Anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent, and cosmetics

Abstract

To provide an anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent having at least any action selected from the group consisting of excellent anti-aging action, antioxidant action, anti-inflammatory action, and whitening action and having high safety; and to provide a cosmetic having at least one action selected from the group consisting of an excellent anti-aging action, antioxidant action, anti-inflammatory action, and whitening action and having high safety.SOLUTION: Provided are an anti-aging agent containing as an active ingredient at least any of the fermentation liquids of Artemisia plant fermentation liquid, Thymus vulgaris fermented liquid, Melissa officinalis fermented liquid, and Centaurea cyanus L. fermented liquid; an antioxidant containing as an active ingredient at least any of the fermentation liquids of Artemisia plant fermentation liquid, Thymus vulgaris fermented liquid, Melissa officinalis fermented liquid, and Centaurea cyanus L. fermented liquid; an anti-inflammatory agent containing as an active ingredient at least either of the fermentation liquids of Artemisia plant fermentation liquid and Melissa officinalis fermented liquid; and a whitening agent containing as an active ingredient at least either of the fermented liquids of Melissa officinalis fermented liquid and Centaurea cyanus L. fermented liquid; as well as a cosmetic.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an anti-aging agent, an antioxidant, an anti-inflammatory agent, and a whitening agent containing a fermentation broth of Aspergillus of plants as an active ingredient, and a cosmetic containing at least any one of the anti-inflammatory agent, the antioxidant, the anti-inflammatory agent, and the whitening agent.

Background Art

[0002] In recent years, reactive oxygen species have attracted attention as factors that oxidize biological components, and their adverse effects on the living body have become a problem. Reactive oxygen species are generated in the energy metabolism process in living cells, and include superoxide [i.e., superoxide anion (·O2 - ) generated by one-electron reduction of oxygen molecules, hydrogen peroxide (H2O2), singlet oxygen ( 1 O2), hydroxyl radical (·OH)], etc. Such reactive oxygen species are essential for the bactericidal mechanism of phagocytes and play an important role in the removal of viruses and cancer cells.

[0003] However, the excessive generation of the reactive oxygen species attacks biomolecules that constitute membranes and tissues in the living body and induces various diseases. Superoxide, which is produced in the living body and also serves as a starting material for other reactive oxygen species, is usually sequentially eliminated by the catalytic action of superoxide dismutase (SOD) contained in cells. However, when the production of superoxide is excessive or the action of SOD is reduced, the elimination of superoxide becomes insufficient and the superoxide concentration increases. This is considered to be one of the causes of tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataract, freckles, chloasma, wrinkles, diabetes, arteriosclerosis, stiff shoulders, cold sensitivity, and skin aging.

[0004] Among these, the skin is an organ where superoxide is easily generated because it is directly exposed to environmental factors such as ultraviolet rays. Therefore, due to an increase in the superoxide concentration, for example, it decomposes, denatures, or crosslinks biological tissues such as collagen, or oxidizes fats to produce lipid peroxides that damage cells, causing skin wrinkles, aging such as a decrease in skin elasticity, inflammation, and skin pigmentation problems (see Non-Patent Document 1). Therefore, by inhibiting and suppressing the generation of reactive oxygen species and in vivo radicals, it is considered possible to prevent, treat, or improve skin aging such as wrinkle formation and elasticity reduction, tissue damage such as rheumatoid arthritis and Behçet's disease, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, cold sensitivity, and other various disorders involving reactive oxygen species.

[0005] Therefore, attempts have been made to obtain active oxygen scavenging substances, radical scavenging substances, hydrogen peroxide scavenging substances, etc. from natural products that are advantageous in terms of safety. The extracts of plants belonging to the genus Brassica of the Brassicaceae family (see Patent Document 1), the extracts of plants belonging to the genus Orostachys of the Crassulaceae family (see Patent Document 2), the extracts of the tobacco butterfly (see Patent Document 3), the extracts of Arisaema heterophyllum Blume (see Patent Document 4), etc. have been confirmed to be effective.

[0006] In addition, the causes and pathogenesis of inflammatory diseases, such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other various skin diseases accompanied by rough skin, are diverse. As the cause, it is mainly known to be due to the enhanced activity of hyaluronidase.

[0007] Hyaluronidase is a hydrolytic enzyme of hyaluronic acid. Hyaluronate, which maintains affinity for body tissues, is decomposed by ultraviolet rays, oxygen, etc. in an aqueous system, and the water retention effect decreases as the molecular weight decreases. In addition, hyaluronic acid exists as an intercellular tissue in the living body and is also involved in vascular permeability. Furthermore, hyaluronidase exists in mast cells, but is released by degranulation caused by its activation and acts as an inflammatory chemical mediator. Therefore, by inhibiting the activity of hyaluronidase, strengthening of moisture retention and prevention / reduction of inflammation are expected. Examples of substances having such an inhibitory action on hyaluronidase activity include extracts of plants of the genus Osbeckia (see Patent Document 5), extracts of Ampelopsis grossedentata (see Patent Document 6), and the like.

[0008] In the skin, melanin also plays a role in protecting the living body from ultraviolet rays. However, excessive production or uneven accumulation of melanin causes skin darkening and freckles. Generally, melanin is formed from tyrosine through the action of the enzyme tyrosinase biosynthesized in melanocytes, changing from tyrosine to dopa, from dopa to dopaquinone, and then through intermediates such as 5,6-dihydroxyindophenol. Therefore, in order to prevent, treat, or improve skin darkening (skin pigmentation disorder), freckles, chloasma, etc., it is conceivable to inhibit the activity of tyrosinase involved in melanin production or to suppress melanin production.

[0009] Conventionally, for the prevention, treatment, or improvement of skin pigmentation disorders, freckles, chloasma, etc., a treatment of applying an external whitening agent containing a chemically synthesized product such as hydroquinone as an active ingredient has been carried out. However, chemically synthesized products such as hydroquinone may have side effects such as skin irritation and allergies. Therefore, the development of a whitening agent containing a highly safe natural raw material as an active ingredient is desired. Examples of substances having an inhibitory action on tyrosinase activity include extracts of Ampelopsis grossedentata (see Patent Document 7), extracts of Saxifraga stolonifera (see Patent Document 8), and the like. Examples of substances having an inhibitory action on melanin production include extracts from the roots of Sesamum indicum (see Patent Document 9), extracts from plants belonging to the genus Saussurea (see Patent Document 10), and the like.

[0010] In addition, the epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and an extracellular matrix such as collagen outside these cells that supports the skin structure. In young skin, the interaction of these skin tissues maintains homeostasis, ensuring water retention, flexibility, elasticity, etc., and the skin is maintained in a firm, shiny, and fresh state both externally. However, when there are certain external factors such as irradiation with ultraviolet rays (UV-A, UV-B), marked drying of the air, excessive skin cleansing, etc., or when aging progresses, the production amount of collagen, which is a major component of the extracellular matrix, decreases and elastic force decreases due to crosslinking. As a result, the skin's moisturizing function and elasticity decrease, and the cutin begins abnormal peeling. Therefore, the skin loses its firmness and luster, becomes rough, and exhibits aging symptoms such as wrinkles. There are various factors involved in the changes associated with skin aging, namely wrinkles, dullness, disappearance of texture, decrease in elasticity, etc. However, a decrease and denaturation of extracellular matrix components such as collagen, hyaluronic acid, and elastin are involved. Therefore, it is considered that skin aging can be prevented and improved by promoting the production of collagen, hyaluronic acid, etc.

[0011] Therefore, attempts have been made to obtain substances having a collagen production promoting effect from natural products that are advantageous in terms of safety. As substances having a collagen production promoting effect, for example, starfruit leaf extract (see Patent Document 11), kuzunoha gashiwa extract (see Patent Document 12), etc. have been confirmed.

[0012] In addition, amino acids, which are the main components of natural moisturizing factors, are produced when filaggrin derived from keratohyalin granules is decomposed in the stratum corneum. This filaggrin is expressed as profilaggrin in epidermal keratinocytes present in the granular layer immediately below the stratum corneum. Immediately afterwards, it is phosphorylated and accumulated in keratohyalin granules, and is decomposed into filaggrin through dephosphorylation and hydrolysis, migrates to the stratum corneum, and has been reported to enhance the aggregation efficiency of keratin filaments and be involved in the internal construction of keratinocytes (see 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 the synthetic ability of filaggrin decreases under conditions such as drying, 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 profilagrin mRNA in epidermal keratinocytes and thereby promoting the synthesis of filaggrin, the amount of amino acids in the stratum corneum can be increased, and the moisture environment of the stratum corneum can be essentially improved.

[0013] As a natural product-derived filaggrin synthesis promoter, for example, licorice extract (see Patent Document 13), liquiritin known as a flavonoid glycoside contained in natural plants (see Patent Document 14), or as at least one of a natural product-derived profilagrin and filaggrin protein production promoter, plant extracts belonging to the genus Citrus or yeast extracts (see Patent Document 15) have been proposed.

[0014] The epidermis has a defensive function of protecting the skin from various external stimuli by constantly producing new keratinocytes through the division and subsequent differentiation of keratinocytes. In particular, during the differentiation process of keratinocytes, proteins such as involucrin are expressed from the spinous layer to the granular layer and cross-linked by the action of the enzyme transglutaminase-1 to form a cornified envelope (hereinafter abbreviated as "CE"), which is an insoluble cell membrane-like structure that envelops keratinocytes and contributes to the stability of the cytoskeleton and structure of keratinocytes. However, when the production amount of transglutaminase-1 in the epidermis decreases due to various factors, the formation of CE becomes incomplete and keratinization cannot proceed normally. As a result, it is considered that the keratin barrier function and the skin moisturizing function decrease, and skin symptoms such as rough skin and dry skin appear. From the above, by increasing the production of transglutaminase-1 in the epidermis of keratinocytes, promoting the formation of CE, and normalizing keratinization, it is considered possible to suppress the decrease in the skin barrier function associated with external stimuli such as dryness and ultraviolet rays, and to prevent and improve various skin symptoms such as skin dryness and rough skin.

[0015] As a natural product-derived transglutaminase-1 production promoter, noni extract (see Patent Document 16), royal jelly extract (see Patent Document 17), etc. have been proposed.

[0016] In skin cells, aquaporin, known as a water channel, is expressed on the cell membrane and is known to play a role in taking up low-molecular substances such as water in the intercellular space into the cell. In humans, the existence of 13 types of aquaporin (AQP0 to AQP12) is known. In epidermal cells, mainly aquaporin 3 (Aquaporin 3; AQP3) exists, and it is considered to play a role in taking up not only water but also low-molecular compounds such as glycerol and urea involved in the water retention function.

[0017] However, since it has been suggested that AQP3 decreases with aging and this is one of the causes of the decline in the water retention function, it is considered possible to control the water retention function and barrier function due to aging by promoting the expression of AQP3 (see Non-Patent Document 4). As those having an AQP3 expression promoting effect, for example, an extract from the leaf part of star fruit (see Patent Document 18) and the like are known.

