JP2026014469A - Vitamin D metabolism promoter - Google Patents

Abstract

The object of the present invention is to provide an active ingredient that is derived from a natural product, has excellent biological safety, promotes vitamin D metabolism in the epidermis, and allows vitamin D to function more effectively.
The present invention uses an extract of Lilium concolor or a subspecies thereof as an active ingredient.
[Selection diagram] None

Description

The present invention relates to an agent that promotes vitamin D metabolism in the epidermis and stimulates epidermal cell differentiation.

Vitamin D, an essential component for the body, is broadly divided into two types: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). Vitamin D3 is known to be ingested orally from food and produced in skin exposed to sunlight, and is therefore used as an ingredient in nutritional supplements.

However, it is known that much of the vitamin D3 used in nutritional supplements is in an inactive form. It has been shown that this inactive vitamin D3 (cholecalciferol) becomes physiologically active when it is hydroxylated in the liver or other organs and converted to calcifediol or calcitriol in the body (Non-Patent Document 1). Therefore, there has been a need for technology to more efficiently convert this vitamin D3 into its active form so that it retains its physiological activity in the body.

Toshiyuki Sakaki, Vitamins (Japan), 93 (11), 469-477 (2019)

The inventors conducted extensive research in light of these problems with the prior art. As a result, they discovered that extracts of Lilium caryophyllus or its subspecies have the effect of activating enzymes (CYP2R1, CYP27B1, CYP27A1) that convert vitamin D3 into its active form, the effect of promoting vitamin D metabolism in the epidermis, and the effect of promoting epidermal cell differentiation.

The present invention is a vitamin D metabolism promoter containing an extract of Lilium concolor or a subspecies thereof as an active ingredient.
The present invention relates to an epidermal cell differentiation promoter containing an extract of Lilium concolor or a subspecies thereof as an active ingredient.

The present invention promotes vitamin D metabolism in the epidermis by applying to the skin a preparation containing an extract of Lilium concolor or one of its subspecies as an active ingredient. As a result, vitamin D can be more efficiently utilized in the epidermis. Furthermore, the present invention suggests that promoting vitamin D metabolism in the epidermis promotes epidermal cell differentiation, promoting, for example, the formation of the cornified envelope of the stratum corneum (expression of involucrin, transglutaminase, etc.), NMF synthesis (expression of peptidylarginine deiminases), the secretion of antimicrobial peptides, and the expression of stratum corneum desquamating enzymes (expression of kallikreins, etc.). Therefore, the present invention is expected to maintain a strong barrier function while regulating cell turnover and promoting stratum corneum exfoliation, which restores the epidermis to a normal state, thereby restoring the epidermis to a normal state.

The present invention uses as an active ingredient an extract of Lilium concolor or a subspecies thereof, which belongs to the genus Lilium and belongs to the family Liliaceae. Examples of subspecies of Lilium include Lilium concolor Salisb. var. mutsuanum Makino, Lilium concolor var. pulchellum, Lilium concolor f. coridion, and Lilium concolor.

The parts of the lily of the valley that can be extracted in this invention include the whole plant, leaves, stems, roots, flowers, buds, and new shoots.

To prepare the extract, first, the part to be used for extraction is washed with water if necessary to remove any foreign matter, then either left as is or dried, and then shredded or crushed as necessary, and then brought into contact with the extraction solvent for extraction. Extraction can be carried out by contacting the part with the extraction solvent using standard methods such as the immersion method, but supercritical extraction methods can also be used instead of the immersion method.

Extraction solvents include water; lower alcohols such as methanol, ethanol, and propanol; polyhydric alcohols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, and glycerin; esters such as ethyl acetate, butyl acetate, and methyl propionate; ketones such as acetone and methyl ethyl ketone; ethers such as ethyl ether and isopropyl ether; and hydrocarbon solvents such as n-hexane, toluene, and chloroform, which may be used alone or in combination.

Among the above extraction solvents, hydrophilic solvents such as water, lower alcohols, or polyhydric alcohols are preferred in the present invention from the standpoint of the effectiveness of the resulting extract, as well as skin irritation, and because they can be widely applied to topical skin preparations (cosmetics, quasi-drugs, topical pharmaceuticals, etc.). Preferred examples of hydrophilic solvents include the use of water, lower alcohols (particularly ethanol), or polyhydric alcohols (particularly 1,3-butylene glycol and glycerin) alone, or a mixed solvent of water and lower alcohols (particularly ethanol), or a mixed solvent of water and polyhydric alcohols (particularly 1,3-butylene glycol and glycerin).

When using a mixed solvent, for example, if the solvent is water and a lower alcohol or polyhydric alcohol, the mixing ratio is preferably 1:5 to 25:1 by volume (same below).

When preparing the extract, there are no particular limitations on its pH, but it is generally preferable to keep it in the range of 3 to 9. In this sense, if necessary, the extraction solvent may be blended with an alkalinity adjuster such as sodium hydroxide, sodium carbonate, or potassium hydroxide, or an acidity adjuster such as citric acid, hydrochloric acid, phosphoric acid, or sulfuric acid to adjust the pH to the desired level.

