Composition, method for producing same, and cosmetic composition containing said composition

Abstract

To provide a novel anthocyanin-derived composition, a method for producing the same, and a cosmetic composition containing said composition.　Provided is a composition containing anthocyanin derived from a plant containing anthocyanin or an Aspergillus oryzae fermentation extract of a plant containing anthocyanin and any one or more of 1,3-propanediol, 1,3-butanediol, glycerin, and ethanol (however, excluding when the anthocyanin derived from a plant containing anthocyanin or Aspergillus oryzae fermentation extract of a plant containing anthocyanin is anthocyanin derived from a plant containing anthocyanin and the solvent contains ethanol).　Also provided is a method for producing an anthocyanin-derived composition that includes a step for extracting an anthocyanin-containing plant or an Aspergillus oryzae fermentation extract thereof with any one or more of 1,3-propanediol, 1,3-butanediol, glycerin, and ethanol (excluding methods including a step for extracting an anthocyanin-containing plant that has not been fermented by Aspergillus oryzae with a solvent containing ethanol).

Description

Composition, method for producing the same, and cosmetic composition containing the composition

[0001] This disclosure relates to a composition, a method for producing the same, and a cosmetic composition containing the composition.

[0002] Natural colorants extracted from plants are considered safer than colorants synthesized from chemicals and are widely used in food, beverages, and cosmetics. Among them, anthocyanin pigments are red and purple pigment components and are used as functional materials due to their antioxidant properties. Patent Document 1 discloses that purple corn pigment can be extracted under pH 1-4 conditions, and that the extract has anti-obesity and anti-diabetic effects. Patent Document 2 discloses that purple corn pigment can be extracted with an acidic aqueous solution containing 20% ​​by volume ethanol. Patent Document 3 discloses that a liquid extracted from purple corn with a 30% ethanol aqueous solution has therapeutic effects on skin diseases. Furthermore, Non-Patent Document 1 discloses that an ethanol aqueous solution extract of a solid fermented product of rice bran by Aspergillus oryzae has higher antioxidant power than an extract of the unfermented product.

[0003] Japanese Patent Publication No. 2003-252766, International Publication No. 2003-064538, Japanese Patent Publication No. 2019-532941

[0004] Foods 2021, 10, 70.

[0005] Anthocyanin pigments have poor stability at neutral to alkaline pH levels, and extraction under acidic conditions, as described in Patent Documents 1 and 2, has been investigated. However, when used as antioxidants, anti-aging agents, enzyme inhibitors, anti-inflammatory agents, DNA damage inhibitors, reactive oxygen species scavengers, or cell activators, for example in cosmetics, acidic extracts have limited formulation options, thus creating a need for anthocyanin pigment extracts that are stable at neutral to weakly alkaline pH levels. While extracting anthocyanin pigments with an ethanol aqueous solution, as described in Patent Document 3, results in an extract with a neutral pH, extracts using ethanol aqueous solutions suffer from a long-term decrease in color development due to the anthocyanin pigment. In addition, ethanol can cause irritation when applied to the skin, limiting its use in cosmetics and restricting its application to cosmetics, antioxidants, anti-aging agents, enzyme inhibitors, anti-inflammatory agents, DNA damage inhibitors, reactive oxygen species scavengers, and cell activators.

[0006] Therefore, this disclosure aims to provide a novel anthocyanin-derived composition, a method for producing the same, and a cosmetic composition containing the said composition.

[0007] This disclosure aims to provide, in one embodiment, an anthocyanin pigment extract that is stable for a long period of time at around neutral pH, a method for producing the same, and a cosmetic composition containing a plant extract containing the extract, as well as an antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, and cell activator.

[0008] Furthermore, in one embodiment, this disclosure aims to provide a plant-derived extract containing anthocyanin pigments with high antioxidant activity, a method for producing the same, a cosmetic composition containing the extract, an antioxidant, and a wrinkle formation inhibitor.

[0009] The present inventors conducted various studies to achieve the above objectives and found that a composition comprising anthocyanin derived from plants containing anthocyanins and a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin can exist stably for a long period of time at a neutral pH (e.g., pH 5 to 8), and a composition comprising an anthocyanin fermentation extract of Aspergillus oryzae and a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol exhibits excellent antioxidant activity, and that both are useful as raw materials for cosmetic compositions, leading to the invention of this disclosure.

[0010] In other words, in one aspect of this disclosure, a plant extract can be produced by solvent extracting a plant containing anthocyanins from a plant containing anthocyanins, using an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin as the extraction solvent. The plant extract can be used in cosmetic compositions, as an antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, and cell activator. In one preferred aspect, the plant extract is characterized by containing anthocyanins derived from corn and a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin. In a further preferred aspect, the plant extract is characterized by containing anthocyanins derived from corn, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water.

[0011] In another aspect, this disclosure provides a method for producing a solid fermented product by fermenting a plant containing anthocyanins with Aspergillus oryzae, and then solvent-extracting the solid fermented product using an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin as the extraction solvent, thereby obtaining an Aspergillus oryzae fermented extract of anthocyanins-containing plants. This Aspergillus oryzae fermented extract can be used, for example, in cosmetic compositions, as antioxidants, and as wrinkle-forming inhibitors.

[0012] In other words, one aspect of the present disclosure includes: [1] A method for producing an anthocyanin-derived composition, comprising the step of extracting a plant containing anthocyanins or a fermented product thereof with Aspergillus oryzae with a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol (except in cases where the step involves extracting an anthocyanin-containing plant that has not been fermented by Aspergillus oryzae with a solvent containing ethanol).

[0013] Examples of means for providing the above-mentioned manufacturing method include the following means A' and means B'.

[0014] (A') Method A' [2] The method for producing a plant extract, wherein a plant containing anthocyanins is used as a raw material, and an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin is used as the extraction solvent. [3] The method for producing a plant extract, wherein the plant extract has a pH of 5 to 8. [4] The method for producing a plant extract, wherein the plant containing anthocyanins is corn.

[0015] (B') Method B' [5] A method for producing an Aspergillus oryzae fermentation extract, comprising a fermentation step of fermenting a plant containing anthocyanins with Aspergillus oryzae, and a step of extracting the Aspergillus oryzae ferment product obtained in the fermentation step with the solvent, as described in [1]. [6] A method for producing an Aspergillus oryzae fermentation extract, as described in [5], wherein the solvent is an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin.

[0016] One aspect of the present disclosure also includes: [7] A composition comprising an anthocyanin derived from a plant containing anthocyanins, or a plant fermentation extract of Aspergillus oryzae containing anthocyanins, and a solvent comprising one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol (except when the anthocyanin derived from a plant containing anthocyanins, or the plant fermentation extract of Aspergillus oryzae containing anthocyanins, is an anthocyanin derived from a plant containing anthocyanins, and the solvent comprises ethanol).

[0017] Examples of means for providing the aforementioned composition include the following means A and means B.

[0018] (A) Means A [8] The composition according to [7], which is a plant extract characterized by comprising anthocyanin derived from corn, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water. [9] An antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator comprising the composition according to [8].

[10] A cosmetic composition comprising the composition according to [8], or the antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator according to [9].

[0019] (B) Means B

[11] The composition according to [7], which is a fermentation extract of a plant containing anthocyanins by Aspergillus.

[12] The composition according to

[11] , which comprises an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin.

[13] The composition according to

[11] or

[12] , wherein the Aspergillus is a fungus of the genus Aspergillus.

[14] The composition according to

[13] , wherein the fungus of the genus Aspergillus is Aspergillus oryzae, Aspergillus sojae, or Aspergillus luchuensis.

[15] The composition according to any one of

[11] to

[14] , wherein the plant containing anthocyanins is corn.

[16] An antioxidant or wrinkle-forming inhibitor comprising the composition according to any one of

[11] to

[15] .

[0020] This disclosure provides a novel anthocyanin-derived composition, a method for producing the same, and a cosmetic composition containing the same.

[0021] According to this disclosure, in one embodiment, a plant extract containing anthocyanins having a pH of 5 to 8 can be produced. Such a plant extract has excellent long-term storage stability, and furthermore, it has the effects of an anti-aging agent, enzyme inhibitor, anti-inflammatory agent, anti-aging agent, DNA damage inhibitor, reactive oxygen species scavenger, and cell activator, and can be suitably used as a functional cosmetic composition.

[0022] According to this disclosure, in one embodiment, a plant fermentation extract containing anthocyanins can be produced. Such a fermentation extract has excellent long-term storage stability, and furthermore, it has antioxidant effects and inhibits wrinkle formation by suppressing the enzymatic activity of collagenase, an enzyme involved in wrinkle formation. For example, it can be suitably used as a functional cosmetic composition.

[0023] Fluorescence images of nuclear staining, intracellular hydrogen peroxide, DNA damage markers, and cellular senescence markers (P21 and SA-β-gal) from anti-aging tests, DNA damage inhibition tests, and reactive oxygen species scavenging tests using hydrogen peroxide addition. Fluorescence images of nuclear staining, intracellular hydrogen peroxide, and DNA damage markers from anti-inflammatory tests, DNA damage inhibition tests, and reactive oxygen species scavenging tests using UVB treatment.

[0024] The present disclosure will be described in detail below. (A) Means A <Plants containing anthocyanins> The plants containing anthocyanins in the present disclosure are preferably plants that contain a large amount of anthocyanins, which are a type of flavonoid (polyphenol) and are known as red or purple pigments. Anthocyanins are abundant in purple plants, and examples of purple plants include fruits such as grapes, apples, blueberries, and cherries, and vegetables such as eggplants, perilla, purple onions, purple cabbage, purple corn, black glutinous corn, black beans, and black rice.

