Plant-derived extract composition

Abstract

Provided, according to one embodiment, is a plant-derived extract composition capable of increasing the expression of transglutaminase (TGM-1) and loricrin (LOR).　The present disclosure relates to, according to one embodiment, a plant-derived extract composition that comprises rosmarinic acid and has a mass ratio of betulinic acid and rosmarinic acid (betulinic acid / rosmarinic acid) of 0.25 or less.

Description

Plant-derived extract composition

[0001] The present disclosure relates to plant-derived extract compositions.

[0002] To improve the skin's moisture retention ability, prevent and improve rough skin, and prevent and improve skin aging such as the formation of wrinkles and reduction of texture, it is believed that it is effective to act on the epidermal cells of the skin, promote keratinization of the epidermal cells, encourage the formation of a healthy stratum corneum, and improve the stratum corneum barrier function to protect the body from external stimuli and other factors.

[0003] It is said that maintaining the stratum corneum's moisture content and flexibility is important for the health of this barrier function. Aging, dryness, ultraviolet rays, and other factors can disrupt the cell turnover process, leading to abnormalities in the formation of stratum corneum cells and the structure of intercellular lipids. This is known to lead to various skin disorders and skin problems, such as rough skin.

[0004] Here, turnover refers to the continuous repetition of keratinocyte (epidermal keratinocyte) proliferation in the basal layer, the cornification process, and the peeling of the stratum corneum. These keratinocytes form each layer by cornifying sequentially outward from basement membrane cells, spinous cells, granular cells, and corneocytes. Proteins constituting the cornified envelope (CE) are synthesized from the upper spinous layer to the granular layer. Furthermore, in the process of reaching the stratum corneum, matrix proteins such as involucrin (IVL), loricrin (LOR), and cystatin bind to the cell membrane of keratinocytes by transglutaminase (TGM), forming insolubilized CE. Furthermore, it is known that ceramides and the like covalently bind to the insolubilized CE, forming the basis of the stratum corneum barrier function.

[0005] Traditionally, skin problems such as rough skin caused by a decrease in the stratum corneum barrier function have been resolved by supplementing the stratum corneum barrier function with creams containing ceramides, etc. However, this improvement in stratum corneum cell CE is insufficient, and ingredients (agents, compositions, etc.) that can promote the formation of healthy CE are needed. For example, International Publication No. 2020 / 158912 (Patent Document 1) proposes a composition for improving skin viscoelasticity, which includes an inhibitor of extracellular ATP concentration increase, in which the content ratio of (A) rosmarinic acid to (B) luteolin glucuronide is (A):(B) = 1:0.2 to 1.5 by weight. Japanese Patent Application Laid-Open No. 2010-90037 (Patent Document 2) proposes a filaggrin production promoter containing rosmarinic acid and / or eriodictyol 7-O-rutinoside as active ingredients, as an agent for promoting mRNA expression of profilaggrin, a precursor of filaggrin involved in improving the skin's moisturizing function. Japanese Patent Laid-Open Publication No. 2007-277149 (Patent Document 3) proposes an involucrin expression promoter and a stratum corneum formation promoter as agents capable of maintaining healthy skin barrier function, which contain one or more plants or extracts thereof selected from Japanese pepper, peppermint, eucalyptus, saxifrage, rosemary, asparagus, comfrey, shiitake mushroom, calamus, thyme, burnet, gambir, ginkgo, kudzu, Chinese quince, gardenia, loquat, cinchona, hawthorn, clove, mugwort, and Phellodendron amurense.

[0006] In one aspect, the present disclosure relates to a plant-derived extract composition containing rosmarinic acid, wherein the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less.

[0007] In one aspect, the present disclosure relates to a composition containing a plant extract component, the composition containing rosmarinic acid, and the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) being 0.25 or less.

[0008] In one aspect, the present disclosure relates to a method for producing a plant-derived extract composition, which includes an extraction step of extracting so that the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less.

[0009] In one aspect, the present disclosure relates to a cosmetic product containing the plant-derived extract composition of the present disclosure.

[0010] As described above, transglutaminase (TGM) is known to catalyze the cross-linking reaction of basic proteins such as involucrin (IVL) and loricrin (LOR), thereby constructing the cornified envelope (CE). There is a need for an extract composition that can increase the expression levels of transglutaminase (TGM) and loricrin (LOR) more than conventional plant extract compositions and promote the formation of a healthy CE.

[0011] Therefore, the present disclosure provides a plant-derived extract composition that can increase the expression levels of transglutaminase 1 (TGM-1) and loricrin (LOR), which are representative transglutaminases (TGM).

[0012] According to one aspect of the present disclosure, a plant-derived extract composition that can increase the expression levels of transglutaminase (TGM-1) and loricrin (LOR) can be provided.