Extraction conditions such as extraction temperature and extraction time vary depending on the type and pH of the solvent used. For example, if the solvent is water alone or a mixture of water and a lower alcohol or polyhydric alcohol, the extraction temperature is preferably in the range of 0°C to 80°C, and the extraction time is preferably in the range of 1 to 168 hours (1 hour to 1 week), and more preferably in the range of 1 to 120 hours (1 hour to 5 days).

To remove impurities and improve stability, the extract of the present invention may be subjected to one or a combination of two or more of the following treatments to remove impurities other than the active ingredients: activated carbon treatment, ion exchange resin treatment, synthetic adsorbent treatment, silica gel treatment, and recrystallization treatment.

The extract prepared as described above may be used as an ingredient in a composition for topical skin application as is after adjusting the pH to 3-8, or may be used at the desired concentration after concentrating under reduced pressure. The extract may also be dried using standard methods such as spray drying.

The extract of the present invention can also be incorporated into topical skin preparations (cosmetics, quasi-drugs, and topical pharmaceuticals), and examples of such preparations include, but are not limited to, emulsions, creams, lotions, essences, packs, lipsticks, foundations, sheet masks, liquid foundations, makeup press powders, blushers, face powders, facial cleansers, body shampoos, cleansing cosmetics such as soaps, hair growth products, and even bath additives.

When incorporated into a composition, the amount of the extracts of the present invention is generally in the range of 0.00001 to 5.0% by weight in terms of solid content of each extract.

When the extract of the present invention is incorporated into topical skin preparations (cosmetics, quasi-drugs, topical pharmaceuticals, etc.), ingredients used in the composition, such as oily ingredients, surfactants (synthetic and natural), moisturizers, thickeners, emulsifiers or emulsifier aids, preservatives/bactericides, powder ingredients, UV absorbers, antioxidants, sequestering agents, pigments, fragrances, anti-wrinkle agents, anti-pigmentation agents, and other physiologically active ingredients, can be incorporated as needed.

Oily ingredients include, for example, olive oil, jojoba oil, castor oil, soybean oil, rice oil, rice germ oil, coconut oil, palm oil, cocoa oil, meadowfoam oil, shea butter, tea tree oil, avocado oil, macadamia nut oil, bergamot oil, lavender oil, rose oil, bergamot oil, chamomile oil, and other plant-derived oils such as squalane; vitamin A oil; animal-derived oils such as mink oil and turtle oil; waxes such as beeswax, carnauba wax, rice wax, and lanolin; liquid paraffin, petrolatum, and paraffin wax. and squalane; fatty acids such as myristic acid, palmitic acid, stearic acid, oleic acid, isostearic acid, and cis-11-eicosenoic acid; higher alcohols such as lauryl alcohol, cetanol, pantothenyl alcohol, and stearyl alcohol; synthetic esters and synthetic triglycerides such as isopropyl myristate, isopropyl palmitate, butyl oleate, 2-ethylhexyl glyceride, and higher fatty acid octyldodecyl (e.g., octyldodecyl stearate).

Examples of surfactants include nonionic surfactants such as polyoxyethylene alkyl ethers, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene hydrogenated castor oil, and polyoxyethylene sorbitol fatty acid esters; fatty acid salts, alkyl sulfates, alkylbenzene sulfonates, polyoxyethylene alkyl ether sulfates, polyoxyethylene fatty amine sulfates, polyoxyethylene alkylphenyl ether sulfates, polyoxyethylene alkyl ether phosphates, α-sulfonated fatty acid alkyl ester salts, polyoxyethylene Examples of surfactants that can be used include anionic surfactants such as ethylene alkyl phenyl ether phosphates; cationic surfactants such as quaternary ammonium salts, primary to tertiary fatty amine salts, trialkylbenzylammonium salts, alkylpyridinium salts, 2-alkyl-1-alkyl-1-hydroxyethylimidazolinium salts, N,N-dialkylmorpholinium salts, and polyethylenepolyamine fatty acid amide salts; and amphoteric surfactants such as N,N-dimethyl-N-alkyl-N-carboxymethylammoniobetaine, N,N,N-trialkyl-N-alkyleneammoniocarboxybetaine, and N-acylamidopropyl-N',N'-dimethyl-N'-β-hydroxypropylammoniosulfobetaine.

Emulsifiers and emulsifier aids that can be used include, for example, stevia derivatives such as enzyme-treated stevia, saponin or its derivatives, casein or its salts (sodium, etc.), sugar and protein complexes, sucrose or its esters, lactose, soy-derived water-soluble polysaccharides, soy-derived protein and polysaccharide complexes, lanolin or its derivatives, cholesterol, stevia derivatives (enzyme-treated stevia, etc.), silicates (aluminum, magnesium, etc.), carbonates (calcium, sodium, etc.), saponin and its derivatives, lecithin and its derivatives (hydrogenated lecithin, etc.), lactic acid bacteria-fermented rice, lactic acid bacteria-fermented germinated rice, lactic acid bacteria-fermented grains (wheat, beans, millet, etc.), etc.