[0018] Conventionally, it was considered that only the stratum corneum was responsible for the barrier function of the skin. However, since the barrier function of the skin collapses when the constituent proteins of the tight junction (hereinafter abbreviated as "TJ") present in the granular layer of the epidermis are deficient at the gene level, in recent years, TJ is also considered to play an important role in the barrier function of the skin (see Non-Patent Document 5). TJ is a binding device that not only adheres adjacent cells tightly but also seals the gaps between cells to control the permeation of substances. Claudin and occludin, which are cell membrane proteins, constitute TJ, and these proteins are considered to form the backbone of the TJ strand and control the barrier function of TJ (see Non-Patent Document 6). From the above, when the expression of claudin or occludin decreases for some reason, a structural disruption of TJ occurs, and it no longer functions as a permeation barrier for substances, which is expected to cause skin symptoms such as dry skin, rough skin, atopic dermatitis, and various infectious diseases.

[0019] Therefore, it is considered that by promoting the production of claudin and occludin in the epidermis to promote the formation of TJs in epidermal keratinocytes, the barrier function and moisture retention function of the skin can be enhanced, and the skin symptoms can be prevented or improved. Based on such an idea, as substances that improve the skin barrier function through the action of promoting TJ formation, extracts derived from natural products such as turmeric extract (see Patent Document 19) and thuja extract (see Patent Document 20) have been proposed.

[0020] In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as a factor inducing changes associated with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1) is known as an enzyme that degrades collagen, a major component of the extracellular matrix of the dermis of the skin. Its expression is greatly increased by ultraviolet irradiation, which is considered to be a contributing factor to the decrease and denaturation of collagen, and a major factor in the formation of wrinkles and loss of elasticity in the skin. Therefore, inhibiting the activity of MMP-1 is important for preventing and improving skin aging symptoms.

[0021] As those having such an MMP-1 inhibitory action, for example, extracts from Prunus jamasakura (see Patent Document 21), extracts from Zingiber cassumunar or Ficus nerifolia (see Patent Document 22), etc. are known.

[0022] On the other hand, although fermentation broths obtained by fermenting plants of the Artemisia genus with Aspergillus (see Patent Document 23) and fermentation broths obtained by fermenting Tatarian aster with Aspergillus (see Patent Document 24) are known, it is not known that these plant fermentation broths have effects such as anti-aging action, antioxidant action, anti-inflammatory action, and whitening action.

Prior Art Documents

Patent Documents

[0023]

Patent Document 1

Patent Document 22

Patent Document 23

Patent Document 24

Non-Patent Document

[0024]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Summary of the Invention

Problems to be Solved by the Invention

[0025] An object of the present invention is to solve the above-described conventional problems and achieve the following objects. That is, an object of the present invention is to provide an anti-aging agent having an excellent anti-aging effect and high safety, an antioxidant having an excellent antioxidant effect and high safety, an anti-inflammatory agent having an excellent anti-inflammatory effect and high safety, and a whitening agent having an excellent whitening effect and high safety. In addition, an object of the present invention is to provide a cosmetic having at least one action selected from the group consisting of excellent anti-aging action, antioxidant action, anti-inflammatory action, and whitening action, and having high safety.

Means for Solving the Problems

[0026] Means for solving the above problems are as follows. That is, <1> An anti-aging agent characterized by containing, as an active ingredient, at least one of the fermentation liquids of mugwort plants by Aspergillus, the fermentation liquid of Tachijakousousou by Aspergillus, the fermentation liquid of Kosuihakka by Aspergillus, and the fermentation liquid of Yagurumagiku by Aspergillus. <2> An antioxidant characterized by containing, as an active ingredient, at least one of the fermentation liquids of mugwort plants by Aspergillus, the fermentation liquid of Tachijakousousou by Aspergillus, the fermentation liquid of Kosuihakka by Aspergillus, and the fermentation liquid of Yagurumagiku by Aspergillus. <3> An anti-inflammatory agent characterized by containing, as an active ingredient, at least one of the fermentation liquids of mugwort plants by Aspergillus and the fermentation liquid of Kosuihakka by Aspergillus. <4> A whitening agent characterized by containing, as an active ingredient, at least one of the fermentation liquids of mugwort plants by Aspergillus, the fermentation liquid of Kosuihakka by Aspergillus, and the fermentation liquid of Yagurumagiku by Aspergillus. <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

[0027] According to the present invention, the above-mentioned various problems in the prior art can be solved, the above object can be achieved, an anti-aging agent having an excellent anti-aging action and high safety, an antioxidant having an excellent antioxidant action and high safety, an anti-inflammatory agent having an excellent anti-inflammatory action and high safety, and a whitening agent having an excellent whitening action and high safety can be provided. In addition, the present invention can provide a highly safe cosmetic having at least one action selected from the group consisting of excellent anti-aging action, antioxidant action, anti-inflammatory action, and whitening action.

Brief Description of the Drawings

[0028]

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Embodiments for Carrying Out the Invention

[0029] (Anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent) <Anti-aging agent> The anti-aging agent of the present invention contains, as an active ingredient, at least any one of the fermented liquids of mugwort genus plants by Aspergillus (hereinafter sometimes referred to as "mugwort genus plant fermented liquid"), the fermented liquid of Actinidia polygama by Aspergillus (hereinafter sometimes referred to as "Actinidia polygama fermented liquid"), the fermented liquid of Artemisia lactiflora by Aspergillus (hereinafter sometimes referred to as "Artemisia lactiflora fermented liquid"), and the fermented liquid of Youngia japonica by Aspergillus (hereinafter sometimes referred to as "Youngia japonica fermented liquid"), and, if necessary, further contains other components.

[0030] The mugwort plant fermentation broth, the tall butterbur fermentation broth, the sweet basil fermentation broth, and the Jerusalem artichoke fermentation broth have at least one action selected from the group consisting of matrix metalloproteinase-1 (MMP-1) activity inhibitory action, hyaluronan synthase 3 (HAS3) mRNA expression promoting action, type I collagen production promoting action, claudin-1 mRNA expression promoting action, claudin-4 mRNA expression promoting action, occludin mRNA expression promoting action, transglutaminase-1 (TGM-1) mRNA expression promoting action, aquaporin 3 (AQP3) mRNA expression promoting action, and filaggrin mRNA expression promoting action, and using these actions, they can be used as an active ingredient of an anti-aging agent.

[0031] Therefore, the anti-aging agent has at least one action selected from the group consisting of MMP-1 activity inhibitory action, hyaluronan synthase 3 mRNA expression promoting action, type I collagen production promoting action, claudin-1 mRNA expression promoting action, claudin-4 mRNA expression promoting action, occludin mRNA expression promoting action, transglutaminase-1 mRNA expression promoting action, aquaporin 3 mRNA expression promoting action, and filaggrin mRNA expression promoting action.

[0032] Regarding the substances that exhibit at least any one of the MMP-1 activity inhibitory action, hyaluronic acid synthase 3 mRNA expression promoting action, type I collagen production promoting action, claudin-1 mRNA expression promoting action, claudin-4 mRNA expression promoting action, occludin mRNA expression promoting action, transglutaminase-1 mRNA expression promoting action, aquaporin 3 mRNA expression promoting action, and filaggrin mRNA expression promoting action, which are possessed by the mugwort fermented liquid, the fermented liquid of *Petasites japonicus* var. *giganteus*, the fermented liquid of *Mentha spicata* L., and the fermented liquid of *Cirsium japonicum* DC., although the details are unknown, it has not been known at all conventionally that the mugwort fermented liquid, the fermented liquid of *Petasites japonicus* var. *giganteus*, the fermented liquid of *Mentha spicata* L., and the fermented liquid of *Cirsium japonicum* DC. have such excellent actions and are useful as anti-aging agents, which is a new finding by the present inventors.

[0033] <Antioxidant> The antioxidant of the present invention contains, as an active ingredient, at least any one of the fermented liquid of mugwort by Aspergillus (mugwort fermented liquid), the fermented liquid of *Petasites japonicus* var. *giganteus* by Aspergillus (*Petasites japonicus* var. *giganteus* fermented liquid), the fermented liquid of *Mentha spicata* L. by Aspergillus (*Mentha spicata* L. fermented liquid), and the fermented liquid of *Cirsium japonicum* DC. by Aspergillus (*Cirsium japonicum* DC. fermented liquid), and further contains other components as necessary.

[0034] The mugwort fermented liquid, the fermented liquid of *Petasites japonicus* var. *giganteus*, the fermented liquid of *Mentha spicata* L., and the fermented liquid of *Cirsium japonicum* DC. have a diphenyl-p-picrylhydrazyl (DPPH) radical scavenging action, and using this action, they can be used as an active ingredient of an antioxidant. Therefore, the antioxidant has a DPPH radical scavenging action.

[0035] Although the details of the substances that exhibit DPPH radical scavenging activity in the mugwort fermented liquid, the butterbur fermented liquid, the peppermint fermented liquid, and the Jerusalem artichoke fermented liquid are unknown, it is a new finding by the present inventors that the mugwort fermented liquid, the butterbur fermented liquid, the peppermint fermented liquid, and the Jerusalem artichoke fermented liquid have such excellent effects and are useful as antioxidants, which has never been known before.

[0036] <Anti-inflammatory agent> The anti-inflammatory agent of the present invention contains, as an active ingredient, at least one of the fermented liquids of mugwort by Aspergillus (mugwort fermented liquid) and the fermented liquid of peppermint by Aspergillus (peppermint fermented liquid), and further contains other components as necessary.

[0037] The mugwort fermented liquid and the peppermint fermented liquid have an inhibitory effect on hyaluronidase activity, and using this effect, they can be used as an active ingredient of an anti-inflammatory agent. Therefore, the anti-inflammatory agent has an inhibitory effect on hyaluronidase activity.

[0038] Although the details of the substances that exhibit the inhibitory effect on hyaluronidase activity in the mugwort fermented liquid and the peppermint fermented liquid are unknown, it is a new finding by the present inventors that the mugwort fermented liquid and the peppermint fermented liquid have such excellent effects and are useful as anti-inflammatory agents, which has never been known before.

[0039] <Whitening agent> The whitening agent of the present invention contains, as an active ingredient, at least one of the fermented liquids of mugwort by Aspergillus (mugwort fermented liquid), the fermented liquid of peppermint by Aspergillus (peppermint fermented liquid), and the fermented liquid of Jerusalem artichoke by Aspergillus (Jerusalem artichoke fermented liquid), and further contains other components as necessary.

[0040] The mugwort plant fermentation broth, Artemisia princeps Pamp. fermentation broth, and Hieracium pilosella L. fermentation broth have at least one of the effects of inhibiting tyrosinase activity and suppressing melanin production, and using this effect, they can be used as an active ingredient of a whitening agent. Therefore, the whitening agent has at least one of the effects of inhibiting tyrosinase activity and suppressing melanin production.

[0041] Details of the substance that exhibits at least any one of the tyrosinase activity inhibitory action and melanin production inhibitory action possessed by the mugwort plant fermentation broth, Artemisia princeps Pamp. fermentation broth, and Hieracium pilosella L. fermentation broth are unknown, but the fact that the mugwort plant fermentation broth and the Hieracium pilosella L. fermentation broth have such excellent effects and are useful as a whitening agent has never been known conventionally and is a new finding by the present inventors.

[0042] <<Mugwort plant fermentation broth>> The mugwort plant fermentation broth is a fermentation broth of a plant belonging to the genus Artemisia (hereinafter sometimes referred to as "mugwort plant") by Aspergillus.

