JP7626587B2 - Autophagy inducers - Google Patents

Description

The present invention relates to agents that are useful for inducing autophagy.

Autophagy is a cellular metabolic function that plays a role in breaking down unnecessary proteins and organelles within cells.

And so, the search is on for ingredients that are effective in inducing such autophagy. For example, Patent Document 1 discloses an autophagy inducer that contains cyclic glucooligosaccharide as an active ingredient.

International Publication No. 2018-173653

The object of the present invention is to provide a novel agent that is useful for inducing autophagy, etc.

As a result of intensive research into achieving the above-mentioned object, the present inventors discovered that quercetin glycosides (particularly glycosides containing quercetin-7-glucoside) and safflower extract are useful for inducing (improving, enhancing, and expressing) autophagy and for anti-aging (e.g., preventing and improving skin aging), and have thus completed the present invention.

That is, the present invention relates to the following inventions, etc.
[1]
Contains quercetin glycoside (as an active ingredient) and is an autophagy inducer (autophagy activator, autophagy expresser).
[2]
Contains quercetin glycoside (as an active ingredient), an anti-aging agent.
[3]
Contains safflower ingredients (as active ingredients) and is an autophagy inducer (autophagy activator, autophagy expresser).
[4]
Contains safflower ingredients (as an active ingredient), an anti-aging agent.
[5]
The agent according to [3] or [4], wherein the safflower component is a safflower extract containing quercetin glycoside.
[6]
The agent according to any one of [1] to [5], wherein the quercetin glycoside or safflower component contains quercetin-7-glucoside.
[7]
The agent according to any one of [3] to [6], wherein the safflower component is a safflower extract containing at least safflower leaves.
[8]
The agent according to any one of [3] to [7], wherein the safflower component is a safflower extract in a solvent containing at least alcohol (such as ethanol).
[9]
The agent according to any one of [3] to [8], wherein the safflower component contains quercetin-7-glucoside in an amount of 0.03% by mass or more.
[10]
The agent according to any one of [1] to [9], which is used for preventing and/or improving skin aging.
[11]
A composition comprising the agent (quercetin glycoside, safflower component) according to any one of [1] to [10].
[12]
The composition according to [11], which is used for preventing and/or improving skin aging.
[13]
The composition according to [11] or [12], containing quercetin-7-glucoside in an amount of 0.00003% by mass or more.
[14]
The composition according to any one of [11] to [13], which is for use in cosmetics.
[15]
The agent or composition according to any one of [1] to [14], for inducing autophagy in epidermal keratinocytes and/or fibroblasts.

The present invention provides novel agents and the like that are useful for inducing autophagy.

FIG. 1 is a graph showing the results obtained in Example 1. FIG. 2 is a graph showing the results obtained in Example 2. FIG. 3 is a graph showing the results obtained in Example 3.

[Agent]
(First aspect)
A first aspect of the present invention provides an agent containing (containing as an active ingredient) a quercetin glycoside (a glycoside of quercetin).

Quercetin is the compound shown below, and quercetin glycoside is a compound in which a sugar is substituted (bonded) to the hydroxyl group of quercetin (this compound).

In quercetin glycosides, the sugar is not particularly limited, but may be, for example, a monosaccharide (monosaccharides), an oligosaccharide, or a polysaccharide (polysaccharides). The sugar (oligosaccharide or polysaccharide) may be either a homopolysaccharide or a heteropolysaccharide.

The number of carbon atoms in a monosaccharide is not particularly limited, and may be, for example, a pentose or a hexose. A monosaccharide may also be, for example, a furanose or a pyranose.

Specific sugars include monosaccharides [e.g., pentoses (e.g., ribose, arabinose, xylose, deoxyribose, etc.), hexoses (e.g., fructose, tagatose, allose, altrose, glucose, mannose, galactose, rhamnose, glucosamine, galactosamine, glucuronic acid, etc.)], and oligosaccharides or polysaccharides formed by combining these monosaccharides [e.g., disaccharides (e.g., maltose, kojibiose, cellobiose, isomaltose, gentibiose, lactose, rutinose, neohesperidose, etc.)], etc.

Among these, representative sugars include monosaccharides (e.g., hexoses such as glucose and galactose).

The sugar may be D-, L- or a mixture of both.

In quercetin glycosides, the substitution (bond) position of the sugar is not particularly limited and may be any of the hydroxyl groups of quercetin. In addition, the number of sugars (quercetin or hydroxyl groups of quercetin) [number of substitutions (bonds)] in quercetin glycosides is also not particularly limited and may be 1 or 2 or more (e.g., 2-5, 2-4, 2-3, 2, etc.).

When the number of sugars is two or more, the sugars may be the same or different.

Quercetin glycosides may be either the α form (α anomer, α-glycoside (O-glycoside)), the β form (β anomer, β-glycoside (O-glycoside)), or a mixture of these.

