degrading enzymes, thereby making it possible to maintain or improve the health of the skin condition by improving stratum corneum peeling. DETAILED DESCRIPTION OF THE INVENTION

[0014] In the present invention, improving stratum corneum desquamation means preventing delayed stratum corneum desquamation or facilitating delayed stratum corneum desquamation to adjust the texture and appearance of the skin and maintain or improve the condition of the skin; more specifically, maintaining or improving the condition of the skin by preventing or improving protein modification of stratum corneum cell adhesion factors.

[0015] Structural changes in stratum corneum adhesion factors include aggregation or polymerization of the stratum corneum adhesion factor proteins, changes in the three-dimensional structure of the constituent proteins, and changes in the properties of the constituent proteins and amino acid functional groups due to chemical reactions or modifications, making it difficult for degrading enzymes to function properly on the stratum corneum adhesion factors, making it difficult for the stratum corneum adhesion factors to be degraded by degrading enzymes. This means that smooth stratum corneum desquamation is hindered.

[0016] Aggregation and polymerization of stratum corneum cell adhesion factors refer to the formation of polymerized aggregates containing protein molecules or complexes, including those that progress to the point of forming visible precipitates. Protein polymerized aggregates include those composed of single or multiple proteins, and the polymerization form can be formed by covalent bonds such as intermolecular crosslinking, or non-covalent bonds such as ionic bonds, hydrophobic interactions, and van der Waals forces. Aggregated / polymerized products, or aggregated / polymerized substances, refer to proteins that exhibit the above state. For example, if the molecular weight or molecular size of a stratum corneum cell adhesion factor increases due to protein modification, the increased molecular weight or molecular size of the protein can be referred to as an aggregated / polymerized product.

[0017] Protein modification refers to various chemical modifications of proteins, such as glycation, carbonylation, or nitration of proteins that are factors that contribute to stratum corneum cell adhesion.

[0018] The term "agent for inhibiting or improving structural changes in a stratum corneum cell adhesion factor" as used herein refers to an agent that has at least either an inhibitory effect or an improving effect on structural changes in a stratum corneum cell adhesion factor. Depending on the state of the structural change, the agent may function as an inhibitor that inhibits structural changes in a stratum corneum cell adhesion factor, or as an improver that improves structural changes in a stratum corneum cell adhesion factor.

[0019] Inhibition of structural changes in stratum corneum cell adhesion factors means inhibiting structural changes in the stratum corneum cell adhesion factor proteins due to the causes mentioned above, and refers to a state in which the degradation of stratum corneum cell adhesion factors by degradative enzymes is not inhibited.

[0020] "Improving the structural change of a stratum corneum cell adhesion factor" refers to changing a state in which the stratum corneum cell adhesion factor is difficult to decompose by enzymes due to a structural change in the protein, to a state in which the stratum corneum cell adhesion factor is easily decomposed by enzymes. For example, in the case of a structural change in a stratum corneum cell adhesion factor due to protein modification, not only is the modified protein or its constituent amino acids lost through digestion or cleavage, but also the characteristic functional groups undergo oxidation-reduction or other chemical reactions to be converted into different functional groups or substances, making the stratum corneum cell adhesion factor more easily decomposed by enzymes.

[0021] The stratum corneum cell adhesion factor of the present invention is not particularly limited, and examples thereof include desmosome components DSG1, 2, 3, 4, DSC1, 2, 3, CDSN, plakoglobin, plakophilin, desmoplakin 1, 2, etc.

[0022] The enzymes that degrade stratum corneum cell adhesion factors are not particularly limited, and examples of enzymes that degrade stratum corneum cell adhesion factors include KLK5, KLK7, and KLK14, which are expressed in the skin.

[0023] The tarragon (Artemisia dracunculus) used in the present invention may be the whole plant, and may be used at any stage.

[0024] The petals (outer and inner perianth) of the maiden lily (Lilium rubellum) used in the present invention can be those that have bloomed from buds, but it is preferable to use petals that have fully bloomed. However, for reasons of industrial production and operation, it is acceptable for some parts other than the petals (for example, filaments, anthers, stigma, style, ovary, and pollen) to be included in the preparation process.

