TWI857919B - Anti-inflammatory agents and cosmetics - Google Patents

Abstract

An anti-aging agent containing at least any one of Artemisia plant fermentation liquid, thyme (Thymus vulgaris) fermentation liquid, Melissa officinalis fermentation liquid, and cornflower (Centaurea cyanus) fermentation liquid as an active ingredient; an antioxidant containing at least any one of Artemisia plant fermentation liquid, thyme fermentation liquid, Melissa officinalis fermentation liquid, and cornflower fermentation liquid as an active ingredient; an anti-inflammatory agent containing at least any one of Artemisia plant fermentation liquid and Melissa officinalis fermentation liquid as an active ingredient; a whitening agent containing at least any one of Artemisia plant fermentation liquid, Melissa officinalis fermentation liquid, and cornflower fermentation liquid as an active ingredient; and cosmetics.

Description

Anti-inflammatory agents and cosmetics

The present invention relates to an anti-aging agent, an antioxidant, an anti-inflammatory agent, and a whitening agent containing a fermented liquid obtained from a plant by yeast as an effective ingredient, and a cosmetic containing at least any one of the anti-inflammatory agent, the antioxidant, the anti-inflammatory agent, and the whitening agent.

In recent years, active oxygen has attracted attention as a factor that causes oxidation of biological components, and its adverse effects on organisms have become a problem. Active oxygen is produced in the process of energy metabolism in biological cells, including superoxide (superoxide anion (・O ₂ ^- ) produced by the reduction of oxygen molecules with one electron), hydrogen peroxide (H ₂ O ₂ ), singlet oxygen ( ¹ O ₂ ), hydroxyl radical (・OH) etc. This type of active oxygen is essential for the bactericidal mechanism of phagocytes and plays an important role in the removal of viruses and cancer cells.

However, the excessive generation of the aforementioned active oxygen will attack the biological molecules that constitute the membranes and tissues in the body and induce various diseases. Usually, superoxide, which is generated in the body and is the starting material of other active oxygen, is gradually eliminated by the catalysis of superoxide dismutase (SOD) contained in the cells. However, in the case of excessive superoxide generation or reduced SOD function, the elimination of superoxide becomes insufficient and the superoxide concentration becomes high. This is considered to be one of the causes of tissue damage such as rheumatoid arthritis, Behcet's disease, myocardial infarction, stroke, cataracts, spots, freckles, wrinkles, diabetes, arteriosclerosis, stiff shoulders, cold sensitivity, skin aging, etc.

Among these, the skin is directly stimulated by environmental factors such as ultraviolet rays and is an organ that easily produces superoxide. Therefore, when the concentration of superoxide increases, for example, biological tissues such as collagen are decomposed, denatured, or cross-linked, or fats are oxidized to produce lipid peroxides that damage cells, forming wrinkles on the skin and causing problems such as aging, inflammation, and skin pigmentation, such as reduced skin elasticity (see non-patent document 1). Therefore, it is generally believed that by blocking or inhibiting the generation of active oxygen or free radicals in the body, it is possible to prevent, treat or improve various injuries involving active oxygen, such as wrinkle formation, decreased elasticity and other skin aging, tissue damage such as rheumatoid arthritis or Behcet's disease, myocardial infarction, stroke, cataracts, diabetes, arteriosclerosis, shoulder stiffness, cold sensitivity, etc.

Therefore, attempts have been made to obtain active oxygen scavenging substances, free radical scavenging substances, hydrogen peroxide scavenging substances, etc. from natural products that are advantageous from the viewpoint of safety, and the effectiveness has been confirmed in extracts of plants of the genus Brassica of the family Brassicaceae (see Patent Document 1), extracts of plants of the genus Kalanchoe of the family Crassulaceae (see Patent Document 2), extracts of Psoralea corylifolia (see Patent Document 3), and extracts of young leaves of sweet potato (see Patent Document 4).

Inflammatory diseases, such as contact dermatitis (rash), tinea versicolor, pemphigus vulgaris, and other skin diseases accompanied by rough skin, have various causes and pathogenesis. As for the cause, it is known that it is mainly caused by the hyperactivity of hyaluronidase.

Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid. Hyaluronic acid salts, which retain affinity for biological tissues, are decomposed by ultraviolet rays, oxygen, etc. in a water-containing system, and the water-retaining effect is reduced as the molecular weight decreases. In addition, hyaluronic acid exists as an intercellular tissue in organisms and is also involved in vascular permeability. Furthermore, hyaluronidase exists in obese cells (mast cells), but is released by degranulation caused by its activation, and acts as an inflammatory chemical mediator. Based on this, it is expected that by inhibiting the activity of hyaluronidase, moisturizing can be enhanced and inflammation can be prevented and reduced. As for those having such hyaluronidase activity inhibitory effects, for example, extracts of Osbeckia plants (see Patent Document 5) and Ampelopsis grossedentata extracts (see Patent Document 6) are known.

In the skin, melanin also has the function of protecting the body from ultraviolet damage, but excessive production or uneven accumulation causes skin darkening or spots. Generally speaking, melanin is formed by the action of the enzyme tyrosinase synthesized in pigment cells, through the transformation from tyrosine to dopa, from dopa to dopaquinone, and then to intermediates such as 5,6-dihydroxyindole. Therefore, in order to prevent, treat or improve skin darkening (skin pigmentation), spots, freckles, etc., it is considered to hinder the activity of tyrosinase related to the production of melanin, or inhibit the production of melanin.

In the past, for the prevention, treatment or improvement of skin pigmentation, spots, freckles, etc., whitening agents with chemical synthetic products such as hydroquinone as active ingredients have been used for external use. However, chemical synthetic products such as hydroquinone have the risk of side effects such as skin irritation and allergies. Therefore, the development of whitening agents with highly safe natural raw materials as active ingredients is desired. For those with tyrosinase activity inhibitory effects, for example, rattan tea extract (see patent document 7) and Polygonum hydropiper extract (see patent document 8) are known. In addition, as for those having the effect of inhibiting melanin production, for example, extracts derived from the root of Ricinus communis (see Patent Document 9) and extracts derived from plants belonging to the genus Saussurea (see Patent Document 10) are known.

In addition, the epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrix such as collagen located outside these cells to support the skin structure. In young skin, by maintaining the constancy of the interaction of these skin tissues, moisture retention, softness, elasticity, etc. are ensured, and the skin is kept tight, shiny, fresh and young in appearance. However, due to the influence of certain external factors such as ultraviolet rays (UV-A, UV-B), significantly dry air, and excessive skin washing, as aging progresses, the production of collagen, which is the main component of the extracellular matrix, decreases, causing a decrease in elasticity due to cross-linking. As a result, the skin's moisturizing function or elasticity decreases, the keratin begins to peel abnormally, the skin loses its firmness or luster, and becomes rough, wrinkled, and other aging symptoms. The changes that accompany skin aging, such as wrinkles, dullness, loss of texture, and reduced elasticity, can be attributed to a variety of factors, but are related to the reduction and denaturation of extracellular matrix components such as collagen, hyaluronic acid, and elastin. Therefore, it is generally believed that skin aging can be improved by promoting the production of collagen, hyaluronic acid, etc.

Therefore, substances having the effect of promoting collagen production have been tried to be obtained from favorable natural substances from the point of view of safety. For example, carambola leaf extract (see Patent Document 11) and Mallotus philippinensis extract (see Patent Document 12) have been confirmed to have the effect of promoting collagen production.

In addition, the amino acids that are the main components of Natural Moisturizing Factors are produced by the decomposition of filaggrin from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilaggrin in the epidermal keratinocytes of the stratum granulosum just below the stratum corneum. Afterwards, it is immediately phosphorylated and accumulated in the keratin granules. After dephosphorylation and hydrolysis, it is decomposed into filaggrin, which migrates to the stratum corneum, improves the aggregation efficiency of keratin filaments, and participates in the internal structure of keratinocytes (refer to non-patent document 2). In recent years, it has been reported that this filaggrin is very important and indispensable for skin moisture retention, and that due to dry conditions, the synthesis of filaggrin decreases, and the amount of amino acids in the stratum corneum decreases (see non-patent document 3). Therefore, it is expected that by promoting the expression of profilaggrin mRNA in epidermal keratinocytes, the amount of amino acids in the stratum corneum can be increased by promoting the synthesis of filaggrin, thereby essentially improving the moisture environment of the stratum corneum.

As for the naturally derived filaggrin synthesis promoter, for example, licorice extract (see Patent Document 13), glycyrrhizin (liquiritin) known as a flavanone glycoside contained in natural plants (see Patent Document 14), and a plant extract or yeast extract belonging to the genus Citrus as at least one of profilaggrin and filaggrin protein production promoters derived from naturally derived substances (see Patent Document 15), etc. have been proposed.

The epidermis continuously produces new keratinocytes through the division and subsequent differentiation of keratinocytes, and has the defensive function of protecting the skin from various external stimuli. In particular, during the differentiation process of keratinocytes, proteins such as involucrin are expressed from the stratum spinosum to the stratum granulosum, and are cross-linked by the enzyme transglutaminase-1 to form an insoluble cell membrane-like structure called cornified envelope (hereinafter abbreviated as "CE") that encapsulates keratinocytes, which helps the stability of the keratinocyte cytoskeleton and structure. However, due to various factors, when the amount of transglutaminase-1 produced in the epidermis decreases, CE formation becomes incomplete and keratinization proceeds abnormally. It is generally believed that the result is a decrease in the barrier function of keratin and the moisturizing function of the skin, and skin symptoms such as rough skin or dry skin. Therefore, by increasing the production of transglutaminase-1 in the epidermis of keratinocytes and promoting the formation of CE to normalize keratinization, it is possible to inhibit the decrease in the skin barrier function associated with dryness or external stimuli such as ultraviolet rays, and prevent and improve various skin symptoms such as dry skin and rough skin.

As a naturally derived transglutaminase-1 production promoter, Morinda citrifolia extract (see Patent Document 16), royal jelly extract (see Patent Document 17), and the like have been proposed.

In skin cells, aquaporins, which are known as water channels, are expressed on the cell membrane and are responsible for taking in low molecular weight substances, including water, from the intercellular space into the cell. In humans, 13 types of aquaporins (AQP0 to AQP12) are known to exist. It is generally believed that aquaporin 3 (Aquaporin 3; AQP3) is mainly present in epidermal cells, and in addition to taking in water, it is also responsible for taking in low molecular weight compounds such as glycerol or urea that participate in the water retention function.

However, since AQP3 is suggested to decrease with aging and is a factor in the reduction of water retention function, it is generally believed that by promoting the expression of AQP3, the water retention function or barrier function caused by aging can be controlled (see Non-Patent Document 4). For example, an extract from the leaves of carambola is known as an agent having an AQP3 expression promoting effect (see Patent Document 18).

In the past, it was generally believed that the barrier function of the skin was only the responsibility of the stratum corneum. However, since the skin barrier function would collapse if the constituent proteins of the tight junction (hereinafter abbreviated as "TJ") present in the stratum granulosum of the epidermis were missing at the genetic level, in recent years, it has been believed that TJ also plays an important role in the barrier function of the skin (see non-patent document 5). TJ is a junction device that not only makes adjacent cells adhere to each other, but also controls the permeability of substances by sealing the gaps between cells. TJs are composed of cell membrane proteins claudin and occludin. It is generally believed that these proteins form the skeleton of the TJ strand and control the barrier function of TJ (see non-patent document 6). As mentioned above, if the expression of claudin and occludin decreases for some reason, the TJ structure will be damaged and lose its function as a barrier for substances to pass through. Therefore, it is expected to become the cause of skin symptoms such as dry skin, rough skin, atopic dermatitis and various infections.

Therefore, it is generally believed that by promoting the production of claudin and nexin in the epidermis and promoting the formation of TJs in epidermal keratinocytes, the barrier function and moisture retention function of the skin can be improved, and the aforementioned skin symptoms can be prevented or improved. Based on this idea, in terms of improving the skin barrier function by promoting the formation of TJs, Coptis Rhizome extract (see Patent Document 19) and hondo spruce extract (see Patent Document 20) derived from natural products have been proposed.

In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as a factor that induces changes with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1) is known to be an enzyme that decomposes collagen, the main component of the extracellular matrix of the dermis of the skin, but it is believed that its expression is greatly increased by ultraviolet irradiation, becoming a factor in the reduction and denaturation of collagen, and is a major factor in the formation of skin wrinkles and reduced elasticity. Therefore, inhibiting the activity of MMP-1 is important in preventing and improving the symptoms of skin aging.

Known substances having such MMP-1 inhibitory effects include, for example, extracts from Yunnan plum (Prunus cerasoides) (see Patent Document 21), extracts from Zingiber cassumunar of the Zingiberaceae family, or extracts from Ficus neriifolia of the Moraceae family (see Patent Document 22), and the like.

On the other hand, although it is known that there are fermented liquids of Artemisia plants fermented with koji (see Patent Document 23) and fermented liquids of Thymus vulgaris fermented with koji (see Patent Document 24), it is not known that these plant fermented liquids have anti-aging effects, antioxidant effects, anti-inflammatory effects, whitening effects, etc.

[Prior technical literature] [Patent literature] [Patent literature 1]    Japanese Patent Publication No. 2003-81848 [Patent literature 2]   Japanese Patent Publication No. 2005-29483 [Patent literature 3]   Japanese Patent Publication No. 2006-321730 [Patent literature 4]    Japanese Patent Publication No. 2007-8902 [Patent literature 5]    Japanese Patent Publication No. 2003-55242 [Patent literature 6]    Japanese Patent Publication No. 2003-12532 [Patent literature 7]    Japanese Patent Publication No. 2002-370962 [Patent literature 8]    Japanese Patent Publication No. 2004-83488 [Patent Document 9]    Patent Publication No. 2001-213757 [Patent Document 10]  Patent Publication No. 2002-201122 [Patent Document 11]  Patent Publication No. 2002-226323 [Patent Document 12]  Patent Publication No. 2003-146837 [Patent Document 13]  Patent Publication No. 2002-363054 [Patent Document 14]  Patent Publication No. 2003-146886 [Patent Document 15]  Patent Publication No. 2001-261568 [Patent Document 16]  Patent Publication No. 2010-090093 [Patent Document 17] Japanese Patent Publication No. 2009-184955 [Patent Document 18] Japanese Patent Publication No. 2009-191039 [Patent Document 19] Japanese Patent Publication No. 2007-176830 [Patent Document 20] Japanese Patent Publication No. 2007-176835 [Patent Document 21] Japanese Patent Publication No. 2003-176232 [Patent Document 22] Japanese Patent Publication No. 2003-176230 [Patent Document 23] Japanese Patent Publication No. 2011-140453 [Patent Document 24] Japanese Patent Publication No. 2011-130689

[Non-patent document] [Non-patent document 1] "FRAGRANCE JOURNAL" Temporary Supplement No.14, p156, 1995 [Non-patent document 2] FRAGRANCE JOURNAL Temporary Supplement, vol.17, pp.14-19(2000) [Non-patent document 3] Arch.Dermatol.Res., vol.288, pp.442-446(1996) [Non-patent document 4] "FRAGRANCE JOURNAL", 2006, Vol.34, No.10, p.19-23 [Non-patent document 5] J.Cell Biol., vol.156, pp.1099-1111(2002) [Non-patent document 6] Journal of the Japanese Society of Perfumery and Fragrance Sciences, vol.31, pp.296-301(2007)

[Problems to be solved by the invention] The purpose of the present invention is to solve the above-mentioned problems in the past and achieve the following purposes. That is, the purpose of the present invention is to provide an anti-aging agent with excellent anti-aging effect and high safety; an antioxidant agent with excellent anti-oxidation effect and high safety; an anti-inflammatory agent with excellent anti-inflammatory effect and high safety; and a whitening agent with excellent whitening effect and high safety. In addition, the purpose of the present invention is to provide a cosmetic product with at least one effect selected from the group consisting of excellent anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect, and high safety.