Specific examples of quercetin glycosides include quercetin glucoside (e.g., quercetin-3-glucoside, quercetin-7-glucoside), quercetin galactoside (e.g., quercetin-3-galactoside), quercetin rhamnoside (e.g., quercetin-3-rhamnoside), quercetin glucuronide (e.g., quercetin-3-glucuronide), and quercetin rutinoside (e.g., quercetin-3-rutinoside).

Quercetin glycosides may be used alone or in combination of two or more types.

Of these, quercetin glucoside and/or quercetin-7-glycoside are preferred, and quercetin-7-glucoside is particularly preferred. Therefore, quercetin glycoside may contain at least such a glycoside (e.g., quercetin-7-glucoside).

In such cases, the quercetin glycoside may contain quercetin glucoside and/or quercetin-7-glycoside in a proportion of, for example, 10% by mass or more, 30% by mass or more, 50% by mass or more, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, relative to the total quercetin glycoside, or may contain substantially 100% by mass.

In particular, when the quercetin glycoside contains quercetin-7-glucoside, the ratio of quercetin-7-glucoside to the total quercetin glycoside may be, for example, 1% by mass or more (e.g., 5% by mass or more), preferably 10% by mass or more (e.g., 15% by mass or more), more preferably 20% by mass or more, and particularly preferably 30% by mass or more.

Quercetin glycosides may be commercially available products or may be synthesized by conventional methods. They may also be derived from plants (the plants themselves or products separated or extracted from plants), such as safflower components described below.

(Second Aspect)
In a second aspect of the present invention, there is provided an agent containing a safflower component (containing the safflower component as an active ingredient).
Examples of safflower components include safflower, safflower extract, etc., and these may be used in combination. As the safflower component, safflower extract is typically used in many cases.

As for safflower, parts such as flowers, leaves, seeds (seeds and seed coats), stems, and roots may be used, or a combination of these may be used, or the whole plant may be used.

As safflower, in particular safflower including at least the leaves may be used.

Safflower extract can be purchased commercially or can be obtained by extracting safflower.

In the extraction, the solvent (extraction solvent) can be selected depending on the part of the safflower to be used, etc., and examples include water, alcohols [e.g., alkanols (e.g., C _1-4 alkanols such as methanol, ethanol, isopropanol, n-butanol, etc.), polyols (e.g., ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, etc.)], ketones (e.g., acetone, methyl ethyl ketone, etc.), ethers (e.g., diethyl ether, etc.), esters (e.g., ethyl acetate, etc.), etc.
The extraction solvents may be used alone or in combination of two or more kinds.

Typical extraction solvents include water, alcohols (such as ethanol), and mixtures of these.

In particular, the solvent may be one that contains at least a polar solvent (e.g., alcohols), particularly one that contains ethanol [e.g., a solvent that contains ethanol at 30% by mass or more, preferably 50% by mass or more, and more preferably 70% by mass or more (e.g., 90% by mass or more)]. By using such a solvent, it is easy to efficiently obtain a safflower extract that contains quercetin glycosides (e.g., quercetin-7-glucoside).

There are no particular limitations on the extraction temperature or time, and these can be selected appropriately depending on the extraction solvent and extraction method.

When extracting, the safflower may be subjected to appropriate processing (such as crushing, heating, or drying) as necessary. The extract may also be further concentrated or dried (such as drying at room temperature or freeze-drying).

The form of the safflower component is not particularly limited, and may be, for example, a liquid (e.g., an extract containing an extraction solvent), a powder (or granular powder), etc.

The safflower component may contain, in particular, quercetin glycoside. Such safflower components containing quercetin glycoside can be efficiently obtained by selecting the part of the safflower to be used and, in the case of an extract, the extraction conditions, etc.

Quercetin glycosides include the same glycosides as those exemplified in the first embodiment, and the preferred embodiments thereof are also the same.

For example, the safflower component may contain at least quercetin glucoside and/or quercetin-7-glycoside, in particular quercetin-7-glucoside.

Safflower components that contain quercetin glycoside may also contain other components (components other than quercetin glycoside). Such other components may include, for example, quercetin, luteolin, and luteolin glycosides (e.g., luteolin-7-glucoside), depending on the part of the safflower and the extraction conditions.

The other ingredients may be contained in the safflower component either alone or in combination of two or more.

When a safflower component (e.g., safflower extract) contains quercetin glycoside, the proportion of quercetin glycoside in the safflower component (proportion to the total solids content when a solvent is included) may be, for example, 0.01 mass% or more (e.g., 0.03 mass% or more), preferably 0.1 mass% or more (e.g., 0.3 mass% or more), and more preferably 1 mass% or more (e.g., 3 mass% or more).
The upper limit of the proportion of quercetin glycoside in the safflower components (proportion to the total solids when a solvent is included) is not particularly limited, and may be, for example, 0.3 mass%, 1 mass%, 3 mass%, 5 mass%, 10 mass%, 20 mass%, 30 mass%, etc.