Humectants include, for example, glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, sorbitol, xylitol, sodium pyrrolidone carboxylate, etc., as well as sugars such as trehalose and raffinose, mucopolysaccharides (e.g., hyaluronic acid and its derivatives, hyaluronic acid fermentation liquid, chondroitin and its derivatives, heparin and its derivatives, etc.), elastin and its derivatives, collagen and its derivatives, collagen peptides, NMF-related substances, lactic acid, urea, higher fatty acid octyldodecyl, seaweed extract, estradiol, and various amino acids and their derivatives.

Thickeners include, for example, ingredients derived from brown algae, green algae, or red algae, such as alginic acid, agar, carrageenan, and fucoidan; polysaccharides such as pectin, pullulan, and aloe polysaccharide; gums such as tragacanth gum, locust bean gum, xanthan gum, and guar gum; cellulose derivatives such as carboxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose; synthetic polymers such as carboxyvinyl polymer, alkyl-modified carboxyvinyl polymer, polyvinyl alcohol, polyvinylpyrrolidone, and acrylic acid/methacrylic acid copolymer; hyaluronic acid and its derivatives; polyglutamic acid and its derivatives, and polyacrylic acid.

Examples of anti-inflammatory agents include allantoin, dipotassium glycyrrhizinate, monoammonium glycyrrhizinate, β-glycyrrhetinic acid, stearyl glycyrrhetinate, ε-aminocaproic acid, d-camphor, dl-camphor, zinc oxide, panthenol, pyridoxine hydrochloride, and riboflavin or its derivatives.

Examples of antiseptics and disinfectants include urea; benzoic acid or its salts, parahydroxybenzoic acid esters such as methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, and butyl parahydroxybenzoate; phenoxyethanol, dichlorophene, hexachlorophene, chlorhexidine hydrochloride, benzalkonium chloride, salicylic acid, sodium salicylate, zinc pyrithione, benzalkonium chloride, ethanol, undecylenic acid, phenols, and ammonium bromide. These include chylisoquinolinium, resorcinol, jamal (imidazolidinyl urea), isopropyl methylphenol, triclosan, trichlorocarbanide, trichlorohydroxydiphenol ether, hinokitiol, 1,2-pentanediol, propanediol, concentrated benzalkonium chloride solution 50, essential oils such as peppermint oil and eucalyptus oil, bark dry distillate, radish fermented liquid, ethanol derived from plants such as sugar cane and corn, or 1,3-butylene glycol.

Examples of cell activators include pantothenyl alcohol, menthol, dl-menthol, and gamma-oryzanol.

Anti-acne agents include, for example, sulfur, salicylic acid or its salts, photosensitizer No. 201, and pyridoxine dicaprylate.

Examples of powder ingredients include sericite, titanium oxide, talc, kaolin, bentonite, zinc oxide, magnesium carbonate, magnesium oxide, zirconium oxide, barium sulfate, silicic anhydride, mica, nylon powder, polyethylene powder, silk powder, cellulose-based powder, powder of grains (rice, wheat, corn, millet, etc.), and powder of beans (soybeans, adzuki beans, etc.).

Examples of ultraviolet absorbers include ethyl para-aminobenzoate, ethylhexyl para-dimethylaminobenzoate, amyl salicylate and its derivatives, 2-ethylhexyl para-methoxycinnamate, octyl cinnamate, oxybenzone, 2,4-dihydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-tert-butyl-4-methoxybenzoylmethane, 2-(2-hydroxy-5-methylphenyl)benzotriazole, urocanic acid, ethyl urocanate, and aloe extract.

Examples of antioxidants include butylhydroxyanisole, butylhydroxytoluene, propyl gallate, carotenoids such as astaxanthin, vitamin E and its derivatives (e.g., tocopherol acetate, tocopherol nicotinate), vitamin A and its derivatives (retinol palmitate, etc.), etc.

Examples of sequestering agents include disodium edetate, trisodium edetate, tetrasodium edetate, etidronic acid or its salts (e.g., tetrasodium etidronate), gluconic acid or its salts (e.g., sodium gluconate), diethylenetriaminepentaacetic acid or its salts (e.g., pentasodium diethylenetriaminepentaacetate), hydroxyethanediphosphonic acid or its salts (e.g., tetrasodium hydroxyethanediphosphonate), and phytic acid or its salts (e.g., sodium phytate).

Examples of anti-pigmentation agents include one or more selected from kojic acid or its derivatives, ascorbic acid or its derivatives, hydroquinone or its derivatives, ellagic acid and its derivatives, resorcinol derivatives, 4-methoxysalicylic acid potassium salt, vitamin E or its derivatives, nicotinic acid or its derivatives, magnolignan (5,5'-dipropyl-biphenyl-2,2'-diol), hydroxybenzoic acid and its derivatives, vitamin E and its derivatives, alpha-hydroxy acid, AMP (adenosine monophosphate, adenosine monophosphate), placenta extract, and linoleic acid.

