External skin barrier function repairing agent

Abstract

The present invention relates to a novel external skin barrier function repairing agent which not only promotes skin barrier repair of damaged skin, but also is expected to have a repair effect on skin barrier dysfunction caused by the use of glucocorticoid and / or psychological stress. The skin barrier function repairing agent achieves skin barrier repairing through any promoting effect of promoting proliferation of keratinocytes, promoting migration of the keratinocytes, promoting expression of HAS-2 of the keratinocytes, and promoting expression of HBD-2 of the keratinocytes.

Description

External use skin barrier function repairing agent Technical Field

[0001] The present invention relates to a novel topical skin barrier function repairing agent that not only promotes the repair of damaged skin barriers but is also expected to improve skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress. The skin barrier function repairing agent achieves skin barrier repair by promoting the proliferation of keratinocytes, promoting the migration of keratinocytes, promoting the expression of hyaluronan synthase-2 (HAS-2) in keratinocytes, and promoting the expression of human β-defensin-2 (HBD-2) in keratinocytes. Background Art

[0002] The skin is the first protective barrier between the body and the external environment, and it has the important function of preventing the rapid loss of water from the body and blocking the invasion of harmful substances from the environment. This barrier function is largely provided by the epidermis. The epidermis is located in the outer layer of the skin and is a stratified epithelial tissue that continuously renews itself. The keratinocytes (also known as basal cells) in the innermost basal layer of the epidermis continuously divide and proliferate to produce daughter cells to renew the epidermis. The newly generated daughter cells gradually differentiate into spinous and granular layer cells during their migration to the outer layers, and finally differentiate into keratinocytes. The outermost stratum corneum is continuously shed through the process of desquamation, and the underlying cells migrate up to form a new stratum corneum. Therefore, maintaining the homeostasis of the epidermal barrier requires a balance between the proliferation, migration, and differentiation activities of keratinocytes.

[0003] As the outermost barrier of the body, the skin is attacked by harmful factors from the external environment every day, which may lead to tissue damage and barrier breakdown. After skin damage, a series of repair processes are usually immediately activated to restore the skin barrier function. The repair of skin damage is a highly coordinated process involving multiple types of cells, extracellular matrix molecules and regulatory factors. As the main cell type of the epidermis, keratinocytes play a key role in wound healing and repair of epidermal barrier function. Once a wound occurs, keratinocytes at the edge of the wound are immediately activated, accelerating migration and proliferation to achieve re-epithelialization of the skin, which is crucial for rapid wound healing.

[0004] In addition to accelerating proliferation and migration, keratinocytes also promote barrier repair and maintenance of healthy skin homeostasis by increasing the expression of multiple genes with important functions, such as hyaluronic acid synthase (HAS) and human β-defensin (HBD). HAS is a key enzyme for skin cells to synthesize hyaluronic acid (HA). Keratinocytes can express three subtypes of HAS, namely HAS-1, HAS-2, and HAS-3. Among them, HAS-2 has the highest expression abundance and is the most important synthase for keratinocytes to synthesize HA. HA is the most abundant extracellular matrix component of skin tissue and is involved in processes such as tissue homeostasis maintenance, hydration, and wound repair. Sufficient research evidence has shown that HA plays an important role in the wound healing process. During wound healing, a large amount of HA is produced locally on the wound surface. The local HA and its decomposition products (medium and low molecular weight HA) selectively activate the CD44 receptor on the surface of keratinocytes, mediate specific signal transduction pathways, regulate the proliferation, migration and differentiation of keratinocytes, and ultimately affect wound healing. Studies have found that overexpression of HAS-2 in keratinocytes accelerates cell migration, while knocking down HAS-2 inhibits keratinocyte migration. Epidermal growth factor (EGF), keratinocyte growth factor (KGF) and all-trans retinoic acid (RA) can all increase the expression level of HAS-2 in keratinocytes, thereby increasing the synthesis of HA. This HAS-2-dependent increase in HA synthesis is directly related to the effects of EGF, KGF, and RA in promoting keratinocyte migration, proliferation and wound healing. It can be seen that HA not only participates in the formation of the epidermal extracellular matrix, but also plays an important role in the proliferation, migration and differentiation of keratinocytes. Therefore, increasing the expression of HAS-2 helps to quickly repair skin damage and maintain the homeostasis of barrier function.

[0005] Human β-defensins (HBDs) are a group of cationic antimicrobial peptides consisting of 12-50 amino acids, produced by epithelial tissues such as the skin and respiratory tract. They possess antimicrobial and antiviral activities, thus contributing to the first line of defense of the innate immune system. Four HBDs have been identified in human skin: HBD-1, HBD-2, HBD-3, and HBD-4. HBD-2 expression in skin tissue is localized to the uppermost layer of the epidermis, with expression levels significantly increased in differentiated keratinocytes. Recent studies have also demonstrated that HBD-2 mediates IL-1β-induced protection of keratinocytes against proteases secreted by Staphylococcus aureus, suggesting that promoting endogenous HBD-2 expression may be beneficial for skin barrier repair and protection against S. aureus colonization in inflammatory skin diseases. A growing body of research indicates that the effects of antimicrobial peptides extend beyond antimicrobial activity; they also participate in wound repair and maintenance of skin barrier homeostasis through multiple pathways. After trauma, local epidermal cells increase the expression of HBD under the stimulation of cytokines, growth factors or bacteria, and the increased HBD then activates signaling pathways such as MAPK, AKT, STAT1 and STAT3, and activates intracellular Ca 2+ Signaling and EGFR phosphorylation promote the proliferation and migration of keratinocytes, thereby accelerating the healing of skin wounds. In addition to its effects on the proliferation and migration of keratinocytes, HBD also stimulates angiogenesis, promotes collagen production by skin fibroblasts, downregulates the expression of matrix metalloproteinase-1, and promotes extracellular matrix deposition to accelerate wound healing. Other studies have found that HBD can improve the skin barrier function by increasing the expression of tight junction proteins in epidermal cells, and can selectively activate Lgr6. + Defensins are involved in the maintenance of skin barrier homeostasis. Therefore, increasing HBD expression in keratinocytes may help improve the speed and quality of skin wound repair and regeneration.

[0006] Under normal circumstances, the skin possesses a robust self-repair capacity, rapidly repairing minor injuries and restoring structural and functional homeostasis. However, the skin's repair process is often affected by various adverse factors, which can disrupt the normal repair process, delay repair, and lead to skin barrier dysfunction, potentially inducing or exacerbating various skin diseases. Stress and the use of glucocorticoids are two common adverse factors that can affect skin repair. Stress is a nonspecific response of the body to excessive or harmful stimuli. It is caused by various physiological factors (such as trauma and pain) and psychological factors (stress) and can be categorized as physiological or psychological. With the accelerated pace of life and increasing competitive pressures, psychological stress has become an unavoidable and recurring event in modern life. Psychological stress can lead to a wide range of skin health issues, including reduced wound healing, barrier dysfunction, abnormal skin immune function and decreased resistance to infection, and even premature skin aging. Stress triggers a complex series of physiological responses, primarily manifested by increased activity of the sympathetic-adrenomedullary system (SAM) and the hypothalamic-pituitary-adrenal axis (HPA axis). SAM activation leads to a rapid increase in levels of stress hormones such as epinephrine and norepinephrine. Increased HPA axis activity, in turn, stimulates the adrenal glands to secrete glucocorticoids (GCs) through the release of corticotropin-releasing hormone (CRH) from the hypothalamus and adrenocorticotropic hormone (ACTH) from the pituitary gland. In recent years, increasing evidence indicates that the skin is a key target organ for these stress hormones. Excessive production of epinephrine and norepinephrine under stress can directly act on β-2-adrenergic receptors on the surface of epidermal keratinocytes, inhibiting keratinocyte migration and inducing oxidative stress and DNA damage, leading to skin barrier disruption, slowed wound healing, and skin aging. Excessive production of glucocorticoids (GC) under stress can also directly act on skin cells, inhibiting the expression of hyaluronic acid and lipid synthases, reducing the content of hyaluronic acid and lipids in skin tissue, negatively impacting the epidermal barrier, reducing stratum corneum hydration, and increasing transepidermal water loss. GC can also inhibit the proliferation and differentiation of epidermal keratinocytes, disrupting the integrity and adhesion of the stratum corneum, and impairing epidermal barrier function. Furthermore, GC inhibits the expression of antimicrobial peptides such as HBD by keratinocytes, impairing the skin's ability to resist infection. The use of exogenous glucocorticoids can produce adverse effects similar to those of endogenous GC overproduced during stress.Epidermal dysfunction is common in patients who use glucocorticoids for a long time. Skin atrophy and barrier dysfunction are recognized adverse reactions of topical glucocorticoids. Studies have found that the glucocorticoid drug dexamethasone can significantly inhibit the expression of HAS-2 and the synthesis of HA in skin fibroblasts and HaCaT keratinocytes at very low doses (1.5nM, 150nM and 1.5μM) and a very short duration of action (3h). Consistent with this, human trials have found that topical dexamethasone ointment is sufficient to induce a significant decrease in HA levels in skin tissue in a very short treatment period (3 days). In contrast, GC treatment requires 3 weeks to observe a decrease in the synthesis of type I and type III collagen in skin tissue. This suggests that the downregulation of HAS-2 expression and the resulting reduction in HA synthesis may be the most critical initial step in GC-induced skin atrophy.

[0007] Natural α-ionone is a secondary metabolite of plants with a violet floral scent. It is widely found in a variety of flowers and fruits, such as cherries, raspberries, and grapes, as well as tea and various plant essential oils. Synthetic α-ionone is widely used as a fragrance ingredient in the food and daily chemical industries. In the "National Food Safety Standard for the Use of Food Additives" (GB2760-2014), promulgated in 2014, α-ionone is classified as a synthetic flavoring for food, and the US FDA has also approved its use as a flavoring. In addition to its use as a food additive, α-ionone is widely used as a flavoring agent in cosmetics, perfumes, shampoos, soaps, and other toiletries, as well as household cleaners and detergents. According to statistics, the global consumption of α-ionone is approximately 100 to 1,000 metric tons per year.

[0008] Our bodies and skin are often exposed to α-ionone, but we know very little about its biological functions. A study reported that α-ionone may have anti-photoaging effects on the skin. The study found that α-ionone can activate the TGF-β-SMAD pathway in UV-irradiated Hs-68 human fibroblasts, induce collagen expression, but inhibit the MAPK-AP-1 signaling pathway and inhibit the expression of MMP1, MMP3 and MMP9 (non-patent document 1, Molecules 2019, 24 (9), 1804). In another related study, α-ionone can stimulate myogenesis in vitro, reduce palmitic acid-induced skeletal muscle myotube atrophy, increase myotube diameter and length, fusion index and cell protein content. This study suggests that α-ionone is a candidate drug that can enhance the quality and strength of skeletal muscle. The above studies show that in addition to its fragrance-giving function, α-ionone may also have biological functions that are beneficial to physical health (non-patent document 2, Food Funct. This journal is The Royal Society of Chemistry 2019, published on February 5, 2019).