[0013] Rosmarinic acid has attracted attention as a component for improving (promoting) keratinization. Rosmarinic acid is a type of polyphenol found in large amounts in Lamiaceae plants such as rosemary. A known example of an extract (plant-derived extract composition) containing rosmarinic acid is rosemary extract extracted from rosemary (Japanese name: Mannenrou). Rosemary extract contains rosmarinic acid as a major component and typically also contains other components such as betulinic acid. After extensive research, the present inventors have found that the coexistence of rosmarinic acid and betulinic acid in a plant-derived extract composition such as rosemary extract inhibits the rosmarinic acid-mediated promotion of transglutaminase (TGM-1) and loricrin (LOR) expression. Furthermore, the present inventors have found that by extracting so that the mass ratio of betulinic acid to rosmarinic acid is reduced to a predetermined value or less, the inhibitory effect of betulinic acid on the expression of transglutaminase (TGM-1) and loricrin (LOR) can be reduced and the amount of promotion of these expressions by the extract can be increased.

[0014] That is, in one aspect, the present disclosure relates to a plant-derived extract composition (hereinafter also referred to as the "extract composition of the present disclosure") that contains rosmarinic acid and has a mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) of 0.25 or less.

[0015] Although the details of the mechanism by which the effects of the present disclosure are manifested are not clear, it is speculated as follows. It is believed that betulinic acid contained in plant-derived extract compositions such as rosemary extract inhibits rosmarinic acid. In the present disclosure, by setting the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) to 0.25 or less, the influence of betulinic acid on the inhibition of the action of rosmarinic acid can be drastically reduced, which is thought to lead to an increase in the expression levels of transglutaminase (TGM-1) and loricrin (LOR). However, the present disclosure need not be interpreted as being limited to these mechanisms.

[0016] In one or more embodiments, the plant in the present disclosure may be a plant containing rosmarinic acid. The type of plant containing rosmarinic acid is not particularly limited as long as it contains rosmarinic acid, and examples thereof include Lamiaceae plants. Examples of Lamiaceae plants include herb plants of the Lamiaceae family, such as rosemary (Rosmarinus officinalis), perilla, lemon balm, and common sage. The plant part used for extraction may be the whole plant or a part of the plant, and can be appropriately selected depending on the purpose. Suitable plant parts for extraction include leaves, stems, flowers, fruits, pericarp, pericarp, aboveground parts, or mixtures thereof. Among these, leaves are preferred from the viewpoint of efficient extraction of rosmarinic acid.

[0017] [Rosmarinic Acid] In one or more embodiments, the extraction composition of the present disclosure contains rosmarinic acid. The concentration of rosmarinic acid in the extraction composition of the present disclosure can be appropriately set. In one or more embodiments, the concentration of rosmarinic acid (content, ppm) in the extraction composition of the present disclosure is preferably 0.5 ppm or more, more preferably 0.7 ppm or more, even more preferably 0.9 ppm or more, and even more preferably 1 ppm or more, from the viewpoint of improving the expression levels of TGM-1 and LOR. In the present disclosure, "ppm" is based on mass. Note that 1 ppm is 0.0001 mass% (the same applies hereinafter).

[0018] [Betulinic Acid] In one or more embodiments, the extraction composition of the present disclosure may contain betulinic acid, or in one or more other embodiments, may not contain betulinic acid. From the viewpoint of improving the expression levels of TGM-1 and LOR, the concentration of betulinic acid (content, ppm) in the extraction composition of the present disclosure preferably satisfies the following mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid). In one or more embodiments, the concentration is preferably 0.25 ppm or less, more preferably 0.20 ppm or less, even more preferably 0.10 ppm or less, even more preferably 0.09 ppm or less, and even more preferably 0.005 ppm or less.

[0019] [Mass Ratio (Betulinic Acid / Rosmarinic Acid)] The mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) in the extract composition of the present disclosure is 0.25 or less, preferably 0.20 or less, more preferably 0.15 or less, even more preferably 0.10 or less, even more preferably 0.09 or less, and even more preferably 0.005 or less, from the viewpoint of improving the expression levels of TGM-1 and LOR. In the present disclosure, the concentrations of rosmarinic acid and betulinic acid can be measured using high-performance liquid chromatography (HPLC), for example, by the method described in the Examples.

[0020] [Method for Producing Extract Composition] In one or more embodiments, the extract composition of the present disclosure can be obtained by extraction from a plant. Accordingly, in one aspect, the present disclosure relates to a method for producing an extract composition (hereinafter also referred to as the "method for producing the extract composition of the present disclosure"), which includes a step of obtaining an extract composition from a plant (an extraction step). In one or more embodiments, the method for producing an extract composition of the present disclosure is a method for producing a plant-derived extract composition, which includes an extraction step in which betulinic acid and rosmarinic acid are extracted so that the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less. Since rosmarinic acid and betulinic acid are relatively hydrophilic and hydrophobic components, respectively, the mass ratio of betulinic acid to rosmarinic acid can be controlled within a predetermined range by utilizing this difference. In one or more embodiments, the content of rosmarinic acid in the extract composition extracted in the extraction step is preferably 0.5 ppm or more. The preferred values ​​for the rosmarinic acid content, betulinic acid content (ppm), and betulinic acid to rosmarinic acid mass ratio in the extract composition extracted in the extraction step are the same as those for the extract composition of the present disclosure described above. Examples of plants used for extraction include those mentioned above, with rosemary being preferred. Examples of plant parts used for extraction include those mentioned above, with leaves being preferred.