[0043] - Mugwort plant - The mugwort plant used as the fermentation raw material is a perennial herb belonging to the family Asteraceae ( Compositae ) and the genus Artemisia ( Artemisia ), and has been used as a raw material for food and medicine since ancient times. It grows wild or is cultivated widely in Japan, such as Hokkaido, Honshu, Shikoku, and Kyushu, and is easily available from these regions.

[0044] The type of the mugwort plant is 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 Examples include Waldst. et kit. etc. These may be used alone or in combination of two or more. There are no particular restrictions on the method for obtaining the Artemisia plants, and they can be appropriately selected according to the purpose. They may be collected from nature or commercial products may be used.

[0045] There are no particular restrictions on the part of the Artemisia plant used as the fermentation raw material, and it can be appropriately selected according to the purpose. For example, above-ground parts such as flowers, buds, fruits, pericarp, seeds, seed coats, stems, leaves, branches, and branch leaves; underground parts such as roots and rhizomes, etc. may be mentioned. These may be used alone or in combination of two or more. Among these, the above-ground part is preferred as the part of the Artemisia plant used.

[0046] There are no particular restrictions on the size of the Artemisia plant used as the fermentation raw material as long as it is of a size that allows the cultivation of Aspergillus koji, and it can be appropriately selected according to the purpose. For example, the size as collected, the desired size after cutting, the size pulverized into fine powder (powder), etc. may be mentioned.

[0047] There are no particular restrictions on the state of the Artemisia plant used as the fermentation raw material as long as it is in a state that allows the cultivation of Aspergillus koji, and it can be appropriately selected according to the purpose. For example, the state as collected, the dried state, the pulverized state, the juiced state, the extract state, etc. may be mentioned. Among these, in terms of the ease of action of Aspergillus koji, the state as collected, the pulverized state, the juiced state, and the extract state are preferred, and the state as collected and the pulverized state are more preferred.

[0048] There are no particular restrictions on the method for drying the Artemisia plant, and it can be appropriately selected according to the purpose. For example, methods such as drying in the sun and drying using a commonly used dryer may be mentioned.

[0049] As a method for making the mugwort plant into the pulverized state, there is no particular limitation, and it can be appropriately selected according to the purpose. For example, methods such as pulverizing with a mixer, sugar mill, power mill, jet mill, impact crusher, etc. can be mentioned.

[0050] As a method for making the mugwort plant into the juice-squeezed state, there is no particular limitation, and it can be appropriately selected according to the purpose. For example, squeezing and the like can be mentioned.

[0051] As a method for making the mugwort plant into the extract state, there is no particular limitation, and methods generally used for plant extraction can be appropriately selected according to the purpose.

[0052] The mugwort plant used as the fermentation raw material is preferably sterilized before inoculation with the koji mold. As a means for sterilizing the mugwort plant, there is no particular limitation, and it can be appropriately selected from known methods.

[0053] -Koji mold- There is no particular limitation on the koji mold for fermenting the mugwort plant, and it can be appropriately selected according to the purpose. For example, Aspergillus oryzae ( Aspergillus oryzae ), Aspergillus sojae ( Aspergillus sojae ) and other yellow koji molds; Aspergillus luchuensis ( Aspergillus luchuensis ) and other black koji molds; Aspergillus kawachii ( Aspergillus kawauchii ) and other white koji molds; mutants thereof and the like can be mentioned. These may be used alone or in combination of two or more. Among these, as the koji mold, Aspergillus oryzae ( Aspergillus oryzae ) is preferable in that it is excellent in at least one of the anti-aging action, antioxidant action, anti-inflammatory action, and whitening action. As for the method of obtaining the Aspergillus oryzae, there are no particular restrictions, and it can be appropriately selected according to the purpose. It may be collected from nature or commercial products may be used. Also, as the Aspergillus oryzae, a mother culture using rice or the like as a raw material may be used, a mother culture of Artemisia plants described later may be used, or Aspergillus oryzae cultured in a medium (agar medium, liquid medium, etc.) may be used. Among these, it is preferable to use the mother culture of Artemisia plants in terms of being excellent in at least any one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect.

[0054] Regarding the inoculation amount of the Aspergillus oryzae to the Artemisia plant used as the fermentation raw material, there are no particular restrictions as long as it is an amount capable of fermenting the Artemisia plant, and it can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, 1×10 3 cells / mL to 1×10 8 cells / mL are preferable, and when the fermentation raw material is in a solid state, 1×10 3 cells / g to 1×10 8 cells / g are preferable.

[0055] When inoculating the Aspergillus oryzae to the Artemisia plant, it is preferable to add water. The amount of water added has no particular restrictions and can be appropriately selected according to the purpose. However, it is preferably added in an amount of 500 parts by mass to 5,000 parts by mass, more preferably 1,000 parts by mass to 4,000 parts by mass, and particularly preferably 1,500 parts by mass to 3,000 parts by mass with respect to 100 parts by mass of the Artemisia plant.

[0056] Regarding the temperature of the fermentation (cultivation), there are no particular restrictions as long as it is within the temperature range capable of fermentation by the Aspergillus oryzae, and it can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the fermentation temperature is less than 20°C, the Artemisia plant cannot be sufficiently fermented, and at least any one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient. In addition, if it exceeds 50°C, the Aspergillus oryzae may not be able to grow.

[0057] The time of the fermentation (cultivation) is not particularly limited and can be appropriately selected according to the purpose. However, 10 hours to 40 hours is preferred, and 20 hours to 30 hours is more preferred. If the fermentation time is less than 10 hours, the mugwort plants cannot be sufficiently fermented, and at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient.

[0058] The method for stopping the fermentation (cultivation) is not particularly limited and can be appropriately selected according to the purpose. Examples include heating. The heating temperature for stopping the fermentation is not particularly limited as long as it is a temperature at which the Aspergillus cannot grow, and can be appropriately selected according to the purpose. However, 50°C or higher is preferred, 70°C or higher is more preferred, and 100°C to 130°C is particularly preferred. If the heating temperature is less than 50°C, the fermentation may not be stopped. If it exceeds 130°C, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient. The heating time for stopping the fermentation is not particularly limited as long as the Aspergillus can be made unable to grow, and can be appropriately selected according to the purpose. However, 5 minutes or more is preferred, and 10 minutes to 20 minutes is more preferred. 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 effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient.

[0059] Note that the fermented liquid of the mugwort plants after stopping the fermentation is preferably cooled. The cooling method is not particularly limited and can be appropriately selected according to the purpose. Examples include leaving it to stand at room temperature, in a refrigerator, etc.

[0060] The number of times of fermentation of the mugwort plants by the Aspergillus, that is, the number of times of performing the fermentation, is not particularly limited and can be appropriately selected according to the purpose. It may be once or multiple times.

[0061] When the fermentation is carried out multiple times, the Aspergillus may be inoculated only for the first time, only for several times, or for all times, but it is preferable to inoculate only for the first time. When the fermentation is carried out multiple times, the fermentation temperature and fermentation time may be different from each other or the same.

[0062] --Artemisia Species Koji-- The Artemisia species koji is obtained by using the Artemisia plant as a koji raw material, inoculating the koji raw material with Aspergillus, and allowing a sufficient amount of spores to grow on the Artemisia plant. By using this in the fermentation, it is advantageous in that an Artemisia plant fermentation broth with excellent skin compatibility can be obtained more efficiently and easily.

[0063] The Artemisia plant used as the koji raw material can be the same as that described in the --Artemisia plant--, and the aspects such as the used part, size, and state of the Artemisia plant are also the same.

[0064] As the Aspergillus used in the production of the Artemisia species koji, the same ones as those described in the --Aspergillus-- can be used.

[0065] The inoculation amount of the Aspergillus to the Artemisia plant used as the koji raw material is not particularly limited and can be appropriately selected according to the purpose. However, with respect to 100 parts by mass of the Artemisia plant, Aspergillus suspended in sterilized water (1×10 3 cells / mL to 1×10 8 cells / mL) is preferably inoculated in an amount of 5 parts by mass to 100 parts by mass, more preferably 10 parts by mass to 50 parts by mass, and particularly preferably 20 parts by mass to 30 parts by mass. If the inoculation amount of the Aspergillus with respect to 100 parts by mass of the Artemisia plant is less than 5 parts by mass, a sufficient amount of spores may not be able to grow on the Artemisia plant, and if it exceeds 100 parts by mass, abnormal growth may occur due to excessive moisture.

[0066] When inoculating the mugwort genus plant used as the starter koji raw material with the koji mold, it is preferable to add water. The amount of water added is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably added in an amount of 10 parts by mass to 250 parts by mass, more preferably 20 parts by mass to 200 parts by mass, and particularly preferably 30 parts by mass to 150 parts by mass based on 100 parts by mass of the mugwort genus plant. If the amount of water added based on 100 parts by mass of the mugwort genus plant is less than 10 parts by mass, a sufficient amount of spores may not be able to germinate on the mugwort genus plant.

[0067] The temperature of the culture is not particularly limited as long as it is within the range in which the koji mold can grow and can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the culture is less than 20°C, a sufficient amount of spores may not be able to germinate on the mugwort genus plant. In addition, if it exceeds 50°C, the koji mold may not be able to grow.

[0068] The time of the culture is not particularly limited and can be appropriately selected according to the purpose. However, 80 hours to 210 hours is preferable, 100 hours to 190 hours is more preferable, and 120 hours to 170 hours is particularly preferable. If the time of the culture is less than 80 hours, a sufficient amount of spores may not be able to germinate on the mugwort genus plant, and if it exceeds 210 hours, the germination rate of the spores may decrease.

[0069] The mugwort genus plant fermentation broth may contain the cells of the koji mold or may be one from which the cells of the koji mold have been removed, but it is preferably one from which the cells of the koji mold have been removed.

[0070] The state of the mugwort fermented liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it may be the mugwort fermented liquid itself, a purified product of the mugwort fermented liquid, a concentrated product of the mugwort fermented liquid, a diluted product of the mugwort fermented liquid, etc. Further, the mugwort fermented liquid may be a product obtained by remixing or dissolving the dried product of the mugwort fermented liquid in a solvent such as water or a hydrophilic solvent again.

[0071] The purified product of the mugwort fermented liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it includes products from which solids in the mugwort fermented liquid (such as the plant body of the mugwort, the cells of Aspergillus oryzae, koji, etc.) are removed. The means for the removal is not particularly limited and can be appropriately selected according to the purpose. For example, filtration, etc. can be mentioned. The method of the filtration is not particularly limited and can be appropriately selected from known methods according to the purpose.

[0072] The diluted product of the mugwort fermented liquid and the concentrated product of the mugwort fermented liquid are not particularly limited and can be appropriately selected according to the purpose. For example, it includes products in which the mugwort fermented liquid is adjusted to a desired concentration. The means for the dilution is not particularly limited and can be appropriately selected from known methods according to the purpose. The means for the concentration is not particularly limited and can be appropriately selected according to the purpose. For example, vacuum concentration, etc. can be mentioned.

[0073] The dried product of the mugwort fermented liquid is not particularly limited and can be appropriately selected according to the purpose. For example, it includes products obtained by drying the mugwort fermented liquid. The means for the drying is not particularly limited and can be appropriately selected according to the purpose. For example, freeze-drying, etc. can be mentioned.