[0009] In addition, in Food Industry Science and Technology 2022; 43(20): 481-488 (Non-Patent Document 3), there is a summary of the known biological activities of α-ionone.

[0010] Prior art literature

[0011] Non-patent literature:

[0012] 1.Molecules 2019,24(9),1804

[0013] 2.Food Funct.This journal is The Royal Society of Chemistry 2019, published on February 5, 2019

[0014] 3. Journal of Food Industry Science and Technology, 2022; 43(20): 481-488

[0015] Summary of the Invention

[0016] Technical problem to be solved by the invention

[0017] Based on the above knowledge, it is known that skin barrier function is closely related to the proliferation and migration of keratinocytes and the expression of HAS-2 and HBD-2 in keratinocytes. Based on the above knowledge, the inventors of the present invention set out to find substances that regulate the proliferation, migration, and expression of HAS-2 and HBD-2 of keratinocytes, and conducted in-depth research, aiming to find new topical skin barrier function repair agents for improving the barrier function of damaged skin and repairing skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress.

[0018] Means of solving technical problems

[0019] In order to find a substance that can promote the proliferation and migration of keratinocytes and promote the expression of HAS-2 and HBD-2 in keratinocytes, the applicant conducted in-depth research and extensive experiments and unexpectedly discovered that α-ionone can promote the proliferation and migration of keratinocytes and promote the expression of HAS-2 and HBD-2 in keratinocytes. Adding α-ionone as an active ingredient in a skin barrier function repair agent in skin topical preparations can significantly promote the repair of the skin barrier. Based on this discovery, the present invention was completed.

[0020] Specifically, the present invention includes the following solutions:

[0021] 1. A skin barrier function repairing agent in an external skin preparation, characterized in that it contains α-ionone as an active ingredient that acts on keratinocytes.

[0022] 2. The skin barrier function repairing agent according to 1 above, wherein the α-ionone acts as a keratinocyte migration promoter.

[0023] 3. The skin barrier function repairing agent according to 1 above, wherein the α-ionone acts as a keratinocyte proliferation promoter.

[0024] 4. The skin barrier function repairing agent according to 1 above, characterized in that the α-ionone acts as an expression promoter of HAS-2 in keratinocytes.

[0025] 5. The skin barrier function repairing agent according to item 1 above, wherein the α-ionone acts as an expression promoter of HBD-2 in keratinocytes.

[0026] 6. The skin barrier function repairing agent according to item 1 above, characterized in that it is used as a repairing agent for skin barrier function impairment caused by the use of glucocorticoids and / or psychological stress.

[0027] 7. The skin barrier function repairing agent according to item 1 above, wherein the external skin preparation is a cosmetic.

[0028] 8. Use of α-ionone in the preparation of cosmetics for promoting skin barrier repair, wherein the promotion of skin barrier repair is achieved by at least one promoting effect selected from the group consisting of promoting keratinocyte migration, promoting keratinocyte proliferation, promoting HAS-2 expression in keratinocytes, and promoting HBD-2 expression in keratinocytes.

[0029] 9. Use of α-ionone in the preparation of a cosmetic suitable for skin suffering from skin atrophy and / or skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress.

[0030] 10. A keratinocyte proliferation promoter comprising α-ionone as an active ingredient.

[0031] 11. Use of α-ionone as a keratinocyte proliferation promoter.

[0032] 12. Use of α-ionone in the preparation of a skin external preparation for promoting the proliferation of skin keratinocytes.

[0033] 13. A keratinocyte migration promoter comprising α-ionone as an active ingredient.

[0034] 14. Use of α-ionone as a keratinocyte migration promoter.

[0035] 15. Use of α-ionone in the preparation of a skin external preparation for promoting the migration of skin keratinocytes.

[0036] 16. An agent for promoting HAS-2 expression in keratinocytes, comprising α-ionone as an active ingredient.

[0037] 17. Use of α-ionone as an expression promoter of HAS-2 in keratinocytes.

[0038] 18. Use of α-ionone in the preparation of a skin external preparation for promoting HAS-2 expression in skin keratinocytes.

[0039] 19. An agent for promoting HBD-2 expression in keratinocytes, comprising α-ionone as an active ingredient.

[0040] 20. Use of α-ionone as an agent for promoting HBD-2 expression in keratinocytes.

[0041] 21. Use of α-ionone in the preparation of a skin external preparation for promoting HBD-2 expression in skin keratinocytes.

[0042] Effects of the Invention

[0043] The inventors of the present invention discovered for the first time that α-ionone can act as a keratinocyte migration promoter, a keratinocyte proliferation promoter, a keratinocyte HAS-2 expression promoter, and a keratinocyte HBD-2 expression promoter. It can achieve skin barrier repair by promoting any one or more of the following effects: promoting the proliferation of keratinocytes, promoting the migration of keratinocytes, promoting the expression of HAS-2 in keratinocytes, and promoting the expression of HBD-2 in keratinocytes. It can not only repair common skin damage, but also repair skin atrophy and / or skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] FIG1 is a graph showing the effect of α-ionone in promoting the proliferation of HaCaT cells.

[0045] FIG2 is a graph showing the effect of α-ionone on HaCaT cell migration.

[0046] FIG3 is a graph showing the effect of α-ionone on the inhibition of HaCaT cell migration by norepinephrine (NE).

[0047] FIG4 is a graph showing the effect of α-ionone on the expression of HAS-2 in HaCaT cells.

[0048] FIG5 is a graph showing the effect of α-ionone on hyaluronic acid (HA) synthesis in HaCaT cells.

[0049] FIG6 is a graph showing the effect of α-ionone on the inhibition of HAS-2 expression by dexamethasone (DEX).

[0050] FIG. 7 is a graph showing the effect of α-ionone on the expression of HBD-2 in HaCaT cells.

[0051] FIG8 is a graph showing the effect of α-ionone on the secretion of HBD-2 from HaCaT cells.

[0052] FIG9 is a graph showing the effect of α-ionone on the inhibition of HBD-2 expression by dexamethasone (DEX).

[0053] FIG. 10 is a graph showing the transepidermal water loss (TEWL value) of the test area of ​​each test group.

[0054] FIG11 is a graph showing the moisture content of the stratum corneum of the skin in the test area of ​​each test group.

[0055] FIG. 12 is a graph showing the barrier repair rate of each test group.

[0056] FIG. 13 is a graph showing the measured values ​​of the moisture content of the stratum corneum of the skin of the subjects.

[0057] FIG. 14 is a graph showing changes in the moisture content of the stratum corneum of the test subjects compared to before peeling.

[0058] FIG. 15 is a graph showing TEWL measurements of the test areas of the subjects.

[0059] FIG. 16 is a graph showing the TEWL repair rate of the subjects. DETAILED DESCRIPTION

[0060] The present invention will be described in detail below, but the present invention is not limited to the specific embodiments listed below, and various modifications can be made to the present invention without departing from the spirit of the present invention.

[0061] The "skin external preparation" mentioned in the present invention refers to any preparation applied to the skin from the outside, including but not limited to cosmetics and perfumes, fragrances, solid perfumes, etc., for example, aqueous solution preparations such as toner, softener, lotion, and repair water; spray preparations such as moisturizing spray, anti-wrinkle spray, and repair spray; emulsion preparations such as moisturizing lotion, anti-wrinkle lotion, repair lotion, body lotion, and hand lotion; essence preparations such as moisturizing essence, anti-wrinkle essence, and repair essence; gel preparations such as moisturizing gel, anti-wrinkle gel, and repair gel; creams or ointments such as face cream, eye cream, moisturizing cream, anti-wrinkle cream, repair cream, massage cream, hand cream, and body cream; powders such as setting powder, loose powder, pressed cake, and talcum powder, etc.

[0062] Ionone, with the structural formula C 13 H 20 O, and the molecular weight is 192.3g / mol. It contains three isomers, α-ionone, β-ionone, and γ-ionone. α-ionone is also known as (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one ((3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one), cyclocitrylideneacetone, irisone, etc. α-ionone is a light yellow transparent liquid component with a melting point of -49°C and a boiling point of 126-128°C. The structural formula is as follows:

[0063] Ionone is an aroma component mainly contained in raspberry (Rubus idaeus), yellow carrot (Daucus carota subsp. Sativus), sweet almond (Prunus dulcis), and mint (Menta), and mainly produces a floral aroma.

[0064] The method for obtaining α-ionone in the present invention is not particularly limited, and α-ionone may be isolated from plants containing the above-mentioned ionones, chemically synthesized using a known production method, or used as a commercially available product.

[0065] The novel skin barrier repairing agent in the external skin preparation of the present invention is characterized by containing α-ionone as an active ingredient acting on keratinocytes.

[0066] The content of the α-ionone as a skin barrier repair agent in the external skin preparation is 0.0001% to 5% by mass, more preferably 0.001% to 2% by mass, further preferably 0.01% to 1.5% by mass, and particularly preferably 0.1% to 1% by mass.

[0067] In addition to containing α-ionone as an active ingredient of the skin barrier repair agent, the skin external preparation of the present invention may contain other additives such as excipients and carriers, and ingredients commonly added to cosmetics may be appropriately used as needed.

[0068] Ingredients commonly added to cosmetics include, but are not limited to, aqueous solvents, oily ingredients, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, water-soluble polymers, thickeners, preservatives, UV absorbers, metal ion blockers, amino acids, organic amines, polymer emulsions, pH regulators, neutralizers, skin nutrients, vitamins, antioxidants, antioxidant adjuvants, fragrances, etc.

[0069] Examples of the aqueous solvent include water, alcohol, a moisturizer, and a mixture thereof.

[0070] As water, water used in cosmetics, quasi-drugs, etc. can be used, and for example, purified water, ion-exchanged water, tap water, etc. can be used. Depending on the purpose, the aqueous phase may further contain a water-soluble alcohol.

[0071] Examples of the water-soluble alcohol include at least one selected from lower alcohols, polyols, polyol polymers, glycol alkyl ethers, glycol ether esters, glycerol monoalkyl ethers, sugar alcohols, monosaccharides, oligosaccharides, polysaccharides, and derivatives thereof.

[0072] Examples of the lower alcohol include ethanol, propanol, isopropanol, isobutanol, and tert-butanol.