Examples of the kojic acid derivatives include kojic acid esters such as kojic acid monobutyrate, kojic acid monocaprate, kojic acid monopalmitate, and kojic acid dibutyrate, kojic acid ethers, and kojic acid sugar derivatives such as kojic acid glucoside. Examples of the ascorbic acid derivatives include sodium L-ascorbic acid 2-phosphate, magnesium L-ascorbic acid 2-phosphate, sodium L-ascorbic acid 2-sulfate, and L-ascorbic acid 2-sulfate. ascorbic acid ester salts such as ammonium nitrate and magnesium nitrate; ascorbic acid sugar derivatives such as L-ascorbic acid-2-glucoside (2-O-α-D-glucopyranosyl-L-ascorbic acid) and L-ascorbic acid-5-glucoside (5-O-α-D-glucopyranosyl-L-ascorbic acid); 6-acylated products of these ascorbic acid sugar derivatives (the acyl group is a hexanoyl group, an octanoyl group, a decanoyl group, etc.); L-ascorbic acid tetraisopalmitate, ... Examples of the hydroquinone derivatives include arbutin (hydroquinone-β-D-glucopyranoside) and α-arbutin (hydroquinone-α-D-glucopyranoside). Examples of the resorcinol derivatives include 4-n-butylresorcinol and 4-isoamylresorcinol. Examples of 2,5-dihydroxybenzoic acid derivatives include 2,5-diacetoxybenzoic acid, 2-acetoxy-5-hydroxybenzoic acid, and 2-hydroxy-5-propionyloxybenzoic acid; examples of nicotinic acid derivatives include nicotinamide and benzyl nicotinate; examples of vitamin E derivatives include vitamin E nicotinate and vitamin E linoleate; and examples of α-hydroxy acids include lactic acid, malic acid, succinic acid, citric acid, and α-hydroxyoctanoic acid.

Examples of anti-wrinkle agents include vitamin A or its derivatives, vitamin E or its derivatives (tocopherol acetate, etc.), vitamin C or its derivatives (ascorbic acid glucoside, 3-O-ethyl ascorbic acid, ascorbic acid phosphate magnesium salt, etc.), pantothenyl alcohol, tranexamic acid, and niacinamide.

Furthermore, it is also possible to use ingredients derived from natural products such as the following plants or microorganisms in combination. For example, collagen or its hydrolysate, yeast extract or hydrolysate, lactic acid bacteria culture, grasses, cruciferous plants, Theaceae plants, Rosaceae plants, Paeoniaceae plants, Rutaceae plants, Amaranthaceae plants, Zosteraea plants, Leguminosae plants, Asteraceae plants, Fabaceae plants, Malvaceae plants, Gentianaceae plants, Lamiaceae plants, Nelumbosacaceae plants, Cucurbitaceae plants, Araliaceae plants, Solanaceae plants, Bignoniaceae plants, Actinidiaceae plants, Mulberry plants, Iridaceae plants, Campanulaceae plants, Oleaceae plants, Actinidiaceae plants, Mulberry plants, Rhamnaceae plants, Orchidaceae plants, and Anacardiaceae plants. Examples of such extracts include extracts of one or more plants selected from the family Garcinia, Valencaceae, Rutaceae, Myrtaceae, Liliaceae, Crassulaceae, Cupressaceae, Convolvulaceae, and Asparagaceae, or hydrolysates or fermented products thereof; extracts of one or more seaweeds selected from the family Laminaria, Mirrataceae, and Ulvulaceae, or hydrolysates or fermented products thereof; jellyfish (autolyzed products of moon jellyfish, Nomura's jellyfish, etc.); hydrolysates or fermented products of hyaluronic acid; and extracts of royal jelly, or hydrolysates or fermented products thereof.