[Means for Solving the Problem] The means for solving the above-mentioned problem are as follows. That is, <1> An anti-aging agent characterized by containing at least one of the fermentation liquid of Artemisia plant obtained by koji, the fermentation liquid of thyme obtained by koji, the fermentation liquid of Melissa officinalis obtained by koji, and the fermentation liquid of cornflower (Centaurea cyanus) obtained by koji as an active ingredient. <2> An antioxidant, characterized by containing as an active ingredient at least one of a fermentation solution of a plant of the genus Artemisia obtained by koji, a fermentation solution of thyme obtained by koji, a fermentation solution of cypress obtained by koji, and a fermentation solution of cornflower obtained by koji. <3> An anti-inflammatory agent, characterized by containing as an active ingredient at least one of a fermentation solution of a plant of the genus Artemisia obtained by koji, and a fermentation solution of cypress obtained by koji. <4> A whitening agent, characterized by containing as an active ingredient at least one of a fermentation solution of a plant of the genus Artemisia obtained by koji, a fermentation solution of cypress obtained by koji, and a fermentation solution of cornflower obtained by koji. <5> A cosmetic characterized by containing at least one selected from the group consisting of the anti-aging agent described in <1>, the antioxidant described in <2>, the anti-inflammatory agent described in <3>, and the whitening agent described in <4>.

[Effects of the invention] According to the present invention, the above-mentioned problems in the past can be solved, the above-mentioned objectives can be achieved, and an anti-aging agent with excellent anti-aging effect and high safety, an antioxidant agent with excellent anti-oxidation effect and high safety, an anti-inflammatory agent with excellent anti-inflammatory effect and high safety, and a whitening agent with excellent whitening effect and high safety can be provided. In addition, the present invention can provide a cosmetic product with at least one effect selected from the group consisting of excellent anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect, and with high safety.

(Anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent) <Anti-aging agent> The anti-aging agent of the present invention contains at least one of the fermentation liquids of Artemisia spp. obtained by koji (hereinafter, sometimes referred to as "Artemisia spp. fermentation liquid"), thyme fermentation liquid obtained by koji (hereinafter, sometimes referred to as "thyme fermentation liquid"), cypress fermentation liquid obtained by koji (hereinafter, sometimes referred to as "cypress fermentation liquid"), and cornflower fermentation liquid obtained by koji (hereinafter, sometimes referred to as "cornflower fermentation liquid") as an active ingredient, and further contains other ingredients as needed.

The aforementioned Artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned Cypress fermentation liquid, and the aforementioned Cornflower fermentation liquid have an effect selected from the group consisting of matrix metalloproteinase-1 (MMP-1) activity inhibition effect, hyaluronan synthase 3 (HAS3) mRNA expression promotion effect, type I collagen production promotion effect, claudin-1 mRNA expression promotion effect, claudin-4 mRNA expression promotion effect, claudin mRNA expression promotion effect, transglutaminase-1 (TGM-1) mRNA expression promotion effect, aquaporin 3 (Aquaporin 3; AQP3) mRNA expression promoting effect, and polycin mRNA expression promoting effect, and utilizing these effects, it can be used as an effective ingredient of an anti-aging agent.

Therefore, the aforementioned anti-aging agent has at least one effect selected from the group consisting of MMP-1 activity inhibition effect, hyaluronic acid synthase 3 mRNA expression promotion effect, type I collagen production promotion effect, claudin-1 mRNA expression promotion effect, claudin-4 mRNA expression promotion effect, clenbuterol mRNA expression promotion effect, transglutaminase-1 mRNA expression promotion effect, aquaporin 3 mRNA expression promotion effect, and filaggrin mRNA expression promotion effect.

Regarding the inhibitory effect of MMP-1 activity, the promoting effect of hyaluronic acid synthase 3 mRNA expression, the promoting effect of type I collagen production, the promoting effect of claudin-1 mRNA expression, the promoting effect of claudin-4 mRNA expression, the promoting effect of claudin mRNA expression, the promoting effect of transglutaminase-1 mRNA expression, and the aquaporin 3 of the aforementioned artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned cypress fermentation liquid, and the aforementioned cornflower fermentation liquid Although the details of at least one of the substance having the effect of promoting the expression of mRNA and the effect of promoting the expression of fibronectin mRNA are not clear, it is completely unknown in the past that the aforementioned Artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned Cypress fermentation liquid, and the aforementioned Cornflower fermentation liquid have such excellent effects and are useful as anti-aging agents, and this is a new discovery of the inventors.

<Antioxidant> The antioxidant of the present invention contains at least one of fermentation liquid obtained by koji of wormwood (wormwood fermentation liquid), fermentation liquid obtained by koji of thyme (thyme fermentation liquid), fermentation liquid obtained by koji of cypress (cypress fermentation liquid), and fermentation liquid obtained by koji of cornflower (cornflower fermentation liquid) as an active ingredient, and further contains other ingredients as needed.

The aforementioned Artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned Cypress fermentation liquid, and the aforementioned Cornflower fermentation liquid have a diphenyl-p-picrylhydrazyl (DPPH) free radical scavenging effect, and can be used as an active ingredient of an antioxidant by utilizing this effect. Therefore, the aforementioned antioxidant is one that has a DPPH free radical scavenging effect.

Although the details of the substance that exerts the DPPH free radical scavenging effect possessed by the aforementioned Artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned Cypress fermentation liquid, and the aforementioned Cornflower fermentation liquid are not clear, it is completely unknown in the past that the aforementioned Artemisia plant fermentation liquid, the aforementioned thyme fermentation liquid, the aforementioned Cypress fermentation liquid, and the aforementioned Cornflower fermentation liquid have such excellent effects and are useful as antioxidants, and this is a new discovery of the inventors.

<Anti-inflammatory agent> The anti-inflammatory agent of the present invention contains at least one of the fermentation liquid of Artemisia spp. obtained by koji mold (Artemisia spp. fermentation liquid) and the fermentation liquid of Cypress spp. obtained by koji mold (Cypress spp. fermentation liquid) as an active ingredient, and further contains other ingredients as needed.

The aforementioned Artemisia plant fermentation liquid and the aforementioned Cypress Fermentation Liquid have the effect of inhibiting hyaluronidase activity, and this effect can be utilized to use them as the active ingredient of anti-inflammatory agents. Therefore, the aforementioned anti-inflammatory agent has the effect of inhibiting hyaluronidase activity.

Although the details of the substance that inhibits hyaluronidase activity in the aforementioned Artemisia plant fermentation liquid and the aforementioned Cypress grass fermentation liquid are not clear, it is completely unknown in the past that the aforementioned Artemisia plant fermentation liquid and the aforementioned Cypress grass fermentation liquid have such excellent effects and are useful as antioxidants, and this is a new discovery of the inventors.

<Whitening agent> The whitening agent of the present invention contains at least one of the fermentation liquid of Artemisia spp. obtained by koji mold (Artemisia spp. fermentation liquid), the fermentation liquid of Cypress spp. obtained by koji mold (Cypress spp. fermentation liquid), and the fermentation liquid of Cornflower spp. obtained by koji mold (Cornflower fermentation liquid) as an active ingredient, and further contains other ingredients as needed.

The aforementioned Artemisia plant fermentation liquid, Cypress grass fermentation liquid, and Cornflower fermentation liquid have at least one of the effects of inhibiting tyrosinase activity and inhibiting melanin production, and can be used as an active ingredient of a whitening agent by utilizing this effect. Therefore, the aforementioned whitening agent has at least one of the effects of inhibiting tyrosinase activity and inhibiting melanin production.

Although the details of the substance that exerts at least one of the tyrosinase activity inhibitory effect and the melanin production inhibitory effect possessed by the aforementioned Artemisia plant fermentation liquid, the aforementioned Cypress grass fermentation liquid, and the aforementioned Cornflower fermentation liquid are not clear, it is completely unknown in the past that the aforementioned Artemisia plant fermentation liquid, the aforementioned Cypress grass fermentation liquid, and the aforementioned Cornflower fermentation liquid have such excellent effects and are useful as whitening agents, and this is a new discovery of the inventors.

<<Artemisia plant fermentation liquid>> The aforementioned Artemisia plant fermentation liquid is a fermentation liquid obtained by using koji fungi from plants belonging to the genus Artemisia (hereinafter, sometimes referred to as "Artemisia plants").

-Artemisia Plants- The Artemisia plants used as the fermentation raw materials are perennial herbs belonging to the genus Artemisia of the family Compositae , and have been used as food and medicinal raw materials since ancient times. They are widely native or cultivated in Japan, such as Hokkaido, Honshu, Shikoku, and Kyushu, and can be easily obtained from these regions.

The species of the aforementioned Artemisia plants are not particularly limited and can be appropriately selected according to the purpose. For example, Artemisia montana (Nakai) Pamp., Artemisia capillaris Thunbergii, Artemisia princeps Pampan., Artemisia japonica Thunb., Artemisia absinthium L., Artemisia lactiflora Wall., Artemisia maritima L., Artemisia scoparia Waldst.et kit., etc. These can be used alone or in combination of two or more. The method for obtaining the aforementioned Artemisia plants is not particularly limited and can be appropriately selected according to the purpose. They can be collected from nature or commercially available products can be used.

The parts of the wormwood plant used as the fermentation raw material are not particularly limited and can be appropriately selected according to the purpose. For example, the aboveground parts of flowers, buds, fruits, peels, seeds, seed coats, stems, leaves, branches, and branches and leaves; the underground parts of roots, rhizomes, etc. can be used alone or in combination. Among these, the aboveground parts of the wormwood plant are preferred.

As for the size of the aforementioned Artemisia plant used as the aforementioned fermentation raw material, there is no particular limitation as long as it is a size that can be cultivated by the aforementioned koji mold, and it can be appropriately selected according to the purpose, for example, the original size taken, the desired size after cutting, the size of micronized (powdered), etc.

The state of the aforementioned Artemisia plant used as the aforementioned fermentation raw material is not particularly limited as long as it is a state that can be cultured by the aforementioned koji fungus. It can be appropriately selected according to the purpose, for example, the original state taken, the dried state, the crushed state, the juiced state, the state of the extract, etc. Among these, the original state taken, the crushed state, the juiced state, and the state of the extract are preferred, and the original state taken and the crushed state are more preferred, as the koji fungus is easy to act on.

The method for drying the Artemisia plant is not particularly limited and can be appropriately selected according to the purpose, for example, a method of drying in sunlight, a method of drying using a commonly used dryer, etc.

The method for making the aforementioned Artemisia plant into the aforementioned pulverized state is not particularly limited and can be appropriately selected according to the purpose. For example, there can be cited a method of pulverizing using a mixer, a sugar mill, a power mill, a jet mill, an impact mill, etc.

The method of making the aforementioned Artemisia plant into the aforementioned juice state is not particularly limited and can be appropriately selected according to the purpose, and for example, pressing can be cited.

The method for preparing the aforementioned Artemisia plant into the aforementioned extract is not particularly limited, and a method generally used for plant extraction can be appropriately selected according to the purpose.

The wormwood plant used as the fermentation raw material is preferably sterilized before the inoculation of the yeast. The method for sterilizing the wormwood plant is not particularly limited and can be appropriately selected from known methods.

-Koji- The koji used to ferment the wormwood plant is not particularly limited and can be appropriately selected according to the purpose. For example, yellow koji such as Aspergillus oryzae and Aspergillus sojae ; black koji such as Aspergillus luchuensis ; white koji such as Aspergillus kawauchii ; variants thereof, etc. These can be used alone or in combination of two or more. Among these, Aspergillus oryzae is preferred in terms of excellent anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect. As for the method for obtaining the aforementioned koji mold, there is no particular limitation, and it can be appropriately selected according to the purpose, and it can be collected from nature or commercially available. In addition, as the aforementioned koji mold, a seed koji made from rice or the like can be used, a seed koji of the genus Artemisia described below can be used, or a koji mold cultured in a culture medium (agar culture medium, liquid culture medium, etc.) can be used. Among these, the use of the aforementioned koji mold is preferred in terms of the superiority of at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect.

The inoculation amount of the koji mold to the Artemisia plant used as the fermentation raw material is not particularly limited as long as the amount can ferment the Artemisia plant, and can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, it is preferably 1×10 ³ cells/mL to 1×10 ⁸ cells/mL, and when the fermentation raw material is in a solid state, it is preferably 1×10 ³ cells/g to 1×10 ⁸ cells/g.

When the wormwood plant is inoculated with the yeast, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 500 to 5,000 parts by mass, more preferably 1,000 to 4,000 parts by mass, and particularly preferably 1,500 to 3,000 parts by mass are added relative to 100 parts by mass of the wormwood plant.

The temperature of the fermentation (cultivation) is not particularly limited as long as it is within the range of the fermentation temperature of the koji mold, and can be appropriately selected according to the purpose, but 20°C to 40°C is preferred, and 25°C to 35°C is more preferred. When the fermentation temperature is lower than 20°C, the wormwood plant cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect becomes insufficient. In addition, when it exceeds 50°C, the koji mold cannot proliferate.

The fermentation (cultivation) time is not particularly limited and can be appropriately selected according to the purpose, but is preferably 10 to 40 hours, more preferably 20 to 30 hours. If the fermentation time is less than 10 hours, the Artemisia plant cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect becomes insufficient.

There is no particular limitation on the method for stopping the aforementioned fermentation (cultivation), and it can be appropriately selected according to the purpose, for example, a heating method, etc. As for the heating temperature used to stop the aforementioned fermentation, if it is a temperature at which the aforementioned koji cannot grow, it is not particularly limited, and it can be appropriately selected according to the purpose, but it is preferably above 50°C, more preferably above 70°C, and particularly preferably 100°C to 130°C. When the aforementioned heating temperature is lower than 50°C, the aforementioned fermentation may not be stopped, and when it exceeds 130°C, at least one of the anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect becomes insufficient. As for the heating time for stopping the aforementioned fermentation, there is no particular limitation as long as the aforementioned yeast cannot grow, and it can be appropriately selected according to the purpose, but it is preferably more than 5 minutes, and more preferably 10 minutes to 20 minutes. When the aforementioned heating time is less than 5 minutes, the aforementioned fermentation cannot be stopped, and when it exceeds 20 minutes, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect becomes insufficient.

Furthermore, the fermented wormwood liquid is preferably cooled after the fermentation is stopped. The cooling method is not particularly limited and can be appropriately selected according to the purpose. For example, the method of leaving the liquid at room temperature or in a refrigerator can be cited.

The number of times the wormwood plant is fermented using the yeast is not particularly limited and can be appropriately selected according to the purpose, and may be once or multiple times.

When the aforementioned fermentation is performed multiple times, the aforementioned yeast may be inoculated only for the first time, may be inoculated only for a few times, or may be inoculated for all times, but it is preferred to perform inoculation only for the first time. When the aforementioned fermentation is performed multiple times, the fermentation temperature and fermentation time may be different or the same.

--Artemisia spp. seed yeast-- The aforementioned Artemisia spp. seed yeast is prepared by using the aforementioned Artemisia spp. as a seed yeast raw material, inoculating the seed yeast raw material with koji fungi, and allowing a sufficient amount of spores to attach to the Artemisia spp. By using it for the aforementioned fermentation, it is advantageous that an Artemisia spp. fermented liquid with excellent skin affinity can be obtained more efficiently and easily.

The aforementioned Artemisia plant used as the aforementioned koji seed raw material may be the same as that described in the aforementioned "Artemisia plant", and the parts, sizes, states, etc. of the aforementioned Artemisia plant used are also the same.

As for the aforementioned koji mold used in the production of the aforementioned Artemisia plant seed koji, the same ones as those described in the aforementioned "koji mold" can be used.

The amount of the koji fungus inoculated with the wormwood plant used as the koji seed raw material is not particularly limited and can be appropriately selected according to the purpose, but preferably 5 to 100 parts by mass of the koji fungus suspended in sterile water (1×10 ³ cells/mL to 1×10 ⁸ cells/mL) is inoculated with 100 parts by mass of the wormwood plant, more preferably 10 to 50 parts by mass, and particularly preferably 20 to 30 parts by mass. When the inoculation amount of the koji fungus is less than 5 parts by mass relative to 100 parts by mass of the wormwood plant, sufficient spores cannot be attached to the wormwood plant, and when it exceeds 100 parts by mass, it reproduces abnormally due to excessive water.

When the koji mold is inoculated with the wormwood plant used as the koji seed material, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 10 to 250 parts by mass, more preferably 20 to 200 parts by mass, and particularly preferably 30 to 150 parts by mass are added relative to 100 parts by mass of the wormwood plant. When the amount of water added relative to 100 parts by mass of the wormwood plant is less than 10 parts by mass, a sufficient amount of spores may not be attached to the wormwood plant.