[0025] Known methods can be used to obtain the extract. The preparation of the extract is not particularly limited, but for example, the above-mentioned plant can be extracted using various suitable solvents at low to elevated temperatures. Examples of extraction solvents that can be used include one or more of the following: water; lower monohydric alcohols such as methyl alcohol and ethyl alcohol; liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol; ketones such as acetone and methyl ethyl ketone; alkyl esters such as ethyl acetate; hydrocarbons such as benzene and hexane; ethers such as diethyl ether; and halogenated alkanes such as dichloromethane and chloroform. Among these, a mixed solvent of one or more of water, ethyl alcohol, and 1,3-butylene glycol is particularly preferred.

[0026] The extraction method for the extract that can be used in the present invention is not particularly limited. In the present invention, the designated part may be cut into small pieces before extraction without drying, or may be dried and then cut into small pieces, or crushed into powder, and then extracted. For example, in the case of fresh fruit, a 1 to 100-fold weight amount, particularly a 4 to 20-fold weight amount, of solvent is used. For room temperature extraction, extraction is preferably performed at 0°C or higher, particularly 20°C to 40°C, for 1 hour or more, particularly 3 to 7 days. For heated extraction, extraction is preferably performed by heating at 60 to 100°C for 1 hour or more, particularly 4 hours or more. Extraction may also be performed at a temperature below 10°C, at which the extraction solvent does not freeze, for 1 hour or more, particularly 1 to 7 days.

[0027] The extract obtained as described above can be purified as needed, to the extent that the effect is not impaired, using activated carbon or activated clay, a styrene-divinylbenzene synthetic adsorbent (HP-20: manufactured by Mitsubishi Chemical Corporation), octadecylsilane-treated silica (Chromatorex ODS: manufactured by Fuji Silysia Chemical), etc. In some cases, the liquid phase after solid-liquid separation can be solidified by a conventional method such as spray drying or freeze drying, and then, if necessary, pulverized to form a powder before use.

[0028] Here, the extract in this embodiment includes an extract obtained by extracting a plant raw material using a solvent, a diluted or concentrated liquid of this extract, or a dried product obtained by evaporating the solvent of the extract, or a crude or purified product of these.

[0029] Furthermore, when the extract obtained as described above is used as each agent of the present invention, the extract may be used as is or may be mixed with a common base. The final form may be any form, such as liquid, emulsion, gel, solid, powder, or granules, and optional ingredients used in topical skin preparations may be appropriately blended as needed, as long as the effects are not impaired. Examples of optional ingredients include oils, surfactants, powders, coloring materials, water, alcohols, thickeners, chelating agents, silicones, antioxidants, UV absorbers, moisturizers, preservatives, fragrances, various medicinal ingredients, pH adjusters, and neutralizers. Furthermore, examples of the final form include, for example, skin external preparations, basic cosmetics such as emulsions, creams, lotions, essences, gels, packs, and facial cleansers; makeup cosmetics such as lipstick, foundation, liquid foundation, and pressed makeup powder; cleansing cosmetics such as facial cleansers, body shampoos, and soaps; and bath additives, but are not limited to these. The amount of the extract in each agent may be adjusted appropriately depending on the desired effect, but can be any amount from 0.0001% to 100% by weight converted to dry weight, preferably from 0.01% to 20% by weight, and more preferably from 0.1% to 10% by weight.

[0030] The effect of each agent in the present application can be judged to be effective if the rate of inhibition or improvement of structural changes in stratum corneum cell adhesion factors due to the presence of the drug is high. For example, structural changes in stratum corneum cell adhesion factors can be explained by the phenomenon of inhibition of degradation of stratum corneum cell adhesion factors by degradative enzymes. When the effect of inhibiting this phenomenon due to the presence of a drug is expressed as an inhibition rate, it can be appropriately set depending on the degree of desired effect, but if it can inhibit by approximately 20% or more, it can be judged to be effective, and if it can inhibit by 50% or more, it can be judged to have an excellent effect. The same applies to the effect of improving the phenomenon of inhibition of degradation of stratum corneum cell adhesion factors by degradative enzymes due to the presence of a drug. [Example]

[0031] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, the blending amounts are expressed in mass %.