As ingredients derived from plants of the Poaceae family, particularly preferred are rice leaf hydrolysate, rice extract hydrolysate, rice bran extract hydrolysate, germinated brown rice hydrolysate, rice fermentation liquid, sake lees extract derived from sake, and bamboo shoot skin extract from Madake or Moso bamboo. Furthermore, as ingredients derived from plants of the Brassicaceae family, white mustard extract or its hydrolysate, and white mustard fermentation product are preferred. Furthermore, as ingredients derived from plants of the Theaceae family, particularly preferred are green tea (Yabukita, Samidori, Asahi, Goko, Ujimidori, Kyomidori, Ujihikari, Samidori, Benifuuki, etc.) and black tea (Darjeeling, Assam, Ceylon, Earl Grey, honey-scented black tea, etc.). As ingredients derived from plants of the Rosaceae family, preferred are Damask rose flower extract, peach flower, leaf, or immature fruit extract, strawberry flower extract, cherry blossom flower or leaf extract, and apricot fruit or seed extract. Furthermore, as components derived from Paeoniaceae plants, extracts of peony root or flower and peony flower or root are preferred. As components derived from Amaranthaceae plants, Salicornia extract is particularly preferred. As components derived from Zosteraea plants, extracts of Zostera marina or Zostera kohlrabi are particularly preferred. As components derived from Leguminosae plants, extracts of white soybean or black soybean or their hydrolysates, fermented soy milk, adzuki bean extract, red clover extract, and pueraria lobata root extract are particularly preferred. As components derived from Asteraceae plants, burdock root extract, sunflower sprout extract, Arctium gracilis extract, arnica extract, and chamomile flower extract are particularly preferred. As components derived from Malvaceae plants, fermented products of hibiscus, rose of sharon, or hibiscus are preferred. As components derived from Gentianaceae plants, gentian extract is preferred. As components derived from Lamiaceae plants, perilla extract and barberry fruit extract are preferred. As components derived from plants of the Nelumbo family, particularly preferred are lotus flower or lotus seed extracts, or fermented lotus seed products. As components derived from plants of the Cucurbitaceae family, particularly preferred is loofah extract. As components derived from plants of the Araliaceae family, preferred is an extract or fermented product of Panax ginseng. As components derived from plants of the Solanaceae family, extracts of eggplant (long eggplant, water eggplant, rice eggplant, Kamo eggplant, etc.) are exemplified. As components derived from plants of the Bignoniaceae family, preferred is Pau d'Arco bark extract. As components derived from plants of the Actinidiaceae family, preferred is immature kiwi extract. As components derived from plants of the Moraceae family, preferred is Morus alba extract, mulberry fruit extract, or fig fruit or bark extract. As components derived from plants of the Rhamnaceae family, preferred is jujube fruit extract. Furthermore, as components derived from plants of the Iridaceae family, preferred is saffron. As components derived from plants of the Campanulaceae family, preferred is an extract or hydrolysate of Codonopsis globulus root. As components derived from plants of the Anacardiaceae family, preferred is mango fruit extract. As a component derived from a plant of the family Garciniae, mangosteen fruit extract is particularly preferred. Furthermore, as a component derived from a plant of the family Valenciaceae, cherimoya fruit extract is preferred. As a component derived from a plant of the family Rutaceae, Satsuma mandarin, bergamot fruit extract, grapefruit or banpeiyu fruit extract (including immature fruit), extracts containing flavonoids and their glycosides contained in plants such as grapefruit or hassaku, or Japanese pepper seed extract are preferred. As a component derived from a plant of the family Liliaceae, extracts of daylilies, daylilies, Casablanca lilies, Madonna lilies, or Japanese lilies are preferred. As a component derived from a plant of the family Crassulaceae, extracts or fermented products of Rhodiola rosea (Rhodiola rosea). As a component derived from a plant of the family Oleaceae, jasmine flower extract is particularly preferred. As a plant of the family Cupressaceae, juniper fruit extract is particularly preferred. As a component derived from a plant of the family Myrtaceae, guava leaf extract is particularly preferred. As an Orchidaceae plant, an extract of Bletilla striata root (Hakuoki) is particularly preferred. As a component derived from a Convolvulaceae plant, an extract of sweet potato or its fermentation product, or an extract of sweet potato shochu lees or its fermentation product is preferred. As a component derived from a plant of the Asparagaceae family, an extract of asparagus is preferred. As a component derived from seaweed of the Laminaceae family, a kelp extract is particularly preferred, as a component derived from seaweed of the Mirinaceae family, an extract of Katamen-kirinsai is preferred, and as a component derived from seaweed of the Ulva family, an extract of Ulva pertusa is particularly preferred. As a component derived from seaweed of the Funoriaceae family, an extract of Funori is particularly preferred.

Next, the present invention will be explained in more detail using production examples, formulation examples, and test examples, but the present invention is not limited to these. Note that, below, all parts mean parts by weight, and all % means % by weight.

Preparation Example 1. Preparation of extract (1)
A mixed solvent of 500 g of purified water and 500 g of 1.3-butylene glycol was added to 1.0 g of dried leaves and stems of Lilium tricolor (Lilium spp.) of the family Liliaceae, and the mixture was stirred at 80°C for 4 hours. The crude extract was then filtered to obtain 610 g of a pale yellow extract (solid content: 0.10%).

Preparation Example 2. Preparation of extract (2)
1000 g of purified water was added to 1.0 g of dried leaves and stems of Lilium tricolor (Lilium genus, Liliaceae) and stirred at 80°C for 4 hours. The crude extract was then filtered to obtain 600 g of a pale yellow extract (solid content: 0.08%).

Preparation Example 3. Preparation of extract (3)
The same procedure as in Production Example 1 was carried out, except that flowers were used instead of the leaves and stems of Lilium thunbergii, to obtain a pale yellow extract (solid concentration: 0.11%).

Preparation Example 4. Preparation of extract (4)
The same procedure as in Production Example 1 was carried out, except that the leaves and stems of Lilium persicae were used instead of Lilium moniliforme, to obtain a pale yellow extract (solid concentration: 0.09%).

Test Example 1. Evaluation of Gene Expression in Epidermal Keratinocytes Normal epidermal keratinocytes (NHEK) were seeded onto a 24-well plate (1 x ¹⁰ cells/well) and cultured at 37°C under 5% _CO2 and saturated steam. After 24 hours, the extract from Preparation Example 1 was added as a sample solution, and the epidermal keratinocytes were cultured under the same conditions. The sample solution was added to the culture medium to achieve a final concentration of 2.0%. A control group was prepared by adding 30% butylene glycol (hereinafter referred to as "30% BG") at the same concentration instead of the sample solution. After 48 hours of culture, cells from each test group were harvested with 0.5 mL of ISOGEN II reagent (Nippon Gene Co., Ltd.), and total RNA was purified according to standard methods. cDNA was synthesized by reverse transcription using a designated kit (PrimeScript RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara Bio Inc.)). Using the synthesized cDNA as a sample, the expression of various genes and the internal standard G3PDH gene were detected using the Thermal Cycler Dice® Real Time System Single (manufactured by Takara Bio Inc.) and SYBR® Premix Ex Taq™ II (Perfect Real Time) (manufactured by Takara Bio Inc.). The test results were obtained by comparing the expression levels of each gene in each test group when the expression level of the G3PDH gene was kept constant. In this test system, the expression level of each gene in the control group was set to 100, and the relative expression level of that gene in other test groups was calculated. In this test example, the genes encoding enzymes (CYP2R1, CYP27B1, CYP27A1) that convert vitamin D3 to its active form were focused on and their expression rates were evaluated.