When a safflower component (e.g., safflower extract) contains quercetin-7-glucoside, the proportion of quercetin-7-glucoside in the safflower component (the proportion relative to the total solids when a solvent is included) may be, for example, 0.001 mass% or more (e.g., 0.003 mass% or more, 0.005 mass% or more, 0.01 mass% or more), preferably 0.01 mass% or more (e.g., 0.03 mass% or more, 0.05 mass% or more, 0.1 mass% or more), and more preferably 0.1 mass% or more (e.g., 0.3 mass% or more, 0.5 mass% or more, 1 mass% or more).
The upper limit of the proportion of quercetin-7-glucoside in the safflower components (proportion to the total solids content when a solvent is included) is not particularly limited, but may be, for example, 0.1 mass%, 1 mass%, 3 mass%, 5 mass%, 10 mass%, etc.

When a safflower component (e.g., safflower extract) contains luteolin-7-glucoside, the proportion of luteolin-7-glucoside in the safflower component (proportion to the total solids content when a solvent is included) may be, for example, 0.01 mass% or more, preferably 0.1 mass% or more, and more preferably 1 mass% or more.
The upper limit of the proportion of luteolin-7-glucoside in the safflower components (proportion to the total solids when a solvent is included) is not particularly limited, but may be, for example, 0.1 mass%, 1 mass%, 3 mass%, 5 mass%, 10 mass%, etc.

(Use of the agent)
The agents of the present invention (the agent of the first embodiment and the agent of the second embodiment) can be used for specific purposes.

First, the agent of the present invention can be used as an autophagy inducer [agent for inducing (expressing, improving, enhancing, activating) autophagy].
As described above, autophagy (function) plays a role in, for example, breaking down unnecessary proteins and organelles within cells, and the agent of the present invention can induce (express, improve, enhance, activate) such autophagy (function).
Such an agent may induce autophagy in skin cells. Examples of skin cells include epidermal keratinocytes, fibroblasts, etc. In particular, the agent of the present invention can induce autophagy in epidermal keratinocytes and fibroblasts, and can efficiently exert the autophagy function (and the associated skin prevention or improvement function).

The agent of the present invention can also be used as an anti-aging agent.

In particular, the agent of the present invention may be an agent for preventing and/or improving skin aging.

In addition, in anti-aging agents (and also agents for preventing and/or improving skin aging), the anti-aging action/function (and also the action/function for preventing and/or improving skin aging) may be accompanied by the induction of autophagy (function).

The agent of the present invention may also be used as an agent for a specific application, such as for medicine, food and beverages, cosmetics, etc.

[Composition]
The present invention also includes a composition containing the agent (first or second aspect).

Such compositions may also be used for the same functions and purposes as the above-mentioned agents. For example, they may be used for autophagy induction, anti-aging, and prevention and/or improvement of skin aging. They may also be used for pharmaceuticals, food and beverages, cosmetics, etc.

In particular, the composition of the present invention may be suitably used as a cosmetic (composition for cosmetic use) from the viewpoint of having an autophagy-inducing effect and an anti-aging effect. The composition for cosmetic use is not particularly limited, and may be classified as a quasi-drug, such as medicated cosmetics, according to the definition of the Pharmaceutical Affairs Law.

The composition of the present invention only needs to contain the above-mentioned agent (quercetin glycoside or safflower component), and may contain other ingredients depending on the intended use, form, etc.

Such other ingredients include bases or carriers, additives, other active ingredients, etc. The other ingredients and their proportions can be appropriately selected depending on the form, properties, etc. of the composition.