[0032] <Preparation of extract> 5g of tarragon (Artemisia dracunculus) whole plant was added to 50g of distilled water (10 times its weight) and extracted by heating at 60°C for 4 hours. After extraction, the plant matter was removed by filtration, evaporated to dryness, and dissolved in distilled water to a solid evaporation residue of 1%. The resulting extract was used as the test substance. 5g of maiden lily (Lilium rubellum) petals were added with 20g of 50V / V% ethanol solution (four times the weight) and extracted by heating at 60°C for 4 hours. After extraction, the extract was filtered to remove the plant matter, evaporated to dryness, and dissolved in distilled water to a solid evaporation residue of 1%. The resulting extract was used as the test substance. As a comparative example, 5 g each of Marco Polo petals and Longiflorum petals, both of which are lilies, were added with 20 g of 50 V / V% aqueous ethanol (four times the weight of each), and extracted by heating at 60°C for 4 hours. After extraction, the extract was filtered to remove the plant matter, evaporated to dryness, and dissolved in distilled water to a solid evaporation residue of 1%. The resulting extract was used as the test substance.

[0033] <Active oxygen scavenging activity test; DPPH test> [Preparation of DPPH solution] The DPPH solution was prepared by mixing the following components in a volume ratio of (A):(B):(C) = 1:4:3. (A) 5.52 g of MES (2-(N-morpholino)ethanesulfonic acid) was dissolved in 100 mL of water, and the pH was adjusted to 6.1 with 1N NaOH. (B) 15.7 mg of DPPH (1,1-diphenyl-2-picrylhydrazyl) was dissolved in 100 mL of ethanol. (C) Purified water [Preparation of Trolox Solution] 25 mg of Trolox was dissolved in 10 mL of DMSO (dimethyl sulfoxide) to prepare a 10 mM solution. [DPPH test] 10 μL of test substance or Trolox solution (0.078 mM, 0.156 mM, 0.313 mM, 0.625 mM, 1.25 mM) was added to each well of a 96-well plate, followed by rapid addition of 190 μL of DPPH solution and mixing. After 10 minutes, the absorbance of each well was measured at 540 nm. A calibration curve was prepared to determine Trolox concentration versus absorbance at 540 nm, and the Trolox equivalent of the test substance was calculated from the absorbance at 540 nm. If Trolox is taken as 1, delta-tocopherol, which is said to have a high active oxygen scavenging effect, is said to have an equivalent of 1.36, so if the Trolox equivalent is 1.0 or higher, it can be said to have a high active oxygen scavenging effect.

[0034] [Table 1]

[0035] As shown in Table 1, tarragon extract, Marco Polo extract, and Easter lily extract showed Trolox equivalent values ​​higher than 2mM, demonstrating a significantly high active oxygen scavenging effect. However, even though they are all lilies of the same genus, the Trolox equivalents of Maiden lily extract, Marco Polo extract, and Easter lily extract differ greatly, confirming that the strength of the active oxygen scavenging effect differs depending on the variety.

[0036] <Test to evaluate the effect of protein modification in suppressing the degradation of stratum corneum cell adhesion factors> (1) Collection of stratum corneum Stratum corneum samples were collected from human forearm skin washed with a cleanser using a skin checker (Promotool). (2) Inhibition of protein modification in the stratum corneum The skin checkers from which the stratum corneum had been collected as described above were treated under the following conditions. [Glycation modification] A mixture of 1M D-glucose and 100mM D-ribose was mixed with 10% distilled water (Control B) or the test substance. The skin checker from which the stratum corneum had been collected was immersed in the mixture and incubated at 60°C for 72 hours. It was then washed three times with PBS (Phosphate Buffered Saline). [Carbonylation modification] A 0.5% acrolein solution (Fujifilm Wako Pure Chemical Industries, Ltd.) was dissolved in PBS, and 10% distilled water (Control B) or the test substance was added to the reaction solution. The skin checker from which the stratum corneum had been collected was immersed in the solution and incubated at 37°C for 24 hours, after which it was washed three times with PBS. [Nitration modification] 10% distilled water (Control B) or test substance was added to 1M phosphate buffer (pH 7.4). The skin checker with the stratum corneum sample was immersed in the solution, and peroxynitrite solution (DOJINDO) was added to a concentration of 100 μM. The mixture was allowed to react at room temperature for 2 hours. The skin checker was then washed three times with PBS. (3) Enzyme treatment of the stratum corneum [KLK5 stock solution] Kallikrein 5, human, recombinant, carrier-free (R&D) was dissolved in a buffer (100 mM sodium phosphate, 1 mM EDTA, pH 7.4) to a concentration of 25 ng / μL. [Degradation of cell adhesion factors in the stratum corneum by degradative enzymes] The KLK5 stock solution was diluted 10-fold with PBS, and the modified skin checkers were immersed in the solution and incubated at 37°C for 24 hours. They were then washed three times with PBS. As a control, an unmodified skin checker was also treated in the same way (Control C). (4) Immunostaining of the stratum corneum After each treatment, the primary antibody (anti-Desmoglein 1 mouse monoclonal, Dsg1-P124, supernatant (PROGEN)) diluted 1:10 in PBS was added to the stratum corneum-extracted skin checkers and incubated for 1 hour at room temperature. After washing with PBS, a secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 568) preadsorbed (Abcam)) diluted 2:200 in 1% BSA / PBS solution was added and incubated for 1 hour at room temperature. After washing with PBS, bright-field images of the stratum corneum cells and fluorescent images of DSG1 were taken using a fluorescence microscope (BZ-X700, KEYENCE). The stratum corneum cell area and DSG1 fluorescence intensity were measured using image analysis software (ImageJ, open source). Finally, the DSG1 fluorescence intensity per stratum corneum cell area was calculated, and the inhibition rate was calculated. (5) Evaluation of effects The rate at which inhibition of protein modification in the stratum corneum inhibited the degradation of stratum corneum cell adhesion factors by degrading enzymes (inhibition rate) was calculated using the following formula.