The culture temperature is not particularly limited as long as it is within the range of the temperature at which the koji fungus can grow, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the culture temperature is lower than 20°C, sometimes sufficient spores cannot attach to the wormwood plant. Also, when the temperature exceeds 50°C, sometimes the koji fungus cannot proliferate.

The time of the aforementioned culture is not particularly limited and can be appropriately selected according to the purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the aforementioned culture time is less than 80 hours, sometimes a sufficient amount of spores cannot be attached to the aforementioned Artemisia plant, and if it exceeds 210 hours, sometimes the germination rate of the spores will decrease.

The fermented wormwood liquid may contain the bacterial bodies of the koji mold or may be a liquid without the bacterial bodies of the koji mold. However, it is preferably a liquid without the bacterial bodies of the koji mold.

The state of the fermented mugwort liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it can be the fermented mugwort liquid itself, a purified product of the fermented mugwort liquid, a concentrated product of the fermented mugwort liquid, a diluted product of the fermented mugwort liquid, etc. In addition, the fermented mugwort liquid can be a dried product of the fermented mugwort liquid mixed with or dissolved in a solvent such as water or a hydrophilic solvent.

As for the purified product of the fermented wormwood liquid, there is no particular limitation and it can be appropriately selected according to the purpose. For example, it can be a product obtained by removing the solid components (for example, the plant body of the wormwood plant, the body of the koji mold, the residue, etc.) in the fermented wormwood liquid. As for the means of removal, there is no particular limitation and it can be appropriately selected according to the purpose. For example, filtration, etc. can be cited. As for the method of filtration, there is no particular limitation and it can be appropriately selected from the known methods according to the purpose.

There are no particular restrictions on the aforementioned dilution of the fermented wormwood plant and the aforementioned concentrate of the fermented wormwood plant, and they can be appropriately selected according to the purpose. For example, the fermented wormwood plant is prepared to a desired concentration. There are no particular restrictions on the aforementioned dilution method, and they can be appropriately selected from the known methods according to the purpose. There are no particular restrictions on the aforementioned concentration method, and they can be appropriately selected according to the purpose. For example, reduced pressure concentration can be listed.

As for the dried product of the fermented wormwood liquid, there is no particular limitation and it can be appropriately selected according to the purpose, for example, the dried product of the fermented wormwood liquid, etc. As for the drying method, there is no particular limitation and it can be appropriately selected according to the purpose, for example, freeze drying, etc.

The aforementioned Artemisia plant fermentation liquid is not particularly limited as long as it is a fermentation liquid obtained from Artemisia plant by yeast, and can be appropriately selected according to the purpose. However, from the perspective of good skin affinity, it is preferably an Artemisia plant fermentation liquid with a contact angle of 81° or less, and more preferably an Artemisia plant fermentation liquid with a contact angle of 78° or less.

In this manual, the contact angle is the value obtained by dropping 3 μL of the test sample onto the sample stage of the dynamic contact angle and surface tension measuring device (FTA1000 Falcon, manufactured by First Ten Angstroms) and measuring the contact angle θ (°) at 1,000 ms using the θ/2 method at a temperature of 22°C and a relative humidity of 20%. The contact angle is used as an indicator of "wetting" and is defined as "the angle formed by the liquid surface and the solid surface at the junction of the free surface of a stationary liquid and the adjacent solid wall (the angle taken inside the liquid)" (refer to the 4th edition of the Physical and Chemical Dictionary, Iwanami Shoten Co., Ltd.). The aforementioned contact angle depends on the relationship between the cohesive force between liquid molecules and the adhesion force between solid walls. When the liquid wets the solid (adhesion is strong), it is a sharp angle, and when it is not wet, it is a blunt angle. Therefore, the smaller the contact angle, the easier it is to get wet, that is, because it indicates better skin affinity, there is no special limit on the lower limit of the contact angle of the aforementioned Artemisia plant fermented liquid, and it can be appropriately selected according to the purpose.

＜＜Thyme fermented liquid＞＞ The aforementioned thyme fermented liquid is the fermented liquid of thyme obtained by using yeast.

- Thyme - Thymus vulgaris Linne used as the fermentation raw material is a perennial woody plant belonging to the genus Thymus in the mint family ( Labiatae ). It is a kind of herb and has been used as a food and medicinal raw material since ancient times. It is also known as Common thyme. It is native to the Mediterranean coast, but it also grows naturally or is cultivated in Japan, and can be easily obtained from such areas. There is no particular limitation on the method of obtaining the thyme mentioned above. It can be appropriately selected according to the purpose. It can be collected from nature or commercially available products can be used.

The parts of thyme used as the fermentation raw material are not particularly limited and can be appropriately selected according to the purpose. For example, the aboveground parts of flowers, buds, fruits, peels, seeds, seed coats, stems, leaves, branches, bark, trunks, branches and leaves; the underground parts of roots, rhizomes, etc. can be used alone or in combination of two or more. Among these, the aboveground parts of thyme are preferably used.

As for the size of the thyme used as the fermentation raw material, there is no particular limitation as long as it is a size that can be cultured by the koji mold. It can be appropriately selected according to the purpose, for example, the original size taken, the desired size after cutting, the size of micronized (powdered) and the like.

As for the state of the thyme used as the fermentation raw material, as long as it is a state in which the koji fungus can be cultured, it is not particularly limited and can be appropriately selected according to the purpose, for example, the original state of the thyme, the dried state, the crushed state, the juiced state, the state of the extract, etc. Among these, the original state of the thyme, the crushed state, the juiced state, the state of the extract are preferred, and the original state of the thyme and the crushed state are more preferred, as the koji fungus can easily act.

The method for drying the thyme is not particularly limited and can be appropriately selected according to the purpose. For example, a method of drying in sunlight or a method of drying using a commonly used dryer can be cited.

The method for making the thyme into the above-mentioned pulverized state is not particularly limited and can be appropriately selected according to the purpose. For example, there can be cited a method of pulverizing using a mixer, a sugar mill, a powder mill, a jet mill, an impact mill, etc.

The method for making the thyme into the aforementioned juice is not particularly limited and can be appropriately selected according to the purpose, and for example, pressing can be cited.

The method for preparing the thyme into the extract is not particularly limited, and a method generally used for plant extraction can be appropriately selected according to the purpose.

The thyme used as the fermentation raw material is preferably sterilized before the inoculation of the yeast. There is no particular limitation on the method for sterilizing the thyme, and the method can be appropriately selected from known methods.

-Aspergillus- As for the aforementioned koji used to ferment the aforementioned thyme, there is no particular limitation and it can be appropriately selected according to the purpose. For example, those described in the aforementioned <<Artemisia plant fermentation liquid>> can be cited. These can be used alone or in combination of two or more. Among these, as for the aforementioned koji, rice koji ( Aspergillus oryzae ) is preferred because it is excellent in at least one of the anti-aging effect and the antioxidant effect. As for the method for obtaining the aforementioned koji, there is no particular limitation and it can be appropriately selected according to the purpose. It can be collected from nature or a commercial product can be used. In addition, as the koji, a seed koji made from rice or the like can be used, a thyme seed koji described below can be used, or a koji cultured in a culture medium (an agar culture medium, a liquid culture medium, etc.) can be used. Among these, the thyme seed koji is preferably used because it is excellent in at least one of the anti-aging effect and the anti-oxidation effect.

The inoculation amount of the koji mold to the thyme used as the fermentation raw material is not particularly limited as long as the thyme can be fermented. It can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, it is preferably 1×10 ³ cells/mL to 1×10 ⁸ cells/mL, and when the fermentation raw material is in a solid state, it is preferably 1×10 ³ cells/g to 1×10 ⁸ cells/g.

When the koji is inoculated into the thyme, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 500 to 5,000 parts by mass, more preferably 1,000 to 4,000 parts by mass, and particularly preferably 1,500 to 3,000 parts by mass are added relative to 100 parts by mass of the thyme.

The fermentation (cultivation) temperature is not particularly limited as long as it is within the fermentation temperature range of the yeast, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the fermentation temperature is lower than 20°C, the thyme cannot be fully fermented, and at least one of the anti-aging effect and the anti-oxidation effect becomes insufficient.

The fermentation (cultivation) time is not particularly limited and can be appropriately selected according to the purpose, but is preferably 10 to 40 hours, more preferably 20 to 30 hours. If the fermentation time is less than 10 hours, the thyme cannot be fully fermented, and at least one of the anti-aging effect and the anti-oxidation effect becomes insufficient.

There is no particular limitation on the method for stopping the fermentation (cultivation), and it can be appropriately selected according to the purpose. For example, a heating method can be cited. As for the heating temperature for stopping the fermentation, if it is a temperature at which the koji mold cannot grow, it is not particularly limited, and it can be appropriately selected according to the purpose, but it is preferably 50°C or more, more preferably 70°C or more, and particularly preferably 100°C to 130°C. If the heating temperature is lower than 50°C, the fermentation may not be stopped, and if it exceeds 130°C, at least one of the anti-aging effect and the anti-oxidation effect becomes insufficient. As for the heating time for stopping the fermentation, if it can make the koji mold unable to grow, it is not particularly limited, and it can be appropriately selected according to the purpose, but it is preferably 5 minutes or more, and more preferably 10 minutes to 20 minutes. If the heating time is less than 5 minutes, the fermentation may not be stopped. If it exceeds 20 minutes, at least one of the anti-aging and antioxidant effects may become insufficient.

Furthermore, the thyme fermented liquid is preferably cooled after the fermentation is stopped. The cooling method is not particularly limited and can be appropriately selected according to the purpose, for example, the method of leaving it at room temperature or in a refrigerator can be cited.

The number of times the thyme is fermented using the yeast is not particularly limited and can be appropriately selected according to the purpose, and may be once or multiple times.

When the aforementioned fermentation is performed multiple times, the aforementioned yeast may be inoculated only for the first time, may be inoculated only for a few times, or may be inoculated for all times, but it is preferred to perform only the first time. When the aforementioned fermentation is performed multiple times, the fermentation temperature and fermentation time may be different or the same.

--Thyme seed yeast-- The aforementioned thyme seed yeast is prepared by using the aforementioned thyme as a seed yeast raw material, inoculating the koji fungus into the seed yeast raw material, and allowing a sufficient amount of spores to attach to the thyme. By using the aforementioned fermentation, it is advantageous in that a thyme fermentation liquid with excellent skin affinity can be obtained more efficiently and easily.

The aforementioned thyme used as the aforementioned seed yeast raw material may be the same as that described in the aforementioned "Thyme", and the parts, sizes, states, etc. of the aforementioned thyme used are also the same.

As for the aforementioned koji mold used in the production of the aforementioned thyme seed koji, the same ones as those described in the aforementioned "koji mold" can be used.

The amount of the koji fungus inoculated with the thyme used as the koji seed material is not particularly limited and can be appropriately selected according to the purpose. However, it is preferred to inoculate 5 to 100 parts by mass of the suspended koji fungus (1×10 ³ cells/mL to 1×10 ⁸ cells/mL) with sterile water relative to 100 parts by mass of the thyme, more preferably 10 to 50 parts by mass, and particularly preferably 20 to 30 parts by mass. When the inoculation amount of the koji fungus relative to 100 parts by mass of the thyme is less than 5 parts by mass, sometimes sufficient spores cannot be attached to the thyme, and when it exceeds 100 parts by mass, sometimes abnormal reproduction occurs due to excessive water.

When the koji mold is inoculated into the thyme used as the seed koji raw material, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 10 to 250 parts by mass, more preferably 20 to 200 parts by mass, and particularly preferably 30 to 150 parts by mass are added relative to 100 parts by mass of the thyme. When the amount of water added relative to 100 parts by mass of the thyme is less than 10 parts by mass, sometimes a sufficient amount of spores cannot be attached to the thyme.

The culture temperature is not particularly limited as long as it is within the range at which the koji fungus can grow, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the culture temperature is lower than 20°C, a sufficient amount of spores may not attach to the thyme. When the culture temperature exceeds 50°C, the koji fungus may not proliferate.

The time of the aforementioned culture is not particularly limited and can be appropriately selected according to the purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the aforementioned culture time is less than 80 hours, sometimes sufficient spores cannot be attached to the aforementioned thyme, and if it exceeds 210 hours, sometimes the germination rate of the spores will decrease.

The thyme fermented liquid may contain the bacterial bodies of the koji mold or may be a liquid from which the bacterial bodies of the koji mold have been removed, but preferably the liquid from which the bacterial bodies of the koji mold have been removed.

The state of the thyme fermented liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it can be the thyme fermented liquid itself, a purified product of the thyme fermented liquid, a concentrated product of the thyme fermented liquid, a diluted product of the thyme fermented liquid, etc. In addition, the thyme fermented liquid can be a dried product of the thyme fermented liquid mixed or dissolved in a solvent such as water or a hydrophilic solvent.

As for the purified product of the aforementioned thyme fermentation liquid, there is no particular limitation and it can be appropriately selected according to the purpose. For example, the solid components (for example, the plant body of the aforementioned thyme, the body of the koji fungus, the residue, etc.) in the aforementioned thyme fermentation liquid are removed. As for the aforementioned removal means, there is no particular limitation and it can be appropriately selected according to the purpose. For example, filtration, etc. can be cited. As for the aforementioned filtering method, there is no particular limitation and it can be appropriately selected from the known methods according to the purpose.

There are no particular restrictions on the aforementioned dilution of the fermented thyme liquid and the aforementioned concentrate of the fermented thyme liquid, and they can be appropriately selected according to the purpose. For example, the fermented thyme liquid is adjusted to the desired concentration. There are no particular restrictions on the aforementioned dilution means, and they can be appropriately selected from the known methods according to the purpose. There are no particular restrictions on the aforementioned concentration means, and they can be appropriately selected according to the purpose. For example, reduced pressure concentration can be listed.

As for the dried product of the aforementioned thyme fermentation liquid, there is no particular limitation and it can be appropriately selected according to the purpose, for example, the dried product of thyme fermentation liquid, etc. As for the aforementioned drying means, there is no particular limitation and it can be appropriately selected according to the purpose, for example, freeze drying, etc.

The aforementioned thyme fermented liquid is not particularly limited as long as it is a fermented liquid of thyme obtained by yeast, and can be appropriately selected according to the purpose. However, in terms of good skin affinity, a thyme fermented liquid with a contact angle of 87° or less is preferred, and a thyme fermented liquid with a contact angle of 81° or less is more preferred.

＜＜Cypress Fermented Liquid＞＞ The aforementioned Cypress Fermented Liquid is a fermented liquid obtained from Cypress by using koji fungus.

- Melissa officinalis Linne used as the fermentation raw material is a perennial herb belonging to the genus Melissa of the mint family ( Labiatae ). It is a kind of spice plant and has been used as a food and medicinal raw material since ancient times. Other names include lemon balm and mountain mint. It is native to southern Europe and has grown naturally or been cultivated in Japan. It can be easily obtained from these regions. There is no particular limitation on the method of obtaining the above-mentioned Melissa officinalis Linne. It can be appropriately selected according to the purpose. It can be collected from nature or commercially available products can be used.

The part of the cypress grass used as the fermentation raw material is not particularly limited and can be appropriately selected according to the purpose. For example, the aboveground parts of flowers, buds, fruits, peels, seeds, seed coats, stems, leaves, branches, branches and leaves; underground parts of roots, rhizomes, etc. can be used alone or in combination of two or more. Among these, the aboveground parts of the cypress grass are preferably used.

The size of the aforementioned cypress grass used as the aforementioned fermentation raw material is not particularly limited as long as it is a size that can be cultivated in the aforementioned koji mold, and can be appropriately selected according to the purpose, for example, the original size taken, the desired size after cutting, the size after micronization (powdering), etc.

The state of the aforementioned cypress grass used as the aforementioned fermentation raw material is not particularly limited as long as it is a state that can be cultured by the aforementioned koji, and can be appropriately selected according to the purpose, for example, the original state of being taken, the dried state, the crushed state, the juiced state, the state of being an extract, etc. Among these, the original state of being taken, the crushed state, the juiced state, the state of being an extract are preferred, and the original state of being taken and the crushed state are more preferred, as the aforementioned koji can easily act.

The method for drying the cypress grass is not particularly limited and can be appropriately selected according to the purpose. For example, there can be mentioned a method of drying in sunlight, a method of drying using a commonly used dryer, and the like.