[0021] Examples of extraction methods used in the extraction step include soaking, decoction, percolation, reflux extraction, supercritical fluid extraction, ultrasonic extraction, and microwave extraction. A preferred method involves soaking a plant in an extraction solvent. For example, a method for extracting rosmarinic acid involves placing a certain amount of rosemary in a kettle, adding a soaking solvent (e.g., water), and then soaking the mixture at 25°C for 24 hours. Stirring may or may not be required during soaking. Therefore, in one or more embodiments, the extraction step involves soaking a plant in an extraction solvent to obtain an extract composition.

[0022] <Extraction Solvent> The extraction solvent used in the extraction step can be either a polar or nonpolar solvent. Examples of the extraction solvent include water; monohydric alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as 1,3-propanediol, dipropylene glycol, pentanediol, hexanediol, propylene glycol, butylene glycol, and glycerin; and mixtures thereof. Among these, at least one selected from water, ethanol, glycerin, 1,3-propanediol, butylene glycol, and mixtures thereof is preferred from the viewpoint of efficiently extracting rosmarinic acid. In one or more embodiments, water is preferred as the extraction solvent from the viewpoint of efficiently extracting rosmarinic acid. When an aqueous solution of an alcohol (monohydric alcohol and / or polyhydric alcohol) is used as the extraction solvent, its concentration is preferably more than 0% by mass and not more than 20% by mass from the viewpoint of efficiently extracting rosmarinic acid and from the viewpoint of extracting so that the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less.

[0023] In one or more embodiments, the extraction solvent used in the extraction step may further contain other components as needed, such as a preservative to prevent decay and a spreading agent to promote penetration into the plant body.

[0024] From the viewpoint of efficient extraction of rosmarinic acid, the amount of the extraction solvent used in the extraction step is preferably 3 times or more, more preferably 5 times or more, and even more preferably 10 times or more, based on the weight or dry weight of the plant. From the same viewpoint, the amount of the extraction solvent used in the extraction step is preferably 3 to 100 times, more preferably 5 to 75 times, and even more preferably 10 to 50 times.

[0025] <Extraction Temperature> The temperature of the extraction solvent used for extraction (extraction temperature) or the temperature at which the plant is immersed in the extraction solvent (immersion temperature) is preferably 5° C. or higher, more preferably 15° C. or higher, and even more preferably 20° C. or higher, from the viewpoint of efficiently extracting rosmarinic acid. From the same viewpoint, the extraction temperature or the immersion temperature is preferably 5° C. or higher and 35° C. or lower, more preferably 15° C. or higher and 30° C. or lower, and even more preferably 20° C. or higher and 30° C. or lower ...

[0026] <Extraction Time> The extraction time or the time for soaking a plant in an extraction solvent (soaking time) is preferably 5 hours or more, more preferably 12 hours or more, and even more preferably 24 hours or more, from the viewpoint of efficiently extracting rosmarinic acid. From the same viewpoint, it is preferably 120 hours or less, more preferably 72 hours or less, and even more preferably 36 hours or less. From the same viewpoint, the extraction time or the soaking time is preferably 5 hours or more and 120 hours or less, more preferably 12 hours or more and 72 hours or less, and even more preferably 24 hours or more and 36 hours or less.

[0027] In one or more embodiments, the extract composition of the present disclosure can be obtained by filtering using a filter. Thus, in one or more embodiments, the extraction step is a step of immersing a plant in an extraction solvent and then filtering using a filter to obtain an extract composition. Examples of filters used for filtration include filter paper, membrane filters, cartridge filters, and disposable filters.

[0028] <Pre-extraction Treatment> In one or more embodiments, the plant before extraction or the plant before immersion in the extraction solvent may be blanched. Therefore, in one or more embodiments, the extraction step may include a step of blanching rosemary before extraction. As the blanching method, a general method can be used. For example, a method of steaming, boiling, or microwave-treating a plant such as rosemary can be used. The steam blanching temperature can be, for example, 100 to 150°C, and the blanching time can be, for example, 5 to 60 minutes.

[0029] <Drying Treatment> In one or more embodiments, the plant before extraction or the plant before immersion in the extraction solvent may be dried after pre-extraction treatment. Therefore, in one or more embodiments, the extraction step may include a step of blanching hydroponically grown rosemary and then drying it before extraction. A common drying method can be used. Examples include natural drying, hot air drying, freeze drying, reduced pressure drying, and spray drying. The hot air drying temperature is, for example, 30°C to 100°C, and the drying time is, for example, 1 to 24 hours.