[0025] Among them, purple corn (also called purple cod) and black glutinous corn, which contain purple coloration, are particularly desirable as sustainable anthocyanin-containing plant materials because the non-edible cob contains a large amount of anthocyanins. Purple corn is characterized mainly by its red to purple husk. Furthermore, the inside of the kernels and the cob are often similarly red to purple. Various types of purple corn are cultivated in the market through selective breeding and / or genetic modification technology.

[0026] In the purple corn and other pericarp used in this disclosure, it is preferable that the red to purple portion constitutes more than half of the entire pericarp, more preferably 70% or more, and even more preferably 80% or more. In this disclosure, commercially available purple corn and other pericarp can be used.

[0027] <Extraction Solvent> Preferably, the extraction solvent in this disclosure is a solvent used to extract an anthocyanin-containing active ingredient from an anthocyanin-containing plant, and is an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin. The aqueous solution preferably contains one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin at a concentration of 5% by mass or more and 90% by mass or less, and more preferably at a concentration of 40% by mass or more and 60% by mass or less.

[0028] The extraction solvent may include solvents other than 1,3-propanediol, 1,3-butylene glycol, and glycerin, as long as they do not impair the effects of the present disclosure. For example, monohydric alcohols such as ethanol and isopropanol, and polyhydric alcohols such as pentylene glycol, 1,2-hexanediol, ethylhexylglycerin, caprylyl glycol, and polyglycerin can also be used. To avoid impairing the effects of the present disclosure, the amount of these solvents added is 10 parts by mass or less, preferably 5 parts by mass or less, more preferably 3 parts by mass or less, and more preferably 0 parts by mass, per 100 parts by mass of the extraction solvent.

[0029] <Extraction Conditions> When extracting plants containing anthocyanins with an extraction solvent, the mass ratio of the former to the latter (former:latter) is preferably 0.5:9.5 to 3:7, more preferably 0.7:9.3 to 3:7, and even more preferably 1:9 to 2:8. The extraction temperature is preferably 15 to 60°C, and more preferably 20 to 50°C. The extraction time is preferably 1 to 96 hours, and more preferably 2 to 72 hours. A known or conventional stirrer may be used during extraction. After extraction, the separated liquid portion may be used as the plant extract described later by a known or conventional solid-liquid separation method such as decantation or filtration.

[0030] <Plant Extract> A composition according to one aspect of the present disclosure is a plant extract containing anthocyanin obtained by extracting a plant containing anthocyanin with an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin. The amount of the solvent (for example, the aqueous solution) with respect to the entire plant extract (100% by mass) may be 70 to 95% by mass, 70 to 93% by mass, 80 to 90% by mass, etc.

[0031] As one aspect of the present disclosure, there is a plant extract characterized by containing anthocyanin derived from purple corn or the like, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water. The plant extract can be produced by extracting a plant containing anthocyanin with an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin. Alternatively, it can be extracted from a plant containing anthocyanin using water, and then one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin can be added to the extract, and water and one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin can be used as the solvent in the plant extract.

[0032] In this solvent, it is preferable that one or more selected from the group consisting of 1,3 - propanediol, 1,3 - butylene glycol, and glycerin are contained at a concentration of 5% by mass or more and 90% by mass or less. More preferably, the concentration is 20% by mass or more and 70% by mass or less, and even more preferably, the concentration is 40% by mass or more and 60% by mass or less. In this solvent, solvents other than one or more selected from the group consisting of 1,3 - propanediol, 1,3 - butylene glycol, and glycerin may be contained as long as the effects of the present disclosure are not impaired. For example, monohydric alcohols such as ethanol and isopropanol, and polyhydric alcohols such as pentylene glycol, 1,2 - hexanediol, ethylhexyl glycerin, caprylyl glycol, and polyglycerin can also be used. As long as the effects of the present disclosure are not impaired, the addition amount is 10 parts by mass or less, preferably 5 parts by mass or less, even more preferably 3 parts by mass or less, and more preferably 0 parts by mass in 100 parts by mass of the extraction solvent.

[0033] As a pretreatment for extraction with the aqueous solution, plants containing anthocyanins can be dried and / or frozen. By drying and / or freezing, the moisture contained in plants containing anthocyanins can be reduced, and effects such as shortening the time required for extraction and increasing the amount of anthocyanins extracted can be achieved. From the perspective of plant preservation, drying is preferred as a pretreatment. The drying temperature is preferably 30°C to 90°C. The freezing temperature is preferably from -196°C to -5°C. Examples of drying methods include hot air drying and vacuum drying. Examples of freezing methods include slow freezing using a normal refrigerator, rapid freezing that cools the object in a short time, and a freezing method using liquid nitrogen. The extract may be an aqueous solution containing an extraction solvent, or may be a solution or solid from which part or all of the extraction solvent has been removed. Known methods can be used to remove the extraction solvent. For example, it may be evaporation to dryness by heating under normal pressure, vacuum distillation at normal temperature or by heating under reduced pressure, freeze - drying, etc.

[0034] The plant extract in the present disclosure has a pH of 4.5 to 8 and excellent long - term storage stability. More preferably, the pH is 5 to 7 or 5 to 8.

[0035] One aspect of this disclosure also includes a plant extract characterized by containing anthocyanins derived from maize, such as purple maize, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water. By containing anthocyanins derived from purple maize, etc., and one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, the plant extract has excellent long-term storage stability.

[0036] <Antioxidants> The antioxidants in this disclosure include products that have the property of suppressing the activity of reactive oxygen species radicals and are applied to animals, including humans (including parenteral administration such as topical application or oral administration; the same applies hereinafter). For example, when mitochondria in human and non-human animal cells react carbohydrates and lipids with oxygen to produce ATP, reactive oxygen species radicals are generated by so-called oxidative stress, which occurs when human or animal cells are exposed to ultraviolet light and air contaminated with chemical substances. The compositions of this disclosure (for example, the plant extracts) have the effect of suppressing the activity of reactive oxygen species radicals.

[0037] The antioxidants in this disclosure include the compositions of this disclosure (e.g., the plant extracts), and the plant extracts may be used in any proportion. The antioxidants in this disclosure may also include various additives commonly used in antioxidants other than the compositions of this disclosure (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. Examples include components derived from plants that have been reported to have antioxidant effects (e.g., grapes, ginseng, or comfrey), proanthocyanidins, tocopherols and their derivatives, ascorbic acid and its derivatives, hesperidin, glucosyl hesperidin, ergothioneine, sodium bisulfite, erythorbic acid and its salts, flavonoids, glutathione, glutathione peroxidase, glutathione-S-transferase, catalase, superoxide dismutase, thioredoxin, taurine, thiotaurine, or hypotaurine.

[0038] <Anti-aging agents> The anti-aging agents in this disclosure include products that are applied to animals, including humans, and that play a role in slowing down the progression of cellular aging, suppressing the appearance of senescent cells, or reducing the effects of cellular aging. For example, when human or non-human animal cells are subjected to so-called oxidative stress, such as exposure to ultraviolet light or air polluted with chemicals, DNA damage occurs, inducing cellular aging and accelerating the process of aging. Senescent cells are known to exhibit senescence-related acid β-galactosidase activity. It is also known that senescent cells have increased production of p21 protein. The compositions of this disclosure (e.g., the plant extracts) have the effect of reducing or decreasing senescence-related acid β-galactosidase activity and p21 protein expression.

[0039] The anti-aging agent in this disclosure includes the composition of this disclosure, and the composition may be used in any proportion. The anti-aging agent in this disclosure may also include various additives commonly used in anti-aging agents other than the composition of this disclosure (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. For example, it may include flavonoids, saponins, polysaccharides, alkaloids, retinoids, various plant extracts, etc., which have been reported to have anti-aging effects. Specifically, these include astragaloside, icariside, tetrahydrocurcumin, quercetin, butein, berberine, catechin, curcumin, epigallocatechin gallate, gastrodin, 6-gingerol, glaucarbinone, ginsenoside Rg1, luteolin, icariside II, naringenin, resveratrol, theaflavin, carnosic acid, catalpol, chrysophanol, cycloastragenol, emodin, galangin, echinacoside, ferulic acid, huperzine, honokiol, isolencinin, phycocyanin, proanthocyanidin, rosmarinic acid, oxymatrin, pisaid, puerarin, and salvianolic acid B.

[0040] <DNA Damage Inhibitors> The DNA damage inhibitors in this disclosure include products that have the property of suppressing intracellular DNA damage and are applicable to animals, including humans. The causes of DNA damage can be broadly classified into two categories: those originating inside the cell and those originating outside the cell. Specifically, intracellular causes include DNA damage caused by reactive oxygen species generated by metabolism, such as mitochondria. Extracellular causes include DNA damage caused by ultraviolet light, compounds in the environment, cigarette smoke, and environmental pollution. The compositions of this disclosure (for example, the plant extracts) have the effect of reducing or decreasing DNA damage.

[0041] The DNA damage inhibitors in this disclosure include the compositions of this disclosure, and these compositions can be used in any proportion. The DNA damage inhibitors in this disclosure may also include various additives commonly used in DNA damage inhibitors other than the compositions of this disclosure (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. Examples of such additives include cinchona extract, comfrey extract, coffee tree extract, kudzu extract, burdock extract, Coptis japonica extract, Sophora flavescens extract, Chlorella extract, lavender extract, evening primrose extract, rose extract, Gynostemma pentaphyllum extract, Enmeisou extract, Lamium album extract, carrot extract, linden extract, citronella extract, lotus extract, tea extract, and okra extract, which have been reported to have DNA damage inhibitory effects.

[0042] <Reactive Oxygen Species Removers> The reactive oxygen species removers in this disclosure include products that have the property of removing reactive oxygen species from cells and are applicable to animals, including humans. Reactive oxygen species are known to be generated in the energy metabolic process in mitochondria, and reactive oxygen species include superoxide anion radicals (•O ２ － ), hydrogen peroxide (H ２ O ２ ), hydroxyl radical (•OH), and hydroxide ion (OH) －) and others are included. When mitochondrial dysfunction occurs due to oxidative stress, environmental stress, or cellular aging, excessive reactive oxygen species are generated, causing various types of damage to cells, including DNA damage. The compositions of this disclosure (for example, the plant extracts) have the effect of reducing or removing reactive oxygen species in cells generated by oxidative stress, etc.