The results of Test Example 1 are shown in Table 1.
[Table 1]

As shown in Table 1, it was confirmed that the extract of the present invention significantly increases the expression of genes encoding enzymes (CYP2R1, CYP27B1, CYP27A1) that convert vitamin D3 to its active form. This suggests that the present invention can promote vitamin D metabolism by converting vitamin D to its active form in the epidermis, thereby enabling vitamin D to function more effectively in the epidermis. Furthermore, since vitamin D is required for the differentiation of epidermal keratinocytes, it is suggested that it contributes to maintaining the homeostasis of the epidermal layer of the skin.

Test Example 2. Evaluation of Vitamin D Receptor Transcription Activity Normal human epidermal cells were prepared at a density of 1 x ¹⁰⁵ cells/mL using HuMedia-KG2 (Kurabo Industries, Ltd.) containing a growth additive. 100 μL of the cells were seeded into a 96-well microplate and cultured at 37°C under 5% _CO2 and saturated steam. After 24 hours of culture, the medium was replaced with HuMedia-KB2 (Kurabo Industries, Ltd.). A firefly luciferase reporter vector containing a vitamin D receptor-binding sequence (VDRE) and a Renilla luciferase vector as an internal standard (Cignal HIF Pathway Reporter Assay Kit) (QIAGEN) were then transfected into the cells using ViaFect transfection reagent (Promega). After 24 hours, the extract from Preparation Example 1 was added as a sample solution, and the epidermal keratinocytes were cultured under the same conditions. The sample solution was added to the culture medium at final concentrations of 0.5%, 1.0%, or 2.0%. A control group was prepared by adding 30% BG solution at the same concentration instead of the sample solution. After 24 hours of incubation under the same conditions, cholecalciferol (prepared with HuMedia-KB2) was added to a final concentration of 100 nM. After 24 hours, the cell contents were extracted, and the luciferase activity of the cell extract was measured using a luciferase assay system (Promega) and a luminometer (Promega GloMax-Multi+ Detection System). The firefly luciferase measurement (RARE transcription level) divided by the Renilla luciferase measurement (cell mass) was used to calculate the vitamin A receptor transcription activity per cell. Furthermore, the VDRE transcription activity in the control group (Control) was also calculated. The VDRE transcriptional activity was calculated as a relative value to the control test group, which was set at 100.

The results of Test Example 2 are shown in Table 2.
[Table 2]

As shown in Table 2, it was confirmed that the extract of the present invention significantly enhanced luciferase activity when used in combination with vitamin D (cholecalciferol), which is produced in the skin. In Test Example 2, the transcriptional activity of vitamin D receptors was evaluated using luciferase activity, suggesting that the extract of the present invention has the effect of increasing the transcriptional activity of vitamin D receptors by converting cholecalciferol to calcifediol or calcitriol.

The results of Test Examples 1 and 2 above suggest that the extract of the present invention can promote vitamin D metabolism by converting vitamin D produced in the epidermis due to factors such as exposure to sunlight into active vitamin D, thereby contributing to homeostasis of the epidermal layer. Furthermore, the present invention suggests that promoting vitamin D metabolism in the epidermis promotes differentiation of epidermal cells, thereby promoting, for example, the formation of the cornified envelope of the stratum corneum (expression of involucrin, transglutaminase, etc.), NMF synthesis (expression of peptidylarginine deiminases), secretion of antimicrobial peptides, and expression of stratum corneum desquamating enzymes (expression of kallikreins, etc.). Therefore, the present invention is expected to maintain a strong barrier function while regulating cell turnover and promoting stratum corneum exfoliation, which restores the epidermis to a normal state, thereby restoring the epidermis to a normal state.

Formulation Example 1. Lotion [Ingredients] Parts Extract of Preparation Example 1 0.5
Squalane 0.2
Polyoxyethylene (5.5) Cetyl Alcohol 5.0
Tocopherol acetate 0.02
Dipotassium glycyrrhizinate 0.5
Nicotinamide 0.1
1,3-butylene glycol 5.0
Sodium citrate 0.2
Methylparaben 0.1
Purified water (enough to make the total volume 100 parts)

Formulation Example 2: Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 0.5 parts of the extract from Preparation Example 2 was used in place of 0.5 parts of the extract from Preparation Example 1.

Formulation Example 3. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 0.5 parts of the extract from Production Example 3 was used in place of 0.5 parts of the extract from Production Example 1.

Formulation Example 4. Lotion A lotion was obtained in the same manner as in Formulation Example 1, except that 0.5 parts of the extract from Production Example 4 was used in place of 0.5 parts of the extract from Production Example 1.