[0030] In one or more embodiments, the extraction composition of the present disclosure may be further purified to produce a purified product. Therefore, in one or more embodiments, the method for producing the extraction composition of the present disclosure may further include a step of purifying the extract composition.

[0031] In one or more embodiments, the extract composition of the present disclosure may be used as is, or may be diluted, concentrated, or dried. That is, the extract composition of the present disclosure includes the extract obtained in the extraction step, as well as its diluted product, concentrated product, dried product, or purified product thereof. Examples of the extract composition of the present disclosure include an extract obtained by soaking a plant in water, and then adjusting the extract to have a predetermined concentration of rosmarinic acid and solvent using only an extracting solvent other than water, or an extracting solvent other than water and water.

[0032] [Uses of Extraction Composition] In one or more embodiments, the extraction composition of the present disclosure can be used to promote transglutaminase expression in human epidermal keratinocytes, promote loricrin expression in human epidermal keratinocytes, promote stratum corneum formation, or promote epidermal turnover. Accordingly, in another aspect, the present disclosure relates to a method for promoting transglutaminase expression in human epidermal keratinocytes using the extraction composition of the present disclosure. In another aspect, the present disclosure relates to a method for promoting loricrin expression in human epidermal keratinocytes using the extraction composition of the present disclosure. In another aspect, the present disclosure relates to a method for promoting stratum corneum formation using the extraction composition of the present disclosure. In another aspect, the present disclosure relates to a method for promoting epidermal turnover using the extraction composition of the present disclosure. In one or more embodiments, the extraction composition of the present disclosure can be used to improve uneven redness of the skin. Accordingly, in another aspect, the present disclosure relates to a method for improving uneven redness of the skin using the extraction composition of the present disclosure. In the present disclosure, the epidermis and skin include the epidermis and skin of humans or non-human mammals. In one or more embodiments, the extract composition of the present disclosure can be used as an active ingredient in a composition for promoting transglutaminase expression in human epidermal keratinocytes, promoting loricrin expression in human epidermal keratinocytes, promoting stratum corneum formation, or promoting epidermal turnover, or can be used to manufacture such a composition. In one or more embodiments, the extract composition of the present disclosure can be used as an active ingredient in a composition for reducing uneven redness on the skin, or can be used to manufacture such a composition. Accordingly, in one aspect, the present disclosure relates to a composition containing a plant extract, which contains rosmarinic acid and has a mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) of 0.25 or less. In one or more embodiments, the content of rosmarinic acid in the composition of this aspect is preferably 0.5 ppm or more. Preferred values ​​for the rosmarinic acid content, betulinic acid content, and mass ratio of betulinic acid to rosmarinic acid in the composition of this aspect are the same as those for the extract composition of the present disclosure described above.In one or more embodiments, the composition of this aspect is a composition for promoting transglutaminase expression in human epidermal keratinocytes, promoting loricrin expression in human epidermal keratinocytes, promoting stratum corneum formation, or promoting epidermal turnover. In one or more embodiments, the composition of this aspect is a composition for reducing uneven redness of the skin. In one or more embodiments, the composition of this aspect is a cosmetic, pharmaceutical, or quasi-drug for reducing uneven redness of the skin. In another aspect, the present disclosure relates to a cosmetic, pharmaceutical, or quasi-drug for reducing uneven redness of the skin, comprising a composition of this disclosure. In one or more embodiments, the composition of this aspect can be suitably used as an external product or food. In the present disclosure, examples of external products include cosmetics, pharmaceuticals, quasi-drugs, bath additives, and perfumed products such as toothpaste. In another aspect, the present disclosure relates to an external product or food containing a composition of this disclosure. In one or more embodiments, the extract composition of this disclosure can be suitably used in cosmetics. That is, in one aspect, the present disclosure relates to a cosmetic containing the extract composition of the present disclosure. Examples of the cosmetic include hair cosmetics and skin cosmetics. Examples of product forms of hair cosmetics include hair shampoo, hair conditioner, and hair treatment. Examples of product forms of skin cosmetics include facial cleanser, moisturizer, emulsion, and serum. In another aspect, the present disclosure relates to a cosmetic containing the extract composition of the present disclosure. When the extract composition of the present disclosure or a composition of this aspect is incorporated into a topical skin preparation such as a cosmetic, it is expected to have effects such as improving the skin's moisture retention ability, preventing and alleviating rough skin, reducing wrinkle formation and texture patterns, improving firmness, and alleviating uneven redness of the skin. Examples of the topical product form include cream, liquid lotion, emulsion lotion, spray, moisturizer, moisturizer, and lotion. In one or more embodiments, the content of the extract composition or composition of this aspect of the present disclosure in an external product or food product is preferably 0.0001% or more, more preferably 0.01% or more, even more preferably 1% or more, and preferably 10% or less, more preferably 5% or less.In one or more embodiments, the content of the extract composition or composition of this aspect of the present disclosure in an external product or food product is preferably 0.0001% or more and 10% or less, more preferably 0.01% or more and 5% or less, and even more preferably 1% or more and 5% or less.