[0043] The reactive oxygen species scavenger in this disclosure includes the composition of this disclosure, and the composition can be used in any proportion. The reactive oxygen species scavenger in this disclosure may also include various additives commonly used in reactive oxygen species scavengers other than the composition (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. Examples include retinoids, ascorbic acid, tocopherol, flavonol, isofvonoid, flavanone, flavone, carotenoid, hydroxycinnamic acid, hydroxybenzoic acid, and coenzyme Q10, which have been reported to have reactive oxygen species scavenging effects.

[0044] <Enzyme Inhibitors> The enzyme inhibitors in this disclosure include products that suppress the activity of enzymes and are applicable to animals, including humans. Enzymes act as catalysts that promote chemical reactions in the bodies of animals, including humans, but enzyme inhibitors play a role in suppressing specific reactions by interfering with their function. For example, elastase is an enzyme that breaks down elastin, a major component of connective tissue, and excessive activity can cause tissue damage, inflammation, and wrinkle formation. For example, collagenase is an enzyme that breaks down collagen, a component that makes up the skin, and excessive activity can cause wrinkle formation in the skin. For example, tyrosinase is an enzyme involved in melanin production, and excessive activity can cause age spots and freckles due to melanin. The compositions of this disclosure (e.g., the plant extracts) have the effect of inhibiting the enzyme activity described above and have the effect of reducing the progress of various reactions mediated by enzymes.

[0045] The enzyme inhibitors in this disclosure include the compositions of this disclosure (e.g., the plant extracts), and the compositions may be used in any proportion. The enzyme inhibitors in this disclosure may also include various additives commonly used in enzyme inhibitors other than the compositions (e.g., the plant extracts), as long as they do not impair the effects of this disclosure.

[0046] For example, extracts of pyrroloquinoline quinone, guava, guarana, rosemary, St. John's wort, peony root, hops, peony, mulberry bark and eucalyptus, urlin, etc., which have been reported to have elastase inhibitory activity; extracts of seaweed belonging to the genera Fucus, Ascophyllum, Lessonia or Darvilleia of brown algae; extracts of plants such as birch, cinnamon, winter linden, summer linden, European linden, Japanese linden, loquat, witch hazel, etc.; extracts of plants such as ginger, hydrolyzed almond, Sanguisorba officinalis, clove, rosehip, hawthorn, birch, etc. These include extracts from plants such as cinnamon, St. John's wort, cinnamon, geranium, comfrey, linden, and peony root; extracts from plants such as yew, schisandra, bupleurum, magnolia, angelica, clematis, and forsythia; extracts from plants such as guava, guarana, rosemary, St. John's wort, peony root, hops, peony, mulberry bark, and mocca; extracts from plants such as burnet, Japanese knotweed, Japanese knotweed, giant knotweed, lemon balsam, rosemary, safflower, and ginger; and extracts from plants such as turmeric, yarrow, mallow, thyme, yacon, and rosehip.

[0047] Other substances reported to have collagenase inhibitory activity include buckwheat hull protein extract, Schizonepeta extract, peppermint extract, lactoferrin, caffeic acid or its salts, ascorbic acid, erythorbic acid and mixtures thereof, acetyl polyamide, gallotannin, κ-casein, cocoa hull extract, hydroxamic acid tetrapeptide, and 1,3-bis-substituted-naphthylaminocarbonylphenylurea derivatives.

[0048] <Anti-inflammatory agents> The anti-inflammatory agents in this disclosure include products used to suppress inflammation in animals, including humans. For example, when human or non-human animal cells are exposed to so-called oxidative stress, such as ultraviolet rays or air polluted with chemicals, inflammation may occur, and the production of inflammatory cytokines such as interleukin-8 (IL-8), interleukin-6 (IL-6), and interleukin-1 (IL-1) by macrophages, epithelial cells, airway smooth muscle cells, and vascular endothelial cells increases. Inflammatory cytokines are also included in the SASP (senescence-associated secretory phenotype) factor secreted by senescent cells, and it is known that SASP factors secreted by senescent cells are the cause of various age-related diseases. The composition of this disclosure (for example, the plant extract) suppresses the production of IL-8.

[0049] The anti-inflammatory agents in this disclosure include the compositions of this disclosure (e.g., the plant extracts), and the plant extracts may be used in any proportion. The anti-inflammatory agents in this disclosure may also include various additives commonly used in anti-inflammatory agents other than the compositions of this disclosure (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. Examples include polyphenols, flavonoids, curcumin, ascorbic acid, quercetin, epigallocatechin gallate, dipotassium glycyrrhizinate, ascorbyl tetrahexyldecanoate, and bisabolol, which have been reported to have anti-inflammatory effects.

[0050] <Cell Activators> Cell activators in this disclosure include products used to maintain or enhance the cellular function, cellular activity, and cell division of animals, including humans, which are applied to animals, including humans. For example, cell activators can act on cells to improve their cell division, survival rate, and activity. Cell activators increase the production of ATP (adenosine triphosphate), which is the energy source for cells. The compositions of this disclosure (e.g., the plant extracts) may be used as activators for various cells, but are preferably used as activators for skin cells. Examples of skin cells to which cell activators according to embodiments of this disclosure can be applied include human normal keratinocytes and human normal fibroblasts.

[0051] The cell activators in this disclosure include the compositions of this disclosure (e.g., the plant extracts), and the plant extracts may be used in any proportion. The cell activators in this disclosure may also include various additives commonly used for cell activation other than the compositions of this disclosure (e.g., the plant extracts), as long as they do not impair the effects of this disclosure. Examples include amino acids such as γ-aminobutyric acid and ε-aminopronic acid, which have been reported to have cell activating effects; retinol and its derivatives; vitamins such as thiamine, riboflavin, pyridoxine hydrochloride, and pantothenic acids; α-hydroxy acids such as glycolic acid and lactic acid; tannins, flavonoids, saponins, and allantoin.

[0052] <Cosmetic Compositions> The cosmetic compositions in this disclosure include all cosmetics applied to the skin, hair, etc., of animals, including humans. Since the aqueous solution of the compositions in this disclosure (for example, the plant extracts) containing the extraction solvent preferably has a pH of 5 to 8, pH control is easy when combined with materials used in cosmetics, and they can be used in a wide range of applications such as facial cleansers, basic cosmetics, and makeup cosmetics.

[0053] In the cosmetic compositions of this disclosure, the composition of this disclosure (e.g., the plant extract), an anti-aging agent, enzyme inhibitor, anti-inflammatory agent, anti-aging agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator containing the composition of this disclosure (e.g., the plant extract) may be used in any proportion. In addition, the cosmetic compositions of this disclosure may contain various additives commonly used in cosmetics other than the composition of this disclosure (e.g., the plant extract), an anti-aging agent, enzyme inhibitor, anti-inflammatory agent, anti-aging agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator containing the composition of this disclosure (e.g., the plant extract), as long as they do not impair the effects of this disclosure. Examples include surfactants, thickeners, humectants, pH adjusters, chelating agents, preservatives, colorants, fragrances, whitening agents, UV protection agents, etc.

[0054] (B) Means B <Plants containing anthocyanins> The plants containing anthocyanins in this disclosure are preferably plants that contain a large amount of anthocyanins, which are a type of flavonoid (polyphenol) and are known as red or purple pigments. Anthocyanins are abundant in purple plants, and examples of purple plants include fruits such as grapes, apples, blueberries, and cherries, and vegetables such as eggplants, perilla, purple onions, purple cabbage, purple corn, black glutinous corn, black beans, and black rice.

[0055] Among them, purple corn (also called purple cod) and black glutinous corn, which contain purple coloration, are particularly desirable as sustainable anthocyanin-containing plant materials because the non-edible cob contains a large amount of anthocyanins. Purple corn is characterized mainly by its red to purple husk. Furthermore, the inside of the kernels and the cob are often similarly red to purple. Various types of purple corn are cultivated in the market through selective breeding and / or genetic modification technology.

[0056] In the purple corn and other pericarp used in this disclosure, it is preferable that the red to purple portion constitutes more than half of the entire pericarp, more preferably 70% or more, and even more preferably 80% or more. In this disclosure, commercially available purple corn and other pericarp can be used.

[0057] <Aspergillus oryzae fermented product> The aspergillus oryzae fermented product of this disclosure is a product obtained by fermenting plants containing anthocyanins with Aspergillus oryzae. Aspergillus is a fungus belonging to the genus Aspergillus, and species such as Aspergillus oryzae, Aspergillus sojae, Aspergillus kawachii, Aspergillus luchuensis, Aspergillus awamori, Aspergillus niger, Aspergillus glaucus, and Aspergillus tamari can be used. In addition to Aspergillus oryzae, other filamentous fungi of the Monascus genus can also be used, such as Monascus pilosus, Monascus purpureus, and Monascus anka.

[0058] From a safety standpoint, it is preferable to use fungi that have a history of being used in food production, such as in the manufacture of fermented products, and more preferably, fungi of the genus Aspergillus, and even more preferably, Aspergillus oryzae, Aspergillus sojae, or Aspergillus luchuensis.