[0073] Examples of the polyol include diols (e.g., ethylene glycol, propylene glycol, trimethylene glycol, 1,2-butanediol, 1,3-butanediol, tetramethylene glycol, 2,3-butanediol, pentamethylene glycol, 2-butene-1,4-diol, hexanediol, octanediol, etc.); triols (e.g., glycerol, trimethylolpropane, etc.); tetraols (e.g., pentaerythritol, etc.); pentaols (e.g., xylitol, etc.); and hexaols (e.g., sorbitol, mannitol, etc.).

[0074] Examples of the polyol polymer include diethylene glycol, dipropylene glycol, triethylene glycol, polypropylene glycol, tetraethylene glycol, diglycerol, triglycerol, and tetraglycerol.

[0075] Examples of the glycol alkyl ethers include ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono-2-methylhexyl ether, ethylene glycol isopentyl ether, ethylene glycol benzyl ether, ethylene glycol isopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, and ethylene glycol dibutyl ether; and diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol butyl ether, diethylene glycol methyl ethyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, and dipropylene glycol butyl ether.

[0076] Examples of the glycol ether esters include ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether acetate, ethylene glycol monophenyl ether acetate, ethylene glycol diadipate, ethylene glycol disuccinate, diethylene glycol monoethyl ether acetate, diethylene glycol monobutyl ether acetate, propylene glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate, propylene glycol monopropyl ether acetate, and propylene glycol monophenyl ether acetate.

[0077] Examples of the glycerol monoalkyl ether include squalane alcohol, selachyl alcohol, and batyl alcohol.

[0078] Examples of sugar alcohols include sorbitol, maltitol, maltotriose, mannitol, sucrose, erythritol, glucose, fructose, amylolytic sugars, maltose, xylose, amylolytic sugar reducing alcohols, tetrahydrofurfuryl alcohol, POE-tetrahydrofurfuryl alcohol, POP-butyl ether, POP·POE-butyl ether, tripolyoxypropylene glycerol ether, POP-glycerol ether, POP-glycerol ether phosphate, and POP·POE-pentaerythritol ether.

[0079] Examples of the monosaccharide include at least one selected from the group consisting of trisaccharides (e.g., D-glyceraldehyde, dihydroxyacetone, etc.), tetrasaccharides (e.g., D-erythrose, D-erythrulose, D-threose, erythritol, etc.), pentoses (e.g., L-arabinose, D-xylose, L-lyxose, D-arabinose, D-ribose, D-ribulose, D-xylulose, L-xylulose, etc.), hexoses (e.g., D-glucose, D-talose, D-psicose, D-galactose, D- fructose, L-galactose, L-mannose, D-tagatose, etc.), heptose (e.g., aldoheptose, heptulose, etc.), octose (e.g., octulose, etc.), deoxy sugars (e.g., 2-deoxy-D-ribose, 6-deoxy-L-galactose, 6-deoxy-L-mannose, etc.), amino sugars (e.g., D-glucosamine, D-galactosamine, sialic acid, muramic acid, etc.), uronic acids (e.g., D-glucuronic acid, L-guluronic acid, D-galacturonic acid, L-iduronic acid, etc.), etc.

[0080] Examples of the oligosaccharide include at least one selected from sucrose, umbelliferyl, lactose, psyllium, and α,α-trehalose.

[0081] Examples of the polysaccharide include at least one selected from the group consisting of cellulose, quince seeds, chondroitin sulfate, starch, galactan, dermatan sulfate, glycogen, gum arabic, heparan sulfate, hyaluronic acid, tragacanth gum, keratan sulfate, chondroitin, xanthan gum, mucin sulfate, guar gum, dextran, locust bean gum, succinoglycan, and caroninic acid.

[0082] Examples of other polyols include at least one selected from polyoxyethylene methyl glucoside (Glucam E-10), polyoxypropylene methyl glucoside (Glucam P-10), and the like.

[0083] Examples of moisturizing agents include propylene glycol, glycerin, 1,3-butylene glycol, dipropylene glycol, xylitol, sorbitol, maltitol, chondroitin sulfate, hyaluronic acid, mucin sulfate, calonic acid, atelocollagen, sodium lactate, bile acid salts, dl-pyrrolidone carboxylate, alkylene oxide derivatives, short-chain soluble collagen, diglycerol (EO) PO adducts, roxburghii extract, yarrow extract, and sweet clover extract.

[0084] Examples of the oily component include liquid oils, solid oils, waxes, hydrocarbons, higher fatty acids, higher alcohols, synthetic ester oils, silicone oils, etc. In this specification, the term "oily component" includes oils and components soluble in oils.

[0085] Examples of the liquid oil include avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, peach kernel oil (almond oil), wheat germ oil, camellia oil, castor oil, linseed oil, safflower oil, cottonseed oil, perilla seed oil, soybean oil, peanut oil, tea seed oil, ginkgo oil, rice bran oil, Chinese tung oil, Japanese tung oil, jojoba oil, germ oil, and triglycerin.

[0086] Examples of solid fats and oils include cocoa butter, coconut oil, horse oil, hydrogenated coconut oil, palm oil, beef tallow, sheep oil, hydrogenated beef tallow, palm kernel oil, lard, beef bone fat (oil), wood wax kernel oil, hydrogenated oil, neatsfoot oil, wood wax, and hydrogenated castor oil.

[0087] Examples of waxes include beeswax, candelilla wax, cotton wax, carnauba wax, bayberry wax, insect wax (white wax), spermaceti wax, montan wax, rice bran wax, lanolin, kapok wax, lanolin acetate, liquid lanolin, sugarcane wax, lanolin fatty acid isopropyl ester, hexyl laurate, reduced lanolin, jojoba wax, hard lanolin, shellac wax, POE lanolin alcohol ether, POE lanolin alcohol acetate, POE cholesterol ether, lanolin fatty acid polyethylene glycol ester, and POE hydrogenated lanolin alcohol ether.

[0088] Examples of the hydrocarbon oil include liquid paraffin, ozokerite, squalane, pristane, paraffin, ceresin, squalene, vaseline, and microcrystalline wax.

[0089] Examples of higher fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, undecenoic acid, tallic acid, isostearic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA).

[0090] Examples of higher alcohols include straight-chain alcohols (e.g., lauryl alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, oleyl alcohol, cetearyl alcohol, etc.); branched-chain alcohols (e.g., monostearyl glyceryl ether (batyl alcohol), 2-decyltetradecyl alcohol, lanolin alcohol, cholesterol, phytosterols, hexyldodecanol, isostearyl alcohol, octyldodecanol, etc.); and the like.

[0091] Examples of the synthetic ester oil include hydrogenated polydecene, isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearate, ethylene glycol di-2-ethylhexanoate, dipentaerythritol fatty acid ester, N-alkyl ethylene glycol monoisostearate, neopentyl glycol dicaprate, diisostearyl malate, glyceryl di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, tetra-2-ethylhexanoate Pentaerythritol esters, tri-2-ethylhexanoin, tricaprylin, triisopalmitin, trimethylolpropane triisostearate, cetyl ethylhexanoate, 2-ethylhexyl palmitate, trimyristin, tri-2-heptylundecanoin, castor oil fatty acid methyl ester, oleyl oleate, acetylated glycerol, 2-heptylundecanoin palmitate, diisobutyl adipate, 2-octyldodecyl N-lauroyl-L-glutamate, di-2-heptylundecanoin, ethyl laurate, di-2-ethylhexyl sebacate, 2-hexyldecyl myristate, 2-hexyldecyl palmitate, 2-hexyldecyl adipate, diisopropyl sebacate, 2-ethylhexyl succinate, triethyl citrate, etc.

[0092] Examples of the silicone oil include polydimethylsiloxane, methyl hydrogen polysiloxane, methylphenyl polysiloxane, stearyloxymethyl polysiloxane, polyether-modified organopolysiloxane, fluoroalkyl / polyoxyalkylene co-modified organopolysiloxane, alkyl-modified organopolysiloxane, terminal-modified organopolysiloxane, fluorine-modified organopolysiloxane, amino-modified organopolysiloxane, PEG-10 polydimethylsiloxane, silicone rubber, acrylic polysiloxane, trimethylsiloxysilicate, and silicone RTV rubber.

[0093] As the oil component used in combination with hydrogenated polyisobutene, it is preferred to have an oil component that has poor compatibility with hydrogenated polyisobutene and is easily volatile. In this way, during application, the volatile oil component will evaporate, and due to its poor compatibility, hydrogenated polyisobutene will remain on the skin to form a film, giving a firming feeling.

[0094] Examples of the anionic surfactant include fatty acid soaps (e.g., sodium laurate, sodium palmitate, etc.); higher alkyl sulfate salts (e.g., sodium lauryl sulfate, potassium lauryl sulfate, etc.); alkyl ether sulfate salts (e.g., triethanolamine POE-lauryl sulfate, sodium POE-lauryl sulfate, etc.); N-acyl sarcosine (e.g., sodium lauroyl sarcosinate, etc.); higher fatty acid amide sulfonates (e.g., sodium N-myristoyl-N-methyl taurate, sodium coconut oil fatty acid methyl taurate, sodium lauryl methyl taurate, etc.); phosphate ester salts (sodium POE-oleyl ether phosphate, POE-stearyl ether phosphate, etc.); sulfosuccinates (e.g., sodium di-2-ethylhexyl sulfosuccinate, sodium monolauroyl monoethanolamide polyoxyethylene sulfosuccinate, lauryl polypropylene glycol sulfonate, etc.); succinate, etc.); alkylbenzene sulfonates (e.g., sodium linear dodecylbenzenesulfonate, triethanolamine linear dodecylbenzenesulfonic acid, linear dodecylbenzenesulfonic acid, etc.); higher fatty acid ester sulfates (e.g., sodium hydrogenated coconut oil fatty acid glyceride sulfate, etc.); N-acyl glutamates (e.g., monosodium N-lauroyl glutamate, disodium N-stearoyl glutamate, monosodium N-myristoyl-L-glutamate, etc.); sulfated oils (e.g., Turkey red oil, etc.); POE-alkyl ether carboxylic acids, POE-alkyl allyl ether carboxylates, α-olefin sulfonates, higher fatty acid ester sulfonates, secondary alcohol sulfates, higher fatty acid alkanolamide sulfates, sodium lauroyl monoethanolamide succinate, di-trolamine N-palmitoyl aspartate, sodium caseinate, potassium cetyl phosphate, etc.