The base or carrier is not particularly limited, but examples thereof include hydrocarbons such as paraffin, liquid paraffin, squalane, white wax, gelling hydrocarbons (such as Plastibase), ozokerite, ceresin, petrolatum, hard fat, microcrystalline wax, α-olefin oligomers, and light liquid paraffin; fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, and isostearic acid; glyceryl tri-2-ethylhexanoate (trioctanoin), tri(caprylic acid), etc. triglycerides such as glyceryl (caprate) and the like; higher alcohols such as cetanol, stearyl alcohol, and behenyl alcohol; methylpolysiloxane, highly polymerized methylpolysiloxane, dimethylsiloxane-methyl(polyoxyethylene)siloxane-methyl(polyoxypropylene)siloxane copolymer, dimethylsiloxane-methyl(polyoxyethylene)siloxane copolymer, dimethylsiloxane-methyl(polyoxypropylene)siloxane copolymer, polyoxyethylene Silicone oils such as methylpolysiloxane copolymer, poly(oxyethylene-oxypropylene)-methylpolysiloxane copolymer, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxysiloxane copolymer, alkyl acrylate copolymer methylpolysiloxane ester, crosslinked methylpolysiloxane, crosslinked methylphenylpolysiloxane, crosslinked polyether modified silicone, crosslinked alkylpolyether modified silicone, crosslinked alkyl modified silicone, decamethylcyclopentasiloxane, ethyltrisiloxane, methyltrimethicone, methylsiloxane network polymer, polyoxyethylene-methylpolysiloxane copolymer, methylhydrogenpolysiloxane, triethoxysilylethyl polydimethylsiloxyethylhexyldimethicone, and dimethylpolysiloxane; ethylene glycol monoacetate, ethylene glycol diacetate, triethylene glycol diacetate, and hexylene glycol diacetate. glycol acetates such as 2-methyl-2-propene-1,1-diol diacetate; glycol esters such as triethylene glycol divalerate, 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate, and 2,2,4-trimethyl-1,3-pentanediol diisobutyrate; glycol acrylates such as ethylene glycol diacrylate, diethylene glycol diacrylate, propylene glycol monoacrylate, 2,2-dimethyl-trimethylene glycol diacrylate, and 1,3-butylene glycol diacrylate; glycol dinitrates such as ethylene glycol dinitrate, diethylene glycol dinitrate, triethylene glycol dinitrate, and propylene glycol dinitrate; 2,2'-[1,4-phenylenedioxy]diethanol; cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose; polyvinylpyrrolidone; carrageenan; Examples of the bases or carriers include polyvinyl butyrate, polyethylene glycol, dioxane, butylene glycol adipic acid polyester, esters such as isopropyl myristate, octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, and pentaerythritol tetra 2-ethylhexanoate, polysaccharides such as dextrin and maltodextrin, lower alcohols such as ethanol and isopropanol, glycol ethers such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monopropyl ether, diethylene glycol monobutyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl ether, dipropylene glycol monoethyl ether, and dipropylene glycol monopropyl ether, and aqueous bases such as water. These bases or carriers can be used alone or in combination of two or more.

Additives are not particularly limited, but examples thereof include antioxidants, surfactants, thickeners, preservatives, pH adjusters, stabilizers, preservatives, colorants, and fragrances. These additives can be used alone or in combination of two or more.

Examples of antioxidants include dibutylhydroxytoluene, butylhydroxyanisole, sorbic acid, sodium sulfite, ascorbic acid, ascorbic acid derivatives (such as ascorbic acid phosphate), tocopherol, tocopherol derivatives, erythorbic acid, and L-cysteine hydrochloride.

Examples of surfactants include sorbitan fatty acid esters such as sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, diglycerol sorbitan penta-2-ethylhexyl acid, and diglycerol sorbitan tetra-2-ethylhexyl acid; propylene glycol fatty acid esters such as propylene glycol monostearate; glycerin alkyl ethers; alkyl glucosides; amines such as stearylamine and oleylamine; silicone surfactants such as polyoxyethylene-methylpolysiloxane copolymer, lauryl PEG-9 polydimethylsiloxyethyl dimethicone, and PEG-9 polydimethylsiloxyethyl dimethicone.

Examples of thickeners include guar gum, locust bean gum, carrageenan, xanthan gum, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, acrylate/methacrylate alkyl copolymer, polyethylene glycol, bentonite, alginic acid, macrogol, and cellulose-based thickeners (methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, carboxyethylcellulose, etc.) and salts thereof.

Examples of preservatives include benzoic acid, sodium benzoate, dehydroacetic acid, sodium dehydroacetate, isobutyl parahydroxybenzoate, isopropyl parahydroxybenzoate, butyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, benzyl parahydroxybenzoate, methyl parahydroxybenzoate, phenoxyethanol, benzyl alcohol, chlorobutanol, sorbic acid and its salts, chlorhexidine gluconate, alkanediols, and glycerin fatty acid esters.

Examples of pH adjusters include inorganic acids (hydrochloric acid, sulfuric acid, etc.), organic acids (lactic acid, sodium lactate, citric acid, sodium citrate, succinic acid, sodium succinate, etc.), inorganic bases (potassium hydroxide, sodium hydroxide, etc.), and organic bases (triethanolamine, diisopropanolamine, triisopropanolamine, etc.).

Examples of stabilizers include sodium polyacrylate, dibutylhydroxytoluene, butylhydroxyanisole, edetate, etc.

Other active ingredients include, but are not limited to, moisturizing ingredients, anti-inflammatory ingredients, antibacterial or germicidal ingredients, vitamins, peptides or their derivatives (esters, amides, etc.), cell activating ingredients, anti-aging ingredients, blood circulation promoting ingredients, keratin softening ingredients, whitening ingredients, astringent ingredients, etc. These active ingredients can be used alone or in combination of two or more.

Moisturizing ingredients include, for example, polymeric compounds such as collagen, elastin, keratin, chitin, and chitosan; natural moisturizing factors such as sodium lactate, urea, and sodium pyrrolidone carboxylate; lipids such as ceramide, cholesterol, and phospholipids; and plant extracts such as chamomile extract, witch hazel extract, tea extract, and perilla extract.