[0059] Aspergillus oryzae fermented products can be produced, for example, by solid-state fermentation, in which Aspergillus oryzae spores are scattered onto plants containing anthocyanins, and the Aspergillus oryzae is grown on the surface. The plants containing anthocyanins used in solid-state fermentation can be dried and / or frozen. From the viewpoint of plant preservation, drying is preferred as a pretreatment. Drying methods include hot air drying and vacuum drying. Freezing methods include slow freezing in a normal freezer, rapid freezing which cools the target in a short time, and freezing using liquid nitrogen. Furthermore, the plants containing anthocyanins can also be crushed. As a pretreatment for adding Aspergillus oryzae, pretreatments commonly used in solid-state fermentation using Aspergillus oryzae can be used. For example, a hydration step can be performed to add water to the plants containing anthocyanins to facilitate Aspergillus oryzae growth, and a steaming step can be performed after the hydration step, in which the plants are heated at atmospheric pressure.

[0060] Fermenting plants containing anthocyanins with Aspergillus oryzae increases the amount of water-soluble components, improving the extraction efficiency in the extraction process using the extraction solvent described later. The process of fermenting plants containing anthocyanins with Aspergillus oryzae is also called the process of fermenting plants containing anthocyanins with Aspergillus oryzae.

[0061] <Extraction Solvent> The extraction solvent in this disclosure is a solvent used to extract the active ingredient containing anthocyanin from the Aspergillus oryzae ferment product, and is preferably an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. The aqueous solution preferably contains 5 to 90 parts by mass of one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin per 100 parts by mass, and more preferably 40 to 60 parts by mass.

[0062] The extraction solvent may include any solvent other than 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin, as long as it does not impair the effects of the present disclosure. For example, monohydric alcohols such as isopropanol and polyhydric alcohols such as pentylene glycol, 1,2-hexanediol, ethylhexylglycerin, caprylyl glycol, and polyglycerin can also be used. To avoid impairing the effects of the present disclosure, the amount of these solvents added is 10 parts by mass or less, preferably 5 parts by mass or less, more preferably 3 parts by mass or less, and more preferably 0 parts by mass, per 100 parts by mass of the extraction solvent.

[0063] <Extraction Conditions> When extracting the Aspergillus oryzae fermented product with an extraction solvent, the mass ratio of the former to the latter (former:latter) is preferably 0.5:9.5 to 3:7, more preferably 0.7:9.3 to 3:7, and even more preferably 1:9 to 2:8. The extraction temperature is preferably 15 to 60°C, and more preferably 20 to 50°C. The extraction time is preferably 1 to 96 hours, and more preferably 2 to 72 hours. A known or conventional stirrer may be used during extraction. After extraction, the separated liquid portion may be used as the Aspergillus oryzae fermentation extract described later by a known or conventional solid-liquid separation method such as decantation or filtration.

[0064] <Aspergillus oryzae fermentation extract> The Aspergillus oryzae fermentation extract in this disclosure is an Aspergillus oryzae fermentation extract containing polyphenols including anthocyanins of plant origin obtained by extracting the Aspergillus oryzae ferment with the extraction solvent, and various physiologically functional components derived from koji. It is also called an extract of Aspergillus oryzae ferment. The extraction solvent is preferably an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. The Aspergillus oryzae fermentation extract can be produced by extracting the Aspergillus oryzae ferment with an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. Alternatively, the Aspergillus oryzae ferment is extracted using water, and then one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin is added to the extract to use water and one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin as solvents in the Aspergillus oryzae fermentation extract.

[0065] In the solvent, it is preferable that one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin be included in an amount of 5 to 90 parts by mass per 100 parts by mass of the solvent. More preferably, it is 20 to 70 parts by mass, and even more preferably 40 to 60 parts by mass. In the solvent, solvents other than one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin may be included as long as they do not impair the effects of the present disclosure. For example, monohydric alcohols such as isopropanol and polyhydric alcohols such as pentylene glycol, 1,2-hexanediol, ethylhexylglycerin, caprylyl glycol, and polyglycerin can also be used. In order not to impair the effects of the present disclosure, the amount of these solvents added is 10 parts by mass or less per 100 parts by mass of the extraction solvent, preferably 5 parts by mass or less, more preferably 3 parts by mass or less, and more preferably 0 parts by mass.

[0066] The extract may be an aqueous solution containing the extraction solvent, or it may be a solution or solid from which some or all of the extraction solvent has been removed. Known methods can be used to remove the extraction solvent. For example, evaporation to dryness by heating under atmospheric pressure, distillation under reduced pressure at room temperature or by heating, or freeze-drying can be used.

[0067] The koji mold fermentation extract in this disclosure, when in aqueous solution, has a pH of 4 to 8 and exhibits excellent long-term storage stability. More preferably, the pH is 4.5 to 7.5. The extraction of a koji mold fermented product obtained by fermenting an anthocyanin-containing plant with koji mold using a solvent is also referred to as the solvent extraction process.

[0068] <Antioxidants> The antioxidants in this disclosure include products that have the property of suppressing the activity of reactive oxygen species radicals and are applicable to animals, including humans. For example, reactive oxygen species radicals are generated when mitochondria in human and non-human animal cells react carbohydrates and lipids with oxygen to produce ATP, or when human and animal cells are exposed to ultraviolet light and air polluted with chemicals, resulting in so-called oxidative stress. The anthocyanin-containing plant koji mold fermentation extract in this disclosure has the effect of suppressing the activity of reactive oxygen species radicals.

[0069] The antioxidants in this disclosure include plant fermentation extracts containing the anthocyanins of this disclosure, and the plant fermentation extracts may be used in any proportion. The antioxidants in this disclosure may also include various additives commonly used in antioxidants other than plant fermentation extracts containing the anthocyanins of this disclosure, as long as they do not impair the effects of this disclosure.

[0070] Examples include components derived from plants that have been reported to have antioxidant effects (e.g., grapes, ginseng, or comfrey), proanthocyanidins, tocopherols and their derivatives, ascorbic acid and its derivatives, hesperidin, glucosyl hesperidin, ergothioneine, sodium bisulfite, erythorbic acid and its salts, flavonoids, glutathione, glutathione peroxidase, glutathione-S-transferase, catalase, superoxide dismutase, thioredoxin, taurine, thiotaurine, or hypotaurine.

[0071] <Wrinkle Formation Inhibitors> The wrinkle formation inhibitors in this disclosure include products that suppress wrinkle formation and are applicable to animals, including humans. Wrinkle formation occurs, for example, when enzymes that break down collagen are activated or when the activity of enzymes that synthesize collagen is suppressed. Therefore, wrinkle formation inhibitors play a role in suppressing the activity of enzymes that break down collagen or the activity of enzymes that synthesize collagen. For example, collagenase enzyme is an enzyme that breaks down collagen, a component that makes up the skin, and excessive activity causes wrinkle formation in the skin. The Aspergillus oryzae fermentation extract of plants containing anthocyanins in this disclosure has the effect of inhibiting the activity of such enzymes, that is, it has the effect of reducing the progress of various reactions by enzymes, and is also called an enzyme inhibitor. Collagenase inhibitors, which are enzyme inhibitors, are preferred as wrinkle formation inhibitors. The wrinkle formation inhibitors in this disclosure include the Aspergillus oryzae fermentation extract of plants containing anthocyanins in this disclosure, and the Aspergillus oryzae fermentation extract can be used in any proportion.

[0072] Furthermore, the wrinkle-forming inhibitor of this disclosure may contain various additives commonly used in wrinkle-forming inhibitors other than the anthocyanin-containing plant fermentation extract of this disclosure, as long as they do not impair the effects of this disclosure. For example, pyrroloquinoline quinone, extracts of guava, guarana, rosemary, St. John's wort, peony root, hops, peony, mulberry bark and eucalyptus, phlox, etc., which have been reported to have elastase inhibitory activity, extracts of seaweed belonging to the genera Fucus, Ascophyllum, Lessonia or Darvilleia of brown algae, extracts of plants such as birch, cinnamon, winter linden, summer linden, European linden, linden, loquat, witch hazel, etc., extracts of plants such as ginger, hydrolyzed almond, Sanguisorba officinalis, clove, rosehip, hawthorn, birch, etc. These include extracts from plants such as cinnamon, St. John's wort, cinnamon, geranium, comfrey, linden, and peony root; extracts from plants such as yew, schisandra, bupleurum, magnolia, angelica, clematis, and forsythia; extracts from plants such as guava, guarana, rosemary, St. John's wort, peony root, hops, peony, mulberry bark, and mocca; extracts from plants such as burnet, Japanese knotweed, Japanese knotweed, giant knotweed, lemon balsam, rosemary, safflower, and ginger; and extracts from plants such as turmeric, yarrow, mallow, thyme, yacon, and rosehip. Other substances reported to have collagenase inhibitory activity include buckwheat hull protein extract, Schizonepeta extract, peppermint extract, lactoferrin, caffeic acid or its salts, ascorbic acid, erythorbic acid and mixtures thereof, acetyl polyamide, gallotannin, κ-casein, cocoa hull extract, hydroxamic acid tetrapeptide, and 1,3-bis-substituted-naphthylaminocarbonylphenylurea derivatives.

[0073] <Cosmetic Compositions> The cosmetic compositions in this disclosure include all cosmetics that can be applied to the skin, hair, etc., of animals, including humans. The anthocyanin-containing plant koji mold fermentation extract in this disclosure has an aqueous solution containing the extraction solvent, preferably with a pH of 4 to 8. Therefore, pH control is easy when combined with materials used in cosmetics, and it can be used in a wide range of applications such as facial cleansers, basic cosmetics, and makeup cosmetics.

[0074] The cosmetic compositions in this disclosure may use plant fermentation extracts containing anthocyanins as disclosed, antioxidants or wrinkle inhibitors containing plant fermentation extracts containing anthocyanins as disclosed, in any proportion. The cosmetic compositions in this disclosure may also contain various additives commonly used in cosmetics, other than plant fermentation extracts containing anthocyanins as disclosed, antioxidants or wrinkle inhibitors containing plant fermentation extracts containing anthocyanins as disclosed, as long as they do not impair the effects of this disclosure. Examples include surfactants, thickeners, moisturizers, pH adjusters, chelating agents, preservatives, colorants, fragrances, whitening agents, UV protection agents, etc.