[0074] The mugwort fermented liquid is not particularly limited as long as it is a fermented liquid of mugwort by Aspergillus, and can be appropriately selected according to the purpose. However, in terms of good skin compatibility, a mugwort fermented liquid with a contact angle of 81° or less is preferable, and a mugwort fermented liquid with a contact angle of 78° or less is more preferable.

[0075] In this specification, the contact angle means a value obtained by dropping 3 μL of a measurement sample onto the sample stage of the above device using a dynamic contact angle / surface tension measurement device (FTA1000 Falcon, manufactured by First Ten Angstroms), and performing measurement by the liquid drop method under the conditions of a temperature of 22°C and a relative humidity of 20%, and obtaining the contact angle θ (°) at 1,000 ms by the θ / 2 method. The contact angle is used as an index representing "wetting", and is defined as "the angle formed between the liquid surface and the solid surface at the place where the free surface of the static liquid contacts the solid wall (taking the angle inside the liquid)" (see the 4th edition of the Rika-Kagaku Jiten, Iwanami Shoten, Ltd.). The contact angle is determined by the magnitude relationship between the cohesive force between liquid molecules and the adhesive force between the solid wall. When the liquid wets the solid (the adhesive force is large), it is an acute angle, and when it does not wet, it is an obtuse angle. Therefore, the smaller the contact angle, the easier it is to wet, that is, the better the skin compatibility. Therefore, there is no particular limitation on the lower limit of the contact angle of the mugwort fermented liquid, and it can be appropriately selected according to the purpose.

[0076] <<Perilla frutescens fermented liquid>> The Perilla frutescens fermented liquid is a fermented liquid of Perilla frutescens by Aspergillus.

[0077] -Perilla frutescens- The Perilla frutescens used as the fermentation raw material ( Thymus vulgaris Linne) belongs to the family Lamiaceae ( Labiatae ) and the genus Perilla frutescens ( Thymus ) and is a perennial woody plant, a type of herb, and has been used as a raw material for food and medicine since ancient times. It has aliases such as common thyme. Its origin is the Mediterranean coast, but it also grows wild or is cultivated in Japan, and is easily available from these regions. There are no particular restrictions on the method for obtaining the *Tachigali* plant, and it can be appropriately selected according to the purpose. It may be collected from nature or commercial products may be used.

[0078] There are no particular restrictions on the part of the *Tachigali* plant used as the fermentation raw material, and it can be appropriately selected according to the purpose. For example, above-ground parts such as flowers, flower buds, fruits, fruit peels, seeds, seed coats, stems, leaves, branches, tree barks, trunks, and branches and leaves; underground parts such as roots and rhizomes, etc. can be mentioned. These may be used alone or in combination of two or more. Among these, the above-ground part is preferred as the part of the *Tachigali* plant used.

[0079] There are no particular restrictions on the size of the *Tachigali* plant used as the fermentation raw material as long as it is of a size that allows the cultivation of the koji mold, and it can be appropriately selected according to the purpose. For example, the size as collected, the desired size after cutting, the size pulverized into fine powder (powder), etc. can be mentioned.

[0080] There are no particular restrictions on the state of the *Tachigali* plant used as the fermentation raw material as long as it is in a state that allows the cultivation of the koji mold, and it can be appropriately selected according to the purpose. For example, the state as collected, the dried state, the pulverized state, the squeezed juice state, the extract state, etc. can be mentioned. Among these, in terms of the ease of action of the koji mold, the state as collected, the pulverized state, the squeezed juice state, and the extract state are preferred, and the state as collected and the pulverized state are more preferred.

[0081] There are no particular restrictions on the method for drying the *Tachigali* plant, and it can be appropriately selected according to the purpose. For example, methods such as drying in the sun and drying using a commonly used dryer can be mentioned.

[0082] As a method for putting the tatijakousou into the pulverized state, there are no particular restrictions, and it can be appropriately selected according to the purpose. For example, methods such as pulverization using a mixer, sugar mill, power mill, jet mill, impact crusher, etc. can be mentioned.

[0083] As a method for putting the tatijakousou into the juice state, there are no particular restrictions, and it can be appropriately selected according to the purpose. For example, squeezing etc. can be mentioned.

[0084] As a method for putting the tatijakousou into the extract state, there are no particular restrictions, and methods generally used for plant extraction can be appropriately selected according to the purpose.

[0085] The tatijakousou used as the fermentation raw material is preferably sterilized before inoculation with the koji mold. As means for sterilizing the tatijakousou, there are no particular restrictions, and it can be appropriately selected from known methods.

[0086] -Koji mold- The koji mold for fermenting the tatijakousou is not particularly limited and can be appropriately selected according to the purpose. For example, those described in the <<Artemisia genus plant fermentation broth>> etc. can be mentioned. These may be used alone or in combination of two or more. Among these, as the koji mold, Aspergillus oryzae ( Aspergillus oryzae ) is preferable in that it is excellent in at least one of the anti-aging action and the antioxidant action. As a method for obtaining the koji mold, there are no particular restrictions, and it can be appropriately selected according to the purpose. It may be collected from nature or commercially available products may be used. Also, as the koji mold, a mother koji using rice etc. as a raw material may be used, a tatijakousou mother koji described later may be used, or a koji mold cultured in a medium (agar medium, liquid medium, etc.) may be used. Among these, it is preferable to use the tatijakousou mother koji in that it is excellent in at least one of the anti-aging action and the antioxidant action.

[0087] As for the inoculation amount of the koji mold to the tachijakousousou used as the fermentation raw material, there is no particular limitation as long as it can ferment the tachijakousousou, and it can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, 1×10 3 cells / mL to 1×10 8 cells / mL are preferred. When the fermentation raw material is in a solid state, 1×10 3 cells / g to 1×10 8 cells / g are preferred.

[0088] When inoculating the koji mold to the tachijakousousou, it is preferable to add water. The amount of water added is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably added in an amount of 500 parts by mass to 5,000 parts by mass, more preferably 1,000 parts by mass to 4,000 parts by mass, and particularly preferably 1,500 parts by mass to 3,000 parts by mass with respect to 100 parts by mass of the tachijakousousou.

[0089] As for the temperature of the fermentation (cultivation), there is no particular limitation as long as it is within the temperature range in which the koji mold can ferment, and it can be appropriately selected according to the purpose. However, 20°C to 40°C is preferred, and 25°C to 35°C is more preferred. If the temperature of the fermentation is less than 20°C, the tachijakousousou cannot be sufficiently fermented, and at least one of the anti-aging effect and the antioxidant effect may be insufficient.

[0090] As for the time of the fermentation (cultivation), there is no particular limitation and it can be appropriately selected according to the purpose. However, 10 hours to 40 hours is preferred, and 20 hours to 30 hours is more preferred. If the time of the fermentation is less than 10 hours, the tachijakousousou cannot be sufficiently fermented, and at least one of the anti-aging effect and the antioxidant effect may be insufficient.

[0091] As for the method of stopping the fermentation (cultivation), there is no particular limitation and it can be appropriately selected according to the purpose. For example, a method of heating can be mentioned. As the heating temperature for stopping the fermentation, there is no particular limitation as long as it is a temperature at which the Aspergillus fungus cannot grow, and it can be appropriately selected according to the purpose. However, 50°C or higher is preferable, 70°C or higher is more preferable, and 100°C to 130°C is particularly preferable. If the heating temperature is less than 50°C, the fermentation may not be stopped. If it exceeds 130°C, at least one of the anti-aging effect and the antioxidant effect may become insufficient. As the heating time for stopping the fermentation, there is no particular limitation as long as the Aspergillus fungus can be made unable to grow, and it can be appropriately selected according to the purpose. However, 5 minutes or more is preferable, and 10 minutes to 20 minutes is more preferable. 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 effect and the antioxidant effect may become insufficient.

[0092] In addition, the fermented liquid of *Tachigajo-koso* after stopping the fermentation is preferably cooled. There is no particular limitation on the cooling method, and it can be appropriately selected according to the purpose. For example, methods such as standing at room temperature or in a refrigerator can be mentioned.

[0093] As for the number of times of fermentation of *Tachigajo-koso* by the Aspergillus fungus, there is no particular limitation on the number of times of performing the fermentation, and it can be appropriately selected according to the purpose. It may be once or multiple times.

[0094] When performing the fermentation multiple times, the Aspergillus fungus may be inoculated only for the first time, only for several times, or for all times. However, it is preferable to inoculate only for the first time. When performing the fermentation multiple times, the fermentation temperature and the fermentation time may be different from each other or the same.

[0095] -- *Tachigajo-koso* starter koji -- The above-mentioned Sachiko Koso koji is prepared by using the Sachiko Koso as koji raw material, inoculating koji mold on the koji raw material, and allowing a sufficient amount of spores to grow on the Sachiko Koso. By using this in the above-mentioned fermentation, it is advantageous in that a Sachiko Koso fermentation broth with excellent skin compatibility can be obtained more efficiently and easily.

[0096] The Sachiko Koso used as the above-mentioned koji raw material can be the same as that described in the above-mentioned - Sachiko Koso -, and the aspects such as the used part, size, and state of the Sachiko Koso are also the same.

[0097] As the koji mold used in the preparation of the above-mentioned Sachiko Koso koji, the same one as that described in the above-mentioned - koji mold - can be used.

[0098] The inoculation amount of the koji mold on the Sachiko Koso used as the above-mentioned koji raw material is not particularly limited and can be appropriately selected according to the purpose. However, with respect to 100 parts by mass of the Sachiko Koso, koji mold suspended in sterilized water (1×10 3 cells / mL to 1×10 8 cells / mL) is preferably inoculated in an amount of 5 parts by mass to 100 parts by mass, more preferably 10 parts by mass to 50 parts by mass, and particularly preferably 20 parts by mass to 30 parts by mass. If the inoculation amount of the koji mold with respect to 100 parts by mass of the Sachiko Koso is less than 5 parts by mass, a sufficient amount of spores may not be able to grow on the Sachiko Koso, and if it exceeds 100 parts by mass, abnormal growth may occur due to excessive moisture.

[0099] When inoculating the koji mold to the *Tachijakokuso* used as the koji starter material, it is preferable to add water. The amount of water added is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably added in an amount of 10 parts by mass to 250 parts by mass, more preferably 20 parts by mass to 200 parts by mass, and particularly preferably 30 parts by mass to 150 parts by mass with respect to 100 parts by mass of the *Tachijakokuso*. If the amount of water added with respect to 100 parts by mass of the *Tachijakokuso* is less than 10 parts by mass, sufficient spores may not be formed on the *Tachijakokuso*.

[0100] The temperature of the culture is not particularly limited as long as it is within the range in which the koji mold can grow and can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the culture is less than 20°C, sufficient spores may not be formed on the *Tachijakokuso*. Note that if it exceeds 50°C, the koji mold may not be able to grow.

[0101] The time of the culture is not particularly limited and can be appropriately selected according to the purpose. However, 80 hours to 210 hours is preferable, 100 hours to 190 hours is more preferable, and 120 hours to 170 hours is particularly preferable. If the time of the culture is less than 80 hours, sufficient spores may not be formed on the *Tachijakokuso*. If it exceeds 210 hours, the germination rate of the spores may decrease.