Anti-inflammatory ingredients include, for example, ingredients derived from plants (e.g., comfrey), allantoin, glycyrrhizic acid or its derivatives, zinc oxide, pyridoxine hydrochloride, tocopherol acetate, salicylic acid or its derivatives, ε-aminocaproic acid, etc.

Examples of antibacterial or germicidal ingredients include ethanol, chlorhexidine, salicylic acid, benzalkonium chloride, acrinol, sulfur, resorcin, benzethonium chloride, adapalene, benzoyl peroxide, clindamycin, cresol, gluconic acid and its derivatives, povidone-iodine, potassium iodide, iodine, isopropylmethylphenol, triclocarban, triclosan, photosensitizer No. 101, photosensitizer No. 201, paraben, phenoxyethanol, 1,2-pentanediol, alkyldiaminoglycine hydrochloride, chlorhexidine gluconate, zinc paraphenolsulfonate, etc.

Examples of vitamins include retinol derivatives such as retinol, retinol acetate, and retinol palmitate, vitamin A such as retinal, retinoic acid, methyl retinoate, ethyl retinoate, retinol retinoic acid, d-δ-tocopheryl retinoate, α-tocopheryl retinoate, and β-tocopheryl retinoate; vitamin E such as dl-α-tocopherol, dl-α-tocopherol acetate, dl-α-tocopherol succinate, and dl-α-tocopherol calcium succinate; riboflavin, flavin mononucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, and riboflavin tetraacetate. Vitamin B2 such as sodium boflavin 5'-phosphate and riboflavin tetranicotinate; nicotinic acids such as dl-α-tocopherol nicotinate, benzyl nicotinate, methyl nicotinate, β-butoxyethyl nicotinate and 1-(4-methylphenyl)ethyl nicotinate; vitamin C such as ascorbigen-A, ascorbyl stearate, ascorbyl palmitate and L-ascorbyl dipalmitate; vitamin D such as methylhesperidin, ergocalciferol and cholecalciferol; vitamin K such as phylloquinone and farnoquinone, γ-oryzanol, dibenzoylthiamine, dibenzoyl Thiamine hydrochloride; vitamin B1 such as thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate ester, thiamine diphosphate ester, thiamine diphosphate ester hydrochloride, thiamine triphosphate ester, thiamine triphosphate ester monophosphate; vitamin B6 such as pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, 5'-pyridoxal phosphate, pyridoxamine hydrochloride; cyanocobalamin, hydroxocobalamin, deoxyadenosylcobalamin, etc. Vitamin B12; folic acids such as folic acid and pteroylglutamic acid; nicotinic acids such as nicotinic acid and nicotinamide; pantothenic acids such as pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-panthesine, D-pantethine, coenzyme A, and pantothenyl ethyl ether; biotins such as biotin and bioticin; ascorbic acid derivatives of vitamin C such as ascorbic acid, sodium ascorbate, dehydroascorbic acid, sodium ascorbyl phosphate, and magnesium ascorbyl phosphate; vitamin-like action factors such as carnitine, ferulic acid, α-lipoic acid, and orotic acid.

Examples of peptides or derivatives thereof include keratin hydrolyzed peptides, hydrolyzed keratin, collagen, fish-derived collagen, atelocollagen, gelatin, elastin, elastin hydrolyzed peptides, collagen hydrolyzed peptides, hydrolyzed collagen, hydroxypropylammonium chloride hydrolyzed collagen, elastin hydrolyzed peptides, conchiolin hydrolyzed peptides, hydrolyzed conchiolin, silk proteolytic peptides, hydrolyzed silk, sodium lauroyl hydrolyzed silk, soybean proteolytic peptides, hydrolyzed soybean protein, wheat protein, wheat proteolytic peptides, hydrolyzed wheat protein, casein hydrolyzed peptides, acylated peptides (palmitoyl oligopeptide, palmitoyl pentapeptide, palmitoyl tetrapeptide, etc.), etc.

Examples of cell activation ingredients include vitamins such as retinol, thiamine, riboflavin, pyridoxine hydrochloride, and pantothenic acids; α-hydroxy acids such as glycolic acid and lactic acid; tannins, flavonoids, saponin, allantoin, and photosensitizer No. 301.

Anti-aging ingredients include pangamic acid, kinetin, ursolic acid, turmeric extract, sphingosine derivatives, silicon, silicic acid, N-methyl-L-serine, mevalonolactone, etc.

Examples of blood circulation-promoting ingredients include ingredients derived from plants (e.g., ginseng, angelica, arnica, ginkgo, fennel, burdock, oak, chamomile, Roman chamomile, carrot, gentian, burdock, rice, Japanese hawthorn, shiitake mushroom, European hawthorn, European juniper, cnidium rhizome, swertia britannica, thyme, clove, tangerine peel, angelica tree, tonin, spruce, carrot, garlic, butcher's broom, grape, peony, horse chestnut, melissa, yuzu, coix seed, rosemary, rose hip, peach, apricot, walnut, corn); glucosyl hesperidin, etc.