[0037]

number

[0038] [Table 2]

[0039] Table 2 shows the percentage inhibition of the degradation of stratum corneum adhesion molecules by each protein modification. Tarragon extract and Maiden lily extract inhibited the structural changes in stratum corneum adhesion molecules caused by protein modifications, i.e., the phenomenon in which degradation is inhibited due to insufficient function of degradative enzymes on stratum corneum adhesion molecules. Both tarragon extract and Maiden lily extract exhibited a highly potent inhibition of almost all degradation inhibition for each protein modification. On the other hand, Marco Polo extract, a different species from Maiden lily, exhibited a similarly potent active oxygen scavenging effect to tarragon, but did not exhibit any inhibition of protein modification in the stratum corneum or inhibition of the degradation of stratum corneum adhesion molecules. Similarly to Maiden lily extract, Longiflorum extract, a member of the Lilium genus, exhibited a highly potent inhibition of the degradation of stratum corneum adhesion molecules due to glycation, but did not exhibit any inhibition of the degradation of protein modifications due to carbonylation or nitration. This confirmed that even if a compound exhibits an effect on one type of protein modification, it does not necessarily have an effect on all protein modifications.

[0040] <Test to evaluate the effect of protein modification on improving the inhibition of degradation of stratum corneum cell adhesion factors> (1) Collection of stratum corneum Stratum corneum samples were collected from human forearm skin washed with a cleanser using a skin checker (Promotool). (2) Protein modification of the stratum corneum The skin checkers from which the stratum corneum had been collected as described above were treated under the following conditions. [Glycation modification] The skin checker from which the stratum corneum had been collected was immersed in a mixture of 1M D-glucose and 100 mM D-ribose and incubated at 60°C for 72 hours, after which it was washed three times with PBS (Phosphate Buffered Saline). [Carbonylation modification] The skin checker from which the stratum corneum had been collected was immersed in a reaction solution prepared by dissolving acrolein (Fujifilm Wako Pure Chemical Industries) in PBS to a concentration of 0.5%, and incubated at 37°C for 24 hours, after which it was washed three times with PBS. [Nitration modification] The skin checker from which the stratum corneum had been collected was immersed in 1M phosphate buffer (pH 7.4), and peroxynitrite solution (DOJINDO) was added to a concentration of 100 μM. The reaction was allowed to proceed at room temperature for 2 hours, after which the skin checker was washed three times with PBS. (3) Application of the test substance to the protein-modified stratum corneum 10% distilled water (control B) or test substance was added to PBS, and the stratum corneum-extracted Skin Checkers that had been modified with each protein were immersed and incubated at 37°C for 24 hours. (4) Enzyme treatment of the stratum corneum [KLK5 stock solution] Kallikrein 5, human, recombinant, carrier-free (R&D) was dissolved in a buffer (100 mM sodium phosphate, 1 mM EDTA, pH 7.4) to a concentration of 25 ng / μL. [Degradation of cell adhesion factors in the stratum corneum by degradative enzymes] The KLK5 stock solution was diluted 10-fold with PBS, and the stratum corneum-extracted skin checkers to which the test substances had been applied after each modification were immersed and incubated at 37°C for 24 hours. They were then washed three times with PBS. As a control, a stratum corneum-extracted skin checker that had not been modified or tested was also treated in the same way (Control C). (5) Immunostaining of the stratum corneum After each treatment, the stratum corneum sample skin checkers were treated with a primary antibody (anti-Desmoglein 1 mouse monoclonal, Dsg1-P124, supernatant (PROGEN)) diluted 10-fold with PBS and incubated at room temperature for 1 hour. After washing with PBS, a secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor(R) 568) preadsorbed (Abcam)) diluted 200-fold with 1% BSA / PBS solution was added and incubated at room temperature for 1 hour. After washing with PBS, Bright-field images of stratum corneum cells and fluorescent images of DSG1 were taken using a fluorescence microscope (BZ-X700, KEYENCE), and the area of ​​stratum corneum cells and the DSG1 fluorescence value were measured using image analysis software (ImageJ, open source). Finally, the DSG1 fluorescence value per stratum corneum cell area was calculated, and the improvement rate was then calculated. (6) Evaluation of effects The rate at which the inhibition of degradation of stratum corneum cell adhesion factors by the degrading enzyme was improved (improvement rate) was calculated using the following formula.