[0033] The present disclosure further relates to one or more of the following embodiments: <1> A plant-derived extract composition containing rosmarinic acid, wherein the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less, 0.20 or less, 0.15 or less, 0.10 or less, 0.09 or less, or 0.005 or less. <2> The extract composition according to <1>, wherein the rosmarinic acid content is 0.5 ppm or more, 0.7 ppm or more, 0.9 ppm or more, or 1 ppm or more. <3> The extract composition according to <1> or <2>, comprising betulinic acid. <4> The extract composition according to any one of <1> to <3>, wherein the betulinic acid concentration (content, ppm) is 0.25 ppm or less, 0.20 ppm or less, 0.10 ppm or less, 0.09 ppm or less, or 0.005 ppm or less. <5> The extract composition according to any one of <1> to <4>, which does not contain betulinic acid. <6> The extract composition according to any one of <1> to <5>, wherein the plant is a plant containing rosmarinic acid. <7> The extract composition according to any one of <1> to <6>, wherein the plant is a plant of the Lamiaceae family. <8> The extract composition according to any one of <1> to <7>, wherein the plant is at least one selected from rosemary, perilla, lemon balm, and common sage. <9> A composition containing a plant extract, which contains rosmarinic acid and has a mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) of 0.25 or less, 0.20 or less, 0.15 or less, 0.10 or less, 0.09 or less, or 0.005 or less. <10> The composition according to <9>, wherein the rosmarinic acid content is 0.5 ppm or more, 0.7 ppm or more, 0.9 ppm or more, or 1 ppm or more. <11> The composition according to <9> or <10>, which contains betulinic acid. <12> The composition according to any one of <9> to <11>, wherein the concentration (content, ppm) of betulinic acid is 0.25 ppm or less, 0.20 ppm or less, 0.10 ppm or less, 0.09 ppm or less, or 0.005 ppm or less. <13> The composition according to <9> or <10>, which does not contain betulinic acid. <14> The composition according to any one of <9> to <13>, for promoting loricrin expression in human epidermal keratinocytes, promoting stratum corneum formation, or promoting epidermal turnover.<15> The composition according to any one of <9> to <13>, for promoting the expression of transglutaminase in human epidermal keratinocytes. <16> The composition according to any one of <9> to <13>, for improving uneven redness of the skin. <17> The composition according to any one of <9> to <16>, wherein the plant is a plant containing rosmarinic acid. <18> The composition according to any one of <9> to <17>, wherein the plant is a plant of the Lamiaceae family. <19> The composition according to any one of <9> to <18>, wherein the plant is at least one selected from rosemary, perilla, lemon balm, and common sage. <20> A method for producing a plant-derived extract composition, comprising an extraction step of extracting so that the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less, 0.20 or less, 0.15 or less, 0.10 or less, 0.09 or less, or 0.005 or less. <21> The method for producing a plant-derived extract composition according to <20>, wherein the extraction step is a step of immersing a plant in an extraction solvent to obtain an extract composition. <22> The method for producing a plant-derived extract composition according to <20> or <21>, wherein the extraction solvent used in the extraction step is at least one selected from polar solvents and non-polar solvents. <23> The method for producing a plant-derived extract composition according to any of <20> to <22>, wherein the extraction solvent is at least one selected from water, monohydric alcohols, polyhydric alcohols, and mixtures thereof, or at least one selected from water, methanol, ethanol, propanol, butanol, 1,3-propanediol, dipropylene glycol, pentanediol, hexanediol, propylene glycol, butylene glycol, glycerin, and mixtures thereof, or at least one selected from water, ethanol, glycerin, 1,3-propanediol, butylene glycol, and mixtures thereof. <24> The method for producing a plant-derived extract composition according to any one of <20> to <23>, wherein the extraction solvent is water. <25> The method for producing a plant-derived extract composition according to any one of <20> to <24>, wherein when an aqueous solution of an alcohol (monohydric alcohol and / or polyhydric alcohol) is used as the extraction solvent, the concentration of the aqueous solution is more than 0 mass % and not more than 20 mass %.<26> The method for producing a plant-derived extract composition according to any one of <20> to <25>, wherein the amount of the extraction solvent used in the extraction step is 3 times or more, 5 times or more, or 10 times or more, and 100 times or less, 75 times or less, or 50 times or less, based on the weight or dry weight of the plant. <27> The method for producing a plant-derived extract composition according to any one of <20> to <26>, wherein the amount of the extraction solvent used in the extraction step is 3 times or more and 100 times or less, 5 times or more and 75 times or less, or 10 times or more and 50 times or less, based on the weight or dry weight of the plant. <28> The method for producing a plant-derived extract composition according to any one of <20> to <27>, wherein the extraction temperature or soaking temperature in the extraction step is 5°C or more, 15°C or more, or 20°C or more, and 35°C or less, or 30°C or less. <29> The method for producing a plant-derived extract composition according to any one of <20> to <28>, wherein the extraction temperature or soaking temperature in the extraction step is 5°C to 35°C, 15°C to 30°C, or 20°C to 30°C. <30> The method for producing a plant-derived extract composition according to any one of <20> to <29>, wherein the extraction time or soaking time in the extraction step is 5 hours or more, 12 hours or more, or 24 hours or more, and 120 hours or less, 72 hours or less, or 36 hours or less. <31> The method for producing a plant-derived extract composition according to any one of <20> to <30>, wherein the extraction time or soaking time in the extraction step is 5 hours or more and 120 hours or less, 12 hours or more and 72 hours or less, or 24 hours or more and 36 hours or less. <32> The method for producing a plant-derived extract composition according to any one of <20> to <31>, wherein the extraction step includes a step of blanching rosemary before extraction. <33> A method for producing an extract composition derived from a plant according to <32>, wherein the blanching treatment is one or more of steam treatment, boiling treatment, and microwave treatment. <34> A method for producing an extract composition derived from a plant according to any one of <20> to <33>, wherein the extraction step includes a step of blanching hydroponically grown rosemary and then drying it before extraction. <35> A method for producing an extract composition derived from a plant according to any one of <20> to <34>, wherein the plant is a plant containing rosmarinic acid. <36> A method for producing an extract composition derived from a plant according to any one of <20> to <34>, wherein the plant is a plant of the Lamiaceae family.<37> A method for producing an extract composition derived from a plant according to any one of <20> to <34>, wherein the plant is at least one selected from rosemary, perilla, lemon balm, and common sage. <38> A cosmetic preparation containing an extract composition derived from a plant according to any one of <1> to <8>. <39> A method for promoting transglutaminase expression in human epidermal keratinocytes, using an extract composition derived from a plant according to any one of <1> to <8>. <40> A method for promoting loricrin expression in human epidermal keratinocytes, using an extract composition derived from a plant according to any one of <1> to <8>. <41> A method for promoting stratum corneum formation, using an extract composition derived from a plant according to any one of <1> to <8>. <42> A method for promoting epidermal turnover, using an extract composition derived from a plant according to any one of <1> to <8>. <43> A method for improving uneven redness of the skin, using an extract composition derived from a plant according to any one of <1> to <8>. <44> A cosmetic, pharmaceutical, or quasi-drug for improving uneven redness of the skin, comprising the composition according to any one of <9> to <19>. <45> An external use product or food product comprising the composition according to any one of <9> to <19>.