The method for making the above-mentioned cypress grass into the above-mentioned pulverized state is not particularly limited and can be appropriately selected according to the purpose. For example, there can be pulverized using a mixer, a sugar mill, a powder mill, a jet mill, an impact mill, etc.

The method of making the aforementioned cypress grass into the aforementioned juice state is not particularly limited and can be appropriately selected according to the purpose, and for example, pressing can be cited.

The method for preparing the aforementioned Herba Lysimachiae into the aforementioned extract is not particularly limited, and a method generally used for plant extraction can be appropriately selected according to the purpose.

The aforementioned cypress grass used as the aforementioned fermentation raw material is preferably sterilized before the aforementioned koji mold is inoculated. There is no particular limitation on the means for sterilizing the aforementioned cypress grass, and it can be appropriately selected from the known methods.

-Koji- As for the aforementioned koji used to ferment the aforementioned cypress grass, there is no particular limitation and it can be appropriately selected according to the purpose. For example, those described in the aforementioned <<Artemisia plant fermentation liquid>> can be cited. These can be used alone or in combination of two or more. Among these, as for the aforementioned koji, the point that it is excellent in at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect is rice koji ( Aspergillus oryzae ). As for the method for obtaining the aforementioned koji, there is no particular limitation and it can be appropriately selected according to the purpose. It can be collected from nature or commercially available products can be used. In addition, as the aforementioned koji, a seed koji made from rice or the like can be used, a koji seed koji described later can be used, or a koji cultured in a culture medium (an agar culture medium, a liquid culture medium, etc.) can be used. Among these, the aforementioned koji seed koji is preferably used because it is excellent in at least one of the anti-aging effect, the anti-oxidation effect, the anti-inflammatory effect, and the whitening effect.

The inoculation amount of the koji mold to the aforementioned cypress grass used as the aforementioned fermentation raw material is not particularly limited as long as the aforementioned cypress grass can be fermented. It can be appropriately selected according to the purpose. However, when the aforementioned fermentation raw material is in a liquid state, it is preferably 1×10 ³ cells/mL to 1×10 ⁸ cells/mL, and when the aforementioned fermentation raw material is in a solid state, it is preferably 1×10 ³ cells/g to 1×10 ⁸ cells/g.

When the koji mold is inoculated into the cypress grass, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 500 to 5,000 parts by mass, more preferably 1,000 to 4,000 parts by mass, and particularly preferably 1,500 to 3,000 parts by mass are added relative to 100 parts by mass of the cypress grass.

The temperature of the fermentation (cultivation) is not particularly limited as long as it is within the temperature range at which the yeast can ferment, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the fermentation temperature is lower than 20°C, the cypress grass cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect becomes insufficient.

The fermentation (cultivation) time is not particularly limited and can be appropriately selected according to the purpose, but is preferably 10 to 40 hours, more preferably 20 to 30 hours. If the fermentation time is less than 10 hours, the cypress grass cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, anti-inflammatory effect, and whitening effect becomes insufficient.

There is no particular limitation on the method for stopping the aforementioned fermentation (cultivation), and it can be appropriately selected according to the purpose. For example, a heating method can be cited. As for the heating temperature used to stop the aforementioned fermentation, if it is a temperature at which the aforementioned koji cannot grow, it is not particularly limited and can be appropriately selected according to the purpose, but it is preferably above 50°C, more preferably above 70°C, and particularly preferably 100°C to 130°C. When the aforementioned heating temperature is lower than 50°C, the aforementioned fermentation may not be stopped. When it exceeds 130°C, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect becomes insufficient. As for the heating time for stopping the aforementioned fermentation, there is no particular limitation as long as it can make the aforementioned yeast unable to grow, and it can be appropriately selected according to the purpose, but it is preferably more than 5 minutes, and more preferably 10 minutes to 20 minutes. If the aforementioned heating time is less than 5 minutes, sometimes the aforementioned fermentation cannot be stopped, and when it exceeds 20 minutes, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect becomes insufficient.

Furthermore, the fermented lycopodiella ciliata liquid is preferably cooled after the fermentation is stopped. The cooling method is not particularly limited and can be appropriately selected according to the purpose. For example, the method of leaving the lycopodiella ciliata liquid at room temperature or in a refrigerator can be cited.

The number of times the fermentation of the cypress grass using the yeast is performed is not particularly limited and can be appropriately selected according to the purpose, and may be once or multiple times.

When the aforementioned fermentation is performed multiple times, the aforementioned yeast may be inoculated only for the first time, may be inoculated only for a few times, or may be inoculated for all times, but it is preferred to perform only the first time. When the aforementioned fermentation is performed multiple times, the fermentation temperature and fermentation time may be different or the same.

--Cypress seed koji-- The aforementioned yew seed koji is obtained by using the aforementioned yew as a seed koji raw material, inoculating koji fungi into the seed koji raw material, and allowing a sufficient amount of spores to attach to the yew. By using it in the aforementioned fermentation, it is advantageous that a yew fermented liquid with excellent skin affinity can be obtained more efficiently and easily.

The aforementioned cypress grass used as the aforementioned seed koji raw material can be the same as that described in the aforementioned "cypress grass", and the used part, size, state, etc. of the aforementioned cypress grass are also the same.

As for the aforementioned koji mold used in the production of the aforementioned cypress grass seed koji, the same ones as those described in the aforementioned "koji mold" can be used.

The amount of the koji fungus inoculated with the cypress grass used as the koji seed material is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably inoculated with 5 to 100 parts by mass of the koji fungus (1×10 ³ cells/mL to 1×10 ⁸ cells/mL) suspended in sterilized water, more preferably 10 to 50 parts by mass, and particularly preferably 20 to 30 parts by mass. When the amount of the koji fungus inoculated with 100 parts by mass of the cypress grass is less than 5 parts by mass, a sufficient amount of spores may not be attached to the cypress grass, and when it exceeds 100 parts by mass, abnormal reproduction may occur due to excessive water.

When the koji mold is inoculated into the cypress grass used as the seed koji raw material, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 10 to 250 parts by mass, more preferably 20 to 200 parts by mass, and particularly preferably 30 to 150 parts by mass are added relative to 100 parts by mass of the cypress grass. When the amount of water added relative to 100 parts by mass of the cypress grass is less than 10 parts by mass, a sufficient amount of spores may not be attached to the cypress grass.

The culture temperature is not particularly limited as long as it is within the range at which the koji mold can grow, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the culture temperature is lower than 20°C, a sufficient amount of spores may not attach to the cypress grass. When the culture temperature exceeds 50°C, the koji mold may not proliferate.

The time of the aforementioned culture is not particularly limited and can be appropriately selected according to the purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the aforementioned culture time is less than 80 hours, sometimes it is not possible to allow a sufficient amount of spores to attach to the aforementioned Cypress Bee Herb, and if it exceeds 210 hours, sometimes the germination rate of the spores will be reduced.

The fermented lycopodiella ciliata liquid may contain the bacterial bodies of the koji mold or may be a liquid in which the bacterial bodies of the koji mold are removed, but preferably the liquid in which the bacterial bodies of the koji mold are removed.

The state of the fermented lycopodiella ciliata liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it can be the fermented lycopodiella ciliata liquid itself, a purified product of the fermented lycopodiella ciliata liquid, a concentrated product of the fermented lycopodiella ciliata liquid, a diluted product of the fermented lycopodiella ciliata liquid, etc. In addition, the fermented lycopodiella ciliata liquid can be a dried product of the fermented lycopodiella ciliata liquid mixed or dissolved in a solvent such as water or a hydrophilic solvent.

As for the purified product of the fermented lycopodiella ciliata, there is no particular limitation and it can be appropriately selected according to the purpose. For example, the solid components (for example, the plant body of the lycopodiella ciliata, the body of the koji mold, the residue, etc.) in the fermented lycopodiella ciliata are removed. As for the means of removal, there is no particular limitation and it can be appropriately selected according to the purpose. For example, filtration, etc. can be cited. As for the method of filtration, there is no particular limitation and it can be appropriately selected from the known methods according to the purpose.

There are no particular restrictions on the dilution of the fermented cypress grass liquid and the concentrate of the fermented cypress grass liquid, and they can be appropriately selected according to the purpose. For example, the fermented cypress grass liquid is prepared to a desired concentration. There are no particular restrictions on the means of dilution, and they can be appropriately selected from known methods according to the purpose. There are no particular restrictions on the means of concentration, and they can be appropriately selected according to the purpose. For example, reduced pressure concentration can be listed.

As for the dried product of the aforementioned fermented cypress grass, there is no particular limitation and it can be appropriately selected according to the purpose, for example, the dried product of the fermented cypress grass, etc. As for the aforementioned drying method, there is no particular limitation and it can be appropriately selected according to the purpose, for example, freeze drying, etc.

The aforementioned fermented liquid of cypress grass is not particularly limited as long as it is a fermented liquid of cypress grass obtained by yeast, and can be appropriately selected according to the purpose. However, in terms of good skin affinity, a fermented liquid of cypress grass with a contact angle of 85° or less is preferred, and a fermented liquid of cypress grass with a contact angle of 79° or less is more preferred.

＜＜Cornflower fermented liquid＞＞ The aforementioned cornflower fermented liquid is a fermented liquid obtained from cornflower using yeast.

-Cornflower- The cornflower ( Centaurea cyanus Linne) used as the fermentation raw material is an annual herb belonging to the genus Centaurea of the family Compositae , and has been used as an edible and medicinal raw material since ancient times. Other names include cornflower, Centaurea, centaurea, etc. Although the origin is Europe, it has been grown naturally or cultivated in Japan and can be easily obtained from such regions. There is no particular limitation on the method of obtaining the cornflower, and it can be appropriately selected according to the purpose. It can be collected from nature or commercially available products can be used.

The parts of the cornflower used as the fermentation raw material are not particularly limited and can be appropriately selected according to the purpose. For example, the aboveground parts of flowers, buds, fruits, peels, seeds, seed coats, stems, leaves, branches, and branches and leaves; the underground parts of roots, rhizomes, etc. can be used alone or in combination of two or more. Among these, the aboveground parts of the cornflower are preferred.

The size of the cornflower used as the fermentation raw material is not particularly limited as long as it is a size that can be cultured with the yeast, and can be appropriately selected according to the purpose, for example, the original size taken, the desired size after cutting, the size after micronization (powdering), etc.

The state of the cornflower used as the fermentation raw material is not particularly limited as long as it is a state that allows the cultivation of the koji, and can be appropriately selected according to the purpose, for example, the original state of the cornflower, the dried state, the crushed state, the juiced state, the state of the extract, etc. Among these, the original state of the cornflower, the crushed state, the juiced state, the state of the extract are preferred, and the original state of the cornflower and the crushed state are more preferred, as the koji can easily act on the cornflower.

The method for drying the cornflower is not particularly limited and can be appropriately selected according to the purpose. For example, there can be mentioned a method of drying in sunlight, a method of drying using a commonly used dryer, and the like.

The method for making the cornflower into the above-mentioned pulverized state is not particularly limited and can be appropriately selected according to the purpose. For example, there can be cited a method of pulverizing using a mixer, a sugar mill, a powder mill, a jet mill, an impact mill, etc.

The method of making the cornflower into the aforementioned juice state is not particularly limited and can be appropriately selected according to the purpose, and for example, pressing can be cited.

The method for preparing the cornflower into the extract is not particularly limited, and a method generally used for plant extraction can be appropriately selected according to the purpose.

The cornflower used as the fermentation raw material is preferably sterilized before the inoculation of the yeast. There is no particular limitation on the method for sterilizing the cornflower, and it can be appropriately selected from known methods.

-Aspergillus- As for the aforementioned koji used to ferment the aforementioned cornflower, there is no particular limitation and it can be appropriately selected according to the purpose. For example, those described in the aforementioned <<Artemisia plant fermentation liquid>> can be cited. These can be used alone or in combination of two or more. Among these, as for the aforementioned koji, the point that it is excellent in at least one of the anti-aging effect, antioxidant effect, and whitening effect is rice Aspergillus oryzae . As for the method for obtaining the aforementioned koji, there is no particular limitation and it can be appropriately selected according to the purpose. It can be collected from nature or commercially available products can be used. In addition, as the aforementioned koji, a seed koji made from rice or the like can be used, or a cornflower seed koji described later can be used, or a koji cultured in a culture medium (an agar culture medium, a liquid culture medium, etc.). Among these, the aforementioned cornflower seed koji is preferably used because it is excellent in at least one of the anti-aging effect, the anti-oxidation effect, and the whitening effect.

The inoculation amount of the aforementioned koji fungus to the aforementioned cornflower used as the aforementioned fermentation raw material is not particularly limited as long as the aforementioned cornflower can be fermented. It can be appropriately selected according to the purpose. However, when the aforementioned fermentation raw material is in a liquid state, it is preferably 1×10 ³ cells/mL to 1×10 ⁸ cells/mL, and when the aforementioned fermentation raw material is in a solid state, it is preferably 1×10 ³ cells/g to 1×10 ⁸ cells/g.

When the koji is inoculated into the cornflower, water is preferably added. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 500 to 5,000 parts by mass, more preferably 1,000 to 4,000 parts by mass, and particularly preferably 1,500 to 3,000 parts by mass are added relative to 100 parts by mass of the cornflower.

The temperature of the fermentation (cultivation) is not particularly limited as long as it is within the temperature range at which the yeast can ferment, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the fermentation temperature is lower than 20°C, the cornflower cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, and whitening effect becomes insufficient.

The fermentation (cultivation) time is not particularly limited and can be appropriately selected according to the purpose, but is preferably 10 to 40 hours, more preferably 20 to 30 hours. If the fermentation time is less than 10 hours, the cornflower cannot be fully fermented, and at least one of the anti-aging effect, anti-oxidation effect, and whitening effect becomes insufficient.

There is no particular limitation on the method for stopping the aforementioned fermentation (cultivation), and it can be appropriately selected according to the purpose. For example, a heating method can be cited. As for the heating temperature used to stop the aforementioned fermentation, if it is a temperature at which the aforementioned koji cannot grow, it is not particularly limited and can be appropriately selected according to the purpose, but it is preferably above 50°C, more preferably above 70°C, and particularly preferably 100°C to 130°C. When the aforementioned heating temperature is lower than 50°C, the aforementioned fermentation may not be stopped. When it exceeds 130°C, at least one of the anti-aging effect, antioxidant effect, and whitening effect becomes insufficient. As for the heating time used to stop the aforementioned fermentation, there is no particular limitation as long as it can make the aforementioned yeast unable to grow, and it can be appropriately selected according to the purpose, but it is preferably more than 5 minutes, and more preferably 10 minutes to 20 minutes. If the aforementioned heating time is less than 5 minutes, sometimes the aforementioned fermentation cannot be stopped, and when it exceeds 20 minutes, at least one of the anti-aging effect, antioxidant effect, and whitening effect becomes insufficient.

Furthermore, the cornflower fermented liquid is preferably cooled after the fermentation is stopped. The cooling method is not particularly limited and can be appropriately selected according to the purpose. For example, the method of leaving the fermented liquid at room temperature or in a refrigerator can be cited.

The number of times the cornflower is fermented using the yeast is not particularly limited and can be appropriately selected according to the purpose, and may be once or multiple times.

When the aforementioned fermentation is performed multiple times, the aforementioned yeast may be inoculated only for the first time, may be inoculated only for a few times, or may be inoculated for all times, but it is preferred to perform only the first time. When the aforementioned fermentation is performed multiple times, the fermentation temperature and fermentation time may be different or the same.

--Cornflower seed yeast-- The aforementioned cornflower seed yeast is prepared by using the aforementioned cornflower as a seed yeast raw material, inoculating the koji fungus into the seed yeast raw material, and allowing a sufficient amount of spores to attach to the cornflower. By using it in the aforementioned fermentation, it is advantageous that a cornflower fermented liquid with excellent skin affinity can be obtained more efficiently and easily.

The aforementioned cornflower used as the aforementioned seed yeast raw material may be the same as that described in the aforementioned "cornflower", and the used part, size, state, etc. of the aforementioned cornflower are also the same.

As for the aforementioned koji mold used in the production of the aforementioned cornflower seed koji, the same ones as those described in the aforementioned "koji mold" can be used.