[0102] The *Tachijakokuso* fermentation broth may contain the cells of the koji mold or may be one from which the cells of the koji mold have been removed. However, it is preferable that the cells of the koji mold have been removed.

[0103] The state of the fermented liquid of *Tatipteris palmata* is not particularly limited and can be appropriately selected according to the purpose. For example, it may be the fermented liquid of *Tatipteris palmata* itself, a purified product of the fermented liquid of *Tatipteris palmata*, a concentrated product of the fermented liquid of *Tatipteris palmata*, a diluted product of the fermented liquid of *Tatipteris palmata*, or the like. Further, the fermented liquid of *Tatipteris palmata* may be a product obtained by mixing or dissolving the dried product of the fermented liquid of *Tatipteris palmata* again in a solvent such as water or a hydrophilic solvent.

[0104] The purified product of the fermented liquid of *Tatipteris palmata* is not particularly limited and can be appropriately selected according to the purpose. For example, it includes a product from which solids in the fermented liquid of *Tatipteris palmata* (for example, the plant body of *Tatipteris palmata*, the cells of Aspergillus, koji, etc.) have been removed. The means for the removal is not particularly limited and can be appropriately selected according to the purpose. For example, filtration and the like are included. The method of the filtration is not particularly limited and can be appropriately selected from known methods according to the purpose.

[0105] The diluted product of the fermented liquid of *Tatipteris palmata* and the concentrated product of the fermented liquid of *Tatipteris palmata* are not particularly limited and can be appropriately selected according to the purpose. For example, it includes a product in which the fermented liquid of *Tatipteris palmata* has been adjusted to a desired concentration. The means for the dilution is not particularly limited and can be appropriately selected from known methods according to the purpose. The means for the concentration is not particularly limited and can be appropriately selected according to the purpose. For example, concentration under reduced pressure and the like are included.

[0106] The dried product of the fermented liquid of *Tatipteris palmata* is not particularly limited and can be appropriately selected according to the purpose. For example, it includes a product obtained by drying the fermented liquid of *Tatipteris palmata*. The means for the drying is not particularly limited and can be appropriately selected according to the purpose. For example, freeze-drying and the like are included.

[0107] The above-mentioned Fermented Liquid of Perilla frutescens var. acuta is not particularly limited as long as it is a fermented liquid by Aspergillus oryzae of Perilla frutescens var. acuta, and can be appropriately selected according to the purpose. However, in terms of good skin compatibility, a Fermented Liquid of Perilla frutescens var. acuta with a contact angle of 87° or less is preferred, and a Fermented Liquid of Perilla frutescens var. acuta with a contact angle of 81° or less is more preferred.

[0108] <<Fermented Liquid of Mentha spicata>> The above-mentioned Fermented Liquid of Mentha spicata is a fermented liquid by Aspergillus oryzae of Mentha spicata.

[0109] -Mentha spicata- The Mentha spicata used as the above-mentioned fermentation raw material ( Melissa officinalis Linne) is a perennial herb belonging to the genus Mentha of the Lamiaceae family ( Labiatae ). It is a kind of herb and has been used as a raw material for food and medicine since ancient times. It has aliases such as Lemon Balm and European Mountain Mint. Its origin is southern Europe, but it also grows wild or is cultivated in Japan and is easily available from these regions. Melissa The method for obtaining the above-mentioned Mentha spicata is not particularly limited and can be appropriately selected according to the purpose. It may be collected from nature or commercial products may be used.

[0110] The part of the above-mentioned Mentha spicata used as the fermentation raw material is not particularly limited and can be appropriately selected according to the purpose. For example, above-ground parts such as flowers, buds, fruits, pericarp, seeds, seed coats, stems, leaves, branches, and branch leaves; underground parts such as roots and rhizomes, etc. These may be used alone or in combination of two or more. Among these, the above-ground part is preferred as the part of the Mentha spicata used.

[0111] The size of the above-mentioned Mentha spicata used as the fermentation raw material is not particularly limited as long as it is a size that allows the culture of the Aspergillus oryzae, and can be appropriately selected according to the purpose. For example, the size as it is collected, the desired size after cutting, the size pulverized into fine powder (powder), etc. may be mentioned.

[0112] As for the state of the perilla mint used as the fermentation raw material, there is no particular limitation as long as it is in a state in which the koji mold can be cultured, and it can be appropriately selected according to the purpose. For example, it can be in the state of being collected as it is, dried, pulverized, juiced, or in the state of an extract. Among these, in terms of the ease of action of the koji mold, the state of being collected as it is, pulverized, juiced, or in the state of an extract is preferable, and the state of being collected as it is and pulverized is more preferable.

[0113] There is no particular limitation on the method of drying the perilla mint, and it can be appropriately selected according to the purpose. For example, methods such as drying in the sun and drying using a commonly used dryer can be mentioned.

[0114] There is no particular limitation on the method of pulverizing the perilla mint, and it can be appropriately selected according to the purpose. For example, methods such as pulverizing with a mixer, sugar mill, power mill, jet mill, impact crusher, etc. can be mentioned.

[0115] There is no particular limitation on the method of juicing the perilla mint, and it can be appropriately selected according to the purpose. For example, pressing can be mentioned.

[0116] There is no particular limitation on the method of making the perilla mint into the state of an extract, and a method generally used for plant extraction can be appropriately selected according to the purpose.

[0117] The perilla mint used as the fermentation raw material is preferably sterilized before inoculation with the koji mold. There is no particular limitation on the means of sterilizing the perilla mint, and it can be appropriately selected from known methods.

[0118] -Koji mold- As the koji mold for fermenting the Artemisia princeps Pamp., there are no particular restrictions, and it can be appropriately selected according to the purpose. For example, those described in the <<Artemisia princeps Pamp. fermentation broth>> can be mentioned. These may be used alone or in combination of two or more. Among these, as the koji mold, Aspergillus oryzae ( Aspergillus oryzae ) is preferable in that it is excellent in at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect. There are no particular restrictions on the method for obtaining the koji mold, and it can be appropriately selected according to the purpose. It may be collected from nature or commercially available products may be used. Also, as the koji mold, a mother koji using rice or the like as a raw material may be used, the Artemisia princeps Pamp. mother koji described later may be used, or a koji mold cultured in a medium (agar medium, liquid medium, etc.) may be used. Among these, it is preferable to use the Artemisia princeps Pamp. mother koji in that it is excellent in at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect.

[0119] The inoculation amount of the koji mold to the Artemisia princeps Pamp. used as the fermentation raw material is not particularly limited as long as it can ferment the Artemisia princeps Pamp., and it can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, 1×10 3 cells / mL to 1×10 8 cells / mL are preferable, and when the fermentation raw material is in a solid state, 1×10 3 cells / g to 1×10 8 cells / g are preferable.

[0120] When inoculating the Artemisia princeps Pamp. with the koji mold, it is preferable to add water. The addition amount of the water is not particularly limited and can be appropriately selected according to the purpose. However, it is preferable to add 500 parts by mass to 5,000 parts by mass, more preferably 1,000 parts by mass to 4,000 parts by mass, and particularly preferably 1,500 parts by mass to 3,000 parts by mass with respect to 100 parts by mass of the Artemisia princeps Pamp.

[0121] As for the temperature of the fermentation (cultivation), there is no particular limitation as long as it is within the temperature range in which the koji mold can ferment, and it can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the fermentation is less than 20°C, the kousui hakka cannot be sufficiently fermented, and at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient.

[0122] As for the time of the fermentation (cultivation), there is no particular limitation, and it can be appropriately selected according to the purpose. However, 10 hours to 40 hours is preferable, and 20 hours to 30 hours is more preferable. If the time of the fermentation is less than 10 hours, the kousui hakka cannot be sufficiently fermented, and at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient.

[0123] As for the method of stopping the fermentation (cultivation), there is no particular limitation, and it can be appropriately selected according to the purpose. For example, a method of heating can be mentioned. As for the heating temperature for stopping the fermentation, as long as it is a temperature at which the koji mold cannot grow, there is no particular limitation, and it can be appropriately selected according to the purpose. However, 50°C or higher is preferable, 70°C or higher is more preferable, and 100°C to 130°C is particularly preferable. If the heating temperature is less than 50°C, the fermentation may not be stopped. If it exceeds 130°C, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient. As for the heating time for stopping the fermentation, as long as the koji mold can be made unable to grow, there is no particular limitation, and it can be appropriately selected according to the purpose. However, 5 minutes or more is preferable, and 10 minutes to 20 minutes is more preferable. 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 effect, antioxidant effect, anti-inflammatory effect, and whitening effect may be insufficient.

[0124] Incidentally, the fermented liquid of *Artemisia princeps* Pamp. after stopping the fermentation is preferably cooled. The cooling method is not particularly limited and can be appropriately selected according to the purpose. For example, methods such as standing at room temperature or in a refrigerator can be mentioned.

[0125] The number of times of fermentation of *Artemisia princeps* Pamp. by the koji mold is not particularly limited as the number of times of performing the fermentation and can be appropriately selected according to the purpose. It may be once or multiple times.

[0126] When performing the fermentation multiple times, the koji mold may be inoculated only for the first time, only for several times, or for all times, but it is preferable to inoculate only for the first time. When performing the fermentation multiple times, the fermentation temperature and fermentation time may be different from each other or the same.

[0127] --*Artemisia princeps* Pamp. starter koji-- The *Artemisia princeps* Pamp. starter koji uses *Artemisia princeps* Pamp. as a starter koji raw material, inoculates the koji mold into the starter koji raw material, and allows a sufficient amount of spores to grow on the *Artemisia princeps* Pamp. By using this in the fermentation, it is advantageous in that a fermented liquid of *Artemisia princeps* Pamp. with excellent skin compatibility can be obtained more efficiently and easily.

[0128] The *Artemisia princeps* Pamp. used as the starter koji raw material can be the same as that described in the --*Artemisia princeps* Pamp.--, and the aspects such as the use part, size, and state of the *Artemisia princeps* Pamp. are also the same.

[0129] The koji mold used in the production of the *Artemisia princeps* Pamp. starter koji can be the same as that described in the --koji mold--.

[0130] The inoculation amount of the koji mold onto the *Artemisia princeps* Pamp. used as the starter koji raw material is not particularly limited and can be appropriately selected according to the purpose. However, relative to 100 parts by mass of the *Artemisia princeps* Pamp., the koji mold suspended in sterilized water (1×103 cells / mL to 1×10 8 It is preferable to inoculate 5 to 100 parts by mass, more preferably 10 to 50 parts by mass, and particularly preferably 20 to 30 parts by mass. If the inoculation amount of the Aspergillus oryzae with respect to 100 parts by mass of the Perilla frutescens is less than 5 parts by mass, a sufficient amount of spores may not be formed on the Perilla frutescens. If it exceeds 100 parts by mass, excessive moisture may cause abnormal growth.

[0131] When inoculating the Aspergillus oryzae onto the Perilla frutescens used as the seed koji raw material, it is preferable to add water. The addition amount of the water is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably 10 to 250 parts by mass, more preferably 20 to 200 parts by mass, and particularly preferably 30 to 150 parts by mass with respect to 100 parts by mass of the Perilla frutescens. If the addition amount of the water with respect to 100 parts by mass of the Perilla frutescens is less than 10 parts by mass, a sufficient amount of spores may not be formed on the Perilla frutescens.