[0041]

number

[0042] [Table 3]

[0043] Table 3 shows the percentage improvement in degradation of stratum corneum adhesion molecules, which had been inhibited by improving the structural changes in the stratum corneum adhesion molecules caused by each protein modification. Tarragon extract and Maiden lily extract showed different improvement rates for each protein modification, but both showed an improvement of more than 25% in the inhibition of degradation. These results demonstrate that tarragon extract and Maiden lily extract have the ability to improve structural changes in stratum corneum adhesion molecules caused by protein modification, i.e., the phenomenon in which degradation is inhibited because degradative enzymes do not function properly on the stratum corneum adhesion molecules. Meanwhile, Marco Polo extract and Longiflorum extract, which are species of Maiden lily and the same genus, showed a high active oxygen scavenging effect comparable to that of tarragon, but did not show any improvement effect on the respective protein modifications.

[0044] With regard to the effects of inhibiting and improving changes in the structure of stratum corneum cell adhesion factors, even when DSG1 is used as an indicator, similar results are obtained with another stratum corneum cell adhesion factor, DSC1 (Patent Document 1). Therefore, it can be said that the present invention is also effective with other stratum corneum cell adhesion factors, not just DSG1.

[0045] Below are examples of formulations of each agent of the present invention. The contents are in mass %. The manufacturing method is a conventional method. The formulations are representative examples, and the present invention is not limited to these formulations. The concentrations of the various extracts in the formulation examples are concentrations based on the dry residue. The effects of the present invention have also been confirmed with the following formulations.

[0046] JPEG0007820193000006.jpg97139

[0047] JPEG0007820193000007.jpg57137

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[0050] JPEG0007820193000010.jpg110138

[0051] JPEG0007820193000011.jpg71138 [Industrial Applicability]

[0052] Structural changes occur in the proteins of stratum corneum cell adhesion factors, which can suppress or improve the phenomenon in which degradation by degrading enzymes is inhibited, and it is expected that this will effectively improve stratum corneum desquamation.

Claims

1. Contains an extract of the petals of the maiden lily (Lilium rubellum) to improve stratum corneum peeling. Agent.

2. Tarragon (Artemisia dracunculus) whole plant extract, Maiden lily (Lilium rubellum) petal extract containing one or more selected from An agent for suppressing or improving structural changes of cell adhesion factors.

3. The structural change of the stratum corneum cell adhesion factor is due to glycation, carbonylation, or nitration of the protein. The inhibitor or ameliorator according to claim 2, which is caused by one or more selected from the following.

4. 4. The inhibitory or anti-inflammatory agent according to claim 2 or 3, wherein the stratum corneum cell adhesion factor is desmoglein 1. is an improver.

5. The agent according to any one of claims 1 to 4, which is a cosmetic or a quasi-drug.

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