[0034] The present disclosure will be described in more detail below with reference to examples, but these are merely illustrative examples and the present disclosure is not limited to these examples.

[0035] (1) Production of Rosemary Extracts 1 to 4 Rosemary Extract 1 Rosemary Extract 1 was produced as follows. [Extraction Pretreatment] Fresh rosemary leaves were blanched (steamed) to obtain a dried plant. <Blanching Treatment> Equipment: Steam convection oven (CSVH-E10-T, Comet Kato) Blanching temperature: 110°C Blanching time: 15 minutes [Drying Treatment] The rosemary leaves after the extraction pretreatment were dried. Equipment: Constant temperature dryer (DRR420DA, ADVANTEC) Drying temperature: 60°C Drying time: 18 hours [Extraction Treatment] The plant after the drying treatment was immersed in water and extracted at 25°C for 24 hours to obtain an extract. The amount of extraction solvent used was 25 times the dry weight of the rosemary. The above extract was prepared with butylene glycol and water so that the rosmarinic acid concentration was 300 ppm and the solvent concentration was 50% (v / v) butylene glycol aqueous solution. This was designated Rosemary Extract 1. The rosemary extract 1 thus obtained contained 300 ppm rosmarinic acid and 1 ppm betulinic acid. Rosemary Extract 2: Rosemary Extract 2 was prepared by immersing the plant, which had undergone the same pre-extraction and drying processes as Rosemary Extract 1, in 50% (v / v) ethanol aqueous solution and extracting at 25°C for 24 hours. The amount of extraction solvent used was 25 times the dry weight of the rosemary. The rosemary extract 2 thus obtained contained 312 ppm rosmarinic acid and 94 ppm betulinic acid. Rosemary Extract 3: Rosemary Extract 2 was diluted with 50% (v / v) ethanol to a betulinic acid concentration of 27 ppm, and reagent rosmarinic acid (Fujifilm Wako Pure Chemical Industries, Ltd.) was added to the diluted solution to a rosmarinic acid concentration of 300 ppm. Rosemary Extract 3 thus obtained contained 300 ppm rosmarinic acid and 27 ppm betulinic acid. Rosemary Extract 4: Rosemary Extract 4 was prepared by diluting Rosemary Extract 2 with 50% (v / v) ethanol to a betulinic acid concentration of 54 ppm, and reagent rosmarinic acid (Fujifilm Wako Pure Chemical Industries, Ltd.) was added to the diluted solution to a rosmarinic acid concentration of 300 ppm.The rosemary extract 4 thus obtained contained 300 ppm of rosmarinic acid and 54 ppm of betulinic acid. The concentrations of rosmarinic acid and betulinic acid were measured as follows.