The inoculation amount of the koji fungus on the cornflower used as the seed koji raw material is not particularly limited and can be appropriately selected according to the purpose, but preferably 5 to 100 parts by mass of the koji fungus (1×10 ³ cells/mL to 1×10 ⁸ cells/mL) suspended in sterile water is inoculated with 100 parts by mass of the cornflower, more preferably 10 to 50 parts by mass, and particularly preferably 20 to 30 parts by mass. When the inoculation amount of the koji fungus is less than 5 parts by mass relative to 100 parts by mass of the cornflower, a sufficient amount of spores may not be attached to the cornflower, and when it exceeds 100 parts by mass, abnormal reproduction may occur due to excessive water.

When the koji fungus is inoculated, water is preferably added when the cornflower used as the seed koji raw material is used. The amount of water added is not particularly limited and can be appropriately selected according to the purpose, but preferably 10 to 250 parts by mass are added relative to 100 parts by mass of the cornflower, more preferably 20 to 200 parts by mass are added, and particularly preferably 30 to 150 parts by mass are added. When the amount of water added relative to 100 parts by mass of the cornflower is less than 10 parts by mass, a sufficient amount of spores may not be attached to the cornflower.

The culture temperature is not particularly limited as long as it is within the range of the temperature at which the aspergillus can grow, and can be appropriately selected according to the purpose, but is preferably 20°C to 40°C, and more preferably 25°C to 35°C. When the culture temperature is lower than 20°C, a sufficient amount of spores may not be attached to the cornflower. When the culture temperature exceeds 50°C, the aspergillus may not proliferate.

The time of the aforementioned culture is not particularly limited and can be appropriately selected according to the purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the aforementioned culture time is less than 80 hours, sometimes it is not possible to allow a sufficient amount of spores to attach to the aforementioned cornflower, and if it exceeds 210 hours, sometimes the germination rate of the spores will decrease.

The cornflower fermented liquid may contain the bacterial bodies of the koji mold or may be a liquid from which the bacterial bodies of the koji mold have been removed, but preferably a liquid from which the bacterial bodies of the koji mold have been removed.

The state of the cornflower fermentation liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it can be the cornflower fermentation liquid itself, a purified product of the cornflower fermentation liquid, a concentrated product of the cornflower fermentation liquid, a diluted product of the cornflower fermentation liquid, etc. In addition, the cornflower fermentation liquid can be a dried product of the cornflower fermentation liquid mixed or dissolved in a solvent such as water or a hydrophilic solvent.

As for the purified product of the aforementioned cornflower fermentation liquid, there is no particular limitation and it can be appropriately selected according to the purpose. For example, the solid components (for example, the aforementioned cornflower plant body, yeast body, slag, etc.) in the aforementioned cornflower fermentation liquid can be removed. As for the aforementioned removal means, there is no particular limitation and it can be appropriately selected according to the purpose. For example, filtration can be cited. As for the aforementioned filtering method, there is no particular limitation and it can be appropriately selected from the known methods according to the purpose.

There are no particular restrictions on the aforementioned dilution of cornflower fermented liquid and the aforementioned concentrate of cornflower fermented liquid, and they can be appropriately selected according to the purpose. For example, the cornflower fermented liquid is prepared to a desired concentration. There are no particular restrictions on the aforementioned dilution means, and they can be appropriately selected from known methods according to the purpose. There are no particular restrictions on the aforementioned concentration means, and they can be appropriately selected according to the purpose. For example, reduced pressure concentration can be cited.

There is no particular limitation on the dried product of the cornflower fermented liquid, and it can be appropriately selected according to the purpose. For example, the dried product of the cornflower fermented liquid can be cited. There is no particular limitation on the drying method, and it can be appropriately selected according to the purpose. For example, freeze drying can be cited.

The aforementioned cornflower fermented liquid can be any fermented liquid obtained from cornflower using yeast, and is not particularly limited. It can be appropriately selected according to the purpose. However, in terms of good skin affinity, a cornflower fermented liquid with a contact angle of 85° or less is preferred, and a cornflower fermented liquid with a contact angle of 79° or less is more preferred.

<<Other ingredients>> As for the aforementioned other ingredients in the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent, there is no particular limitation and they can be appropriately selected according to the purpose. For example, they can include shaping agents, moisturizers, preservatives, reinforcing agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, colorants, fragrances, whitening agents, moisturizers, oily ingredients, ultraviolet absorbers, surfactants, thickeners, alcohols, powder ingredients, colorants, aqueous ingredients, water, skin nutrients, etc. These can be used alone or in combination of two or more. As for the content of the aforementioned other ingredients, there is no particular limitation and they can be appropriately selected according to the purpose.

-Application- The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention have at least one of excellent anti-aging, antioxidant, anti-inflammatory, and whitening effects, and can therefore be suitably used as, for example, medicines, quasi-drugs, cosmetics, beverages, etc., and their blending amount, usage, and dosage form can be appropriately selected according to their purpose of use.

The amount of the mixture can be appropriately adjusted according to the physiological activity of the fermentation liquid, etc. In addition, the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent can be the fermentation liquid itself.

The aforementioned usage is not particularly limited and can be appropriately selected according to the purpose, for example, oral use, parenteral use, external use, etc. Among these, external use is preferred.

The dosage form is not particularly limited and can be appropriately selected according to the purpose. For example, oral administration forms such as tablets, powders, capsules, granules, extracts, and syrups; parenteral administration forms such as injections, drips, and suppositories; external preparations such as lotions, emulsions, creams, ointments, beauty essences, lotions, facial masks, gels, lip balms, lipsticks, foundations, bath preparations, soaps, shower gels, astringents, scalp toners, hair creams, hair lotions, hair waxes, shampoos, and conditioners.

Furthermore, the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention can also be used as a reagent for research related to the mechanism of action of the anti-aging effect, antioxidant effect, anti-inflammatory effect, or whitening effect.

The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention are suitable for use in humans, but as long as their respective effects are produced, they can also be used in animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.).

(Cosmetics) The cosmetics of the present invention contain at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention, and further contain other ingredients as needed.

<Anti-aging agent, antioxidant, anti-inflammatory agent, whitening agent> The content of at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent in the aforementioned cosmetic is not particularly limited and can be appropriately selected according to the purpose, but is preferably 5 volume % or more, and more preferably 20 volume % or more relative to the total volume of the aforementioned cosmetic. When the content of at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent is less than 5 volume %, at least any one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect becomes insufficient. Furthermore, the content of at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent is better, and there is no particular restriction on the upper limit, which can be appropriately selected according to the purpose. Furthermore, the aforementioned cosmetic can be at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent.

<Other ingredients> The aforementioned cosmetics may be further added with various main agents, auxiliary agents, and other ingredients commonly used in the manufacture of cosmetics as needed, within the scope of not impairing the purpose and effect of the present invention. As for the aforementioned other ingredients, there is no particular limitation and they can be appropriately selected according to the purpose. For example, astringents, bactericides, antibacterial agents, ultraviolet absorbers, cell activators, oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc. These may be used alone or in combination of two or more. When these ingredients are used together with at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent, it is possible that they will work synergistically and bring about the excellent effects that are usually expected. As for the content of the aforementioned other ingredients in the aforementioned cosmetics, there is no particular limitation as long as it does not damage the effects of the present invention, and it can be appropriately selected according to the purpose.

<Application> There is no particular limitation on the application of the aforementioned cosmetics, and they can be appropriately selected from general cosmetics, such as toners, lotions, creams, ointments, beauty serums, lotions, facial masks, gels, lip balms, lipsticks, foundations, bath agents, soaps, shower gels, etc. for skin cosmetics; astringents, scalp toners, hair creams, hair lotions, hair waxes, shampoos, conditioners, etc. for scalp and hair cosmetics, etc.

The aforementioned cosmetic may be at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent, mixed with any cosmetic in a manner that does not hinder the activity thereof, or may be a cosmetic having at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent as a main component. Furthermore, the aforementioned cosmetic may be at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent itself.

The cosmetic of the present invention is suitable for use on humans, but can also be used on animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective effects are produced.

The cosmetic of the present invention contains at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent, and is therefore useful in exerting at least one of excellent anti-aging effects, antioxidant effects, anti-inflammatory effects, and whitening effects when used on the skin.

[Example] The present invention is specifically described with the following manufacturing examples and test examples, but the present invention is not limited to these test examples at all.

<Production Example 1: Preparation of Artemisia oryzae fermentation liquid 1> -First koji preparation steps- Take the koji ( Aspergillus oryzae , strain name: AOK1714, manufactured by Akita Konno Shoten Co., Ltd.) from Platinum Ear and suspend it in 50 mL of sterile water to prepare a koji solution. The bacterial count of the koji solution was calculated using a Thoma hemacytometer (manufactured by EKDS), and the result was 1.0×10 ⁵ cells/mL. Then, 10 g of Artemisia oryzae (manufactured by ALBION Co., Ltd.) cut into 0.5 cm~5 cm pieces was placed in a conical flask, sterilized by pressure, and 2 mL of the above-mentioned koji solution was inoculated therein, and cultured statically at 30°C for 168 hours. After the incubation, the mixture was dried at 45°C for 24 hours to obtain the Artemisia selengensis seed yeast.

- Fermentation step - Use a grinder (sugar mill) to grind Artemisia oleracea (manufactured by ALBION Co., Ltd.) and pass it through a 2 mm mesh to obtain Artemisia oleracea grind. Add 1,000 mL of water to 50 g of this Artemisia oleracea grind, mix, and inoculate 20 mL of the Artemisia oleracea seed koji obtained in the above seed koji preparation step (bacterial count: about 1.0×10 ⁶ /mL). Then, incubate at 25°C for 22 hours. Filter the obtained fermentation liquid using diatomaceous earth to obtain "Artemisia oleracea fermentation liquid 1".

<Production Example 2: Preparation of Artemisia oryzae Fermentation Liquid 2>"Artemisia oryzae Fermentation Liquid 2" was obtained in the same manner as in Production Example 1, except that the Artemisia oryzae seed koji was replaced with a seed koji made from rice ( Aspergillus oryzae , Shirakami white koji, Akita Konno Shoten Co., Ltd.) (hereinafter sometimes referred to as "rice seed koji").

<Comparative Production Example 1: Preparation of Artemisia annua extract> Artemisia annua (manufactured by ALBION Co., Ltd.) was crushed using a crusher (sugar mill) and passed through a 2 mm mesh to obtain an Artemisia annua crushed product. 1,000 mL of water was added to 50 g of the Artemisia annua crushed product, mixed, and stirred at 25°C for 22 hours. Then, the obtained mixture was filtered using diatomaceous earth to obtain an "Artemisia annua extract".

(Test Example A-1: Measurement of contact angle) The Artemisia annua fermented liquid 1 obtained in Production Example 1, the Artemisia annua fermented liquid 2 obtained in Production Example 2, and the Artemisia annua extract obtained in Comparative Production Example 1 were used as test samples, and the contact angle was measured by the following method. Specifically, a dynamic contact angle and surface tension measuring device (FTA1000 Falcon, manufactured by First Ten Angstroms) was used, and 3 μL of each test sample was dripped onto the sample stand (made of aluminum) of the aforementioned device, and the measurement was performed by the drop method at a temperature of 22°C and a relative humidity of 20%. The contact angle θ (°) at 1,000 ms was obtained by the θ/2 method. The contact angle was measured 3 times, and the average value was obtained. The results are shown in Table 1 below. In addition, one example of a droplet during the measurement of the contact angle of each test sample is shown in Figures 1A to 1C.

[Table 1] Test sample Fermentation Aspergillus Contact angle θ(°) Manufacturing Example 1 Artemisia annua fermented liquid 1 have Artemisia arborescens yeast 77.44 Manufacturing Example 2 Artemisia annua fermented liquid 2 have Rice Kernel 80.02 Comparative Manufacturing Example 1 Artemisia annua extract without - 81.57

Compared with the Artemisia annua extract obtained in Preparation Example 1, the Artemisia annua fermented liquid 1 obtained in Preparation Example 1 and the Artemisia annua fermented liquid 2 obtained in Preparation Example 2 both have a small contact angle of 81° or less, and are excellent in skin affinity. Furthermore, the Artemisia annua fermented liquid 1 obtained in Preparation Example 1 has a contact angle of 78° or less, and is more excellent in skin affinity.

(Test Example 1-1: Test for inhibitory effect on matrix metalloproteinase-1 (MMP-1) activity) The Artemisia sutchuenensis fermentation liquid 1 obtained in Preparation Example 1, the Artemisia sutchuenensis fermentation liquid 2 obtained in Preparation Example 2, and the Artemisia sutchuenensis extract obtained in Comparative Preparation Example 1 were used as test samples, and the inhibitory effect on matrix metalloproteinase-1 (MMP-1) activity was tested by the following test method in which the Wunsch and Heidrich method was partially modified.

In a test tube with a cap, each test sample was dissolved in a Tris-HCl buffer (pH 7.1) containing 20 mmol/L calcium chloride and 0.1 mol/L. Next, 50 μL of the above test sample solution, 50 μL of MMP-1 (collagenase (COLLAGENASE) type IV, from Clostridium histolyticum, manufactured by Sigma) solution, and 400 μL of Pz-peptide (Pz-Pro-Leu-Gly-Pro-D-Arg-OH, manufactured by BACHEM Feinchemikalien AG) solution were mixed, and reacted at 3°C for 30 minutes, and then 1 mL of 25 mmol/L citric acid solution was added to stop the reaction. In addition, the final concentration of the above test sample at this time was the concentration shown in Table 2 below. Then, 5 mL of ethyl acetate was added and the mixture was shaken vigorously. The mixture was centrifuged at 1,600 × g for 10 minutes and the absorbance of the ethyl acetate layer was measured at a wavelength of 320 nm.

In addition, as a blank control, the same operation and absorbance measurement as above were performed except that the MMP-1 solution (enzyme solution) was changed to 0.1 mol/L Tris-HCl buffer (pH 7.1).

As a control group, the same operation and absorbance measurement were performed as above except that the solution of the test sample was changed to 0.1 mol/L Tris-HCl buffer (pH 7.1) containing 20 mmol/L calcium chloride without the test sample.

The MMP-1 activity inhibition rate was calculated based on the following formula 1 from the measured value of the absorbance obtained. The results are shown in the following Table 2. ＜Formula 1＞ MMP-1 activity inhibition rate (%) = {1-(C-D)/(A-B)} × 100 In the above formula 1, A~D each represent the following. A: Absorbance at a wavelength of 320nm when no test sample is added and enzyme is added B: Absorbance at a wavelength of 320nm when no test sample is added and enzyme is not added C: Absorbance at a wavelength of 320nm when the test sample is added and enzyme is added D: Absorbance at a wavelength of 320nm when the test sample is added and enzyme is not added

[Table 2] Test sample Concentration of the test sample (μg/mL) MMP-1 activity inhibition rate (%) Manufacturing Example 1 Artemisia annua fermented liquid 1 400 7.0 Manufacturing Example 2 Artemisia annua fermented liquid 2 400 4.2 Comparative Manufacturing Example 1 Artemisia annua extract 400 1.5

(Test Example 1-2: Test for promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) Using Artemisia selengensis fermentation liquid 1 obtained in Preparation Example 1, Artemisia selengensis fermentation liquid 2 obtained in Preparation Example 2, and Artemisia selengensis extract obtained in Comparative Preparation Example 1 as test samples, the hyaluronic acid synthase 3 (HAS3) mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration as shown in Table 3 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (Normal Human Epidermal Keratinocytes; NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells reached confluence, and then the cells were collected by trypsin treatment. Use normal human epidermal keratinocyte proliferation medium (HuMedia-KG2) to adjust it to 1.5×10 ⁵ cells/mL. Then, inoculate 2mL of the above-mentioned NHEK (1.5×10 ⁵ cells/mL) into a 35mm dish and culture it overnight at 37℃ and 5% CO _2. After the culture is completed, the medium is replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and cultured for another 24 hours. After the culture is completed, the medium is replaced with 2mL of the above-mentioned medium supplemented with the test sample and cultured for 24 hours at 37℃ and 5% CO ₂ . After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.). The amount of each RNA was measured by a spectrophotometer, and the total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above total RNAs were used as templates to measure the expression levels of hyaluronic acid synthase 3 (HAS3) mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal standard. The detection of mRNA was performed by a two-step RT-PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression level of HAS3 mRNA without and with the addition of the test sample was corrected by the expression level of GAPDH mRNA. Based on this correction value, the HAS3 mRNA expression promotion rate was calculated based on the following formula 2. The results are shown in the following Table 3. <Formula 2> HAS3 mRNA expression promotion rate (%) = A/B × 100 In the above formula 2, A and B each represent the following. A: Calibration value when the test sample is added B: Calibration value when the test sample is not added

[Table 3] Test sample Concentration of the test sample (μg/mL) HAS3 mRNA expression promotion rate (%) Manufacturing Example 1 Artemisia annua fermented liquid 1 5 172.0 Manufacturing Example 2 Artemisia annua fermented liquid 2 5 127.3 Comparative Manufacturing Example 1 Artemisia annua extract 5 125.3

(Test Example 1-3: DPPH free radical elimination test) Using Artemisia selengensis fermentation liquid 1 obtained in Preparation Example 1, Artemisia selengensis fermentation liquid 2 obtained in Preparation Example 2, and Artemisia selengensis extract obtained in Comparative Preparation Example 1 as test samples, the DPPH free radical elimination effect was tested by the following test method.