Examples of keratin softening ingredients include salicylic acid, glycolic acid, fruit acid, phytic acid, sulfur, etc.

Examples of whitening ingredients include ascorbic acid and its derivatives (phosphate esters, etc.), arbutin, tocopherol, etc.

Examples of astringent ingredients include zinc paraphenolsulfonate, zinc oxide, menthol, and ethanol.

The form of the composition of the present invention is not particularly limited and can be appropriately selected depending on the application, etc. Examples of the form include liquid, fluid, semi-solid, etc.
The form of the composition of the present invention (e.g., cosmetic composition) is not particularly limited, and examples thereof include liquids, suspensions, emulsions, creams, ointments, gels, liniments, lotions, aerosols, sheets in which nonwoven fabric is impregnated with a drug solution, etc. Among these, preferred are liquids, suspensions, emulsions, creams, etc.

Specific uses of the composition of the present invention are not particularly limited, but examples thereof include basic cosmetics such as lotion, cosmetic milky lotion, cosmetic oil, gel, cream, beauty serum, sunscreen cosmetic, pack, mask, hand cream, body lotion, and body cream; cleansing cosmetics such as face wash, makeup remover, body shampoo, shampoo, rinse, and treatment; makeup cosmetics such as makeup base, foundation, face powder, water-based face powder, face powder, grease gun, eye shadow base, eye shadow, nose shadow, lip pencil, lipstick, lip gloss, blusher, and various colors; hair care products such as shampoo, dry shampoo, conditioner, rinse, rinse-in shampoo, treatment, hair tonic, hair styling product, hair oil, pomade, and hair coloring agent; dentifrices, lip balms, soaps, and body soaps.
Among these, preferred are lotions, cosmetic emulsions, cosmetic oils, gels, creams, beauty essences, sunscreen cosmetics, etc., and more preferred are lotions, cosmetic emulsions, creams, beauty essences, etc.

Specific uses of the composition of the present invention when it is a quasi-drug are not particularly limited, but examples include medicated cosmetics, medicated soaps, medicated dentifrices, antiperspirants, medicated creams, hair restorers, hair dyes, mouthwashes, energy drinks, mouth fresheners, contact lens wearers, germicides, disinfectants, chilblains, digestive aids, herbal medicine-containing health medicines, vitamin-containing health medicines, bath additives, etc. Among these, medicated cosmetics, medicated creams, hair restorers, etc. are preferred.

The method for producing the composition of the present invention is not particularly limited as long as it uses the agent of the present invention (quercetin glycoside, safflower component), and any conventionally known method can be used depending on the application and properties of the composition. For example, the composition can be produced by blending the agent of the present invention as it is or after emulsification, solubilization or dispersion during production of the composition by a conventional method.

In the composition of the present invention, the proportion of the agent of the present invention can be appropriately selected depending on the type of composition, the blended ingredients, etc., and is not particularly limited. For example, the proportion of quercetin glycoside (proportion to the total solids when a solvent is included) may be, for example, 0.00003 mass% or more, preferably 0.0003 mass% or more, and more preferably 0.003 mass% or more.
The upper limit of the proportion of quercetin glycoside in the composition (proportion relative to the total solids content when a solvent is included) is not particularly limited, and may be, for example, 0.0001 mass%, 0.0003 mass%, 0.001 mass%, 0.003 mass%, 0.01 mass%, 0.03 mass%, etc.

When the composition of the present invention contains quercetin-7-glucoside, the proportion of quercetin-7-glucoside in the composition (the proportion relative to the total solid content when a solvent is contained) may be, for example, 0.00001 mass % or more, preferably 0.0001 mass % or more, and more preferably 0.001 mass % or more.
The upper limit of the proportion of quercetin-7-glucoside in the composition (the proportion relative to the total solids when a solvent is included) is not particularly limited, and may be, for example, 0.0001 mass%, 0.0003 mass%, 0.001 mass%, 0.003 mass%, 0.01 mass%, 0.03 mass%, etc.

Furthermore, when the composition of the present invention contains a safflower component (e.g., safflower extract), the proportion of the safflower component in the composition (the proportion relative to the total solids when a solvent is included) may be, for example, 0.01 mass% or more, preferably 0.1 mass% or more, and more preferably 1 mass% or more.
The upper limit of the proportion of safflower components in the composition (proportion relative to the total solids content when a solvent is included) is not particularly limited, but may be, for example, 0.1 mass %, 1 mass %, 10 mass %, etc.

[Usage, etc.]
The present invention includes various methods using the agent or composition.
Such a method may be, for example, a method of using the agent or composition on a subject to be used (applied).

And such a method may involve the above-mentioned functions and actions (e.g., autophagy induction, anti-aging, prevention and/or improvement of skin aging).