[0132] The temperature of the culture is not particularly limited as long as it is within the range in which the Aspergillus oryzae can grow and can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the culture is less than 20°C, a sufficient amount of spores may not be formed on the Perilla frutescens. If it exceeds 50°C, the Aspergillus oryzae may not be able to grow.

[0133] The time of the culture is not particularly limited and can be appropriately selected according to the purpose. However, 80 to 210 hours is preferable, 100 to 190 hours is more preferable, and 120 to 170 hours is particularly preferable. If the time of the culture is less than 80 hours, a sufficient amount of spores may not be formed on the Perilla frutescens. If it exceeds 210 hours, the germination rate of the spores may decrease.

[0134] The kou sui hakka fermentation broth may contain the cells of the koji mold, or may be the one from which the cells of the koji mold have been removed, but it is preferably the one from which the cells of the koji mold have been removed.

[0135] The state of the kou sui hakka fermentation broth is not particularly limited and can be appropriately selected according to the purpose. For example, it may be the kou sui hakka fermentation broth itself, a purified product of the kou sui hakka fermentation broth, a concentrated product of the kou sui hakka fermentation broth, a diluted product of the kou sui hakka fermentation broth, etc. Further, the kou sui hakka fermentation broth may be a product obtained by remixing or dissolving the dried product of the kou sui hakka fermentation broth in a solvent such as water or a hydrophilic solvent again.

[0136] The purified product of the kou sui hakka fermentation broth is not particularly limited and can be appropriately selected according to the purpose. For example, it includes products from which solids in the kou sui hakka fermentation broth (such as the plant body of the kou sui hakka, the cells of the koji mold, olives, etc.) have been removed. The means for the removal is not particularly limited and can be appropriately selected according to the purpose. For example, filtration and the like can be mentioned. The method of the filtration is not particularly limited and can be appropriately selected from known methods according to the purpose.

[0137] The diluted product of the kou sui hakka fermentation broth and the concentrated product of the kou sui hakka fermentation broth are not particularly limited and can be appropriately selected according to the purpose. For example, it includes products in which the kou sui hakka fermentation broth has been adjusted to a desired concentration. The means for the dilution is not particularly limited and can be appropriately selected from known methods according to the purpose. The means for the concentration is not particularly limited and can be appropriately selected according to the purpose. For example, concentration under reduced pressure and the like can be mentioned.

[0138] The dried product of the kou sui hakka fermentation broth is not particularly limited and can be appropriately selected according to the purpose. For example, it includes products obtained by drying the kou sui hakka fermentation broth. The means of drying is not particularly limited and can be appropriately selected according to the purpose. For example, freeze-drying can be mentioned.

[0139] The Kosui Hakka fermentation broth is not particularly limited as long as it is a fermentation broth of Kosui Hakka by Aspergillus koji, and can be appropriately selected according to the purpose. However, in terms of good skin compatibility, a Kosui Hakka fermentation broth with a contact angle of 85° or less is preferred, and a Kosui Hakka fermentation broth with a contact angle of 79° or less is more preferred.

[0140] <<Yaguruma Kiku Fermentation Broth>> The Yaguruma Kiku fermentation broth is a fermentation broth of Yaguruma Kiku by Aspergillus koji.

[0141] -Yaguruma Kiku- The Yaguruma Kiku used as the fermentation raw material ( Centaurea cyanus Linne) is an annual herb belonging to the Asteraceae ( Compositae ) Yaguruma Kiku genus ( Centaurea ), and has been used as a raw material for food and medicine since ancient times. Its aliases include Yaguruma Sō, Centaurea, and Centaurea. Its origin is Europe, but it also grows wild or is cultivated in Japan, and is easily available from these regions. The method of obtaining the Yaguruma Kiku is not particularly limited and can be appropriately selected according to the purpose. It may be collected from nature or commercial products may be used.

[0142] The part of the Yaguruma Kiku used as the fermentation raw material is not particularly limited and can be appropriately selected according to the purpose. For example, above-ground parts such as flowers, buds, fruits, fruit peels, seeds, seed coats, stems, leaves, branches, and branch leaves; underground parts such as roots and rhizomes can be mentioned. These may be used alone or in combination of two or more. Among these, the above-ground part is preferred as the part of the Yaguruma Kiku used.

[0143] The size of the safflower to be used as the fermentation raw material is not particularly limited as long as it allows for the cultivation of Aspergillus oryzae, and can be appropriately selected according to the purpose. For example, it can be the size as collected, the desired size after cutting, the size pulverized into fine powder (powder), etc.

[0144] The state of the safflower to be used as the fermentation raw material is not particularly limited as long as it allows for the cultivation of Aspergillus oryzae, and can be appropriately selected according to the purpose. For example, it can be the state as collected, the dried state, the pulverized state, the juiced state, the extract state, etc. Among these, in terms of the ease of action of Aspergillus oryzae, the state as collected, the pulverized state, the juiced state, and the extract state are preferred, and the state as collected and the pulverized state are more preferred.

[0145] The method for drying the safflower is not particularly limited and can be appropriately selected according to the purpose. For example, it can be a method of drying in the sun, a method of drying using a commonly used dryer, etc.

[0146] The method for pulverizing the safflower is not particularly limited and can be appropriately selected according to the purpose. For example, it can be a method of pulverizing using a mixer, a sugar mill, a power mill, a jet mill, an impact crusher, etc.

[0147] The method for juicing the safflower is not particularly limited and can be appropriately selected according to the purpose. For example, it can be pressing, etc.

[0148] The method for making the safflower into the extract state is not particularly limited, and the methods generally used for plant extraction can be appropriately selected according to the purpose.

[0149] The Youngia japonica used as the fermentation raw material is preferably sterilized before inoculation with the koji mold. The means for sterilizing the Youngia japonica is not particularly limited and can be appropriately selected from known methods.

[0150] -Koji mold- The koji mold used for fermenting the Youngia japonica is not particularly limited and can be appropriately selected according to the purpose. For example, those described in the <<Fermentation broth of Artemisia plants>> can be mentioned. These may be used alone or in combination of two or more. Among these, as the koji mold, Aspergillus oryzae ( Aspergillus oryzae ) is preferable in that it is excellent in at least one of the anti-aging effect, antioxidant effect, and whitening effect. The method for obtaining the koji mold is not particularly limited and can be appropriately selected according to the purpose. It may be collected from nature or commercially available products may be used. Also, as the koji mold, a seed koji using rice or the like as a raw material may be used, the Youngia japonica seed koji described later may be used, or the koji mold cultured in a medium (agar medium, liquid medium, etc.) may be used. Among these, it is preferable to use the Youngia japonica seed koji in that it is excellent in at least one of the anti-aging effect, antioxidant effect, and whitening effect.

[0151] The inoculation amount of the koji mold to the Youngia japonica used as the fermentation raw material is not particularly limited as long as it can ferment the Youngia japonica and can be appropriately selected according to the purpose. However, when the fermentation raw material is in a liquid state, 1×10 3 cells / mL to 1×10 8 cells / mL are preferable, and when the fermentation raw material is in a solid state, 1×10 3 cells / g to 1×10 8 cells / g are preferable.

[0152] When inoculating the Cirsium japonicum var. australe with the Aspergillus oryzae, it is preferable to add water. The amount of water added is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably added in an amount of 500 parts by mass to 5,000 parts by mass, more preferably 1,000 parts by mass to 4,000 parts by mass, and particularly preferably 1,500 parts by mass to 3,000 parts by mass based on 100 parts by mass of the Cirsium japonicum var. australe.

[0153] The temperature of the fermentation (cultivation) is not particularly limited as long as it is within the range of temperatures at which the Aspergillus oryzae can ferment and can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the fermentation is less than 20°C, the Cirsium japonicum var. australe cannot be sufficiently fermented, and at least one of the anti-aging effect, antioxidant effect, and whitening effect may be insufficient.

[0154] The time of the fermentation (cultivation) is not particularly limited and can be appropriately selected according to the purpose. However, 10 hours to 40 hours is preferable, and 20 hours to 30 hours is more preferable. If the time of the fermentation is less than 10 hours, the Cirsium japonicum var. australe cannot be sufficiently fermented, and at least one of the anti-aging effect, antioxidant effect, and whitening effect may be insufficient.

[0155] The method for stopping the fermentation (cultivation) is not particularly limited and can be appropriately selected according to the purpose. Examples include a method of heating. The heating temperature for stopping the fermentation is not particularly limited as long as it is a temperature at which the Aspergillus oryzae cannot grow and can be appropriately selected according to the purpose. However, 50°C or higher is preferable, 70°C or higher is more preferable, and 100°C to 130°C is particularly preferable. If the heating temperature is less than 50°C, the fermentation may not be stopped, and if it exceeds 130°C, at least one of the anti-aging effect, antioxidant effect, and whitening effect may be insufficient. The heating time for stopping the fermentation is not particularly limited as long as the Aspergillus can no longer grow, and it can be appropriately selected according to the purpose. However, it is preferably 5 minutes or more, and more preferably 10 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 effect, antioxidant effect, and whitening effect may be insufficient.

[0156] In addition, the fermented liquid of Hieracium pilosella after stopping the fermentation is preferably cooled. The cooling method is not particularly limited and can be appropriately selected according to the purpose. For example, methods such as standing at room temperature or in a refrigerator can be mentioned.

[0157] The number of times of fermentation of Hieracium pilosella by the Aspergillus is not particularly limited, and can be appropriately selected according to the purpose. It may be once or multiple times.

[0158] When the fermentation is carried out multiple times, the Aspergillus may be inoculated only for the first time, only for several times, or for all times. However, it is preferably inoculated only for the first time. When the fermentation is carried out multiple times, the fermentation temperature and fermentation time may be different or the same.

[0159] --Hieracium pilosella Seed Koji-- The Hieracium pilosella seed koji is obtained by using Hieracium pilosella as a seed koji raw material, inoculating the seed koji raw material with Aspergillus, and allowing a sufficient amount of spores to grow on the Hieracium pilosella. By using this in the fermentation, it is advantageous in that a Hieracium pilosella fermented liquid with excellent skin compatibility can be obtained more efficiently and easily.

[0160] The Hieracium pilosella used as the seed koji raw material can be the same as that described in the -Hieracium pilosella-, and the aspects such as the use part, size, and state of the Hieracium pilosella are also the same.

[0161] As the koji mold used in the production of the above-mentioned Artemisia princeps var. orientalis koji, those similar to those described in the above-mentioned -koji mold- can be used.

[0162] The inoculation amount of the koji mold onto the above-mentioned Artemisia princeps var. orientalis used as the koji starter material is not particularly limited and can be appropriately selected according to the purpose. However, with respect to 100 parts by mass of the above-mentioned Artemisia princeps var. orientalis, koji mold suspended in sterilized water (1×10 3 cells / mL to 1×10 8 cells / mL) is preferably inoculated in an amount of 5 parts by mass to 100 parts by mass, more preferably 10 parts by mass to 50 parts by mass, and particularly preferably 20 parts by mass to 30 parts by mass. If the inoculation amount of the koji mold with respect to 100 parts by mass of the above-mentioned Artemisia princeps var. orientalis is less than 5 parts by mass, a sufficient amount of spores may not be able to germinate on the above-mentioned Artemisia princeps var. orientalis, and if it exceeds 100 parts by mass, excessive moisture may cause abnormal growth.