Dissolve each test sample in an ethanol solution (manufactured by Fuji Film Wako Pure Chemicals Co., Ltd.) and prepare a test sample solution. Add 3 mL of the test sample solution to 3 mL of a 150 μmol/L DPPH (diphenyl-p-picrylhydrazyl) ethanol solution, immediately seal the container and shake it, and after standing for 30 minutes, measure the absorbance at a wavelength of 520 nm. In addition, the final concentration of the test sample at this time is the concentration shown in Table 4 below.

In addition, as a blank group, the same operation and absorbance measurement as above were performed except that the DPPH ethanol solution was changed to an ethanol solution not containing DPPH.

As a control group, the same operation and absorbance measurement as above were performed except that the test sample solution was changed to an ethanol solution containing no test sample (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.).

The DPPH radical elimination rate was calculated based on the following formula 3 from the measured value of the absorbance obtained. The results are shown in the following Table 4. ＜Formula 3＞ DPPH radical elimination rate (%)＝{A-(B-C)}/A×100 In the above formula 3, A~C each represent the following. A: Absorbance at a wavelength of 520nm without adding the test sample and adding DPPH B: Absorbance at a wavelength of 520nm with adding the test sample and adding DPPH C: Absorbance at a wavelength of 520nm without adding the test sample and adding DPPH

[Table 4] Test sample Concentration of the test sample (μg/mL) DPPH free radical elimination rate (%) Manufacturing Example 1 Artemisia annua fermented liquid 1 12.5 35.0 Manufacturing Example 2 Artemisia annua fermented liquid 2 12.5 32.1 Comparative Manufacturing Example 1 Artemisia annua extract 12.5 29.7

(Test Example 1-4: Hyaluronidase Activity Inhibition Test) Using Artemisia selengensis fermentation liquid 1 obtained in Production Example 1, Artemisia selengensis fermentation liquid 2 obtained in Production Example 2, and Artemisia selengensis extract obtained in Comparative Production Example 1 as test samples, the hyaluronidase activity inhibition effect was tested by the following test method.

Each test sample was dissolved in 0.1 mol/L acetic acid buffer (pH 3.5) to prepare a test sample solution. 0.1 mL of hyaluronidase solution (Type IV-S (from bovine testis), 400 NF units/mL, manufactured by SIGMA) was added to 0.2 mL of the test sample solution, and the mixture was reacted at 37°C for 20 minutes. Then, 0.2 mL of 2.5 mmol/L calcium chloride was added as an activator, and the mixture was further reacted at 37°C for 20 minutes. 0.5 mL of 0.8 mg/mL sodium hyaluronate solution (from rooster comb) (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.) was added thereto, and the mixture was reacted at 37°C for 40 minutes. In addition, the final concentration of the test sample at this time was the concentration shown in Table 5 below. Then, add 0.2 mL of 0.4 mol/L sodium hydroxide to stop the reaction. After cooling, add 0.2 mL of boric acid solution to each reaction solution and boil for 3 minutes. After ice cooling, add 6 mL of p-DABA reagent (dissolve 10 g of p-dimethylaminobenzaldehyde in a mixture of 12.5 mL of 10N hydrochloric acid and 87.5 mL of acetic acid, and dilute 10 times with acetic acid) and react at 37°C for 20 minutes. Then, measure the absorbance at a wavelength of 585 nm.

In addition, as a blank group, the same operation and absorbance measurement as above were performed except that the hyaluronidase solution (enzyme solution) was changed to 0.1 mol/L acetic acid buffer (pH 3.5).

As a control group, the same operation and absorbance measurement as above were performed except that the test sample solution was changed to a 0.1 mol/L acetic acid buffer solution (pH 3.5) containing no test sample.

The hyaluronidase activity inhibition rate was calculated based on the following formula 4 from the measured value of the absorbance obtained. The results are shown in the following Table 5. ＜Formula 4＞ Hyaluronidase activity inhibition rate (%) = {1-(C-D)/(A-B)} × 100 In the above formula 4, A~D represent the following respectively. A: Absorbance at a wavelength of 585nm without adding the test sample and adding the enzyme B: Absorbance at a wavelength of 585nm without adding the test sample and adding the enzyme C: Absorbance at a wavelength of 585nm with adding the test sample and adding the enzyme D: Absorbance at a wavelength of 585nm with adding the test sample and adding the enzyme

[Table 5] Test sample Concentration of the test sample (μg/mL) Hyaluronidase activity inhibition rate (%) IC ₅₀ (µg/mL) Manufacturing Example 1 Artemisia annua fermented liquid 1 400 67.7 320.6 Manufacturing Example 2 Artemisia annua fermented liquid 2 400 66.4 317.2 Comparative Manufacturing Example 1 Artemisia annua extract 400 49.8 ＞400

(Test Example 1-5: Tyrosinase Activity Inhibition Test) Using Artemisia oleracea fermentation liquid 1 obtained in Production Example 1, Artemisia oleracea fermentation liquid 2 obtained in Production Example 2, and Artemisia oleracea extract obtained in Comparative Production Example 1 as test samples, the tyrosinase activity inhibition effect was tested by the following test method.

Each test sample was dissolved in 25% DMSO solution to prepare the test sample solution. In a 48-well plate, 0.2 mL of Mcllvaine buffer (pH 6.8), 0.06 mL of 0.3 mg/mL tyrosine solution, and 0.18 mL of the above test sample solution were added and allowed to stand at 37°C for 10 minutes. 0.02 mL of 800 units/mL tyrosinase solution (manufactured by SIGMA) was added thereto and further reacted at 37°C for 15 minutes. After the reaction was completed, the absorbance at a wavelength of 475 nm was measured. In addition, the final concentration of the above test sample at this time was the concentration shown in Table 6 below.

In addition, as a blank control, the same operation and absorbance measurement as above were performed except that the tyrosinase solution (enzyme solution) was changed to Mcllvaine buffer (pH 6.8).

In addition, as a control group, the same operation and absorbance measurement as above were performed except that the test sample solution was changed to a 25% DMSO solution containing no test sample.

The tyrosinase activity inhibition rate was calculated based on the following formula 5 from the measured value of the absorbance obtained. The results are shown in the following Table 6. ＜Formula 5＞ Tyrosinase activity inhibition rate (%) = {1-(C-D)/(A-B)} × 100 In the above formula 5, A~D represent the following respectively. A: Absorbance at a wavelength of 475nm without adding the test sample and adding the enzyme B: Absorbance at a wavelength of 475nm without adding the test sample and adding the enzyme C: Absorbance at a wavelength of 475nm with adding the test sample and adding the enzyme D: Absorbance at a wavelength of 475nm with adding the test sample and adding the enzyme

[Table 6] Test sample Concentration of the test sample (μg/mL) Tyrosinase activity inhibition rate (%) Manufacturing Example 1 Artemisia annua fermented liquid 1 400 5.7 Manufacturing Example 2 Artemisia annua fermented liquid 2 400 9.5 Comparative Manufacturing Example 1 Artemisia annua extract 400 4.2

<Production Example 3: Preparation of Artemisia capillaris fermented liquid 1> In the seed yeast preparation step of the aforementioned Production Example 1, except that Artemisia capillaris Thunbergii (manufactured by ALBION Co., Ltd.) was replaced with Artemisia capillaris Thunbergii, "Artemisia capillaris seed yeast" was prepared in the same method as the seed yeast preparation step of the aforementioned Production Example 1. In addition, in the fermentation step of the aforementioned Production Example 1, except that Artemisia capillaris Thunbergii (manufactured by ALBION Co., Ltd.) was replaced with Artemisia capillaris Thunbergii, "Artemisia capillaris fermented liquid 1" was obtained in the same method as the fermentation step of the aforementioned Production Example 1.

<Production Example 4: Preparation of Artemisia oryzae Fermentation Liquid 2> In the above-mentioned Production Example 3, except that the Artemisia oryzae seed koji was replaced with rice seed koji ( Aspergillus oryzae , Shirakami white koji, Akita Konno Shoten Co., Ltd.), the "Artemisia oryzae Fermentation Liquid 2" was obtained by the same method as the above-mentioned Production Example 3.

<Comparative Preparation Example 2: Preparation of Artemisia capillaris extract> In the aforementioned Comparative Preparation Example 1, except that Artemisia capillaris Thunbergii (manufactured by ALBION Co., Ltd.) was replaced with Artemisia capillaris Thunbergii, "Artemisia capillaris extract" was obtained in the same manner as in the aforementioned Comparative Preparation Example 1.

(Test Example A-2: Measurement of contact angle) In the above-mentioned Test Example A-1, the contact angle was measured in the same manner as in the above-mentioned Test Example A-1, except that the test samples were changed to Artemisia capillaris fermented liquid 1 obtained in Production Example 3, Artemisia capillaris fermented liquid 2 obtained in Production Example 4, and Artemisia capillaris extract obtained in Comparative Production Example 2. The results are shown in Table 7 below. In addition, an example of the droplet when measuring the contact angle of each test sample is shown in Figures 2A to 2C.

[Table 7] Test sample Fermentation Aspergillus Contact angle θ(°) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 have Artemisia capillaris seed yeast 77.50 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 have Rice Kernel 79.58 Comparative Manufacturing Example 2 Artemisia Capillaris Extract without - 87.75

Compared with the Artemisia capillaris extract obtained in Preparation Example 2, the Artemisia capillaris fermented liquid 1 obtained in Preparation Example 3 and the Artemisia capillaris fermented liquid 2 obtained in Preparation Example 4 both have a small contact angle of 81° or less, and are excellent in skin affinity. Furthermore, the Artemisia capillaris fermented liquid 1 obtained in Preparation Example 3 has a contact angle of 78° or less, and is even more excellent in skin affinity.

(Test Example 2-1: Test for promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) In Test Example 1-2, except that the test samples were changed to Artemisia capillaris fermented liquid 1 obtained in Production Example 3, Artemisia capillaris fermented liquid 2 obtained in Production Example 4, and Artemisia capillaris extract obtained in Comparative Production Example 2, and the final concentration of the test samples was changed to the concentration shown in Table 8 below, the hyaluronic acid synthase 3 (HAS3) mRNA expression promoting effect was tested in the same manner as in Test Example 1-2. The results are shown in Table 8 below.

[Table 8] Test sample Concentration of the test sample (μg/mL) HAS3 mRNA expression promotion rate (%) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 20 38.7 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 20 46.1 Comparative Manufacturing Example 2 Artemisia Capillaris Extract 20 28.3

(Test Example 2-2: Test for promoting the production of type I collagen) Using Artemisia capillaris fermented liquid 1 obtained in Production Example 3, Artemisia capillaris fermented liquid 2 obtained in Production Example 4, and Artemisia capillaris extract obtained in Comparative Production Example 2 as test samples, the effect of promoting the production of type I collagen was tested by the following test method.

Each test sample was dissolved in Dulbecco's MEM (Nissui Pharmaceutical Co., Ltd.) containing 0.25% fetal bovine serum (FBS, Biosera) to a final concentration as shown in Table 9 below, and the test sample-added culture medium was prepared. Normal human fibroblasts (NB1RGB, purchased from RIKEN BRC) were cultured at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. The cell density was adjusted to 1.6×10 ⁵ cells/mL using DMEM containing 10% FBS. Then, 100 μL of the above-mentioned NB1RGB (1.6×10 ⁵ cells/mL) was inoculated into each well of a 96-well microplate and cultured overnight at 37°C and 5% CO _2. After the culture was completed, the culture medium was exchanged with 100 μL of the culture medium to which the test sample was added and cultured for 3 days at 37°C and 5% CO _2. After the culture was completed, the amount of type I collagen in the culture medium of each well was measured by ELISA.

Specifically, 90 μL of the culture supernatant was transferred to an ELISA plate and incubated overnight at 4°C to allow the plate to absorb. The solution was discarded and the plate was washed with phosphate buffer (PBS-T) containing 0.05% Tween-20. Thereafter, a blocking operation was performed with phosphate buffer (PBS-T) containing 1% FBS. The solution was discarded and the plate was washed with phosphate buffer (PBS-T) containing 0.05% Tween-20, and an anti-human collagen type I antibody (rabbit IgG, manufactured by Chemi-Con) was reacted. The solution was discarded, and the sample was washed with phosphate buffered saline (PBS-T) containing 0.05% Tween-20, and reacted with HRP-labeled anti-rabbit IgG antibody, followed by the same washing operation and color development. The amount of type I collagen was calculated by performing the above ELISA using a standard product and making a calibration curve.

As a control group, the same operation and ELISA were performed as above except that the test sample solution was changed to DMEM containing 0.25% FBS without the test sample.

From the obtained measured value, the type I collagen production promotion rate was calculated based on the following formula 6. The results are shown in the following Table 9. ＜Formula 6＞ Type I collagen production promotion rate (%) = A/B × 100 In the above formula 6, A and B each represent the following. A: The amount of type I collagen when the test sample is added B: The amount of type I collagen when the test sample is not added

[Table 9] Test sample Concentration of the test sample (μg/mL) Type I collagen production promotion rate (%) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 100 72.9 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 100 65.5 Comparative Manufacturing Example 2 Artemisia Capillaris Extract 100 56.9

(Test Example 2-3: Claudin-1 mRNA expression promoting effect test) Using Artemisia capillaris fermented liquid 1 obtained in Preparation Example 3, Artemisia capillaris fermented liquid 2 obtained in Preparation Example 4, and Artemisia capillaris extract obtained in Comparative Preparation Example 2 as test samples, the claudin-1 mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration as shown in Table 10 below, and the test sample addition medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. Adjust the medium to 1.5×10 ⁵ cells/mL using normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Then, inoculate 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) in a 35 mm dish and culture overnight at 37°C and 5% CO _2. After the culture is completed, exchange the medium with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and culture for a further 24 hours. After the culture is completed, exchange the medium with 2 mL of the aforementioned medium supplemented with the test sample and culture for 24 hours at 37°C and 5% CO ₂ . After the culture was completed, the culture medium was removed and total RNA was extracted using an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.). The amount of RNA was measured using a spectrophotometer and the total RNA was adjusted to 200 ng/μL using pure water.