[0095] Examples of the cationic surfactant include alkyltrimethylammonium salts (e.g., stearyltrimethylammonium chloride, lauryltrimethylammonium chloride, etc.); alkylpyridinium salts (e.g., cetylpyridinium chloride, etc.); distearyldimethylammonium chloride, dialkyldimethylammonium salts; poly(N,N″-dimethyl-3,5-methylenepiperidinium) chloride; alkyl quaternary ammonium salts; alkyldimethylbenzylammonium salts; alkylisoquinolinium salts; dialkylmorpholinium salts; POE-alkylamines; alkylamine salts; polyamine fatty acid derivatives; amyl alcohol fatty acid ester derivatives; benzalkonium chloride; benzethonium chloride, etc.

[0096] Examples of the amphoteric surfactant include imidazoline-based amphoteric surfactants (e.g., 2-undecyl-N,N,N-(hydroxyethylcarboxymethyl)-2-imidazolinium sodium, 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethoxy disodium salt, etc.); betaine-based surfactants (e.g., 2-heptadecanyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, lauryldimethylaminoacetic acid betaine, alkyl betaine, amide betaine, sulfobetaine, etc.); and the like.

[0097] Examples of the lipophilic nonionic surfactant include sorbitan fatty acid esters (e.g., sorbitan monooleate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan sesquioleate, sorbitan trioleate, sorbitan penta-2-ethylhexanoic acid diglyceryl, sorbitan tetra-2-ethylhexanoic acid diglyceryl, etc.); polyglycerol; Fatty acid glycerides (e.g., cottonseed oil fatty acid glyceride, erucic acid glyceride, sesquioleic acid glyceride, monostearate glyceride, α,α'-oleyl pyroglutamate glyceride, glyceryl malate monostearate, etc.); propylene glycol fatty acid esters (e.g., propylene glycol monostearate, etc.); hydrogenated castor oil derivatives; glyceryl alkyl ethers; PEG-10 polydimethylsiloxane, glyceryl stearate, PPG-13-decyltetradecyl alcohol polyether-24, PEG-5 glyceryl stearate, etc.

[0098] Examples of the hydrophilic nonionic surfactant include: POE-sorbitan fatty acid esters (e.g., POE-sorbitan monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate, POE-sorbitan tetraoleate, etc.); POE-sorbitan fatty acid esters (e.g., POE-sorbitan monolaurate, POE-sorbitan monooleate, POE-sorbitan pentaoleate, POE-sorbitan monostearate, etc.); POE-glycerol fatty acid esters (e.g., POE-glycerol monostearate, etc.); , POE-glyceryl monoisostearate, POE-glyceryl triisostearate, POE-monooleate, etc.); POE-fatty acid esters (for example, POE-distearate, POE-mono-dioleate, ethylene glycol distearate, etc.); POE-alkyl ethers (for example, POE-lauryl ether, POE-oleyl ether, POE-stearyl ether, POE-behenyl ether, POE-2-octyldodecyl ether, POE-cholestanol ether, etc.); Pluronic type (for example, Pluronic, etc.); POE·POP-alkyl ethers (for example, POE·POP- Cetyl ether, POE·POP-2-decyltetradecyl ether, POE·POP-monobutyl ether, POE·POP-hydrogenated lanolin, POE·POP-glycerol ether, etc.); tetra-POE·tetra-POP-ethylenediamine condensate (for example, Tetronic, etc.); POE-castor oil hydrogenated castor oil derivatives (for example, POE-castor oil, POE-hydrogenated castor oil, POE-hydrogenated castor oil monoisostearate, POE-hydrogenated castor oil triisostearate, POE-hydrogenated castor oil monopyroglutamic acid monoisostearate diester, POE-hydrogenated castor oil maleic acid Esters, etc.); POE-beeswax lanolin derivatives (for example, POE-sorbitol beeswax, etc.); alkanolamides (for example, coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropyl alcoholamide, etc.); POE-propylene glycol fatty acid esters; POE-alkylamines; POE-fatty acid amides; sucrose fatty acid esters; alkyl ethoxydimethylamine oxide; trioleyl phosphate; oleth-10, PEG-100 stearate, methoxy PEG / PPG-25 / 4 polydimethicone, polysorbate-60, PEG-40 stearate, sucrose stearate. Examples of natural water-soluble polymers include: plant polymers (e.g., gum arabic, tragacanth gum, galactan, guar gum, carob gum, karaya gum, carrageenan, pectin, agar, quince seeds, algae colloid (brown algae extract), starch (rice, corn, potato, wheat), glycyrrhizic acid); microbial polymers (e.g., xanthan gum, glucan, succinylglucan, pullulan, etc.); animal polymers (e.g., collagen, casein, albumin, gelatin, etc.); and the like.

[0099] Examples of semi-synthetic water-soluble polymers include starch-based polymers (e.g., carboxymethyl starch, methylhydroxypropyl starch, etc.); cellulose-based polymers (methylcellulose, ethylcellulose, methylhydroxypropyl cellulose, hydroxyethyl cellulose, sodium cellulose sulfate, hydroxypropyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, crystalline cellulose, cellulose powder, etc.); alginic acid-based polymers (e.g., sodium alginate, propylene glycol alginate, etc.), PEG-240 / HDI copolymer bis-decyltetradecyl alcohol polyether-20, acrylic acid (esters) / C10-30 alkyl acrylate crosspolymer, etc.

[0100] Examples of the thickener include carbomer, acrylates / C10-30 alkyl acrylate crosspolymer, sodium acrylate / sodium acryloyldimethyl taurate copolymer, acrylates / cetylethoxy (20) itaconate copolymer, acrylates / cetylethoxy (20) methacrylate copolymer, acrylates / tetradecylethoxy (25) acrylate copolymer, acrylates / octadecylethoxy (20) itaconate copolymer, acrylates / octadecylethoxy (20) methacrylate copolymer, acrylates / octadecylethoxy (50) acrylate copolymer, acrylates / VA crosspolymer, PAA (polyacrylic acid), sodium acrylate / ethylene isodecanoate crosspolymer, carbomer (polyacrylic acid) and its sodium salt and other polyacrylic acid thickeners.

[0101] Examples of the preservative include aromatic preservatives such as phenoxyethanol, benzyl alcohol, methylparaben, and p-hydroxyacetophenone.

[0102] Examples of the ultraviolet absorber include benzoic acid ultraviolet absorbers (e.g., para-aminobenzoic acid (hereinafter abbreviated as PABA), PABA monoglyceride, N,N-dipropoxy PABA ethyl ester, N,N-diethoxy PABA ethyl ester, N,N-dimethyl PABA ethyl ester, N,N-dimethyl PABA butyl ester, N,N-dimethyl PABA ethyl ester, etc.); anthranilic acid ultraviolet absorbers (e.g., homomenthyl N-acetylanthranilate, etc.); salicylic acid ultraviolet absorbers (e.g., amyl salicylate, menthyl salicylate, homomenthyl salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropylphenyl salicylate, etc.); ); cinnamic acid ultraviolet absorbers (for example, octyl methoxycinnamate, ethyl 4-isopropylcinnamate, methyl 2,5-diisopropylcinnamate, ethyl 2,4-diisopropylcinnamate, methyl 2,4-diisopropylcinnamate, propyl p-methoxycinnamate, isopropyl p-methoxycinnamate, isoamyl p-methoxycinnamate, octyl p-methoxycinnamate (2-ethylhexyl p-methoxycinnamate), 2-ethoxyethyl p-methoxycinnamate, cyclohexyl p-methoxycinnamate, ethyl α-cyano-β-phenylcinnamate, 2-ethylhexyl α-cyano-β-phenylcinnamate, mono-2-ethylhexanoyl-di-p-methoxycinnamate glyceryl); Benzophenone-based UV absorbers (e.g., 2,4-dihydroxybenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone, 2,2'-dihydroxy-4,4'-dimethoxybenzophenone, 2,2',4,4'-tetrahydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-phenylbenzophenone, 2-ethylhexyl-4'-phenyl-benzophenone-2-carboxylate, 2-hydroxy-4-n-octyloxybenzophenone, 4-hydroxy-3-carboxybenzophenone, etc.); 3-(4'-methylbenzylidene)-d,l-camphor, 3- ... Benzyl-d,l-camphor; 2-phenyl-5-methylbenzoxazole; 2,2'-hydroxy-5-methylphenylbenzotriazole; 2-(2'-hydroxy-5'-tert-octylphenyl)benzotriazole; 2-(2'-hydroxy-5'-methylphenylbenzotriazole; dibenzylhydrazine; dianisolemethane; 4-methoxy-4'-tert-butyldibenzoylmethane; 5-(3,3-dimethyl-2-nitro-norbornyl)-3-pentan-2-one, dimorpholinopyridazinone; 2-ethylhexyl-2-cyano-3,3-diphenylacrylate; 2,4-bis-{[4-(2-ethylhexyloxy)-2-hydroxy]-phenyl}-6-(4-methoxyphenyl)-(1,3,5)-triazine, etc.

[0103] Examples of metal ion blocking agents include 1-hydroxyethane-1,1-diphosphonic acid, 1-hydroxyethane-1,1-diphosphonic acid tetrasodium salt, disodium edetate (2 sodium EDTA), trisodium edetate (3 sodium EDTA), tetrasodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, phosphoric acid, citric acid, ascorbic acid, succinic acid, edetic acid, and trisodium ethylenediaminehydroxyethyltriacetate.

[0104] Examples of amino acids include neutral amino acids (e.g., threonine, cysteine, etc.), basic amino acids (e.g., hydroxylysine, etc.), and amino acid derivatives include sodium acyl sarcosinate (sodium lauroyl sarcosinate), acyl glutamate, sodium acyl β-alanine, glutathione, and pyrrolidone carboxylic acid.

[0105] Examples of the organic amine include monoethanolamine, diethanolamine, triethanolamine, morpholine, triisopropanolamine, 2-amino-2-methyl-1,3-propanediol, and 2-amino-2-methyl-1-propanol.

[0106] Examples of the polymer emulsion include acrylic resin emulsion, polyethyl acrylate emulsion, acrylic resin solution, polyalkyl acrylate emulsion, polyvinyl acetate resin emulsion, and natural rubber latex.

[0107] Examples of the pH adjuster include buffers such as lactic acid-sodium lactate, citric acid-sodium citrate, and succinic acid-sodium succinate.

[0108] Examples of the neutralizing agent include potassium hydroxide, sodium hydroxide, aminomethylpropanol, and arginine.

[0109] Examples of vitamins include vitamins A, B1, B2, B6, C, E and derivatives thereof, pantothenic acid and derivatives thereof, and biotin.

[0110] Examples of the antioxidant include tocopherols, butylated hydroxytoluene, butylated hydroxyanisole, and gallic acid esters.

[0111] Examples of the antioxidant aid include phosphoric acid, citric acid, ascorbic acid, maleic acid, malonic acid, succinic acid, fumaric acid, cephalin, hexametaphosphate, phytic acid, and ethylenediaminetetraacetic acid.