Formulation Example 5. Lotion [Ingredients] Parts Extract of Preparation Example 1 1.0
Glyceryl caprylate 3.0
Polyglyceryl-10 Laurate 3.0
Cetanol 2.0
Behenyl alcohol 2.0
Niacinamide 5.0
Dipotassium glycyrrhizinate 0.1
Stearyl glycyrrhetinate 0.1
Glycerin 1.0
1,3-butylene glycol 1.0
Pentanediol 1.0
Potassium hydroxide 0.5
Purified water (enough to make the total volume 100 parts)

Formulation Example 6. Lotion [Ingredients] Parts Extract of Preparation Example 2 1.0
Jojoba oil 1.0
Polyoxyethylene (5.5) Cetyl Alcohol 5.0
Methylparaben 0.1
Ascorbic acid glucoside 2.0
Niacinamide 3.5
Tranexamic acid 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Sodium citrate 0.2
Sodium metabisulfite 0.2
Purified water (enough to make the total volume 100 parts)

Formulation Example 7. Emulsion [Ingredients] Parts Extract of Preparation Example 1 2.0
Squalane 5.0
Cyclopentanesiloxane 1.0
Hexalan 3.0
Hexyldecyl isostearate 1.0
Caprylic/Capric Triglyceride 1.0
Polyglyceryl-10 Laurate 5.0
Polyglyceryl-10 Isostearate 5.0
Ascorbyl dipalmitate 15.0
Hydrogenated soy lecithin 1.5
Ascorbic acid phosphate magnesium salt 3.0
Arbutin 3.0
Glycerin 3.0
1,3-butylene glycol 2.0
Carboxymethyl cellulose 0.3
Xanthan gum 0.2
Tremella fuciformis polysaccharide 0.2
Sodium hyaluronate 0.01
Tocopherol acetate 0.3
Tocopherol nicotinate 0.1
Dipotassium glycyrrhizinate 0.1
Stearyl glycyrrhetinate 0.1
Isopropylmethylphenol 0.1
Water-soluble collagen 1.0
Hydrolyzed Collagen 1.0
Acetyl hyaluronic acid 0.1
Disodium edetate 0.05
Diethylenetriaminepentaacetic acid pentasodium 0.05
Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts)

Formulation Example 8. Milky lotion A milky lotion was obtained in the same manner as in Formulation Example 7, except that 2.0 parts of L-ascorbic acid-2-glucoside was used in place of 2.0 parts of ascorbic acid phosphate magnesium salt.

Formulation Example 9. Milky lotion A milky lotion was obtained in the same manner as in Formulation Example 7, except that 2.0 parts of ascorbyl tetrahexyldecanoate was used in place of 2.0 parts of magnesium ascorbyl phosphate.

Formulation Example 10. Milky Lotion A milky lotion was obtained in the same manner as in Formulation Example 7, except that 2.0 parts of ascorbic acid phosphate magnesium salt was replaced with 3.0 parts of 3-O-ethyl ascorbic acid.

Formulation Example 11. Milky Lotion An emulsion was obtained in the same manner as in Formulation Example 7, except that 2.0 parts of ascorbic acid phosphate magnesium salt and 4.0 parts of niacinamide were used in place of potassium hydroxide.

Formulation Example 12. Cream [Ingredients] Parts Extract of Preparation Example 1 2.0
Olive oil 5.0
Squalane 5.0
Jojoba oil 5.0
Jojoba wax 1.0
Behenyl alcohol 1.0
Stearyl alcohol 1.0
Candelilla Wax 1.0
Lactic acid bacteria fermented rice 2.0
Soybean-derived hydrogenated lecithin 0.5
Carboxyvinyl polymer 1.0
Sodium alginate 1.0
Glycerin 2.0
Pentanediol 1.0
pH adjuster (appropriate amount) Purified water (enough to make the total amount 100 parts)

Formulation Example 13. Cream [Ingredients] Parts Extract of Preparation Example 2 1.0
Olive oil 5.0
Jojoba oil 5.0
Squalane 5.0
Hexyldecyl isostearate 5.0
Di(octyldodecyl/phytosteryl/behenyl) lauroyl glutamate 5.0
Glyceryl caprylate 1.0
Glyceryl stearate 1.0
Isostearyl glyceryl 3.0
γ-oryzanol 0.1
Behenyl alcohol 2.0
Palmitic acid 2.5
D-pantothenyl alcohol 3.0
Allantoin 0.1
Riboflavin 0.01
Resorcinol 0.1
Benzalkonium chloride 0.05
Urea 3.0
β-glycyrrhetinic acid 0.1
Stearyl glycyrrhetinate 0.1
Ammonium glycyrrhizinate 0.1
Niacinamide 5.0
Lactic acid bacteria fermented rice 2.0
Hydrogenated lecithin 0.5
Hydrogenated lysolecithin 0.5
Hydrolyzed Collagen 1.0
Xanthan gum 1.0
Zinc oxide 0.5
dl-camphor 0.3
l-menthol 0.5
Purified water (enough to make the total volume 100 parts)