[0163] When inoculating the above-mentioned Artemisia princeps var. orientalis used as the koji starter material with the koji mold, it is preferable to add water. The addition amount of the water is not particularly limited and can be appropriately selected according to the purpose. However, with respect to 100 parts by mass of the above-mentioned Artemisia princeps var. orientalis, it is preferably added in an amount of 10 parts by mass to 250 parts by mass, more preferably 20 parts by mass to 200 parts by mass, and particularly preferably 30 parts by mass to 150 parts by mass. If the addition amount of the water with respect to 100 parts by mass of the above-mentioned Artemisia princeps var. orientalis is less than 10 parts by mass, a sufficient amount of spores may not be able to germinate on the above-mentioned Artemisia princeps var. orientalis.

[0164] The temperature of the above-mentioned culture is not particularly limited as long as it is within the temperature range in which the koji mold can grow, and can be appropriately selected according to the purpose. However, 20°C to 40°C is preferable, and 25°C to 35°C is more preferable. If the temperature of the above-mentioned culture is less than 20°C, a sufficient amount of spores may not be able to germinate on the above-mentioned Artemisia princeps var. orientalis. In addition, if it exceeds 50°C, the koji mold may not be able to grow.

[0165] The culturing time is not particularly limited and can be appropriately selected according to the purpose. However, 80 to 210 hours is preferred, 100 to 190 hours is more preferred, and 120 to 170 hours is particularly preferred. If the culturing time is less than 80 hours, a sufficient amount of spores may not be able to germinate on the Echinops sphaerocephalus, and if it exceeds 210 hours, the germination rate of the spores may decrease.

[0166] The Echinops sphaerocephalus fermentation broth may contain the cells of the Aspergillus oryzae or may be the one from which the cells of the Aspergillus oryzae have been removed. However, it is preferred that the cells of the Aspergillus oryzae have been removed.

[0167] The state of the Echinops sphaerocephalus fermentation broth is not particularly limited and can be appropriately selected according to the purpose. For example, it may be the Echinops sphaerocephalus fermentation broth itself, a purified product of the Echinops sphaerocephalus fermentation broth, a concentrated product of the Echinops sphaerocephalus fermentation broth, a diluted product of the Echinops sphaerocephalus fermentation broth, etc. Further, the Echinops sphaerocephalus fermentation broth may be a product obtained by remixing or dissolving the dried product of the Echinops sphaerocephalus fermentation broth in a solvent such as water or a hydrophilic solvent again.

[0168] The purified product of the Echinops sphaerocephalus fermentation broth is not particularly limited and can be appropriately selected according to the purpose. For example, it includes products from which solids in the Echinops sphaerocephalus fermentation broth (such as the plant body of the Echinops sphaerocephalus, the cells of the Aspergillus oryzae, olives, etc.) have been removed. The means for removal is not particularly limited and can be appropriately selected according to the purpose. For example, filtration etc. can be mentioned. The filtration method is not particularly limited and can be appropriately selected from known methods according to the purpose.

[0169] The diluted product of the Echinops sphaerocephalus fermentation broth and the concentrated product of the Echinops sphaerocephalus fermentation broth are not particularly limited and can be appropriately selected according to the purpose. For example, it includes products in which the Echinops sphaerocephalus fermentation broth has been adjusted to a desired concentration. The means of dilution is not particularly limited and can be appropriately selected from known methods according to the purpose. The means of concentration is not particularly limited and can be appropriately selected according to the purpose, and examples thereof include concentration under reduced pressure.

[0170] The dried product of the Hieracium pilosella fermentation broth is not particularly limited and can be appropriately selected according to the purpose, and examples thereof include a product obtained by drying the Hieracium pilosella fermentation broth. The means of drying is not particularly limited and can be appropriately selected according to the purpose, and examples thereof include freeze-drying.

[0171] The Hieracium pilosella fermentation broth is not particularly limited as long as it is a fermentation broth by Aspergillus of Hieracium pilosella and can be appropriately selected according to the purpose. However, in terms of good skin compatibility, a Hieracium pilosella fermentation broth having a contact angle of 85° or less is preferable, and a Hieracium pilosella fermentation broth having a contact angle of 79° or less is more preferable.

[0172] <<Other components>> The other components in the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent are not particularly limited and can be appropriately selected according to the purpose. Examples thereof include excipients, moisture-proof agents, preservatives, strengthening agents, thickening agents, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, coloring agents, fragrances, whitening agents, moisturizing agents, oily components, ultraviolet absorbers, surfactants, thickening agents, alcohols, powder components, colorants, aqueous components, water, skin nutrients, and the like. These may be used alone or in combination of two or more. The content of the other components is not particularly limited and can be appropriately selected according to the purpose.

[0173] - Use - 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. Therefore, for example, they can be suitably used as pharmaceuticals, quasi-drugs, cosmetics, food and beverages, etc., and their blending amounts, usage methods, and dosage forms can be appropriately selected according to the purpose of use.

[0174] The blending amount can be appropriately adjusted according to the physiological activity of the fermentation broth, etc. Further, the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent may be the fermentation broth itself.

[0175] The usage method is not particularly limited and can be appropriately selected according to the purpose. For example, usage methods such as oral, parenteral, and topical can be mentioned. Among these, topical use is preferred.

[0176] The dosage form is not particularly limited and can be appropriately selected according to the purpose. For example, oral dosage forms such as tablets, powders, capsules, granules, extracts, and syrups; parenteral dosage forms such as injections, infusions, and suppositories; topical agents such as lotions, emulsions, creams, ointments, beauty liquids, lotions, packs, jellies, lip creams, lipsticks, foundations, bath agents, soaps, body shampoos, astringents, hair tonics, hair creams, hair liquids, pomades, shampoos, and rinses, etc. can be mentioned.

[0177] Further, the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention can also be used as reagents for research on the mechanism of action of anti-aging, antioxidant, anti-inflammatory, or whitening effects.

[0178] The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention are suitably applicable to humans, but as long as their respective effects are exhibited, they can also be applied to animals other than humans (for example, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.).

[0179] (Cosmetics) The cosmetic of the present invention contains at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent of the present invention, and may further contain other components as needed.

[0180] <Anti-aging agent, antioxidant, anti-inflammatory agent, skin-whitening agent> The content of at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent in the cosmetic is not particularly limited and can be appropriately selected according to the purpose. However, it is preferably 5% by volume or more, more preferably 20% by volume or more, based on the total amount of the cosmetic. If the content of at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent is less than 5% by volume, at least one of the anti-aging effect, antioxidant effect, anti-inflammatory effect, and skin-whitening effect may be insufficient. Note that the content of at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent is preferably as large as possible, and the upper limit is not particularly limited and can be appropriately selected according to the purpose. Also, the cosmetic may be at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent itself.

[0181] <Other components> The cosmetic can further add various main agents, auxiliary agents, and other components usually used in the manufacture of cosmetics, as needed, within a range that does not impair the purpose and effects of the present invention. The other components are not particularly limited and can be appropriately selected according to the purpose. For example, astringents, bactericides, antibacterial agents, ultraviolet absorbers, cell activators, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc. can be mentioned. These can be used alone or in combination of two or more. When these components are used in combination with at least one selected from the group consisting of the anti-aging agent, antioxidant, anti-inflammatory agent, and skin-whitening agent, they may act synergistically to bring about excellent effects beyond the normal expectation. The content of the other components in the cosmetic is not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected according to the purpose.

[0182] <Use> The use of the cosmetic is not particularly limited, and can be appropriately selected from general cosmetics. For example, skin cosmetics such as lotion, emulsion, cream, ointment, beauty essence, lotion, pack, jelly, lip cream, lipstick, foundation, bath agent, soap, body shampoo, etc.; scalp and hair cosmetics such as astringent, hair tonic, hair cream, hair liquid, pomade, shampoo, rinse, etc.

[0183] The cosmetic may be one in which at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent is blended into an arbitrary cosmetic so as not to interfere with its activity, or may be a cosmetic having at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent as a main component. Further, the cosmetic may be at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent itself.

[0184] The cosmetic of the present invention is preferably applicable to humans, but can also be applied to animals other than humans (for example, mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective effects are exhibited.

[0185] Since the cosmetic of the present invention contains at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent, it is useful in that it exhibits at least one of excellent anti-aging, antioxidant, anti-inflammatory, and whitening effects when used on the skin.

Examples

[0186] The present invention will be specifically described below by way of production examples and test examples, but the present invention is not limited to these test examples in any way.

[0187] <Production Example 1: Preparation of mugwort fermentation liquid 1> - Seed koji preparation process - Koji mold ( Aspergillus oryzae , strain name: AOK1714, manufactured by Akita Konno Shoten Co., Ltd.) was picked up with a platinum loop and suspended in 50 mL of sterilized water to prepare a koji mold solution. When the number of bacteria in the koji mold solution was calculated using a Thoma hemocytometer (manufactured by EKDS), it was 1.0×10 5 cells / mL. Next, 10 g of mugwort (manufactured by Albion Co., Ltd.) cut into 0.5 cm to 5 cm pieces was placed in an Erlenmeyer flask, autoclaved, and 2 mL of the koji mold solution was inoculated therein, followed by static culture at 30 °C for 168 hours. After completion of the culture, it was dried at 45 °C for 24 hours to obtain "mugwort seed koji".

[0188] - Fermentation process - Mugwort (manufactured by Albion Co., Ltd.) was ground using a pulverizer (sugar mill) and passed through a 2 mm mesh screen to obtain a ground mugwort product. 1,000 mL of water was added to 50 g of this ground mugwort product and mixed, and then 20 mL of the mugwort seed koji (number of bacteria: about 1.0×10 6 cells / mL) obtained in Preparation Example 1 was inoculated. Next, it was pre-cultured at 25 °C for 22 hours. The obtained fermentation liquid was filtered using diatomaceous earth to obtain "mugwort fermentation liquid 1".

[0189] <Production Example 2: Preparation of mugwort fermentation liquid 2> In the above Production Example 1, "mugwort fermentation liquid 2" was obtained in the same manner as in Production Example 1, except that the mugwort seed koji was changed to a seed koji using rice as a raw material ( Aspergillus oryzae , Shirakami white koji, manufactured by Akita Konno Shoten Co., Ltd.) (hereinafter sometimes referred to as "rice seed koji").

[0190] <Comparative Production Example 1: Preparation of mugwort extract> The mugwort (manufactured by Albion Co., Ltd.) was pulverized using a pulverizer (sugar mill), passed through a 2 mm mesh screen, and a pulverized mugwort product was obtained. 1,000 mL of water was added to 50 g of this pulverized mugwort product, mixed, and then stirred at 25°C for 22 hours. Next, the resulting stirred product was filtered using diatomaceous earth to obtain a "mugwort extract".