[0112] Test example

[0113] Experimental Example 1. Effect of α-ionone on the proliferation of HaCaT cells

[0114] The CCK8 (Cell Counting Kit-8) assay confirmed the effect of α-ionone on promoting the proliferation of HaCaT cells. HaCaT cells are immortalized human keratinocytes derived from normal adult skin. They possess biological properties similar to primary keratinocytes and retain epidermal differentiation capacity. They are often used as an in vitro surrogate model for keratinocyte-related epidermal biology research.

[0115] HaCaT cells were collected at 5×10 3 Cells were seeded at a density of 100 cells / well in a 96-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (1-400 μM) were then added for 24 hours (Figure 1, top) or 48 hours (Figure 1, bottom). After the culture period, 10 μl of CCK-8 solution was added to each well, and the culture plate was incubated in the incubator for 1 hour. After the incubation period, the absorbance OD value of each well at 450 nm was measured using a microplate reader. The proliferation rate of HaCaT cells was calculated based on the percentage of the OD value of each experimental group relative to the OD value of the control group. The results are shown in Figure 1. The data in the figure are Mean ± SD from three repeated experiments. ** p<0.01vs blank control group.

[0116] As shown in Figure 1, α-ionone can effectively promote HaCaT cell proliferation within a dose range of 1-200 μM, with the strongest pro-proliferative effect occurring at a dose of 50 μM. At doses above 100 μM, the pro-proliferative effect is not further enhanced but rather weakened (Figure 1). Therefore, α-ionone can directly act on HaCaT cells to promote cell proliferation, and its effective dose range is 1-200 μM.

[0117] Experimental Example 2. Effect of α-ionone on promoting HaCaT cell migration

[0118] The cell scratch assay was used to confirm the effect of α-ionone in promoting HaCaT cell migration.

[0119] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 10 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator. When the cells grew to 90% confluence, the culture medium was aspirated and a 20 μl sterile pipette tip was used to scratch perpendicularly to the cell plane to create a "scratch". After washing with PBS, 2 ml of complete culture medium containing different concentrations of α-ionone (1, 10, 25, 50 μM) was added and cultured for another 24 h. The cells were observed and photographed under an inverted microscope at different times after scratching (0, 12, 24 h). The scratch area was calculated using Image J software, and the cell migration rate was calculated according to the following formula:

[0120] Cell migration rate (%) = (initial scratch area - scratch area after culture) / initial scratch area × 100%,

[0121] The results are shown in Figure 2. The data in the figure are Mean ± SD (n = 4). * p<0.05vs vehicle control group.

[0122] According to the results in Figure 2, α-ionone promoted HaCaT cell migration in a dose-dependent manner within the dose range of 10-50 μM. Although the cell migration rate increased to a certain extent at the lower dose of 1 μM, it did not reach a statistically significant difference compared with the blank control group, suggesting that α-ionone is basically ineffective at doses below 1 μM. Compared with the 25 μM dose group, the cell migration rate of the 50 μM dose group of α-ionone did not increase significantly, but instead decreased slightly at 24 hours, suggesting that further increasing the dose of α-ionone did not further enhance its effect of promoting cell migration (Figure 2). Therefore, α-ionone can promote HaCaT cell migration, and its effective dose range is 10-50 μM.

[0123] Experimental Example 3. α-ionone effectively attenuates the inhibitory effect of norepinephrine (NE) on HaCaT cell migration

[0124] The stress hormone norepinephrine (NE) is known to inhibit keratinocyte migration, thereby delaying skin wound healing. To examine the effect of α-ionone on NE inhibition of HaCaT cell migration, confluent HaCaT cell monolayers were scratched and then treated with 10 μM NE or NE (10 μM) + α-ionone (25, 50 μM). The cells were cultured for 24 hours and observed under an inverted microscope and photographed at different times after scratching (0, 12, and 24 hours). The scratch area and cell migration rate were calculated using Image J software. The results are shown in Figure 3. Data in the figure are Mean ± SD (n = 4). * p<0.05vs blank control group, # p<0.05vs NE alone group.

[0125] According to the results in Figure 3, NE (10 μM) alone can significantly inhibit the migration of HaCaT cells. The cell migration rate of the NE+α-ionone (25, 50 μM) experimental group was significantly higher than that of the NE alone group, indicating that α-ionone can effectively weaken the inhibitory effect of NE on cell migration.

[0126] Experimental Example 4. Effect of α-ionone on HAS-2 expression in HaCaT cells

[0127] The effect of α-ionone alone on HAS-2 gene expression was confirmed in normal cultured HaCaT cells.

[0128] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 10 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Then, different concentrations of α-ionone (1, 10, 25, 50, 100 μM) were added for 9 hours. The expression level of HAS-2 mRNA in the cells was determined by real-time quantitative PCR (qPCR). The results are shown in Figure 4. The data in the figure are Mean ± SD from three repeated experiments. ** p<0.01.

[0129] The results in Figure 4 show that α-ionone increased HAS-2 gene expression levels in a dose-dependent manner within the 10-50 μM dose range. At a higher dose of 100 μM, its pharmacological activity was not further enhanced but weakened. Therefore, α-ionone has a promoting effect on HAS-2 expression in HaCaT cells, and its effective dose range is 10-50 μM.

[0130] Experimental Example 5. Effect of α-ionone on promoting HA synthesis in HaCaT cells

[0131] In this experiment, the inventors quantitatively measured the HA level in the culture supernatant of HaCaT cells after 24 hours of stimulation with α-ionone.

[0132] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 100 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (25, 50 μM) were added for 24 hours. The cell culture supernatant was collected and the HA concentration in the cell culture supernatant was determined by ELISA. The results are shown in Figure 5. The data in the figure are Mean ± SD from three repeated experiments. ** p<0.01.

[0133] According to the results in Figure 5, α-ionone (25, 50 μM) can significantly increase the HA level in the HaCaT cell culture supernatant after 24 hours, indicating that α-ionone promotes the synthesis of HA by increasing the expression of hyaluronan synthase HAS-2. Therefore, this result further confirms the pharmacological activity of α-ionone in inducing the expression of hyaluronan synthase.

[0134] Experimental Example 6. Effect of α-ionone on the inhibition of HAS-2 expression by dexamethasone (DEX)

[0135] Glucocorticoids reduce HA synthesis by inhibiting HAS-2 expression in keratinocytes. This is one of the primary pathological mechanisms by which glucocorticoids and psychological stress lead to skin atrophy and skin barrier dysfunction. DEX is a potent synthetic glucocorticoid. Because it is more active and stable than endogenous glucocorticoids, it is often used as a tool to study the effects of glucocorticoids. DEX-stimulated HaCaT cells are often used as a cell model for skin stress. This study investigated the effect of α-ionone on DEX-induced inhibition of HAS-2 expression in a DEX-induced skin stress cell model.

[0136] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 10 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (1, 10, 25, 50, 100 μM) were added for 3 hours, followed by 1 μM DEX for 6 hours. The expression level of HAS-2 mRNA in the cells was determined by qPCR. The results are shown in Figure 6. The data in the figure are Mean ± SD from three repeated experiments. * p<0.05, ** p<0.01.

[0137] According to the results in Figure 6, DEX (1 μM) can significantly inhibit the expression of HAS-2 in HaCaT cells, and α-ionone can effectively reduce the inhibitory effect of DEX on HAS-2 expression in the dose range of 10-50 μM, indicating that it can increase the HAS-2 expression level of keratinocytes under the action of stress hormones. Therefore, α-ionone has an antagonistic effect on the inhibition of HAS-2 expression by DEX, and its effective dose range is 10-50 μM.

[0138] Experimental Example 7. Effect of α-ionone on the expression of HBD-2 in HaCaT cells

[0139] The effect of α-ionone alone on HBD-2 gene expression was confirmed in normal cultured HaCaT cells.

[0140] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 10 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (1, 10, 25, 50, 100 μM) were added for 9 hours, and the mRNA expression level of HBD-2 in the cells was determined by qPCR. The results are shown in Figure 7. The data in the figure are Mean ± SD from three repeated experiments. ** p<0.01.

[0141] According to the results in FIG7 , α-ionone can effectively increase the expression level of HBD-2 gene in the dose range of 10-50 μM. These results confirm the pharmacological activity of α-ionone in inducing HBD-2 expression.

[0142] Experimental Example 8. Effect of α-ionone on HBD-2 secretion by HaCaT cells

[0143] In this experiment, the inventors quantitatively measured the HBD-2 level in the culture supernatant of HaCaT cells after 24 hours of stimulation with α-ionone.

[0144] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 100 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (25, 50 μM) were added for 24 hours. The cell culture supernatant was collected and the concentration of HBD-2 in the cell culture supernatant was determined by ELISA. The results are shown in Figure 8. The data in the figure are Mean ± SD from three repeated experiments. * p<0.05.

[0145] According to the results in Figure 8, α-ionone (25, 50 μM) can significantly increase the level of HBD-2 in the culture supernatant of HaCaT cells after 24 hours of treatment, indicating that α-ionone promotes the synthesis and secretion of HBD-2 through the expression of HBD-2. Therefore, this result further confirms the pharmacological activity of α-ionone in inducing HBD-2 expression.

[0146] Experimental Example 9. Effect of α-ionone on the inhibition of HBD-2 expression in HaCaT cells by dexamethasone (DEX)

[0147] Glucocorticoids inhibit HBD-2 expression in keratinocytes, which is one of the key pathological mechanisms by which glucocorticoid use and psychological stress increase susceptibility to microbial infection and impair epidermal barrier function. This study further validated the effect of α-ionone on DEX-induced inhibition of HBD-2 expression in a DEX-induced skin stress cell model.

[0148] HaCaT cells were collected at 2×10 5 Cells were seeded at a density of 10 cells / well in a 6-well cell culture plate and cultured in a cell culture incubator for 24 hours. Different concentrations of α-ionone (1, 10, 25, 50, 100 μM) were added for 3 hours, followed by 1 μM DEX for 6 hours. The mRNA expression level of HBD-2 in the cells was determined by qPCR. The results are shown in Figure 9. The data in the figure are Mean ± SD from three repeated experiments. ** p<0.01.

[0149] As shown in Figure 9, DEX (1 μM) significantly inhibits HBD-2 expression in HaCaT cells. α-ionone can effectively alleviate the inhibitory effect of DEX on HBD-2 expression within a dose range of 10-50 μM and increase HBD-2 expression in keratinocytes exposed to stress hormones. This result indicates that α-ionone antagonizes the inhibitory effect of DEX on HBD-2 expression, with an effective dose range of 10-50 μM.

[0150] Example

[0151] Example 1. Lotion

[0152] According to the following formula, a lotion containing α-ionone as an active ingredient for skin barrier repair is prepared using conventional cosmetic manufacturing processes.