[0036] [Measurement of Rosmarinic Acid and Betulinic Acid Concentrations] The concentrations of rosmarinic acid and betulinic acid were measured using high-performance liquid chromatography (HPLC) under the following measurement conditions. <Measurement Conditions> Apparatus: Ultimate 3000 (HPLC), Corona Ultra (detector) (manufactured by Dionex Corporation) Column: InertSustain ODS-3 5 μm 3.0 × 150 mm (manufactured by GL Sciences) Eluent: 0.1% formic acid (Solution A) and acetonitrile (Solution B) Gradient conditions: Gradient from 95:5 parts of Solution A to 10:90 parts of Solution A and Solution B Flow rate: 0.5 mL / min Column temperature: 40°C Detector: UV (quantitation of rosmarinic acid), CAD (quantitation of betulinic acid)

[0037] (2) Analysis of Transglutaminase 1 (TGM-1) and Loricrin (LOR) Expression (Examples 1 to 3, Comparative Examples 1 and 2) [Preparation of Additives] Additives containing rosmarinic acid and betulinic acid at the concentrations shown in Table 1 below (Examples 1 to 3, Comparative Examples 1 and 2) were prepared using rosemary extracts 1 to 4 or an aqueous ethanol solution (50% (v / v)).

[0038] [Experimental Cell Culture] Normal human epidermal keratinocytes (derived from neonatal foreskin; Kurabo Industries) were used. Cell proliferation was performed using EpiLife Medium with 60 μM calcium (Thermo Fisher) supplemented with HKGS (Thermo Fisher). For material supplementation, EpiLife Medium with 60 μM calcium supplemented with HKGS Kit (BPE- and EGF-free; Thermo Fisher) was used. Cell culture was performed according to standard procedures at 37°C in a 5% CO2 atmosphere.

[0039] [Protein extraction and transglutaminase 1 (TGM-1) and loricrin (LOR) expression analysis] Normal human epidermal keratinocytes were cultured in a 6-well plate (collagen-coated; Corning) at a concentration of 1.5 × 105Cells were seeded at a density of 1000 cells / well and cultured in growth medium for 1 day. The medium was then replaced with medium supplemented with the ingredients and cultured for another day. The following day, the additives listed in Table 1 were added and the cells were cultured for 48 hours. After washing once with PBS, the cells were lysed in RIPA buffer (SIGMA) supplemented with a 1:100 dilution of Protease / Phosphatase Inhibitor Cocktail (Cell Signaling) and the cell suspension was collected. The cell suspension was homogenized (sonicated) and centrifuged (15,000 × g, 10 min), and the supernatant was collected. Protein quantification was then performed using the Pierce® BCA Protein Assay Kit (Thermo Fisher). Detection required 8-10 μg of protein. The samples were separated by electrophoresis using NuPAGE® 4-12% SDS (sodium dodecyl sulfate)-polyacrylamide gels (Thermo Fisher). After transfer to a PVDF membrane, the membrane was blocked for 1 hour with TBS-T (Tween 20 0.1% (w / v)) containing 5% (w / v) skim milk and then incubated overnight at 4°C with primary antibodies (anti-transglutaminase 1 antibody; Novus; anti-loricrin antibody; Abcam) diluted 2000-fold in blocking solution. After three 15-minute washes with PBS-T, the membrane was incubated for 1 hour at room temperature with secondary antibodies (anti-rabbit IgG antibody (HRP-conjugated); Dako) diluted 4000-fold in blocking solution. After three 10-minute washes with PBS-T, the membrane was imaged using the SuperSignal® West Dura Extended Duration substrate (Thermo Fisher) and a detector (Amersham Imager 600). Band density was quantified using the Amersham Imager 600. The measurement data (mean, SD) were evaluated relative to the control group value of 100, and the protein expression levels of transglutaminase 1 (TGM-1) and loricrin (LOR) were evaluated. The results are shown in Table 1.In addition, since the expression level of LOR was not evaluated for Example 3, it is shown as "-" in Table 1.

[0040]

[0041] As shown in Table 1, when rosemary extracts 1, 3, and 4 having a mass ratio (betulinic acid / rosmarinic acid) of 0.25 or less (Examples 1 to 3) were used, the expression level of transglutaminase (TGM-1) was increased compared to when rosemary extract 2 having a mass ratio (betulinic acid / rosmarinic acid) of 0.3 was used (Comparative Example 2). Also, as shown in Table 1, when rosemary extracts 1 and 3 having a mass ratio (betulinic acid / rosmarinic acid) of 0.25 or less (Examples 1 and 2) were used, the expression level of loricrin (LOR) was increased compared to when rosemary extract 2 having a mass ratio (betulinic acid / rosmarinic acid) of 0.3 was used (Comparative Example 2).