[0075] <Method for Producing the Composition> (A') Method A' This disclosure provides a method for producing a plant extract, using a plant containing anthocyanins as a raw material and an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin as an extraction solvent. In the method for producing the plant extract, it is preferable to have a step of extracting the plant containing anthocyanins with an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, using a plant containing anthocyanins as a raw material and an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin as an extraction solvent. For the plant containing anthocyanins, the extraction solvent, the extraction conditions, and the plant extract in the method for producing the plant extract, refer to the descriptions of them in (A) Method A above.

[0076] (B') Means B' The present disclosure provides a method for producing an Aspergillus oryzae fermentation extract, comprising a fermentation step of fermenting a plant containing anthocyanins with Aspergillus oryzae, and a step of extracting the Aspergillus oryzae ferment product obtained in the fermentation step with a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol. Preferably, the solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol is an aqueous solution containing one or more of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. For the plant containing anthocyanins, Aspergillus oryzae ferment product (including fermentation conditions), extraction solvent, extraction conditions, and Aspergillus oryzae fermentation extract in the method for producing the plant extract, refer to the descriptions of them in (B) Means B.

[0077] The present disclosure will be further described below with reference to examples, but the present disclosure is not limited to these examples. Unless otherwise specified, "parts" means "parts by mass" and "%" means "percent mass".

[0078] (A) Method A <pH measurement> The pH of the extract was measured using a pH meter F-55 (manufactured by Horiba, Ltd.).

[0079] <Storage Stability Test> The extract was added to a 96-well plate (Thermo Fisher Scientific) at a rate of 100 μL / well, and the absorbance spectrum at wavelengths of 380-780 nm was measured using a microplate reader SH-9000Lab (Hitachi High-Tech Science Corporation). From the transmittance data at each wavelength, the a* value was calculated using UVPC color measurement Personal Spectroscopy Software (Shimadzu Corporation). The a* value (initial value) of the extracts of the examples and comparative examples described later was measured, and the Δa* value after 4°C / 1 month storage and the Δa* value after 50°C / 1 month storage were calculated by subtracting the a* value after 4°C / 1 month storage and the Δa* value after 50°C / 1 month storage, respectively. The Δa* value is a value that reflects the red color and mainly measures the degree of change of the red component contained in anthocyanins.

[0080] <Antioxidant Activity Measurement> Using an H-ORAC analysis kit (manufactured by FUJIFILM Wako Pure Chemical Corporation), the high antioxidant activity value of the extract (plant extract) was measured. For the stability of antioxidant activity over time, the values before and after storage at 50°C for 1 month were measured, and the value after storage at 50°C for 1 month was calculated when the antioxidant activity value before storage was set to 100. <Measurement of Polyphenol Content> The polyphenol content in the extract (plant extract) was measured by the method described in "Micro-titer plate assay for measurement of total phenolic and total flavonoid contents in medicinal plant extract. Arabian Journal of Chemistry Volume 16, Issue 9, September 2023".

[0081] <Measurement of Anthocyanin Content> Using an Anthocyanins assay kit (manufactured by Fujigen Corporation), the anthocyanin content in the extract (plant extract) was measured.

[0082] <Anti-aging Test, DNA Damage Inhibition Test, and Reactive Oxygen Species Removal Test by Hydrogen Peroxide Addition> Human normal keratinocytes derived from neonatal foreskin (manufactured by Kurabo Industries Ltd.) were seeded at 2×10 ４ cells / cm ２ in a Sensoplate (trademark) 24-well plate (manufactured by Greiner Japan Co., Ltd.) using HuMedia-KG2 medium (manufactured by Kurabo Industries Ltd.) at a culture solution volume of 1 mL / well. After culturing for 24 hours in an atmosphere of 37°C and 5% CO ２ , the HuMedia-KG2 medium (manufactured by Kurabo Industries Ltd.) was removed. HuMedia-KB2 medium (manufactured by Kurabo Industries Ltd.) added with the extract (plant extract) to a final concentration of 0.55% or 1.1% was added to a 24-well plate at 1 mL / well, and incubated at 37°C and 5% CO ２After culturing for 24 hours under atmospheric conditions, the HuMedia-KB2 medium was removed. For the group without extract, only HuMedia-KB2 medium was added at a rate of 1 mL / well. The cells were washed with 1 mL / well of D-PBS(-) buffer (Fujifilm Wako Pure Chemical Industries, Ltd.). After removing the D-PBS(-) buffer, 30% hydrogen peroxide solution (Merck KGaA) was added to the HuMedia-KB2 medium to a final concentration of 170 μM as an oxidative stress agent. This hydrogen peroxide-added HuMedia-KB2 medium was then added to 24-well plates at a rate of 0.5 mL / well and incubated at 37°C under 5% CO2. ２ The cells were incubated for 1 hour under controlled atmosphere. For comparison, the untreated oxidative stress group was cultured in HuMedia-KB2 medium without added hydrogen peroxide. The hydrogen peroxide-added HuMedia-KB2 medium was removed, and the cells were washed twice with 1 mL / well of D-PBS(-) buffer. HuMedia-KB2 medium to which extracts (plant extracts) were added to a final concentration of 0.55% or 1.1% was added at 1 mL / well to 24-well plates, and the cells were incubated at 37°C in 5% CO2. ２The cells were cultured for 24 hours under atmospheric conditions. For comparison, only HuMedia-KB2 medium was added to the cells without the addition of extract (plant extract). After washing the cells cultured for 24 hours with D-PBS(-) buffer (manufactured by Fujifilm Wako Pure Chemical Corporation), the reactive oxygen species scavenging ability was confirmed by visualizing intracellular hydrogen peroxide using Hyp-Stamp™ (manufactured by Funakoshi Co., Ltd.), and the DNA damage suppression ability was confirmed by visualizing DNA damage markers using DNA Damage Detection Kit-γH2AX-Red- (manufactured by Dojin Chemical Laboratories Co., Ltd.). Nuclear staining was performed using CellCover (Funakoshi Co., Ltd.) as the cell fixative, DAKO protein block serum free (Agilent Technologies, Inc.) as the blocking reagent, and DAPI solution (Thermo Fisher Scientific) as the counterstain. Cell permeabilization was performed using 1% TritonX-100-added D-PBS(-) buffer. Immunostaining was performed according to the protocol provided with Hyp-Stamp™, DNA Damage Detection Kit-γH2AX-Red-, and fluorescence images were taken using a fluorescence microscope. Furthermore, cells cultured for 24 hours were washed with D-PBS(-) buffer (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), and then stained with senescence-associated β-galactosidase (SA-β-gal), one of the cellular senescence markers, using Cellular Senescence Detection Kit-SPiDER-βGal (manufactured by Dojin Chemical Laboratories, Ltd.). In addition, the p21 protein, another cellular senescence marker, was stained using Anti-p21 antibody (manufactured by Abcam), to confirm anti-aging activity. Nuclear staining was performed using CellCover (Funakoshi Co., Ltd.) as the cell fixative, DAKO protein block serum free (Agilent Technologies, Inc.) as the blocking reagent, and DAPI solution (Thermo Fisher Scientific) as the counterstain. Cell permeabilization was performed using 1% Triton X-100-added D-PBS(-) buffer. After immunostaining, fluorescence images were taken using a fluorescence microscope.

[0083] <Collagenase Activity Inhibition Test> The collagenase inhibitory activity of the extract (plant extract) was measured using the method described in Quinic acid esters from Plucea indexa with collagenase, MMP-2 and MMP-9 inhibitory activities. Phytother Res. 2008 Feb;22(2):264-266, and the 50% inhibitory concentration (IC50) was calculated.

[0084] <Elastase Activity Inhibition Test> The elastase inhibitory activity of the extract (plant extract) was measured using the method described in "Functionality of Vegetables, mainly Kyoto Vegetables: Hyaluronidase, Elastase, and Collagenase Inhibitory Activity" Kinki University Faculty of Agriculture Bulletin No. 50, pp. 41-45 (2016), and the 50% inhibitory concentration (IC50) was calculated.

[0085] <Tyrosinase Activity Inhibition Test> The tyrosinase inhibitory activity of the extract (plant extract) was measured using the method described in "Functionality of Vegetables, mainly Kyoto Vegetables: Hyaluronidase, Elastase, and Collagenase Inhibitory Activity," Kinki University Faculty of Agriculture Bulletin No. 50, pp. 41-45 (2016), and the 50% inhibitory concentration (IC50) was calculated.