[0153] Example 2. Lotion

[0154] According to the following formula, a lotion containing α-ionone as an active ingredient for skin barrier repair is prepared using conventional cosmetic manufacturing processes.

[0155] Example 3. Emulsion

[0156] According to the following formula, a lotion containing α-ionone as a skin barrier repair active ingredient is prepared using conventional cosmetic manufacturing processes.

[0157] Example 4. Emulsion

[0158] According to the following formula, a lotion containing α-ionone as a skin barrier repair active ingredient is prepared using conventional cosmetic manufacturing processes.

[0159] Example 5. Facial Cream

[0160] According to the following formula, a facial cream containing α-ionone as a skin barrier repair active ingredient is prepared using conventional cosmetic manufacturing processes.

[0161] Example 6. Facial Cream

[0162] According to the following formula, a facial cream containing α-ionone as a skin barrier repair active ingredient is prepared using conventional cosmetic manufacturing processes.

[0163] Example 7. Hydrogel

[0164] According to the following formula, a hydrogel containing α-ionone as a skin barrier repair active ingredient was prepared.

[0165] Take an appropriate amount of deionized water and heat it to 85°C in a water bath. Slowly add carbomer U20 powder to the water and stir to dissolve evenly. Then, add 10% arginine to neutralize it. Weigh p-hydroxyacetophenone and 1,2-hexanediol, add them to an appropriate amount of deionized water, slightly heat to dissolve, and then add them to the carbomer U20 gel and stir evenly to obtain a hydrogel matrix. When the hydrogel matrix is ​​cooled to below 40°C, add α-ionone and stir evenly to obtain α-ionone hydrogel.

[0166] Example 8. Hydrogel

[0167] According to the following formulation and the manufacturing method described in Example 7, a hydrogel containing α-ionone as a skin barrier repair active ingredient was prepared.

[0168] Effect test example

[0169] 1. Test Principle

[0170] In human clinical trials, subjective evaluation and objective skin index evaluation are mainly used to analyze the improvement effect of the subjects' skin condition before and after using the test samples. The results are more intuitive and suitable for the efficacy evaluation of finished or semi-finished cosmetics.

[0171] This experiment recruited healthy people to form a test group, created a model of damaged skin barrier on the flexor side of the arm through tape stripping, and measured the changes in physiological indicators such as skin stratum corneum moisture content and transepidermal water loss rate (TEWL) before and after the application of cosmetics on the flexor side of the arm with damaged skin barrier, so as to evaluate the efficacy of cosmetics in promoting skin barrier repair.

[0172] 2. Materials and Instruments

[0173] 2.1 Test samples:

[0174] How to use the test sample: Apply the test product to the area of ​​the arm where the tape has been peeled, once a day, approximately 2 mg / cm 2 An area where the tape was stripped but no product was applied was set up as a model control.

[0175] 2.2 Instruments

[0176] Stratum corneum moisture content tester (Corneometer CM 825, C&K, Germany); transepidermal water loss rate tester (Vapometer, Delfin, Finland).

[0177] 3. Test methods

[0178] 3.1 Subjects

[0179] According to the Declaration of Helsinki, the selection of subjects must adhere to medical and ethical standards for human testing. All testing must be voluntary and the subjects must sign an informed consent form before testing. Before signing the informed consent form, the tester must inform the subject of the purpose of the test, possible benefits, potential risks and issues, and related rights and obligations.

[0180] 3.1.1 Number of subjects

[0181] Target number of subjects: 32

[0182] 3.1.2 Inclusion criteria:

[0183] Age 18 to 65 years old;

[0184] The skin on the curved side of the forearm is healthy, with even skin tone, and without scars or other conditions that may affect the test;

[0185] Able to return for visits and undergo skin testing as required.

[0186] 3.1.3 Exclusion criteria:

[0187] During pregnancy or breastfeeding;

[0188] Have a history of allergies to skin care products or cleaning products, or are allergic to certain ingredients in the test products and supporting samples;

[0189] People with serious systemic diseases and other skin diseases, such as lupus erythematosus, psoriasis, etc.;

[0190] The test area has received cosmetic treatment;

[0191] Participated in other clinical trials currently or before the start of this study.

[0192] 3.1.4 Exit (fall-off) criteria:

[0193] The subject was lost to follow-up or voluntarily requested to withdraw;

[0194] Poor compliance, failure to use samples on time and in the correct amount, and failure to make return visits as required;

[0195] Other cosmetics similar to the test samples were used during the research;

[0196] The emergence of new diseases throughout the study period that directly affect the assessment of clinical status, including skin diseases;

[0197] Those who developed serious illness during the entire study period;

[0198] Subjects who became pregnant during the study;

[0199] Those who cannot tolerate this study;

[0200] Serious adverse reactions (SAEs) occurred during the entire study period.

[0201] 3.2 Test indicators

[0202] 3.2.1 Water content of the skin stratum corneum

[0203] The moisture content of the skin's stratum corneum is measured using the Corneometer CM 825 skin moisture probe in the German CK Multifunctional Skin Tester MPA 580. This probe measures the moisture content of the skin's surface layer using the capacitance method. The dielectric constant of water in the skin's surface layer is much higher than that of other substances. Changes in the dielectric constant of the skin's surface layer are primarily due to changes in its moisture content. By measuring the dielectric constant of the skin's surface layer, the moisture content of the skin's surface layer can be analyzed. The measured value is a relative value expressed in CU (Corneometer Units). A higher value indicates a higher moisture content in the skin's surface layer.

[0204] 3.2.2 Transepidermal water loss rate (TEWL)

[0205] Transepidermal water loss (TEWL) is tested using a Vapometer from Delfin, Finland. The top of the vapometer has a circular hole that forms a closed cavity when in contact with the skin. A high-precision humidity sensor in the cavity records changes in the air humidity within the cavity. The rate of change in air humidity over time within a specific period of time after contact with the skin can be used to calculate the rate of water loss through the skin. The unit of measurement is g / (m 2 The lower the value, the lower the rate of water loss from the skin surface and the better the skin barrier function.

[0206] 3.3 Measurement location

[0207] Four 3cm*3cm areas were selected on the flexor side of the arm as test areas, with at least 1cm between each area. From Day 0 to Day 2, tape was applied five times daily for three consecutive days. According to the randomization table, three of the areas served as sample areas and were treated with the test product once daily. The other area served as a model control and was not treated with the test product.

[0208] 3.4 Test Environment

[0209] The subjects exposed the skin of the flexed side of their arms and the skin was measured after equilibration in an environment with a temperature of 20-22°C and a humidity of 40-60% for 20 minutes.

[0210] 3.5 Testing Process

[0211] On day 0, the subjects signed the informed consent form and were screened. Qualified subjects were enrolled. Four 3cm*3cm areas were marked on the curved side of the arm as test areas. The test areas were kept in a constant temperature and humidity environment for 20 minutes. The moisture content and TEWL of the stratum corneum of the test areas were measured. The test areas were then stripped with tape, with each area stripped 5 times. On days 1-2, the test areas were also stripped with tape every day, with each area stripped 5 times. After the tape stripping was completed, the product was applied to the sample area at approximately 2mg / cm 2 , the model control area does not use the product. On days 3-7, the product is applied to the sample area at approximately 2mg / cm2 per day. 2 The model control area did not use the product. The stratum corneum moisture content and TEWL of the test area were measured after tape stripping on the second day and on the seventh day after equilibration in a constant temperature and humidity environment for 20 minutes.

[0212] 3.6 Adverse Reactions

[0213] If any adverse reaction occurs, volunteers should immediately stop using the test product and visit the research center for examination by a physician. The physician will then determine whether to terminate sample use based on the circumstances. Any adverse reactions and other related events that occur during the test should be recorded and included in the report.

[0214] 3.7 Data Analysis

[0215] The mean and standard deviation of each test index for all subjects at each time point were calculated. The barrier repair rate of each test group was calculated based on the change rate of the TEWL value of each test group after the end of treatment (day 7) relative to the TEWL value on day 0 (baseline TEWL value) and the TEWL value after tape stripping on day 2. The calculation formula is as follows: Barrier repair rate (%) = (T2-T7) / (T2-T0)×100%,

[0216] Wherein, T2 is the TEWL value measured after tape stripping on day 2, T7 is the TEWL value of each experimental group after the completion of treatment on day 7, and T0 is the baseline TEWL value before tape application measured on day 0.

[0217] SPSS 22.0 software was used for statistical analysis of the data sets, using one-way ANOVA followed by Dunnett's post hoc test for pairwise comparisons.

[0218] 4 Test results

[0219] This trial recruited 32 eligible subjects, including 3 males and 29 females, aged 24 to 58 years (mean age 41.2 ± 10.7 years). No adverse reactions were reported during the trial. The results of transepidermal water loss (TEWL) and stratum corneum moisture content in the test area of ​​each trial group are shown below.

[0220] 4.1 Transepidermal water loss rate (TEWL value) of each test group

[0221] The changing trend of TEWL values ​​in the test areas of each test group is shown in Figure 10:

[0222] ( # Compared with the normal skin control group, p<0.05, * Compared with the blank hydrogel matrix group (p<0.05)

[0223] The test results of TEWL values ​​of the test areas of each test group showed that TEWL values ​​of each test area increased from 5 g / m2 to 10 g / m2 after 3 consecutive days of tape peeling. 2 The baseline value of h increased to ~13g / m2 after tape stripping on the second day 2 h, indicating that tape stripping caused significant damage to the epidermal barrier function. After using the test product once daily for seven days after tape stripping, the TEWL values ​​of the test areas in each test group were significantly lower than those after tape stripping on the second day, indicating that the damaged epidermal barrier had been repaired to a certain extent. The TEWL values ​​in the areas treated with the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" decreased to a greater extent, and these TEWL values ​​were statistically significantly different from those in the test group using the "blank hydrogel matrix" and the model group that only underwent tape stripping but did not use the test samples. This suggests that the use of the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" can accelerate the repair of the epidermal barrier damaged by tape adhesion.