Example 14. Pack [Ingredients] Parts Extract of Production Example 1 1.0
Dipropylene glycol 5.0
Polyoxyethylene (60) hydrogenated castor oil 5.0
Cetyl alcohol 3.0
Behenyl alcohol 3.0
Allantoin 0.1
Dipotassium glycyrrhizinate 0.1
Ammonium glycyrrhizinate 0.1
β-glycyrrhetinic acid 0.1
Stearyl glycyrrhetinate 0.1
Salicylic acid 0.1
Tocopherol acetate 0.5
Tocopherol nicotinate 0.1
D-pantothenyl alcohol 0.3
Resorcinol 0.1
Sulfur 2.0
Estradiol 0.002
Water-soluble collagen 1.0
Xanthan gum 2.0
Polyglyceryl-6 myristate 1.0
Potassium cocoyl glutamate 1.0
Hydrogenated lecithin 3.0
Hydroxylated lecithin 3.0
Purified water (enough to make the total volume 100 parts)

Formulation Example 15. Hair Shampoo [Ingredients] Parts Extract of Preparation Example 1 2.0
Sodium laureth sulfate 10.0
Glyceryl monostearate 1.0
Coconut oil fatty acid diethanolamide 2.0
Polyoxyethylene (40) hydrogenated castor oil 0.5
Benzalkonium chloride 1.0
Stearyl alcohol 2.0
Behenyl alcohol 2.0
Dimethicone 3.0
Allantoin 0.1
Dipotassium glycyrrhizinate 0.1
Salicylic acid 0.1
Sodium salicylate 0.1
Tocopherol acetate 0.1
Pyrithione zinc 0.3
Benzoic acid 0.2
Triclosan 0.2
Citric acid 0.1
Propylene glycol 2.0
Purified water (enough to make the total volume 100 parts)

Formulation Example 16: Hair conditioner [Ingredients] Parts Extract of Production Example 1 1.0
Polyoxyethylene (10) hydrogenated castor oil 1.0
Distearyldimethylammonium chloride 1.5
Stearyltrimethylammonium chloride 2.0
Glyceryl 2-ethylhexanoate 1.0
Benzalkonium chloride 1.0
Cetyl alcohol 3.0
Stearyl alcohol 1.0
Allantoin 0.1
Isopropylmethylphenol 0.1
Dipotassium glycyrrhizinate 0.1
Salicylic acid 0.1
Sulfur 0.5
Alkylisoquinolinium bromide solution (75%) 0.06
Pyrithione zinc 0.3
Methylparaben 0.1
Triclosan 0.2
Resorcinol 0.1
Purified water (enough to make the total volume 100 parts)

Formulation Example 17. Cleansing cosmetic [Ingredients] Parts Extract of Production Example 1 2.0
Potassium cocoyl glycinate 5.0
Glycerin 10.0
Glyceryl caprylate 1.0
Sodium lauroyl aspartate 10.0
Water-soluble collagen 5.0
Cetyl alcohol 3.0
Myristyl alcohol 3.0
Isopropyl methyl alcohol 0.1
Allantoin 0.1
Sulfur 0.5
Glycyrrhizic acid 0.1
Dipotassium glycyrrhizinate 0.1
Monoammonium glycyrrhizinate 0.1
β-glycyrrhetinic acid 0.05
Stearyl glycyrrhetinate 0.1
Salicylic acid 0.2
Tocopherol acetate 0.2
Triclosan 0.1
Trichlorocarbanide 0.5
Trichlorohydroxydiphenyl ether 0.2
Concentrated benzalkonium chloride solution 50 0.2
Benzalkonium chloride 0.1
Purified water (enough to make the total volume 100 parts)

Formulation Example 18. Sheet Mask A sheet mask is obtained by impregnating a nonwoven fabric with the following ingredients.
[Ingredients] Parts Extract of Production Example 1 2.0
Niacinamide 1.0
Citric acid 0.1
Sodium citrate 0.3
Xanthan gum 0.1
Water-soluble collagen 0.1
Sodium hyaluronate 0.01
Glycerin 1.0
1,3-butylene glycol 2.0
Pentanediol 1.0
Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts)

Formulation Example 19. Serum [Ingredients] Parts Extract of Preparation Example 1 2.0
Sodium hyaluronate 1.0
Water-soluble collagen 1.0
Tranexamic acid 0.1
Ethanol 2.0
Glycerin 5.0
1,3-butylene glycol 5.0
Methylparaben 0.1
Citric acid 0.3
Sodium citrate 0.6
Eucheuma muscaria extract 5.0
Purified water (enough to make the total volume 100 parts)

Formulation Example 20. Cream [Ingredients] Parts Extract of Preparation Example 4 2.0
Olive oil 5.0
Squalane 5.0
Jojoba oil 5.0
Jojoba wax 1.0
Shea Butter 2.0
Behenyl alcohol 1.0
Stearyl alcohol 1.5
Candelilla wax 0.5
Niacinamide 1.0
Lactic acid bacteria fermented rice 3.0
Hydrogenated lecithin 2.0
Eucheuma muscaria extract 2.0
Carboxyvinyl polymer 0.3
Sodium alginate 0.2
Glycerin 4.0
Potassium hydroxide (appropriate amount) Purified water (enough to make the total amount 100 parts)

Claims (2)

A vitamin D metabolism promoter whose active ingredient is an extract of Lilium concolor or one of its subspecies. An epidermal cell differentiation improver containing an extract of Lilium concolor or its subspecies as an active ingredient.