[0086] <Anti-inflammatory test, DNA damage inhibition test, and reactive oxygen species removal test using UVB treatment> 2 x 10⁶ human normal keratinocytes derived from neonatal foreskin (manufactured by Kurabo Industries Ltd.) ４ cells / cm ２ Seeds were seeded in 24-well SensoPlate® containers (manufactured by Greiner Japan Co., Ltd.) using HuMedia-KG2 medium at a culture volume of 1 mL / well, and incubated at 37°C and 5% CO2. ２ After culturing for 24 hours under atmospheric conditions, the HuMedia-KG2 medium was removed to obtain a 24-well plate containing the cultured cells. HuMedia-KB2 medium supplemented with extract (plant extract) was added to the 24-well plate at a rate of 1 mL / well, and the plate was incubated at 37°C and 5% CO2. ２After further incubation for 24 hours under atmospheric conditions, the HuMedia-KB2 medium was removed. For comparison with the absence of extract (plant extract), only the HuMedia-KB2 medium was added. D-PBS(-) buffer (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was added at a rate of 1 mL / well to the 24-well plate, and UVB 13 mJ / cm² was applied as oxidative stress. ２ The samples were treated with UVB. For comparison, the untreated oxidative stress group was not treated with UVB. After removing the D-PBS(-) buffer, HuMedia-KB2 medium containing the extract (plant extract) was added to the 24-well plate at a rate of 1 mL / well and the samples were heated at 37°C under 5% CO2. ２The cells were cultured for 24 hours under controlled conditions. For comparison with the control of the control without the plant extract, only HuMedia-KB2 medium was added. After culturing, the medium was collected and used for IL-8 quantification experiments, and normal human keratinocytes derived from neonatal foreskin were collected and used for immunostaining experiments with intracellular hydrogen peroxide, etc. The amount of IL-8 produced in the medium collected after the anti-inflammatory test was measured using Human CXCL8 / IL-8 ELISA Kit (Proteintech). The amount of IL-8 in the untreated oxidative stress group and the HuMedia-KB2 medium-added group was set to 1, and the relative value of IL-8 production in each group was calculated. Human normal keratinocytes derived from neonatal foreskin, collected after anti-inflammatory testing, were washed with D-PBS(-) buffer. Hydrogen peroxide in the cells was visualized using Hyp-Stamp™ (manufactured by Funakoshi Co., Ltd.) to confirm reactive oxygen species scavenging ability. Furthermore, DNA damage suppression ability was confirmed by visualizing DNA damage markers using DNA Damage Detection Kit-γH2AX-Red- (manufactured by Dojin Chemical Research Institute Co., Ltd.). Nuclear staining was performed using CellCover (manufactured by Funakoshi Co., Ltd.) as the cell fixative, DAKOprotein block serum free (manufactured by Agilent Technologies, Inc.) as the blocking reagent, and DAPI solution (manufactured by Thermo Fisher Scientific) as the counterstain. Cell permeabilization was performed using D-PBS(-) buffer with 1% Triton X-100 added. Immunostaining was performed according to the protocol provided with Hyp-Stamp™, DNA Damage Detection Kit-γH2AX-Red-, and fluorescence images were taken using a fluorescence microscope.

[0087] <Cell activation test using human normal keratinocytes> Human normal keratinocytes derived from neonatal foreskin (manufactured by Kurabo Industries Ltd.) were used in a 2.5 × 10⁶ test. ４ cells / cm ２ Seeds were seeded in 96-well plates (Corning) using HuMedia-KG2 medium at a culture volume of 0.1 mL / well. The cultures were incubated at 37°C and 5% CO2. ２After incubation for 24 hours under atmospheric conditions, the HuMedia-KG2 medium (manufactured by Kurabo Industries Ltd.) was removed. HuMedia-KB2 medium with added extract (plant extract), or HuMedia-KB2 medium at 0.1 mL / well, was added to a 96-well plate and incubated at 37°C in 5% CO2. ２ The cells were cultured under atmospheric conditions for 48 hours. Using the CellTiter-Glo® 2.0 Cell Viability Assay kit (manufactured by Promega), the ATP levels were measured according to the protocol provided with the kit, and the relative values ​​were calculated with the ATP levels of the Humedia KB2 medium-added group (without the addition of the extract (plant extract)) set to 100.

[0088] <Cell activation test using human normal fibroblasts> Human normal fibroblasts derived from neonatal foreskin (manufactured by Kurashiki Spinning Co., Ltd.) were used in a 1.5 × 10⁶ test. ４ cells / cm ２ Seeds were seeded in 96-well plates (Corning) using FibroLife S2 Comp kit medium (manufactured by Kurabo Industries Ltd.) at a culture medium volume of 0.1 mL / well. The culture was maintained at 37°C and 5% CO2. ２ After incubation for 24 hours under atmospheric conditions, the FibroLife S2 Comp kit medium (manufactured by Kurabo Industries Ltd.) was removed. DMEM medium (manufactured by Fujifilm Wako Pure Chemical Industries Ltd.) with added extract (plant extract), or DMEM medium, was added to a 96-well plate at 0.1 mL / well, and incubated at 37°C in 5% CO2. ２ The cells were incubated under atmospheric conditions for 48 hours. Using the CellTiter-Glo® 2.0 Cell Viability Assay kit (manufactured by Promega), the amount of ATP was measured according to the protocol provided with the kit, and the relative value was calculated with the amount of ATP in the DMEM medium-added group (without the addition of extract (plant extract)) set to 100.

[0089] <Preparation and Storage Stability of Extract (Plant Extract)> [Example 1] After removing the bracts and silks from the purple corn variety Yamato Rouge (Hiruzen Farm (Okayama Prefecture)), the corn was dried at 50°C for 2 days to obtain a dried purple corn product with a moisture content of 10% or less. The corn was ground into pieces smaller than 10 mm using a food processor to obtain a purple corn pulverized product (dried). The purple corn pulverized product (dried) and a 50 wt% aqueous solution of 1,3-propanediol were mixed in a weight ratio of 2:8, and the mixture was stirred at 50°C and 240 rpm for 3 hours to obtain a purple corn extract. Solid plant residue was removed from the extract, and the mixture was further filtered using a 0.45 μm filter system (Corning Corporation) to prepare a 1,3-propanediol extract (plant extract) of purple corn. The pH of the extract (plant extract) was 6.0. The initial a* value in the storage stability test was 28.71, which was 26.2 after storage at 4°C for 1 month and 22.2 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 2.51, and the Δa* value after storage at 50°C for 1 month was 6.51.

[0090] [Example 2] After removing the bracts and silks from the purple corn variety Yamato Rouge (Hiruzen Farm (Okayama Prefecture)), the corn was frozen at -20°C for more than 24 hours, and then ground into pieces smaller than 10 mm using a food processor to obtain purple corn pulverized material (frozen). The purple corn pulverized material (frozen) was mixed with a 50 wt% aqueous solution of 1,3-propanediol in a weight ratio of 1:9, and then stirred at 50°C and 240 rpm for 2 hours to obtain purple corn extract. Solid plant residue was removed from the extract, and it was further filtered using a 0.45 μm filter system (Corning Corporation) to prepare purple corn 1,3-propanediol extract (plant extract). The pH of the extract (plant extract) was 6.3. The initial a* value in the storage stability test was 1.82, 1.19 after storage at 4°C for 1 month, and -1.35 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 0.63, and the Δa* value after storage at 50°C for 1 month was 3.17.

[0091] [Example 3] A 1,3-butanediol extract (plant extract) of purple corn was prepared by following the same procedure as in Example 1, except that a 50 wt% aqueous solution of 1,3-butanediol was used instead of a 50 wt% aqueous solution of 1,3-propanediol, and the purple corn pulverized material (dried) and the 50 wt% aqueous solution of 1,3-butanediol were mixed in a weight ratio of 1:9. The pH of the extract (plant extract) was 6.3. The initial a* value in the storage stability test was 13.11, 11.51 after storage at 4°C for 1 month, and 3.20 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 1.60, and the Δa* value after storage at 50°C for 1 month was 9.91.

[0092] [Example 4] The same procedure as in Example 3 was followed to prepare a 1,3-butanediol extract (plant extract) of purple corn, except that the purple corn powder (frozen product) from Example 2 was used instead of the purple corn powder (dried product). The pH of the extract (plant extract) was 7.0. The initial a* value in the storage stability test was 2.03, 1.2 after storage at 4°C for 1 month, and -0.43 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 0.83, and the Δa* value after storage at 50°C for 1 month was 2.46.

[0093] [Example 5] A glycerin extract (plant extract) of purple corn was prepared by following the same procedure as in Example 1, except that a 50 wt% aqueous solution of glycerin was used instead of a 50 wt% aqueous solution of 1,3-propanediol, and the purple corn pulverized material (dried) and the 50 wt% aqueous solution of glycerin were mixed in a weight ratio of 1:9. The pH of the extract (plant extract) was 5.9. The initial a* value in the storage stability test was 14.05, 13.29 after storage at 4°C / 1 month, and 10.02 after storage at 50°C / 1 month. The Δa* value after storage at 4°C / 1 month was 0.76, and the Δa* value after storage at 50°C / 1 month was 4.03.

[0094] [Example 6] A glycerin extract of purple corn (plant extract) was prepared by following the same procedure as in Example 5, except that the purple corn powder (frozen product) from Example 2 was used instead of the purple corn powder (dried product). The pH of the extract (plant extract) was 6.4. The initial a* value in the storage stability test was 1.84, 1.31 after storage at 4°C for 1 month, and -1.45 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 0.53, and the Δa* value after storage at 50°C for 1 month was 3.29.

[0095] [Comparative Example 1] An ethanol extract of purple corn was prepared by the same procedure as in Example 1, except that a 30 wt% aqueous solution of ethanol was used instead of a 50 wt% aqueous solution of 1,3-propanediol. The pH of the extract was 5.9. The initial a* value in the storage stability test was 28.52, 25.01 after storage at 4°C for 1 month, and 7.77 after storage at 50°C for 1 month. The Δa* value after storage at 4°C for 1 month was 3.51, and the Δa* value after storage at 50°C for 1 month was 20.75. <pH and storage stability of extracts (plant extracts) from Examples 1 to 6 and comparative example 1 extract> The pH of the extracts (plant extracts) from Examples 1 to 6 was all between 5.9 and 7.0, which is near neutral. The Δa* value of the extracts did not change much at both 4°C and 50°C compared to the Δa* value of the extract from Comparative Example 1. Therefore, it can be seen that the extracts (plant extracts) from Examples 1 to 6 all have excellent storage stability at near neutral pH.

[0096] <Antioxidant Activity Test of Example 1 and Comparative Example 1> The measured antioxidant activity of Example 1 was 23808 μmol-TE / L, and the value after storage at 50°C for 1 month was 23998 μmol-TE / L. When the antioxidant activity value before storage was set to 100, the value after storage at 50°C for 1 month was 100.8. On the other hand, the measured antioxidant activity of Comparative Example 1 was 26609 μmol-TE / L, and the value after storage at 50°C for 1 month was 22662 μmol-TE / L, and the value after storage at 50°C for 1 month was 85.2 when the antioxidant activity value before storage was set to 100. Both have antioxidant activity, but the antioxidant activity of the extract of Example 1 (plant extract) was maintained even after storage at 50°C for 1 month, compared to the decrease in antioxidant activity of the extract of Comparative Example 1.