In addition, as a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test samples was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test samples, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNA was used as a template to measure the expression level of claudin-1 mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was carried out by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression level of claudin-1 mRNA without and with the addition of the test sample was corrected by the expression level of GAPDH mRNA. Based on this correction value, the claudin-1 mRNA expression promotion rate was calculated based on the following formula 7. The results are shown in the following Table 10. <Formula 7> Claudin-1 mRNA expression promotion rate (%) = A/B × 100 In the above formula 7, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 10] Test sample Concentration of the test sample (μg/mL) Claudin-1 mRNA expression promotion rate (%) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 20 739.6 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 20 690.5 Comparative Manufacturing Example 2 Artemisia Capillaris Extract 20 662.8

(Test Example 2-4: Claudin-4 mRNA expression promoting effect test) Using Artemisia capillaris fermented liquid 1 obtained in Preparation Example 3, Artemisia capillaris fermented liquid 2 obtained in Preparation Example 4, and Artemisia capillaris extract obtained in Comparative Preparation Example 2 as test samples, the claudin-4 mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Co., Ltd.) to a final concentration as shown in Table 11 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurashiki Co., Ltd.) under 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. The concentration was adjusted to 1.5×10 ⁵ cells/mL using a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Next, 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) was inoculated in a 35 mm dish and cultured overnight at 37°C and 5% CO _2. After the culture was completed, the culture medium was exchanged with normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) and cultured for a further 24 hours. After the culture was completed, the culture medium was exchanged with 2 mL of the aforementioned culture medium supplemented with the test sample and cultured for 24 hours at 37°C and 5% CO ₂ . After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer. The total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNA was used as a template to measure the expression level of claudin-4 mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was carried out by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression level of claudin-4 mRNA without and with the addition of the test sample was corrected by the expression level of GAPDH mRNA. Based on this correction value, the claudin-4 mRNA expression promotion rate was calculated based on the following formula 8. The results are shown in the following Table 11. <Formula 8> Claudin-4 mRNA expression promotion rate (%) = A/B × 100 In the above formula 8, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 11] Test sample Concentration of the test sample (μg/mL) Claudin-4 mRNA expression promotion rate (%) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 20 674.0 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 20 734.6 Comparative Manufacturing Example 2 Artemisia Capillaris Extract 20 487.6

(Test Example 2-5: Test for promoting the expression of nexin mRNA) Using the Artemisia capillaris fermented liquid 1 obtained in Preparation Example 3, the Artemisia capillaris fermented liquid 2 obtained in Preparation Example 4, and the Artemisia capillaris extract obtained in Comparative Preparation Example 2 as test samples, the nexin mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration as shown in Table 12 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. The cell density was adjusted to 1.5×10 ⁵ cells/mL using normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Then, 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) was inoculated in a 35 mm dish and cultured overnight at 37°C and 5% CO _2. After the culture was completed, the medium was exchanged with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and cultured for a further 24 hours. After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer. The total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNAs were used as templates to measure the expression levels of nexin mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was performed by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression levels of nexin mRNA without and with the addition of the test sample were corrected with the expression level of GAPDH mRNA. Based on this correction value, the nexin mRNA expression promotion rate was calculated based on the following formula 9. The results are shown in the following Table 12. <Formula 9> Nexin mRNA expression promotion rate (%) = A/B × 100 In the above formula 9, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 12] Test sample Concentration of the test sample (μg/mL) Connexin mRNA expression promotion rate (%) Manufacturing Example 3 Artemisia capillaris fermented liquid 1 20 707.9 Manufacturing Example 4 Artemisia capillaris fermented liquid 2 20 635.7 Comparative Manufacturing Example 2 Artemisia Capillaris Extract 20 621.0

<Production Example 5: Preparation of thyme fermented liquor 1> In the seed yeast preparation step of the aforementioned production example 1, except that the artemisia vulgaris was replaced with thyme ( Thymus vulgaris Linne) (manufactured by ALBION Co., Ltd.), "thyme seed yeast" was prepared in the same method as the seed yeast preparation step of the aforementioned production example 1. In addition, in the fermentation step of the aforementioned production example 1, except that the artemisia vulgaris was replaced with thyme ( Thymus vulgaris Linne) (manufactured by ALBION Co., Ltd.), "thyme fermented liquor 1" was obtained in the same method as the fermentation step of the aforementioned production example 1.

<Production Example 6: Preparation of Thyme Fermentation Liquid 2> In the above-mentioned Production Example 5, except that the thyme koji was replaced with rice koji ( Aspergillus oryzae , Shirakami white koji, manufactured by Akita Konno Shoten Co., Ltd.), the same method as the above-mentioned Production Example 5 was used to obtain "Thyme Fermentation Liquid 2".

<Comparative Preparation Example 3: Preparation of Thyme Extract> In the aforementioned Comparative Preparation Example 1, except that Artemisia vulgaris was replaced with Thyme ( Thymus vulgaris Linne) (manufactured by ALBION Co., Ltd.), the same method as the aforementioned Comparative Preparation Example 1 was used to obtain "thyme extract".

(Test Example A-3: Measurement of contact angle) In the above-mentioned Test Example A-1, the contact angle was measured in the same manner as in the above-mentioned Test Example A-1, except that the test samples were changed to thyme fermentation liquid 1 obtained in Production Example 5, thyme fermentation liquid 2 obtained in Production Example 6, and thyme extract obtained in Comparative Production Example 3. The results are shown in Table 13 below. In addition, an example of the droplet during the contact angle measurement of each test sample is shown in Figures 3A to 3C.

[Table 13] Test sample Fermentation Aspergillus Contact angle θ(°) Manufacturing Example 5 Thyme fermented liquid 1 have Thyme yeast 78.32 Manufacturing Example 6 Thyme fermented liquid 2 have Rice Kernel 82.77 Comparative Manufacturing Example 3 Thyme Extract without - 88.59

Compared with the thyme extract obtained in Preparation Example 3, the thyme fermented liquid 1 obtained in Preparation Example 5 and the thyme fermented liquid 2 obtained in Preparation Example 6 both have a small contact angle of 87° or less, and are excellent in skin affinity. Furthermore, the thyme fermented liquid 1 obtained in Preparation Example 5 has a contact angle of 81° or less, and is even more excellent in skin affinity.

(Test Example 3-1: Test for promoting the expression of transglutaminase-1 (TGM-1) mRNA) Using the thyme fermentation liquid 1 obtained in Preparation Example 5, the thyme fermentation liquid 2 obtained in Preparation Example 6, and the thyme extract obtained in Comparative Preparation Example 3 as test samples, the expression promoting effect of transglutaminase-1 (TGM-1) mRNA was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration shown in Table 14 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. Adjust the medium to 1.5×10 ⁵ cells/mL using normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Then, inoculate 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) in a 35 mm dish and culture overnight at 37°C and 5% CO _2. After the culture is completed, exchange the medium with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and culture for a further 24 hours. After the culture is completed, exchange the medium with 2 mL of the aforementioned medium supplemented with the test sample and culture for 24 hours at 37°C and 5% CO ₂ . After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer. The total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNAs were used as templates to measure the expression levels of transglutaminase-1 (TGM-1) mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was performed by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression levels of TGM-1 mRNA without and with the addition of the test sample were corrected by the expression level of GAPDH mRNA. Based on this correction value, the TGM-1 mRNA expression promotion rate was calculated based on the following formula 10. The results are shown in the following Table 14. <Formula 10> TGM-1 mRNA expression promotion rate (%) = A/B × 100 In the above formula 10, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 14] Test sample Concentration of the test sample (μg/mL) TMG-1 mRNA expression promotion rate (%) Manufacturing Example 5 Thyme fermented liquid 1 20 625.0 Manufacturing Example 6 Thyme fermented liquid 2 20 586.4 Comparative Manufacturing Example 3 Thyme Extract 20 515.7

(Test Example 3-2: Aquaporin 3 (AQP3) mRNA expression promoting effect test) Using thyme fermentation liquid 1 obtained in Preparation Example 5, thyme fermentation liquid 2 obtained in Preparation Example 6, and thyme extract obtained in Comparative Preparation Example 3 as test samples, the aquaporin 3 (AQP3) mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration shown in Table 15 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. Adjust the medium to 1.5×10 ⁵ cells/mL using normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Then, inoculate 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) in a 35 mm dish and culture overnight at 37°C and 5% CO _2. After the culture is completed, exchange the medium with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and culture for a further 24 hours. After the culture is completed, exchange the medium with 2 mL of the aforementioned medium supplemented with the test sample and culture for 24 hours at 37°C and 5% CO ₂ . After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer. The total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNAs were used as templates to measure the expression levels of aquaporin 3 (AQP3) mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was performed by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression levels of AQP3 mRNA without and with the addition of the test sample were corrected with the expression level of GAPDH mRNA. Based on this correction value, the AQP3 mRNA expression promotion rate was calculated based on the following formula 11. The results are shown in the following Table 15. <Formula 11> AQP3 mRNA expression promotion rate (%) = A/B × 100 In the above formula 11, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 15] Test sample Concentration of the test sample (μg/mL) AQP3 mRNA expression promotion rate (%) Manufacturing Example 5 Thyme fermented liquid 1 20 196.4 Manufacturing Example 6 Thyme fermented liquid 2 20 160.5 Comparative Manufacturing Example 3 Thyme Extract 20 145.7

(Test Example 3-3: Test for promoting the expression of nexin mRNA) In Test Example 2-5, except that the test samples were changed to thyme fermentation liquid 1 obtained in Production Example 5, thyme fermentation liquid 2 obtained in Production Example 6, and thyme extract obtained in Comparative Production Example 3, and the final concentration of the test samples was changed to the concentration shown in Table 16 below, the nexin mRNA expression promoting effect was tested in the same manner as in Test Example 2-5. The results are shown in Table 16 below.

[Table 16] Test sample Concentration of the test sample (μg/mL) Connexin mRNA expression promotion rate (%) Manufacturing Example 5 Thyme fermented liquid 1 20 795.2 Manufacturing Example 6 Thyme fermented liquid 2 20 1023.7 Comparative Manufacturing Example 3 Thyme Extract 20 562.9

(Test Example 3-4: DPPH free radical elimination test) In Test Example 1-3, except that the test samples were changed to thyme fermentation liquid 1 obtained in Production Example 5, thyme fermentation liquid 2 obtained in Production Example 6, and thyme extract obtained in Comparative Production Example 3, and the final concentration of the test samples was changed to the concentration shown in Table 17 below, the DPPH free radical elimination effect was tested in the same manner as in Test Example 1-3. The results are shown in Table 17 below.

[Table 17] Test sample Concentration of the test sample (μg/mL) DPPH free radical elimination rate (%) Manufacturing Example 5 Thyme fermented liquid 1 12.5 41.9 Manufacturing Example 6 Thyme fermented liquid 2 12.5 39.9 Comparative Manufacturing Example 3 Thyme Extract 12.5 36.4

<Production Example 7: Preparation of Melissa officinalis fermented liquor 1> In the seed koji preparation step of the aforementioned production example 1, except that the artemisia was replaced with Melissa officinalis Linne (manufactured by ALBION Co., Ltd.), "Melissa officinalis seed koji" was prepared in the same method as the seed koji preparation step of the aforementioned production example 1. In addition, in the fermentation step of the aforementioned production example 1, except that the artemisia was replaced with Melissa officinalis Linne (manufactured by ALBION Co., Ltd.), "Melissa officinalis fermented liquor 1" was obtained in the same method as the fermentation step of the aforementioned production example 1.

<Production Example 8: Preparation of Aspergillus oryzae fermented liquid 2> In the above-mentioned Production Example 7, except that the Aspergillus oryzae koji was replaced with rice koji ( Aspergillus oryzae , Shirakami white koji, Akita Konno Shoten Co., Ltd.), the "Aspergillus oryzae fermented liquid 2" was obtained by the same method as the above-mentioned Production Example 7.

<Comparative Preparation Example 4: Preparation of Melissa officinalis Extract> In the aforementioned Comparative Preparation Example 1, except that Artemisia officinalis was replaced with Melissa officinalis Linne (manufactured by ALBION Co., Ltd.), the "Melissa officinalis extract" was obtained in the same manner as in the aforementioned Comparative Preparation Example 1.

(Test Example A-4: Measurement of contact angle) In the above-mentioned Test Example A-1, the contact angle was measured in the same manner as in the above-mentioned Test Example A-1, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4. The results are shown in Table 18 below. In addition, an example of the droplet during the contact angle measurement of each test sample is shown in Figures 4A to 4C.

[Table 18] Test sample Fermentation Aspergillus Contact angle θ(°) Manufacturing Example 7 Cypress Fermented Liquid 1 have Cypress bee grass yeast 77.09 Manufacturing Example 8 Cypress Fermented Liquid 2 have Rice Kernel 80.16 Comparative Manufacturing Example 4 Cypress extract without - 86.53

Compared with the cypress extract obtained in Preparation Example 4, the cypress fermented liquid 1 obtained in Preparation Example 7 and the cypress fermented liquid 2 obtained in Preparation Example 8 both have a small contact angle of less than 85°, and are excellent in skin affinity. Furthermore, the cypress fermented liquid 1 obtained in Preparation Example 7 has a contact angle of more than 79°, and is even more excellent in skin affinity.

(Test Example 4-1: Test for promoting the production of type I collagen) In Test Example 2-2, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4, and the final concentration of the test samples was changed to the concentration shown in Table 19 below, the type I collagen production promotion effect was tested in the same manner as in Test Example 2-2. The results are shown in Table 19 below.

[Table 19] Test sample Concentration of the test sample (μg/mL) Type I collagen production promotion rate (%) Manufacturing Example 7 Cypress Fermented Liquid 1 1.56 94.4 Manufacturing Example 8 Cypress Fermented Liquid 2 1.56 89.2 Comparative Manufacturing Example 4 Cypress extract 1.56 77.7

(Test Example 4-2: Aquaporin 3 (AQP3) mRNA expression promoting effect test) In Test Example 3-2, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4, and the final concentration of the test samples was changed to the concentration shown in Table 20 below, the aquaporin 3 (AQP3) mRNA expression promoting effect was tested in the same manner as in Test Example 3-2. The results are shown in Table 20 below.

[Table 20] Test sample Concentration of the test sample (μg/mL) AQP3 mRNA expression promotion rate (%) Manufacturing Example 7 Cypress Fermented Liquid 1 20 105.4 Manufacturing Example 8 Cypress Fermented Liquid 2 20 135.7 Comparative Manufacturing Example 4 Cypress extract 20 97.7

(Test Example 4-3: DPPH free radical elimination test) In Test Example 1-3, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4, and the final concentration of the test samples was changed to the concentration shown in Table 21 below, the DPPH free radical elimination effect was tested in the same manner as in Test Example 1-3. The results are shown in Table 21 below.

　 Tested sample Concentration of the tested sample (μg/mL) DPPH free radical elimination rate (%) Manufacturing example 7 Cypress Fermented Liquid 1 12.5 73.2 Manufacturing example 8 Cypress Fermented Liquid 2 12.5 72.2 Comparative Manufacturing Example 4 Cypress extract 12.5 69.6

(Test Example 4-4: Hyaluronidase Activity Inhibition Test) In Test Example 1-4, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4, and the final concentration of the test samples was changed to the concentration shown in Table 22 below, the hyaluronidase activity inhibition effect was tested in the same manner as in Test Example 1-4. The results are shown in Table 22 below.

[Table 22] Test sample Concentration of the test sample (μg/mL) Hyaluronidase activity inhibition rate (%) Manufacturing Example 7 Cypress Fermented Liquid 1 100 71.9 Manufacturing Example 8 Cypress Fermented Liquid 2 100 67.7 Comparative Manufacturing Example 4 Cypress extract 100 63.9

(Test Example 4-5: Tyrosinase Activity Inhibition Test) In Test Example 1-5, except that the test samples were changed to the fermented solution 1 of the cypress grass obtained in Production Example 7, the fermented solution 2 of the cypress grass obtained in Production Example 8, and the cypress grass extract obtained in Comparative Production Example 4, and the final concentration of the test samples was changed to the concentration shown in Table 23 below, the tyrosinase activity inhibition effect was tested in the same manner as in Test Example 1-5. The results are shown in Table 23 below.

[Table 23] Test sample Concentration of the test sample (μg/mL) Tyrosinase activity inhibition rate (%) Manufacturing Example 7 Cypress Fermented Liquid 1 100 3.6 Manufacturing Example 8 Cypress Fermented Liquid 2 100 2.6 Comparative Manufacturing Example 4 Cypress extract 100 -2

<Production Example 9: Preparation of Cornflower Fermentation Liquid 1> In the seed yeast preparation step of the aforementioned Production Example 1, except that Artemisia serrata was replaced with Cornflower ( Centaurea cyanus Linne) (manufactured by ALBION Co., Ltd.), "Cornflower Seed Yeast" was prepared in the same manner as the seed yeast preparation step of the aforementioned Production Example 1. In addition, in the fermentation step of the aforementioned Production Example 1, except that Artemisia serrata was replaced with Cornflower ( Centaurea cyanus Linne) (manufactured by ALBION Co., Ltd.), "Cornflower Fermentation Liquid 1" was obtained in the same manner as the fermentation step of the aforementioned Production Example 1.