The method of using (applying) the agent or composition of the present invention is not particularly limited and can be appropriately selected depending on the subject of use (application), etc. For example, when used as a cosmetic, the method can be appropriately selected depending on the skin condition, age, sex, etc., and for example, an appropriate amount (e.g., about 0.005 to 0.5 g) may be applied (sprayed, sprayed, pasted, etc.) to the skin several times a day (e.g., 1 to 5 times a day, preferably 1 to 3 times a day). It can also be applied to any skin, such as the face, neck, hands, feet, fingers, trunk, scalp, etc.

The amount of the agent or composition of the present invention to be used (applied) is also not particularly limited and may vary appropriately depending on the type and components of the composition, but may be, for example, preferably about 0.001 to 10 mg/cm ² (skin area) and more preferably about 0.01 to 1 mg/cm ² (skin area) of quercetin glycoside (particularly quercetin glycoside containing quercetin-7-glucoside) or safflower component per day for an adult.

The present invention will be described in detail below with reference to examples, but the present invention is not limited to these.

[Quantitative Analysis of Quercetin-7-glucoside and Quercetin Glycoside]
The samples were analyzed according to a known high performance liquid chromatography (HPLC) method, and the peak area of quercetin-7-glucoside was calculated by comparing it with the peak area of a standard sample.
The calculation was also carried out in the same manner for quercetin glycosides other than quercetin-7-glucoside.

Synthesis example (safflower extract)
10 g of safflower leaf powder was suspended in 100 mL of 90% by weight aqueous ethanol solution and stirred at room temperature for 1 hour. The extract was separated by filtration, evaporated under reduced pressure, and then freeze-dried to obtain 1 g of safflower extract.

Example 1
Human epidermal fibroblasts (derived from a 51-year-old Caucasian female) (hereinafter referred to as NHDF) were seeded onto a 6-well plate and cultured overnight at 37°C in 5% CO2 in DMEM medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% _L -glutamine.
Safflower extract was added to the medium at 10 and 100 μg/mL and cultured for 24 hours. At the same time, for autophagy flux analysis, safflower extract was added in the same manner and cultured for 8 hours, after which lysosomal inhibitor chloroquine (hereinafter, CQ) was added at 5 μM and cultured for an additional 16 hours. After culture, the cells were collected with RIPA buffer (Millipore), and samples for SDS-PAGE were prepared using SDS sample buffer.
The amount of LC3-II, a marker protein for autophagy, was measured by Western blotting using an LC3 antibody (manufactured by Cell Signaling Technology, Inc.) Band intensity was measured using image analysis software ImageJ.
When comparatively quantifying the band intensity of LC3-II, GAPDH, which is produced in a constant amount in vivo, was used as an internal standard, and the ratio of the band intensity of LC3-II to that of GAPDH was calculated and standardized. In the autophagy flux analysis, if the following two conditions are met: the band intensity of LC3-II increases upon treatment with the test substance, and the band intensity further increases upon treatment with the addition of a lysosomal inhibitor, the test substance is judged to have an autophagy-inducing effect.
The expression level of LC3-II normalized with GAPDH as an internal standard is shown in Fig. 1. The expression level of LC3-II in untreated cells was set to "1", and the relative values of various cells were calculated.
1, the LC3-II band intensity increased in the groups to which 10 and 100 μg/mL safflower extract was added, compared to the control group. It was also confirmed that the band intensity increased further by the addition of CQ.

Example 2
Normal human epidermal keratinocytes (derived from a 47-year-old Caucasian female) (hereinafter referred to as NHEK) were seeded onto a 6-well plate and cultured overnight at 37°C under 5% CO2 in Epilife medium (Thermo Fisher Scientific) supplemented with human keratinocyte growth factor ( _HKGS ).
Safflower extract was added to the medium at 1, 10, and 100 μg/mL and cultured for 24 hours. At the same time, for autophagy flux analysis, safflower extract was added in the same manner and cultured for 8 hours, after which lysosomal inhibitor chloroquine (hereinafter, CQ) was added at 5 μM and cultured for an additional 16 hours. After culture, the cells were collected with RIPA buffer (Millipore), and samples for SDS-PAGE were prepared using SDS sample buffer.
The amount of LC3-II, a marker protein for autophagy, was measured by Western blotting using an LC3 antibody (manufactured by Cell Signaling Technology). Image analysis software ImageJ was used to measure band intensity. When comparatively quantifying the band intensity of LC3-II, GAPDH, which is produced in a constant amount in vivo, was used as an internal standard, and the ratio of the band intensity of LC3-II to that of GAPDH was calculated and standardized.
The expression level of LC3-II normalized with GAPDH as an internal standard is shown in Fig. 2. The expression level of LC3-II in untreated cells was set to "1", and the relative values of various cells were calculated.
2, the LC3-II band intensity increased in the groups to which 1, 10, and 100 μg/mL safflower extract was added, compared to the control group. It was also confirmed that the band intensity increased further by the addition of CQ.