[0097] <Measurement of total polyphenols and total anthocyanins in Example 1> The total polyphenol content in the extract (plant extract) of Example 1 was 1381 μg-gallic acid / mL, and the total anthocyanin content was 18.7 mg / L; calculated as cyanidin 3-glucoside. The total polyphenol and total anthocyanin content in the extract (plant extract) of Example 1 is high.

[0098] <Anti-aging test, DNA damage inhibition test, and reactive oxygen species removal test by hydrogen peroxide addition in Example 1> Figure 1 shows fluorescence images of nuclear staining, intracellular hydrogen peroxide, DNA damage markers, and cellular senescence markers (P21 and SA-β-gal) from the anti-aging test, DNA damage inhibition test, and reactive oxygen species removal test by hydrogen peroxide addition using the extract (plant extract) of Example 1. From the nuclear staining images, it can be confirmed that there is no oxidative stress and that there is no significant difference in the number of cells in any of the hydrogen peroxide addition groups. In addition, when comparing intracellular hydrogen peroxide, DNA damage marker γH2AX, and cellular senescence markers in cells that are not treated with hydrogen peroxide or have the extract (plant extract) added with intracellular hydrogen peroxide, DNA damage marker γH2AX, and cellular senescence markers in cells that are treated with hydrogen peroxide but have not had the extract (plant extract) added, it can be confirmed that hydrogen peroxide treatment promotes cellular senescence, increases intracellular hydrogen peroxide, and increases DNA damage. Cells treated with hydrogen peroxide after being given 0.55% or 1.1% of the extract (plant extract) from Example 1 showed a decrease in the detection regions of intracellular hydrogen peroxide, the DNA damage marker γH2AX, and the cellular senescence marker as the concentration of the extract (plant extract) increased, compared to cells treated with hydrogen peroxide without the addition of the extract (plant extract). This indicates that the extract (plant extract) has the effect of reducing cellular senescence caused by hydrogen peroxide treatment, removing reactive oxygen species such as hydrogen peroxide from cells, and suppressing DNA damage caused by hydrogen peroxide treatment.

[0099] <Enzyme activity inhibition test of Example 1> Collagenase activity inhibition: 1.70% Elastase activity inhibition: 0.58% Tyrosinase activity inhibition: 3.42%

[0100] <Anti-inflammatory test, DNA damage inhibition test, and reactive oxygen species removal test by UVB treatment of Example 1> Add 0.55% or 1.1% of the extract (plant extract) from Example 1 and UVB 13 mJ / cm ２ The relative values ​​of IL-8 production in the treated culture medium were 5.7 and 4.3, respectively, while the relative value of IL-8 production in the UVB-treated culture medium without the addition of the extract (plant extract) was 7.6. Since the relative value of IL-8 production decreased as the concentration of the added extract increased, it can be seen that the extract (plant extract) has an anti-inflammatory effect on cells. Furthermore, the relative value of IL-8 in the culture medium without UVB treatment or the addition of the extract (plant extract) was 1.0, confirming that UVB treatment causes inflammation in the culture medium. Figure 2 shows fluorescence images of nuclear staining, intracellular hydrogen peroxide, and DNA damage markers from the anti-inflammatory test, DNA damage inhibition test, and reactive oxygen species removal test using the extract (plant extract) from Example 1. From the nuclear staining images, it can be confirmed that there was no oxidative stress and no significant difference in cell number between the UVB-treated groups. In addition, when comparing intracellular hydrogen peroxide and the DNA damage marker γH2AX in cells without UVB treatment or the addition of the extract (plant extract) with intracellular hydrogen peroxide and the DNA damage marker γH2AX in cells treated with UVB but without the addition of the extract (plant extract), it can be confirmed that intracellular hydrogen peroxide and DNA damage increase with UVB treatment. The extract (plant extract) from Example 1 was added at 0.55% or 1.1%, and UVB was applied at 13 mJ / cm². ２ Compared to cells treated with UVB without the addition of the extract (plant extract), the detection regions for intracellular hydrogen peroxide and the DNA damage marker γH2AX decreased as the concentration of the extract (plant extract) increased. This indicates that the extract (plant extract) has the effect of removing reactive oxygen species such as hydrogen peroxide generated in cells by UVB treatment and the effect of suppressing DNA damage caused by UVB treatment.

[0101] <Cell Activation Test of Example 1: Human Normal Keratinocytes> The extract (plant extract) of Example 1 was added to final concentrations of 0.005%, 0.014%, or 0.041%. The relative ATP levels at each concentration were 101, 105, and 111, respectively, confirming an improvement in ATP levels. It was confirmed that the extract (plant extract) has a cell-activating effect on human normal keratinocytes.

[0102] <Cell Activation Test of Example 1: Human Normal Fibroblasts> The extract (plant extract) of Example 1 was added to final concentrations of 0.002%, 0.005%, 0.014%, 0.041%, 0.123%, or 0.37%. The relative ATP levels at each addition concentration were 114, 114, 113, 111, 105, and 106, confirming an improvement in ATP levels. It was confirmed that the extract (plant extract) has a cell-activating effect on human normal fibroblasts.

[0103] <Example 7> Using the extract (plant extract) from Example 1, a lotion and a cream were prepared according to the formulations shown in Tables 1 and 2 below.

[0104] Contains lotion

[0105]

[0106] Cream formula

[0107]

[0108] The lotion prepared in Table 1 and the cream prepared in Table 2 exhibit minimal change in reddish-purple color over time due to anthocyanins and have a superior feel.

[0109] (B) Method B <pH measurement> The pH of the extract (Aspergillus oryzae fermentation extract) was measured using a pH meter F-55 (manufactured by Horiba, Ltd.).

[0110] <Antioxidant Activity Measurement> The high antioxidant activity value of the extract (Aspergillus oryzae fermentation extract) was measured using the H-ORAC analysis kit (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). The temporal stability of antioxidant activity was measured before and after storage at 50°C for 1 month, and the value after storage at 50°C for 1 month was calculated with the antioxidant activity value before storage set to 100.

[0111] <Measurement of Polyphenol Content> The polyphenol content in the extract (Aspergillus oryzae fermentation extract) was measured using the method described in "Micro-titer plate assembly for measurement of total phenolic and total flavor contents in medical plant extract. Arabian Journal of Chemistry Volume 16, Issue 9, September 2023."

[0112] <Anthocyanin Content Measurement> The anthocyanin content in the extract (Aspergillus oryzae fermentation extract) was measured using the Anthocyanins Assay Kit (manufactured by Philgen Co., Ltd.).

[0113] <Collagenase Activity Inhibition Test> The collagenase inhibitory activity of the extract (Aspergillus oryzae fermentation extract) was measured using the method described in Quinic acid extracters from Plucea indexa with collagenase, MMP-2 and MMP-9 inhibitory activities. Phytother Res. 2008 Feb;22(2):264-266, and the 50% inhibitory concentration (IC50) was calculated.

[0114] <Manufacturing of extract (fermented extract by Aspergillus oryzae)>

[0115] [Example 8] After removing the bracts and silks from the purple corn variety Yamato Rouge (Hiruzen Farm (Okayama Prefecture)), the corn was shredded using scissors and stored at -20°C for more than 24 hours to obtain frozen purple corn shreds. Aspergillus oryzae (product name: Three Diamond) was purchased from Higuchi Matsunosuke Shoten. Frozen purple corn A. oryzae koji was prepared using the frozen purple corn shreds. The frozen purple corn A. oryzae koji and a 50 wt% aqueous solution of 1,3-propanediol were mixed in a weight ratio of 1:9, and the mixture was extracted by stirring at 45 rpm in a water bath at 50°C for 2 hours. After removing solid plant residue from the extract, it was further filtered using a 0.45 μm filter system (Corning Corporation) to obtain frozen purple corn A. An extract of 1,3-propanediol from oryzae koji (aspergillus fermentation extract) was prepared. The pH of the extract (aspergillus fermentation extract) was 6.7, and the antioxidant activity was 3330 μ-mol-TE / L.

[0116] [Example 9] An extract of 1,3-propanediol from frozen purple corn Aspergillus sojae koji (koji mold fermentation extract) was prepared using the method described in Example 8, except that Aspergillus oryzae (product name: Three Diamond) was replaced with Aspergillus sojae (product name: Sawyer No. 9) (purchased from Higuchi Matsunosuke Shoten). The pH of the extract (koji mold fermentation extract) was 6.2, and the antioxidant activity was 2346 μ-mol-TE / L.

[0117] [Example 10] A 1,3-propanediol extract (koji mold fermentation extract) of frozen purple corn Aspergillus luchuensis koji was prepared in the same manner as described in Example 8, except that Aspergillus oryzae (product name: Three Diamond) was replaced with Aspergillus luchuensis (product name: Authentic Shochu Koji Mold for Barley) (purchased from Higuchi Matsunosuke Shoten). The pH of the extract (koji mold fermentation extract) was 4.6, and the antioxidant activity was 2218 μ-mol-TE / L.

[0118] [Example 11] After removing the bracts and silks from the purple corn variety Yamato Rouge (Hiruzen Farm (Okayama Prefecture)), the corn was dried at 50°C to obtain dried purple corn. The dried corn was then ground using a food processor to obtain ground purple corn. Aspergillus oryzae (trade name: Three Diamond) was purchased from Higuchi Matsunosuke Shoten. Purple corn A. oryzae koji was prepared using the ground purple corn. The purple corn A. oryzae koji and a 50 wt% aqueous solution of 1,3-propanediol were mixed in a weight ratio of 1:9. The mixture was then extracted at 50°C for 2 hours, stirring at 45 rpm using a water bath. After removing solid plant residue from the extract, it was further filtered using a 0.45 μm filter system (Corning Corporation) to prepare a 1,3-propanediol extract (koji mold fermentation extract) of purple corn A. oryzae koji. The pH of the extract (Aspergillus oryzae fermentation extract) was 6.8, the antioxidant activity was 6142 μ-mol-TE / L, the total polyphenol content was 419 μg-gallic acid / mL, and the total anthocyanin content was 0.404 mg / L.