<Production Example 10: Preparation of Cornflower Fermentation Liquid 2> In the above-mentioned Production Example 9, except that the cornflower seed koji was replaced with rice seed koji ( Aspergillus oryzae , Shirakami white koji, Akita Konno Shoten Co., Ltd.), the same method as the above-mentioned Production Example 9 was used to obtain "Cornflower Fermentation Liquid 2".

<Comparative Preparation Example 5: Preparation of Cornflower Extract> In the aforementioned Comparative Preparation Example 1, except that Artemisia cyanus Linne (manufactured by ALBION Co., Ltd.) was replaced with Cornflower ( Centaurea cyanus Linne ) , the same method as the aforementioned Comparative Preparation Example 1 was used to obtain "Cornflower Extract".

(Test Example A-5: Measurement of contact angle) In the above-mentioned Test Example A-1, the contact angle was measured in the same manner as in the above-mentioned Test Example A-1, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5. The results are shown in Table 24 below. In addition, an example of the droplet during the contact angle measurement of each test sample is shown in Figures 5A to 5C.

[Table 24] Test sample Fermentation Aspergillus Contact angle θ(°) Manufacturing Example 9 Cornflower fermented liquid 1 have Cornflower Seed Yeast 78.52 Manufacturing Example 10 Cornflower fermented liquid 2 have Rice Kernel 80.83 Comparative Manufacturing Example 5 Cornflower Extract without - 86.96

Compared with the cornflower extract obtained in Preparation Example 5, the cornflower fermented liquid 1 obtained in Preparation Example 9 and the cornflower fermented liquid 2 obtained in Preparation Example 10 both have a small contact angle of less than 85°, and are excellent in skin affinity. Furthermore, the cornflower fermented liquid 1 obtained in Preparation Example 9 has a contact angle of less than 79°, and is even more excellent in skin affinity.

(Test Example 5-1: Test for promoting the production of type I collagen) In Test Example 2-2, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5, and the final concentration of the test samples was changed to the concentration shown in Table 25 below, the promotion effect of type I collagen production was tested in the same manner as in Test Example 2-2. The results are shown in Table 25 below.

[Table 25] Test sample Concentration of the test sample (μg/mL) Type I collagen production promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 1.56 98.0 Manufacturing Example 10 Cornflower fermented liquid 2 1.56 96.8 Comparative Manufacturing Example 5 Cornflower Extract 1.56 85.3

(Test Example 5-2: Test for promoting the expression of transglutaminase-1 mRNA) In Test Example 3-1, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5, and the final concentration of the test samples was changed to the concentration shown in Table 26 below, the expression of transglutaminase-1 mRNA was tested in the same manner as in Test Example 3-1. The results are shown in Table 26 below.

[Table 26] Test sample Concentration of the test sample (μg/mL) TMG-1 mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 20 178.0 Manufacturing Example 10 Cornflower fermented liquid 2 20 156.3 Comparative Manufacturing Example 5 Cornflower Extract 20 140.6

(Test Example 5-3: Test for promoting the expression of filaggrin mRNA) Using the cornflower fermentation liquid 1 obtained in Preparation Example 9, the cornflower fermentation liquid 2 obtained in Preparation Example 10, and the cornflower extract obtained in Comparative Preparation Example 5 as test samples, the filaggrin mRNA expression promoting effect was tested by the following test method.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurafuki Co., Ltd.) to a final concentration shown in Table 27 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurafuki Co., Ltd.) were cultured in a normal human epidermal keratinocyte proliferation medium (HuMedia-KG2, manufactured by Kurafuki Co., Ltd.) at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. Adjust the medium to 1.5×10 ⁵ cells/mL using normal human epidermal keratinocyte proliferation medium (HuMedia-KG2). Then, inoculate 2 mL of the aforementioned NHEK (1.5×10 ⁵ cells/mL) in a 35 mm dish and culture overnight at 37°C and 5% CO _2. After the culture is completed, exchange the medium with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and culture for a further 24 hours. After the culture is completed, exchange the medium with 2 mL of the aforementioned medium supplemented with the test sample and culture for 24 hours at 37°C and 5% CO ₂ . After the culture was completed, the culture medium was removed, and total RNA was extracted with an RNA extraction reagent (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer. The total RNA was adjusted to 200 ng/μL using pure water.

As a control group, the same operation and absorbance measurement were performed as above, except that the above-mentioned culture medium with added test sample 2 mL was replaced with 2 mL of normal human epidermal keratinocyte basal culture medium (HuMedia-KB2) without test sample, and the total RNA was adjusted to 200 ng/μL by the same method as above.

The above-mentioned total RNAs were used as templates to measure the expression levels of fibronectin mRNA and GAPDH mRNA as an internal standard. The detection of mRNA was performed by a two-step real-time PCR reaction using a real-time PCR device (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Co., Ltd.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Co., Ltd.). The expression levels of fibronectin mRNA without and with the addition of the test sample were corrected by the expression level of GAPDH mRNA. Based on this correction value, the expression promotion rate of fibronectin mRNA was calculated based on the following formula 12. The results are shown in the following Table 27. <Formula 12> Filaggrin mRNA expression promotion rate (%) = A/B × 100 In the above formula 12, A and B each represent the following. A: Correction value when the test sample is added B: Correction value when the test sample is not added

[Table 27] Test sample Concentration of the test sample (μg/mL) Filaggrin mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 20 159.8 Manufacturing Example 10 Cornflower fermented liquid 2 20 136.8 Comparative Manufacturing Example 5 Cornflower Extract 20 122.3

(Test Example 5-4: Aquaporin 3 (AQP3) mRNA expression promoting effect test) In Test Example 3-2, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 28 below, the aquaporin 3 (AQP3) mRNA expression promoting effect was tested in the same method as in Test Example 3-2. The results are shown in Table 28 below.

[Table 28] Test sample Concentration of the test sample (μg/mL) AQP3 mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 20 119.3 Manufacturing Example 10 Cornflower fermented liquid 2 20 98.3 Comparative Manufacturing Example 5 Cornflower Extract 20 78.1

(Test Example 5-5: Test for promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) In Test Example 1-2, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5, and the final concentration of the test samples was changed to the concentration shown in Table 29 below, the hyaluronic acid synthase 3 (HAS3) mRNA expression promoting effect was tested in the same manner as in Test Example 1-2. The results are shown in Table 29 below.

[Table 29] Test sample Concentration of the test sample (μg/mL) HAS3 mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 20 145.0 Manufacturing Example 10 Cornflower fermented liquid 2 20 170.0 Comparative Manufacturing Example 5 Cornflower Extract 20 112.3

(Test Example 5-6: Test for promoting the expression of claudin-1 mRNA) In Test Example 2-3, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Preparation Example 9, cornflower fermentation liquid 2 obtained in Preparation Example 10, and cornflower extract obtained in Comparative Preparation Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 30 below, the claudin-1 mRNA expression promoting effect was tested in the same manner as in Test Example 2-3. The results are shown in Table 30 below.

[Table 30] Test sample Concentration of the test sample (μg/mL) Claudin-1 mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 5 98.2 20 158.3 Manufacturing Example 10 Cornflower fermented liquid 2 5 101.5 20 184.5 Comparative Manufacturing Example 5 Cornflower Extract 5 84.0 20 129.2

(Test Example 5-7: Test for promoting the expression of claudin-4 mRNA) In Test Example 2-4, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Preparation Example 9, cornflower fermentation liquid 2 obtained in Preparation Example 10, and cornflower extract obtained in Comparative Preparation Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 31 below, the claudin-4 mRNA expression promoting effect was tested in the same manner as in Test Example 2-4. The results are shown in Table 31 below.

[Table 31] Test sample Concentration of the test sample (μg/mL) Claudin-4 mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 5 138.8 20 141.2 Manufacturing Example 10 Cornflower fermented liquid 2 5 82.9 20 85.6 Comparative Manufacturing Example 5 Cornflower Extract 5 64.0 20 71.8

(Test Example 5-8: Test for promoting the expression of nexin mRNA) In Test Example 2-5, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Production Example 9, cornflower fermentation liquid 2 obtained in Production Example 10, and cornflower extract obtained in Comparative Production Example 5, and the final concentration of the test samples was changed to the concentration shown in Table 32 below, the nexin mRNA expression promoting effect was tested in the same manner as in Test Example 2-5. The results are shown in Table 32 below.

[Table 32] Test sample Concentration of the test sample (μg/mL) Connexin mRNA expression promotion rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 5 137.9 Manufacturing Example 10 Cornflower fermented liquid 2 5 113.6 Comparative Manufacturing Example 5 Cornflower Extract 5 108.3

(Test Example 5-9: DPPH free radical elimination test) In Test Example 1-3, except that the test samples were changed to cornflower fermentation liquid 1 obtained in Preparation Example 9, cornflower fermentation liquid 2 obtained in Preparation Example 10, and cornflower extract obtained in Comparative Preparation Example 5, and the final concentration of the test samples was changed to the concentration shown in Table 33 below, the DPPH free radical elimination effect was tested in the same manner as in Test Example 1-3. The results are shown in Table 33 below.

[Table 33] Test sample Concentration of the test sample (μg/mL) DPPH free radical elimination rate (%) IC ₅₀ (µg/mL) Manufacturing Example 9 Cornflower fermented liquid 1 200 90.0 107.1 Manufacturing Example 10 Cornflower fermented liquid 2 200 89.4 106.6 Comparative Manufacturing Example 5 Cornflower Extract 200 81.3 123.0

(Test Example 5-10: Test for the inhibitory effect on melanin production of B16 melanoma cells) Using the cornflower fermentation liquid 1 obtained in Production Example 9, the cornflower fermentation liquid 2 obtained in Production Example 10, and the cornflower extract obtained in Comparative Production Example 5 as test samples, the inhibitory effect on melanin production of B16 melanoma cells was tested by the following test method.

Each test sample was dissolved in DMEM (Nissui Pharmaceutical Co., Ltd.) containing 10% FBS (Biosera) and 1mmol/L theophylline (Fuji Film Wako Pure Chemical Industries, Ltd.) to prepare a test sample-added culture medium. B16 melanoma cells were cultured in DMEM containing 10% FBS at 37°C and 5% CO ₂ until the cells became confluent, and then the cells were collected by trypsin treatment. The cell density was adjusted to 2.4×10 ⁵ cells/mL using DMEM containing 10% FBS and 1mmol/L theophylline. Next, 300 μL of the aforementioned B16 melanoma cells (2.4×10 ⁵ cells/mL) were inoculated into each well of a 48-well plate and cultured at 37°C and 5% CO ₂ for 6 hours. After the culture was completed, the medium was exchanged with 100 μL of the medium supplemented with the test sample and cultured at 37°C and 5% CO ₂ for 3 days. After the culture was completed, 300 μL of the medium supplemented with the test sample was added to each well and cultured at 37°C and 5% CO ₂ for 4 days. At this time, the final concentration of the aforementioned test sample was the concentration shown in Table 34 below.

After the culture was completed, the culture medium was removed from each well, 200 μL of 2 mol/L sodium hydroxide solution was added, the cells were disrupted by ultrasonic disruption, and the absorbance at a wavelength of 475 nm was measured. The amount of melanin was calculated based on the measured absorbance value based on a calibration line prepared using synthetic melanin (manufactured by SIGMA).

In addition, in order to measure the cell survival rate, B16 melanoma cells were cultured in the same manner as the above method using the culture medium with the test sample added, and then the culture medium was removed and washed with 400 μL of PBS buffer. Then, 200 μL of neutral red dissolved in DMEM solution containing 10% FBS was added to each well to a final concentration of 0.05 mg/mL, and cultured for 2.5 hours. After the culture was completed, the neutral red solution was removed, and 200 μL of ethanol-acetic acid solution (ethanol: acetic acid: water = 50:1:49 (volume ratio)) was added to each well to extract the pigment. After extraction, the absorbance at a wavelength of 540 nm was measured.

As a control group, the same operation and absorbance measurement as above were performed except that the test sample solution was changed to DMEM containing no test sample and 10% FBS and 1 mmol/L theophylline.

From the obtained measured value, the melanin production inhibition rate (%) corrected by the cell survival rate calculated based on the following formula 13 was calculated based on the following formula 14. The results are shown in the following Table 34. ＜Formula 13＞ Cell survival rate (%) = (D/C) × 100 In the above formula 13, C and D are each represented as follows. C: Absorbance at a wavelength of 540nm without adding the test sample D: Absorbance at a wavelength of 540nm with adding the test sample ＜Formula 14＞ Melanin production inhibition rate (%) = {1-(B/D)/(A/C)} × 100 In the above formula 14, A~D are each represented as follows. A: Melanin content in the unadded test sample B: Melanin content in the added test sample C: Absorbance at 540nm wavelength of the unadded test sample D: Absorbance at 540nm wavelength of the added test sample

[Table 34] Test sample Concentration of the test sample (μg/mL) Melanin production inhibition rate (%) Manufacturing Example 9 Cornflower fermented liquid 1 100 24.3 200 32.7 Manufacturing Example 10 Cornflower fermented liquid 2 100 18.8 200 26.0 Comparative Manufacturing Example 5 Cornflower extract 100 15.7 200 20.5

[Possibility of industrial application] The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention have at least one of excellent anti-aging, antioxidant, anti-inflammatory, and whitening effects, and are natural substances with high safety. Therefore, they can be widely used in fields other than cosmetics, food, and pharmaceuticals. The cosmetics of the present invention contain at least one selected from the group consisting of the aforementioned anti-aging agent, the aforementioned antioxidant, the aforementioned anti-inflammatory agent, and the aforementioned whitening agent, and thus can be used in skin cosmetics such as toners, lotions, creams, ointments, beauty serums, lotions, facial masks, gels, lip balms, lipsticks, foundations, bath agents, soaps, and shower gels; and scalp and hair cosmetics such as astringents, scalp care water, hair creams, hair lotions, hair waxes, shampoos, and conditioners.

without

FIG1A is a photograph showing an example of droplets when the contact angle of Artemisia montana (Nakai) Pamp. fermentation liquid 1 of Production Example 1 is measured. FIG1B is a photograph showing an example of droplets when the contact angle of Artemisia montana (Nakai) Pamp. fermentation liquid 2 of Production Example 2 is measured. FIG1C is a photograph showing an example of droplets when the contact angle of the Artemisia montana extract of Comparative Production Example 1 is measured. FIG2A is a photograph showing an example of droplets when the contact angle of Artemisia capillaris fermentation liquid 1 of Production Example 3 is measured. FIG2B is a photograph showing an example of droplets when the contact angle of Artemisia capillaris fermentation liquid 2 of Production Example 4 is measured. FIG2C is a photograph showing an example of a droplet when measuring the contact angle of the Artemisia capillaris extract of Comparative Manufacturing Example 2. FIG3A is a photograph showing an example of a droplet when measuring the contact angle of the thyme fermented liquid 1 of Manufacturing Example 5. FIG3B is a photograph showing an example of a droplet when measuring the contact angle of the thyme fermented liquid 2 of Manufacturing Example 6. FIG3C is a photograph showing an example of a droplet when measuring the contact angle of the thyme extract of Comparative Manufacturing Example 3. FIG4A is a photograph showing an example of a droplet when measuring the contact angle of the Herba Lycopodii fermented liquid 1 of Manufacturing Example 7. FIG4B is a photograph showing an example of a droplet when measuring the contact angle of the Herba Lycopodii fermented liquid 2 of Manufacturing Example 8. FIG4C is a photograph showing an example of a droplet when measuring the contact angle of the cypress extract of Comparative Example 4. FIG5A is a photograph showing an example of a droplet when measuring the contact angle of the cornflower fermented liquid 1 of Production Example 9. FIG5B is a photograph showing an example of a droplet when measuring the contact angle of the cornflower fermented liquid 2 of Production Example 10. FIG5C is a photograph showing an example of a droplet when measuring the contact angle of the cornflower extract of Comparative Example 5.

without

Claims (2)

A use of a fermentation liquid of Artemisia capillaris Thunbergii or Artemisia montana (Nakai) Pamp. obtained by using a yeast, characterized in that it is used to prepare an anti-inflammatory agent having a hyaluronidase activity inhibitory effect. A use of a fermented liquid of Artemisia capillaris Thunbergii or Artemisia montana (Nakai) Pamp. obtained by using a yeast, characterized in that it is used for preparing a cosmetic containing an anti-inflammatory agent having a hyaluronidase activity inhibitory effect.