[0224] 4.2 Water content of the skin stratum corneum in each experimental group

[0225] The changing trend of moisture content in the stratum corneum of the test area of ​​each test group is shown in Figure 11:

[0226] ( * Compared with the blank hydrogel matrix group, p<0.05, # Compared with the normal skin control group (p<0.05)

[0227] The test results of the stratum corneum water content in the test areas of each test group showed that 3 consecutive days of tape stripping did not lead to significant changes in the stratum corneum water content in the test areas. However, it led to a significant decrease in the stratum corneum water content in the model group (only tape stripping without the use of test samples) on the 7th day. Compared with the baseline level of stratum corneum water content, the average value decreased by 9.13%, indicating that the transepidermal water loss rate caused by tape stripping increased, leading to dry skin. Although the average stratum corneum water content of the skin areas using the test samples "blank hydrogel matrix", "0.1% α-ionone hydrogel", and "1% α-ionone hydrogel" also decreased compared with the baseline value, the decrease was smaller, decreasing by 7.71%, 3.99%, and 2.81%, respectively. The measured values ​​of stratum corneum moisture content in the area using the test sample "blank hydrogel matrix" showed no significant difference compared with the model group (p≥0.05), but the measured values ​​of stratum corneum moisture content in the area using the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" were significantly higher than those in the model group (p<0.05), which further suggests that the use of the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" can accelerate the repair of the epidermal barrier damaged by tape adhesion, thereby increasing the moisture content of the stratum corneum in the test area.

[0228] 4.3 Barrier repair rate of each experimental group

[0229] The barrier repair rate calculated based on the change rate of TEWL values ​​of each test group is shown in Figure 12:

[0230] ( * Compared with the blank hydrogel matrix group, p<0.05, ** Compared with the blank hydrogel matrix group, p<0.01)

[0231] The results of the barrier repair rate calculated based on the rate of change of the TEWL values ​​of each test group further showed that there was no significant difference in the barrier repair rate of the test group using the test sample "blank hydrogel matrix" compared with the model group that did not use the test sample, while the barrier repair rate of the test group using the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" were significantly increased compared with the model group and the "blank hydrogel matrix" group. This result indicates that the use of the test samples "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" can indeed accelerate the repair of the epidermal barrier damaged by tape adhesion, and thus has a repair effect.

[0232] 5 Conclusion

[0233] The test results of this experiment showed that the test sample "blank hydrogel matrix" had no repair effect, while "0.1% α-ionone hydrogel" and "1% α-ionone hydrogel" had repair effect.

[0234] Cosmetic effect testing

[0235] In order to verify the effects of the cosmetics of the present invention, the present inventors conducted effect experiments on the lotions, emulsions, and creams of Examples 1 to 6, as follows.

[0236] A. Test sample:

[0237] B. Test Methods

[0238] B.1 Subjects

[0239] B.1.1 Target number of people: 30.

[0240] B.1.2 Inclusion criteria

[0241] Age 30-50 years old;

[0242] The skin tone of the arm is even, without folliculitis, eczema and other skin problems that may affect the test;

[0243] Agree to have the skin on your arm stripped with tape;

[0244] No other products were used on the test area during the test;

[0245] Informed consent, able to use the product and complete the corresponding tests as required by the researcher.

[0246] B.1.3 Exclusion criteria

[0247] Pregnant or breastfeeding women;

[0248] Those with severe systemic diseases and currently taking medication for systemic diseases;

[0249] The test area has been subjected to skin treatment, cosmetic or other tests that may affect the results;

[0250] People with allergic diseases or those who are allergic to cosmetics;

[0251] Participated in other clinical trials currently or before the start of this study.

[0252] B.1.4 Shedding standard

[0253] The subject was lost to follow-up or voluntarily requested to withdraw;

[0254] Poor compliance, failure to use samples on time and in the correct amount, and failure to make return visits as required;

[0255] Other cosmetics similar to the test samples were used during the research;

[0256] The emergence of new diseases throughout the study period that directly affect the assessment of clinical status, including skin diseases;

[0257] Those who developed serious illness during the entire study period;

[0258] Subjects who became pregnant during the study;

[0259] Those who cannot tolerate this study;

[0260] Serious adverse reactions (SAEs) occurred during the entire study period.

[0261] B.2 Instruments

[0262] Transdermal water loss rate tester (C&K, Germany, Tewameter);

[0263] Skin stratum corneum moisture content tester (Corneometer, C&K, Germany).

[0264] B.3 Test indicators

[0265] B.3.1 Skin stratum corneum moisture content: Skin stratum corneum moisture content is one of the evaluation indicators for the repair efficacy of cosmetics. A higher value indicates a higher stratum corneum moisture content. Following tape stripping to create a model with slightly damaged skin barrier, if the stratum corneum moisture content in the product-treated area is significantly higher than in the untreated area or the control area, this indicates that the product has a repairing effect.

[0266] B.3.2 TEWL (Transepidermal Water Loss) is one of the evaluation indicators of the repair efficacy of cosmetics. The higher the value of this indicator, the faster the rate of water loss from the skin surface and the poorer the skin barrier function. After the skin is subjected to tape stripping to create a model with slightly damaged skin barrier, if the TEWL of the area where the product was applied is significantly lower than that of the area where the product was not applied or the control area, it can be considered that the product has a repair effect.

[0267] B.4 Measurement location

[0268] The measurement locations for skin stratum corneum moisture content and TEWL were both on the inner arm where the skin barrier was slightly damaged by tape stripping.

[0269] B.5 Test Environment

[0270] The subjects exposed their skin at the measurement site and equilibrated it in an environment with a temperature of 20-22°C and a humidity of 40-60% for 20 minutes before skin measurement.

[0271] B.6 Testing Process

[0272] (1) On day 0, the subjects signed the informed consent form and were screened. The qualified subjects were enrolled in the study. The skin on the flexor side of the arm was washed with soapy water and 11 3cm×3cm areas were marked as test areas. The skin was kept in a constant temperature and humidity environment for 20 minutes. The moisture content and TEWL of the stratum corneum of the test areas were measured. Then, 10 of the test areas were peeled with tape. Each area was peeled 10 times. The other area was not peeled with tape as a blank control. After 15 minutes, the moisture content and TEWL of the stratum corneum of the test areas were measured. The product was quantitatively applied to the 9 areas peeled with tape according to the random table, with each area receiving approximately 2mg / cm 2 , one tape-stripped area was left unused with no product as a model control; 6 hours later, the stratum corneum moisture content and TEWL of the test area were measured.

[0273] (2) On days 1-9, apply the product twice a day to the sample area, approximately 2 mg / cm2 each time 2 , no product is used in the model control and blank control areas.

[0274] (3) On the 1st, 3rd, 7th, and 10th days, wash the skin on the flexor side of the arm with soapy water. After equilibration in a constant temperature and humidity environment for 20 minutes, measure the moisture content of the stratum corneum and TEWL of the test area.

[0275] B.7 Adverse Reactions

[0276] If any adverse reaction occurs, volunteers should immediately stop using the test product and visit the research center for examination by a physician. The physician will then determine whether to terminate sample use based on the circumstances. Any adverse reactions and other related events that occur during the test should be recorded and included in the report.

[0277] B.8 Data Analysis

[0278] Calculate the mean and standard deviation of each test index for all subjects at each time point. For the moisture content of the stratum corneum, calculate the change value and change rate relative to before peeling. The calculation formula is as follows: Change value (Δ value) = T n -T0

[0279] Where, T0 is the measured value of the measurement index before peeling

[0280] T n ——Measurement value of the measurement indicator at the nth return visit

[0281] n——number of return visits

[0282] For TEWL, calculate the repair rate for each location using the following formula:

[0283] Where, Tc is the TEWL value immediately after the tape is peeled off.

[0284] Ts——TEWL value after using the product

[0285] Tb——TEWL value before tape peeling

[0286] SPSS 22.0 software was used to perform statistical analysis of the data. One-way analysis of variance was used for inter-group tests, and the LSD method was used for pairwise comparisons. All tests were two-tailed, with an α level of 0.05.

[0287] C test results

[0288] This trial recruited 30 subjects, all with Chinese skin, including 5 males and 25 females, aged 30-50 years (mean age 38.0 ± 5.5 years). All 30 subjects completed all tests. The test results are as follows.

[0289] C.1 Water content of stratum corneum

[0290] The moisture content of the skin's stratum corneum is one of the indicators used to evaluate the repair efficacy of cosmetics. Higher values ​​indicate higher moisture content in the stratum corneum. In a model with mildly damaged skin barrier caused by tape stripping, if the moisture content of the stratum corneum in the product-treated area is significantly higher than in the untreated area or the control area, it indicates that the product has a repairing effect.

[0291] The measured values ​​of the moisture content of the stratum corneum of the subjects' skin are shown in Figure 13; in Figure 13, a: 3 water samples, b: 3 lotion samples, c: 3 cream samples; the measured values ​​of the model control and the blank control were subjected to significance analysis, #p<0.05; the measured values ​​of sample 2, sample 3 and sample 1 were subjected to significance analysis, *p<0.05.

[0292] The changes in the moisture content of the subjects' skin stratum corneum compared with before peeling are shown in Figure 14: In Figure 14, a: 3 water samples, b: 3 lotion samples, c: 3 cream samples; the change values ​​of the model control and the blank control were analyzed for significance, #p<0.05; the change values ​​of sample 2, sample 3 and sample 1 were analyzed for significance, *p<0.05.

[0293] The results of the rate of change of the average value of the moisture content of the skin stratum corneum during the test period relative to that before peeling are shown in Table 1.

[0294] Table 1 Results of the change rate of the average moisture content of the stratum corneum relative to that before peeling

[0295] Result description:

[0296] (1) There was no significant difference in the moisture content of the stratum corneum of all test areas before tape stripping.

[0297] (2) Immediately after tape stripping, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, water 1, water 2, water 3, lotion 1, lotion 2, lotion 3, cream 1, cream 2, and cream 3 areas were -3.4%, 128.6%, 116.9%, 127.5%, 123.3%, 130.4%, 126.4%, 127.5%, 127.9%, 129.1%, and 124.1%, respectively. The moisture content of the skin stratum corneum in the area stripped with tape was significantly increased compared with that in the blank control, and there was no significant difference between the areas stripped with tape.

[0298] (3) After tape stripping and 6 hours after the first use of the product, the change rates of the skin stratum corneum moisture content in the blank control, model control, water 1, water 2, and water 3 areas were -4.6%, 71.0%, 84.0%, 94.3%, and 92.4%, respectively.

[0299] On the first day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, water 1, water 2, and water 3 areas were -4.6%, 29.2%, 25.0%, 33.3%, and 34.7%, respectively.

[0300] On the third day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, water 1, water 2, and water 3 areas were -4.7%, -36.2%, -27.7%, -27.0%, and -23.1%, respectively.

[0301] On the 7th day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, water 1, water 2, and water 3 areas were -2.0%, -24.7%, -18.0%, -12.0%, and -2.1%, respectively; the moisture content of the skin stratum corneum in water 2 was higher than that in water 1, and the moisture content of the skin stratum corneum in water 3 was significantly higher than that in water 1; the change value of the moisture content of the skin stratum corneum in water 2 was higher than that in water 1, and the change value of the moisture content of the skin stratum corneum in water 3 was significantly higher than that in water 1.