[0119] [Example 12] Using the purple corn A. oryzae koji prepared in Example 11, a 1,3-butanediol extract (koji mold fermentation extract) of purple corn A. oryzae koji was prepared in the same manner as in Example 11, except that a 50 wt% aqueous solution of 1,3-butanediol was used instead of the 50 wt% aqueous solution of 1,3-propanediol described in Example 11, and the purple corn A. oryzae koji and the 50 wt% aqueous solution of 1,3-butanediol were mixed in a weight ratio of 1:9. The pH of the extract (koji mold fermentation extract) was 7.0, the antioxidant activity was 6638 μ-mol-TE / L, the total polyphenols were 415 μg-gallic acid / mL, and the total anthocyanins were 0.355 mg / L.

[0120] [Example 13] Using the purple corn A. oryzae koji prepared in Example 11, a purple corn A. oryzae koji glycerin extract (koji mold fermentation extract) was prepared in the same manner as in Example 11, except that a 50 wt% aqueous solution of glycerin was used instead of the 50 wt% aqueous solution of 1,3-propanediol described in Example 11, and the purple corn A. oryzae koji and the 50 wt% aqueous solution of glycerin were mixed in a weight ratio of 1:9. The pH of the extract (koji mold fermentation extract) was 7.0, the antioxidant activity was 6521 μ-mol-TE / L, the total polyphenols were 449 μg-gallic acid / mL, and the total anthocyanins were 0.338 mg / L.

[0121] [Example 114] Using the purple corn A. oryzae koji prepared in Example 11, a purple corn A. oryzae koji ethanol extract (koji mold fermentation extract) was prepared in the same manner as in Example 11, except that a 50 wt% aqueous solution of ethanol was used instead of the 50 wt% aqueous solution of 1,3-propanediol described in Example 11, and the purple corn A. oryzae koji and the 50 wt% aqueous solution of ethanol were mixed in a weight ratio of 1:9. The pH of the extract (koji mold fermentation extract) was 7.1, the antioxidant activity was 5249 μ-mol-TE / L, the total polyphenols were 333 μg-gallic acid / mL, and the total anthocyanins were 0.679 mg / L.

[0122] [Reference Example] After removing the bracts and silks from the purple corn variety Yamato Rouge (Hiruzen Farm (Okayama Prefecture)), the corn was shredded using scissors and stored at -20°C for more than 24 hours to obtain frozen purple corn shreds. The frozen purple corn shreds were mixed with a 50 wt% aqueous solution of 1,3-propanediol in a weight ratio of 1:9, and the mixture was extracted by stirring at 45 rpm in a water bath at 50°C for 2 hours. After removing solid plant residue from the extract, it was further filtered using a 0.45 μm filter system (Corning Corporation) to prepare purple corn 1,3-propanediol extract (Aspergillus oryzae fermentation extract). The pH of the extract (Aspergillus oryzae fermentation extract) was 6.7, and the antioxidant activity was 1111 μ-mol-TE / L.

[0123] It was found that solid-state fermentation using Aspergillus oryzae increases the antioxidant activity of the extract, and that an extract containing high levels of total polyphenols and total anthocyanins can be obtained.

[0124] <Collagenase Activity Inhibition Test> When the collagenase activity inhibition of the extract from Example 8 was measured, the IC50 value was 5.8%. When the collagenase activity inhibition of the extract from the reference example was measured, the IC50 value was 14%. It was confirmed that solid-state fermentation by Aspergillus oryzae improved the collagenase activity inhibitory effect.

[0125] <Example 15> Using the 1,3-propanediol extract (koji mold fermentation extract) of purple corn A. oryzae koji from Example 11, a lotion with the formulation shown in Table 3 and a cream with the formulation shown in Table 4 were prepared.

[0126]

[0127]

[0128] The lotion in Table 3 and the cream in Table 4 exhibit minimal change in reddish-purple coloration due to anthocyanins over time, and also have a superior feel.

[0129] This disclosure includes the following variations: [A1] A method for producing a plant extract using a plant containing anthocyanins as a raw material and an aqueous solution containing one or more of 1,3-propanediol, 1,3-butylene glycol, or glycerin as an extraction solvent. [A2] A method for producing a plant extract according to [A1], wherein the plant extract has a pH of 6 to 8. [A3] A method for producing a plant extract according to [A1] or [A2], wherein the plant containing anthocyanins is corn. [A4] A plant extract characterized by containing anthocyanins derived from corn, one or more of 1,3-propanediol, 1,3-butylene glycol, or glycerin, and water. [A5] An anti-aging agent, enzyme inhibitor, anti-inflammatory agent, anti-aging agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator containing the plant extract according to [A4]. A cosmetic composition comprising the plant extract described in [A6] or [A4], or the anti-aging agent, enzyme inhibitor, anti-inflammatory agent, anti-aging agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator described in [A5].

[0130] [B1] A koji mold fermentation extract using a plant containing anthocyanins. [B2] The koji mold fermentation extract according to [B1], further comprising an aqueous solution containing one or more of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. [B3] The koji mold fermentation extract according to [B1], wherein the koji mold is a fungus of the genus Aspergillus. [B4] The koji mold fermentation extract according to [B1], wherein the fungus of the genus Aspergillus is Aspergillus oryzae, Aspergillus sojae, or Aspergillus luchuensis. [B5] The koji mold fermentation extract according to [B1], wherein the plant containing anthocyanins is corn. [B6] A method for producing a koji mold fermentation extract, comprising the steps of fermenting a plant containing anthocyanins with koji mold and extracting it with a solvent. [B7] A method for producing a koji mold fermentation extract according to [B6], wherein the solvent is an aqueous solution containing one or more of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin. [B8] An antioxidant or wrinkle formation inhibitor containing the koji mold fermentation extract according to [B4].

[0131] Furthermore, this disclosure includes the following variations: [C1] A plant extract characterized by comprising anthocyanins derived from corn, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water, for use as an antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, cell activator, or for the manufacture of cosmetic compositions. [C2] A plant extract characterized by comprising anthocyanins derived from corn, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water, for use as an antioxidant method, anti-aging method, enzyme inhibitory method, anti-inflammatory method, DNA damage inhibitory method, reactive oxygen species scavenger, or cell activator, applied to humans or non-human animals. [C3] A plant extract characterized by comprising anthocyanin derived from corn, used as an antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, cell activator, or for the manufacture of cosmetic compositions, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water. [C4] Use of an anthocyanin-containing plant fermentation extract by Aspergillus oryzae for the manufacture of antioxidants or wrinkle formation inhibitors. [C5] An antioxidant method or a method for wrinkle formation inhibition, applying an anthocyanin-containing plant fermentation extract by Aspergillus oryzae to humans or non-human animals. [C6] An anthocyanin-containing plant fermentation extract by Aspergillus oryzae for use in the manufacture of antioxidants or wrinkle formation inhibitors.

Claims

1. A method for producing an anthocyanin-derived composition, comprising the step of extracting a plant containing anthocyanins or a fermented product thereof with Aspergillus oryzae using a solvent containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol (except in cases where the method includes the step of extracting an anthocyanin-containing plant that has not been fermented by Aspergillus oryzae using a solvent containing ethanol).

2. The method for producing a plant extract according to claim 1, wherein the plant containing anthocyanins is used as a raw material, and an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin is used as the extraction solvent.

3. The manufacturing method according to claim 2, wherein the plant extract has a pH of 5 to 8.

4. The manufacturing method according to claim 2 or 3, wherein the plant containing anthocyanins is corn.

5. The method for producing a koji mold fermentation extract, comprising a fermentation step of fermenting a plant containing anthocyanins with koji mold, and a step of extracting the koji mold fermented product obtained in the fermentation step with the solvent, according to claim 1.

6. The method for producing a koji mold fermentation extract according to claim 5, wherein the solvent is an aqueous solution containing one or more of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin.

7. A composition comprising an anthocyanin derived from a plant containing anthocyanins, or a fermented extract of a plant containing anthocyanins by Aspergillus oryzae, and a solvent comprising one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, glycerin, and ethanol (except in cases where the anthocyanin derived from a plant containing anthocyanins, or the fermented extract of a plant containing anthocyanins, is an anthocyanin derived from a plant containing anthocyanins, and the solvent contains ethanol).

8. The composition according to claim 7, which is a plant extract characterized by comprising anthocyanin derived from corn, one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, and glycerin, and water.

9. An antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator comprising the composition described in claim 8.

10. A cosmetic composition comprising the composition described in claim 8, or the antioxidant, anti-aging agent, enzyme inhibitor, anti-inflammatory agent, DNA damage inhibitor, reactive oxygen species scavenger, or cell activator described in claim 9.

11. The composition according to claim 7, comprising an extract of Aspergillus oryzae fermentation of a plant containing anthocyanins, and an aqueous solution containing one or more selected from the group consisting of 1,3-propanediol, 1,3-butylene glycol, ethanol, and glycerin.

12. The composition according to claim 11, wherein the koji mold is a fungus of the genus Aspergillus.

13. The composition according to claim 12, wherein the Aspergillus fungus is Aspergillus oryzae, Aspergillus sojae, or Aspergillus luchuensis.

14. The composition according to any one of claims 11 to 13, wherein the plant containing the anthocyanin is maize.

15. An antioxidant or wrinkle-inhibiting agent comprising the composition according to any one of claims 11 to 14.

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