[0042] (3) Effect of Improving Stratum Corneum Function (Example 4, Comparative Example 3, Reference Examples 1-2) Moisturizing solutions were prepared with the compositions and blending amounts (active ingredient, mass %) shown in Tables 2-3, and their effects were evaluated. (Production Method) Xanthan gum, 1,3-butylene glycol, and 30% by mass of purified water were heated to 80°C, mixed uniformly with stirring, and then cooled to 25°C. The remaining ingredients (rosemary extract, citric acid, sodium citrate, and phenoxyethanol) were added and mixed uniformly with stirring at 25°C to obtain each moisturizing solution (Example 4, Comparative Example 3, Reference Examples 1-2). (Evaluation Method) The effect of improving stratum corneum function after four consecutive weeks of application was evaluated in 32 healthy female subjects. The subjects' faces were split into two equal parts, with one side receiving a moisturizing solution (Example 4, Comparative Example 3) containing each rosemary extract (Example 1, Comparative Example 2) and the other side receiving a placebo moisturizing solution (Reference Examples 1, 2) not containing rosemary extract, at approximately 0.2 g per application, twice daily. Measurements were taken before and after four weeks of application, and the number of subjects who showed greater improvement was compared between the placebo moisturizer and the rosemary extract-containing moisturizer. Evaluations were based on moisture retention capacity and visual assessment of skin condition. Skin moisture retention capacity was assessed by changes in stratum corneum moisture content. Measurements of stratum corneum moisture content were performed using a corneometer (Courage+Khazaka). Measurements were taken five times on each cheek, and the average of three measurements, excluding the lowest and highest values, was calculated. Measurements before and after four weeks of application were compared, and the greater the change, the greater the improvement. Visual assessment of skin condition was performed by a specialist evaluator engaged in cosmetic development and regular evaluation. Facial images were evaluated before and after four weeks of application, and the number of subjects who were deemed to have experienced greater improvement in skin condition was reported. Skin redness and unevenness were assessed using image analysis. Using a VISIA-CR (Canfield Scientific), facial photographs with surface light removed were taken before and after four weeks of application. Pigment components derived from hemoglobin were separated from the images obtained according to the method described in Patent 6,977,744, and hemoglobin-extracted images (color images obtained by multiplying the amount of hemoglobin component by the hemoglobin vector and then converting it to RGB) were created.Using image analysis software Image J, the standard deviation (StdDev) of red on the cheeks was calculated, and the value after 4 weeks of application was subtracted from the value before application to determine the change in redness unevenness. The smaller the change in redness unevenness, the more the skin's redness unevenness has improved. The results are shown in Tables 2 and 3.

[0043]

[0044]

[0045] The results in Table 2 show that the moisturizer containing rosemary extract of Example 1 (Example 4) was superior in improving stratum corneum function compared to the placebo moisturizer not containing rosemary extract (Reference Example 1). It was also found that the moisturizer of Example 4 had a greater effect on improving uneven redness of the skin compared to the placebo moisturizer (Reference Example 1). On the other hand, the results in Table 3 show that the moisturizer containing rosemary extract of Comparative Example 2 (Comparative Example 3) did not have a greater effect on improving stratum corneum function compared to the placebo moisturizer not containing rosemary extract (Reference Example 2). Furthermore, the moisturizer of Comparative Example 3 did not have a greater effect on improving uneven redness of the skin compared to the placebo moisturizer (Reference Example 2).

[0046] According to one aspect of the present disclosure, a plant-derived extract composition capable of increasing the expression levels of transglutaminase (TGM-1) and loricrin (LOR) can be provided.

Claims

1. A plant-derived extract composition containing rosmarinic acid, in which the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less.

2. The extract composition according to claim 1, wherein the content of rosmarinic acid is 0.5 ppm or more.

3. The extract composition according to claim 1 or 2, wherein the plant is rosemary.

4. A composition containing a plant extract component, which contains rosmarinic acid and has a mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) of 0.25 or less.

5. The composition described in claim 4 for promoting loricrin expression in human epidermal keratinocytes, promoting stratum corneum formation, or promoting epidermal turnover.

6. The composition according to claim 4 or 5 for promoting the expression of transglutaminase in human epidermal keratinocytes.

7. A composition according to any one of claims 4 to 6 for improving uneven redness of the skin.

8. A composition according to any one of claims 4 to 7, wherein the plant is rosemary.

9. A method for producing a plant-derived extract composition, comprising an extraction step in which extraction is performed so that the mass ratio of betulinic acid to rosmarinic acid (betulinic acid / rosmarinic acid) is 0.25 or less.

10. The method of claim 9, wherein the plant is rosemary.

11. A cosmetic preparation containing the plant-derived extract composition according to any one of claims 1 to 3.

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