[0302] On the 10th day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, water 1, water 2, and water 3 areas were -0.6%, -13.5%, -1.6%, 7.2%, and 16.5%, respectively; the moisture content of the skin stratum corneum in water 2 was higher than that in water 1, and the moisture content of the skin stratum corneum in water 3 was significantly higher than that in water 1; the change value of the moisture content of the skin stratum corneum in water 2 was higher than that in water 1, and the change value of the moisture content of the skin stratum corneum in water 3 was significantly higher than that in water 1.

[0303] (4) After tape stripping and 6 hours after the first use of the product, the change rates of the skin stratum corneum moisture content in the blank control, model control, lotion 1, lotion 2, and lotion 3 areas were -4.6%, 71.0%, 91.8%, 98.6%, and 111.1%, respectively.

[0304] On the first day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, lotion 1, lotion 2, and lotion 3 areas were -4.6%, 29.2%, 35.3%, 38.1%, and 39.7%, respectively.

[0305] On the third day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, lotion 1, lotion 2, and lotion 3 areas were -4.7%, -36.2%, -23.7%, -24.9%, and -22.6%, respectively.

[0306] On the 7th day of the experiment, the change rates of the skin stratum corneum moisture content in the blank control, model control, lotion 1, lotion 2, and lotion 3 areas were -2.0%, -24.7%, -6.5%, -2.8%, and 3.9%, respectively; the skin stratum corneum moisture content of lotion 2 was higher than that of lotion 1, and the skin stratum corneum moisture content of lotion 3 was significantly higher than that of lotion 1.

[0307] On the 10th day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, lotion 1, lotion 2, and lotion 3 areas were -0.6%, -13.5%, 15.3%, 18.6%, and 25.9%, respectively; the moisture content of the skin stratum corneum in lotion 2 was higher than that in lotion 1, and the moisture content of the skin stratum corneum in lotion 3 was significantly higher than that in lotion 1.

[0308] (5) After tape stripping and 6 hours after the first use of the product, the change rates of the skin stratum corneum moisture content in the blank control, model control, cream 1, cream 2, and cream 3 areas were -4.6%, 71.0%, 92.3%, 95.8%, and 95.5%, respectively.

[0309] On the first day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, cream 1, cream 2, and cream 3 areas were -4.6%, 29.2%, 36.2%, 38.7%, and 43.5%, respectively.

[0310] On the third day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, cream 1, cream 2, and cream 3 areas were -4.7%, -36.2%, -22.1%, -21.6%, and -14.4%, respectively.

[0311] On the 7th day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, Cream 1, Cream 2, and Cream 3 areas were -2.0%, -24.7%, -1.1%, 3.1%, and 12.8%, respectively; the moisture content of the skin stratum corneum in Cream 2 was higher than that in Cream 1, and the moisture content of the skin stratum corneum in Cream 3 was significantly higher than that in Cream 1. The change value of the moisture content of the skin stratum corneum in Cream 2 was higher than that in Cream 1, and the change value of the moisture content of the skin stratum corneum in Cream 3 was significantly higher than that in Cream 1.

[0312] On the 10th day of the experiment, the change rates of the moisture content of the skin stratum corneum in the blank control, model control, Cream 1, Cream 2, and Cream 3 areas were -0.6%, -13.5%, 21.6%, 25.2%, and 35.7%, respectively; the moisture content of the skin stratum corneum in Cream 2 was higher than that in Cream 1, and the moisture content of the skin stratum corneum in Cream 3 was significantly higher than that in Cream 1. The change value of the moisture content of the skin stratum corneum in Cream 2 was higher than that in Cream 1, and the change value of the moisture content of the skin stratum corneum in Cream 3 was significantly higher than that in Cream 1.

[0313] C.2 Transepidermal water loss rate (TEWL)

[0314] Transepidermal water loss (TEWL) is one of the indicators used to evaluate the repair efficacy of cosmetics. Higher values ​​indicate faster water loss from the skin surface and poorer skin barrier function. In a model where the skin barrier is mildly damaged by tape stripping, if the TEWL in the area treated with the product is significantly lower than that in the untreated area or the control area, it indicates that the product has repaired skin.

[0315] The TEWL measurement values ​​of the test areas of the subjects are shown in Figure 15; in Figure 15, a: 3 water samples, b: 3 lotion samples, c: 3 cream samples; the measurement values ​​of the model control and the blank control were analyzed for significance, #p<0.05; the measurement values ​​of Sample 2, Sample 3 and Sample 1 were analyzed for significance, *p<0.05.

[0316] The TEWL repair rates of the subjects are shown in Figure 16; in Figure 16, a: 3 water samples, b: 3 lotion samples, c: 3 cream samples; the repair rates of sample 2, sample 3 and sample 1 were analyzed for significance, *p<0.05.

[0317] Result description:

[0318] (1) There was no significant difference in TEWL among all test areas before tape stripping.

[0319] (2) Immediately after tape stripping, the TEWL of the tape stripped area was significantly higher than that of the blank control, but there was no significant difference between the tape stripped areas.

[0320] (3) After tape stripping and 6 hours after the first use of the product, the TEWL repair rates of the model control, water 1, water 2, and water 3 were 1.7%, 8.0%, 4.1%, and 2.2%, respectively.

[0321] On the first day of the experiment, the TEWL repair rates of the model control, water 1, water 2, and water 3 were 8.9%, 22.5%, 21.3%, and 19.5%, respectively.

[0322] On the third day of the experiment, the TEWL repair rates of the model control, water 1, water 2, and water 3 were 41.3%, 49.0%, 47.3%, and 51.4%, respectively.

[0323] On the 7th day of the experiment, the TEWL repair rates of the model control, water 1, water 2, and water 3 were 74.4%, 83.2%, 82.5%, and 82.6%, respectively.

[0324] On the 10th day of the experiment, the TEWL repair rates of the model control, water 1, water 2, and water 3 were 79.4%, 84.8%, 87.2%, and 87.2%, respectively.

[0325] (4) After tape stripping and 6 hours after the first use of the product, the TEWL repair rates of the model control, emulsion 1, emulsion 2, and emulsion 3 were 1.7%, 7.4%, 4.5%, and 4.2%, respectively.

[0326] On the first day of the experiment, the TEWL repair rates of the model control, emulsion 1, emulsion 2, and emulsion 3 were 8.9%, 20.0%, 11.4%, and 15.0%, respectively.

[0327] On the third day of the experiment, the TEWL repair rates of the model control, emulsion 1, emulsion 2, and emulsion 3 were 41.3%, 48.5%, 39.3%, and 51.2%, respectively.

[0328] On the 7th day of the experiment, the TEWL repair rates of the model control, emulsion 1, emulsion 2, and emulsion 3 were 74.4%, 75.6%, 76.2%, and 83.3%, respectively.

[0329] On the 10th day of the experiment, the TEWL repair rates of the model control, emulsion 1, emulsion 2, and emulsion 3 were 79.4%, 83.8%, 80.1%, and 84.0%, respectively.

[0330] (5) Six hours after tape stripping and the first application of the product, the TEWL repair rates of the model control, Cream 1, Cream 2, and Cream 3 were 1.7%, 6.9%, -7.6%, and -0.6%, respectively. On the first day of the experiment, the TEWL repair rates of the model control, Cream 1, Cream 2, and Cream 3 were 8.9%, 14.9%, 6.6%, and 13.5%, respectively.

[0331] On the third day of the experiment, the TEWL repair rates of the model control, cream 1, cream 2, and cream 3 were 41.3%, 43.1%, 42.3%, and 44.3%, respectively.

[0332] On the 7th day of the experiment, the TEWL repair rates of the model control, cream 1, cream 2, and cream 3 were 74.4%, 75.8%, 80.0%, and 82.7%, respectively.

[0333] On the 10th day of the experiment, the TEWL repair rates of the model control, cream 1, cream 2, and cream 3 were 79.4%, 80.5%, 81.5%, and 85.9%, respectively.

[0334] D Conclusion

[0335] In summary, the repairing effect of the test sample Water 2 is better than that of Water 1, and Water 3 has a more significant repairing effect than Water 1; the repairing effect of Emulsion 2 is better than that of Emulsion 1, and Emulsion 3 has a more significant repairing effect than Emulsion 1; the repairing effect of Cream 2 is better than that of Cream 1, and Cream 3 has a more significant repairing effect than Cream 1.

[0336] Industrial applicability

[0337] The inventors discovered that α-ionone regulates keratinocyte function, promoting HaCaT cell proliferation and migration, and attenuating the inhibitory effect of norepinephrine on cell migration. α-ionone also induces HAS-2 and HBD-2 gene expression, promoting HA and HBD-2 synthesis in HaCaT cells, and alleviating the inhibitory effect of dexamethasone on HAS-2 and HBD-2 expression. These findings suggest that α-ionone may have the potential to promote the repair of skin damage.

[0338] Therefore, the present invention can use α-ionone as an active ingredient of a skin barrier repair agent in a skin topical preparation, and can be used as a repair agent for skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress. It can be used in skin topical preparations, especially cosmetics, to repair the skin barrier, for example, to moisturize the skin, promote the repair of damaged skin, and promote the repair of damaged skin barrier caused by topical glucocorticoids and stress.

Claims

1. A skin barrier function repairing agent in an external skin preparation, It is characterized in that Contains α-ionone as an active ingredient that acts on keratinocytes.

2. The skin barrier function repairing agent according to claim 1, It is characterized in that The α-ionone acts as a keratinocyte migration promoter.

3. The skin barrier function repairing agent according to claim 1, It is characterized in that The α-ionone acts as a keratinocyte proliferation promoter.

4. The skin barrier function repairing agent according to claim 1, It is characterized in that The α-ionone acts as an expression promoter of hyaluronan synthase-2 of keratinocytes.

5. The skin barrier function repairing agent according to claim 1, It is characterized in that The α-ionone acts as an expression promoter of human β-defensin-2 in keratinocytes.

6. The skin barrier function repairing agent according to claim 1, It is characterized in that Used as a repair agent for skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress.

7. The skin barrier function repairing agent according to claim 1, It is characterized in that The external skin preparation is a cosmetic.

8. Use of α-ionone in the preparation of cosmetics for promoting skin barrier repair, wherein the promotion of skin barrier repair is achieved by at least one promoting effect selected from the group consisting of promoting the migration of keratinocytes, promoting the proliferation of keratinocytes, promoting the expression of HAS-2 of keratinocytes, and promoting the expression of HBD-2 of keratinocytes.

9. Use of α-ionone in the preparation of cosmetics suitable for skin in a state of skin atrophy and / or skin barrier dysfunction caused by the use of glucocorticoids and / or psychological stress.

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