Example 3
Human epidermal fibroblasts (neonatal derived) (hereinafter referred to as NHDF) were seeded onto a 6-well plate and cultured _overnight at 37°C under 5% CO2 in DMEM medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and 1% L-glutamine.
Quercetin-7-glucoside was added to the medium at concentrations of 0.05, 0.5, and 5 μg/mL, and the cells were cultured for 24 hours. At the same time, safflower extract was added in the same manner for autophagy flux analysis, and the cells were cultured for 8 hours, after which chloroquine (hereafter, CQ), a lysosomal inhibitor, was added to a concentration of 5 μM, and the cells were cultured for an additional 16 hours. After the culture, the cells were collected in RIPA buffer (Millipore), and samples for SDS-PAGE were prepared using SDS sample buffer.
The amount of LC3-II, a marker protein for autophagy, was measured by Western blotting using an LC3 antibody (manufactured by Cell Signaling Technology). Image analysis software ImageJ was used to measure band intensity. When comparatively quantifying the band intensity of LC3-II, GAPDH, which is produced in a constant amount in vivo, was used as an internal standard, and the ratio of the band intensity of LC3-II to that of GAPDH was calculated and standardized.
The expression level of LC3-II normalized with GAPDH as an internal standard is shown in Fig. 3. The expression level of LC3-II in untreated cells was set to "1", and the relative values of various cells were calculated.
3, the LC3-II band intensity was increased in the groups to which 0.05, 0.5, and 5 μg/mL quercetin-7-glucoside was added, compared to the control group. It was also confirmed that the band intensity was further increased by the addition of CQ.

Specific examples of formulations of compositions containing the agent of the present invention are given below.

Formulation Example 1: Lotion A lotion having the following composition is prepared according to the following preparation method.
The following ingredients are formulated so that the total is 100% by weight. All ingredients are mixed and stirred at room temperature to obtain a homogeneous solution, and the pH is adjusted to 5.5 to obtain a lotion.
Ingredients Weight %
Concentrated glycerin 4.0
Sorbitol 4.0
Sodium citrate 0.3
Polyoxyethylene hydrogenated castor oil 0.5
Safflower extract 0.1
Ethanol 15.0
Fragrance 0.005
Citric acid (appropriate amount) Purified water (remainder)

Formulation Example 2: Facial Cleansing Cream A facial cleansing cream having the following composition is prepared according to the following preparation method.
Components A to C are formulated in the following proportions so that the total of components A, B and C is 100% by weight. First, the mixture of component A is heated and dissolved, and maintained at 80°C. Next, the mixture of component B, which has been separately heated and dissolved at 80°C, is added to the mixture of component A and thoroughly stirred. After that, the mixture is cooled with stirring, and the mixture of component C is added at 50°C to obtain a facial cleansing cream.
"Component A"
Ingredients Weight %
Myristic acid 14.0
Stearic acid 12.0
Lauric acid 3.5
Oleyl alcohol 1.5
Coconut oil fatty acid amidopropyl betaine 10.5
"Component B"
Ingredients Weight %
Concentrated glycerin 18.0
Potassium hydroxide 7.0
Paraoxybenzoic acid ester Appropriate amount of purified water Remainder "Component C"
Ingredients Weight %
Safflower extract 0.1
Fragrance (appropriate amount)

Formulation Example 3: Milky Lotion A milky lotion having the following composition is prepared according to the following preparation method.
Components D to F are formulated in the following proportions so that the total of components D, E and F is 100% by weight. First, the component D mixture is heated and dissolved and maintained at 80°C. Next, the component E mixture, which has been separately heated and dissolved at 80°C, is added to the component D mixture and thoroughly stirred. After that, the mixture is cooled with stirring, and the component F mixture is added at 50°C to obtain an emulsion.
"Component D"
Ingredients Weight %
Sucrose fatty acid ester 1.0
Carboxyvinyl polymer 0.06
Potassium hydroxide 0.028
Paraoxybenzoic acid ester Appropriate amount of purified water Remaining "ingredient E"
Ingredients Weight %
Olive oil 4.0
Jojoba oil 4.0
Myristyl lactate 2.0
Self-emulsifying glyceryl monostearate 1.5
Lipophilic glyceryl monostearate 1.5
"Component F"
Ingredients Weight %
Safflower extract 0.1
Fragrance 0.2

The agent of the present invention can be used as an autophagy inducer, etc.

Claims (7)

An autophagy inducer containing quercetin -7-glucoside as an active ingredient . An anti-aging agent containing quercetin -7-glucoside as an active ingredient . An autophagy inducer containing safflower components containing quercetin -7-glucoside as an active ingredient . The agent according to claim 3 , wherein the safflower component is a safflower extract containing quercetin-7-glucoside in a proportion of 0.001% by mass or more. The agent according to claim 3 or 4 , wherein the safflower component is a safflower extract containing at least safflower leaves. The agent according to any one of claims 3 to 5 , wherein the safflower component is a safflower extract in a solvent containing at least alcohol. The agent according to any one of claims 3 to 6 , wherein the safflower component contains quercetin-7-glucoside in an amount of 0.03% by mass or more.