[0191] (Test Example A-1: Measurement of Contact Angle) Using the mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 as test samples, the contact angle was measured by the following method. Specifically, using a dynamic contact angle and surface tension measuring device (FTA1000 Falcon, manufactured by First Ten Angstroms), 3 μL of each test sample was dropped onto the sample stage (made of aluminum) of the device, and the measurement was performed by the liquid drop method under the conditions of a temperature of 22°C and a relative humidity of 20%. The contact angle θ (°) at 1,000 ms was determined by the θ / 2 method. The measurement of the contact angle was performed 3 times, and the average value was determined. The results are shown in Table 1 below. Also, an example of the liquid drop at the time of measuring the contact angle of each test sample is shown in FIGS. 1A to 1C.

[0192]

Table 1

[0193] With respect to the mugwort extract obtained in Comparative Production Example 1, both the mugwort fermentation broth 1 obtained in Production Example 1 and the mugwort fermentation broth 2 obtained in Production Example 2 had small contact angles, which were 81° or less, and were excellent in skin affinity. Furthermore, the mugwort fermentation broth 1 obtained in Production Example 1 had a contact angle of 78° or less, and was more excellent in skin affinity.

[0194] (Test Example 1-1: Matrix Metalloproteinase-1 (MMP-1) Activity Inhibition Test) Using the mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 as test samples, the inhibitory effect on matrix metalloproteinase-1 (MMP-1) activity was tested by the following test method with a partial modification of the Wunsch and Heidrich method.

[0195] In a test tube with a lid, each test sample was dissolved in a 0.1 mol / L Tris-HCl buffer solution (pH 7.1) containing 20 mmol / L calcium chloride. Then, 50 μL of the solution of the test sample, 50 μL of an MMP-1 (COLLAGENASE Type IV from Clostridium histolyticum, manufactured by Sigma) solution, and 400 μL of a 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 a 25 mmol / L citric acid solution was added to stop the reaction. The final concentration of the test sample at this time was the concentration shown in Table 2 below. Then, 5 mL of ethyl acetate was added and shaken vigorously. This was centrifuged at 1,600×g for 10 minutes, and the absorbance at a wavelength of 320 nm of the ethyl acetate layer was measured.

[0196] Also, as a blank, the same operations and absorbance measurements as above were performed except that the MMP-1 solution (enzyme solution) was changed to a 0.1 mol / L Tris-HCl buffer solution (pH 7.1).

[0197] Furthermore, as a control, the same operations and absorbance measurements as above were performed except that the solution of the test sample was changed to a 0.1 mol / L Tris-HCl buffer solution (pH 7.1) containing 20 mmol / L calcium chloride without the test sample.

[0198] Based on the measured absorbance values obtained, the MMP-1 activity inhibition rate was calculated according to the following formula 1. The results are shown in Table 2 below. <Formula 1> MMP-1 activity inhibition rate (%) = {1 - (C - D) / (A - B)} × 100 In the above formula (1), A to D each represent the following. A: Absorbance at a wavelength of 320 nm with no test sample added and enzyme added B: Absorbance at a wavelength of 320 nm with no test sample added and no enzyme added C: Absorbance at a wavelength of 320 nm with the test sample added and enzyme added D: Absorbance at 320 nm with the test sample added and no enzyme added

[0199] [Table 2]

[0200] (Test Example 1-2: Test for the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) Using the mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 as test samples, the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA was tested by the following test method.

[0201] Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was the concentration 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 Kuraray Co., Ltd.) were cultured using a normal human epidermal keratinocyte growth medium (HuMedia-KG2, manufactured by Kuraray Co., Ltd.) at 37 °C and 5% CO2 until confluent, and then the cells were collected by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with a normal human epidermal keratinocyte growth medium (HuMedia-KG2). Next, the above NHEK (1.5×10 52 mL of cells / mL were seeded and cultured overnight at 37 °C under 5% CO2 conditions. After the culture was completed, the medium was replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2), and further cultured for 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium, and cultured for 24 hours under the conditions of 37 °C and 5% CO2. After the culture was completed, the culture solution was removed, and total RNA was extracted with a reagent for RNA extraction (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer, and total RNA was prepared to be 200 ng / μL using purified water.

[0202] Also, as a control, the same operations and absorbance measurements as above were performed except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL in the same manner as above.

[0203] Using each of the total RNAs as a template, the expression levels of hyaluronan synthase 3 (HAS3) mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, which is an internal standard, were measured. 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of HAS3 mRNA without and with the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the HAS3 mRNA expression promotion rate was calculated based on the following formula 2. The results are shown in Table 3 below. <Formula 2> HAS3 mRNA expression promotion rate (%) = A / B × 100 In Formula 2 above, A and B represent the following, respectively. A: Corrected value when the test sample is added B: Correction value when no test sample is added

[0204]

Table 3

[0205] (Test Examples 1 - 3: DPPH radical scavenging activity test) The mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 were used as test samples, and the DPPH radical scavenging activity was tested by the following test method.

[0206] Each test sample was dissolved in an ethanol solution (manufactured by Fujifilm Wako Pure Chemical Corporation) to prepare a test sample solution. 3 mL of the test sample solution was added to 3 mL of a 150 μmol / L DPPH (diphenyl - p - picrylhydrazyl) ethanol solution. Immediately, the container was sealed and shaken, and after standing for 30 minutes, the absorbance at a wavelength of 520 nm was measured. The final concentration of the test sample at this time was the concentration shown in Table 4 below.

[0207] Also, as a blank, the same operations and absorbance measurements as above were performed, except that the DPPH ethanol solution was changed to an ethanol solution not containing DPPH.

[0208] Furthermore, as a control, the same operations and absorbance measurements as above were performed, except that the test sample solution was changed to an ethanol solution (manufactured by Fujifilm Wako Pure Chemical Corporation) not containing the test sample.

[0209] Based on the measured absorbance values obtained, the DPPH radical scavenging rate was calculated according to the following formula 3. The results are shown in Table 4 below. <Formula 3> DPPH radical scavenging rate (%) = {A - (B - C)} / A × 100 In the above formula 3, A to C represent the following respectively. A: Absorbance at a wavelength of 520 nm with no test sample added and DPPH added B: Absorbance at 520 nm with test sample added and DPPH added C: Absorbance at 520 nm without test sample added and without DPPH added

[0210]

Table 4

[0211] (Test Examples 1-4: Hyaluronidase Activity Inhibition Test) The mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 were used as test samples, and the hyaluronidase activity inhibition effect was tested by the following test method.

[0212] Each test sample was dissolved in 0.1 mol / L acetic acid buffer (pH 3.5) to prepare a test sample solution. To 0.2 mL of the test sample solution, 0.1 mL of hyaluronidase solution (Type IV-S (from bovine testis), 400 NF units / mL, manufactured by SIGMA) was added, and the reaction was carried out at 37 °C for 20 minutes. Then, 0.2 mL of 2.5 mmol / L calcium chloride as an activator was added, and the reaction was further carried out at 37 °C for 20 minutes. To this, 0.5 mL of 0.8 mg / mL sodium hyaluronate solution (from rooster comb) (manufactured by Fujifilm Wako Pure Chemical Corporation) was added, and the reaction was carried out at 37 °C for 40 minutes. The final concentration of the test sample at this time was the concentration shown in Table 5 below. Then, 0.2 mL of 0.4 mol / L sodium hydroxide was added to stop the reaction. After cooling, 0.2 mL of boric acid solution was added to each reaction solution, and it was boiled for 3 minutes. After ice-cooling, 6 mL of p-DABA reagent (p-dimethylaminobenzaldehyde 10 g dissolved in a mixed solution of 10 N hydrochloric acid 12.5 mL and acetic acid 87.5 mL, diluted 10-fold with acetic acid) was added, and the reaction was carried out at 37 °C for 20 minutes. Then, the absorbance at 585 nm was measured.

[0213] Also, as a blank, the same operations and absorbance measurements as above were performed, except that the hyaluronidase solution (enzyme solution) was changed to 0.1 mol / L acetic acid buffer (pH 3.5).

[0214] Furthermore, as a control, the same operations and absorbance measurements as above were performed, except that the test sample solution was changed to 0.1 mol / L acetic acid buffer (pH 3.5) not containing the test sample.

[0215] Based on the measured absorbance values obtained, the hyaluronidase activity inhibition rate was calculated according to the following formula 4. The results are shown in Table 5 below. <Formula 4> Hyaluronidase activity inhibition rate (%) = {1 - (C - D) / (A - B)} × 100 In the above formula 4, A to D represent the following respectively. A: Absorbance at a wavelength of 585 nm without addition of the test sample and with addition of the enzyme B: Absorbance at a wavelength of 585 nm without addition of the test sample and without addition of the enzyme C: Absorbance at 585 nm with addition of the test sample and with addition of the enzyme D: Absorbance at a wavelength of 585 nm with addition of the test sample and without addition of the enzyme

[0216]

Table 5

[0217] (Test Example 1 - 5: Tyrosinase Activity Inhibition Test) Using the mugwort fermentation broth 1 obtained in Production Example 1, the mugwort fermentation broth 2 obtained in Production Example 2, and the mugwort extract obtained in Comparative Production Example 1 as test samples, the tyrosinase activity inhibition effect was tested by the following test method.

[0218] Each test sample was dissolved in a 25% DMSO solution to prepare a test sample solution. 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 test sample solution were added to a 48-well plate, and the plate was left to stand at 37° C. for 10 minutes. 0.02 mL of 800 unit / mL tyrosinase solution (manufactured by SIGMA) was added to the plate, and the plate was further reacted at 37° C. for 15 minutes. After the reaction, the absorbance at a wavelength of 475 nm was measured. The final concentration of the test sample was as shown in Table 6 below.

[0219] As a blank, the same operations and absorbance measurements were carried out as above, except that the tyrosinase solution (enzyme solution) was changed to McIlvaine buffer (pH 6.8).

[0220] Furthermore, as a control, the same operations and absorbance measurements were carried out as above, except that the test sample solution was changed to a 25% DMSO solution not containing a test sample.

[0221] From the obtained absorbance measurements, the tyrosinase activity inhibition rate was calculated according to the following formula 5. The results are shown in Table 6 below. <Formula 5> Tyrosinase activity inhibition rate (%) = {1-(CD) / (AB)} x 100 In the formula 5, A to D respectively represent the following. A: Absorbance at 475 nm without adding test sample and with adding enzyme B: Absorbance at 475 nm without test sample or enzyme C: Absorbance at 475 nm after addition of test sample and enzyme D: Absorbance at 475 nm with and without the addition of test sample and enzyme

[0222] [Table 6]

[0223] <Production Example 3: Preparation of Artemisia capillaris fermentation liquid 1> In the seed koji preparation step of the above-mentioned Production Example 1, Artemisia japonica was replaced with Artemisia capillaris (Artemisia capillaris "Kawarayomogi seed koji" was prepared in the same manner as the seed koji preparation step of Production Example 1, except that it was changed to (manufactured by Albion Co., Ltd.). Also, in the fermentation step of Production Example 1, mugwort was changed to kawarayomogi ( Artemisia capillaris "Kawarayomogi fermentation broth 1" was obtained in the same manner as the fermentation step of Production Example 1, except that it was changed to (manufactured by Albion Co., Ltd.).

[0224] <Production Example 4: Preparation of Kawarayomogi Fermentation Broth 2> In Production Example 3, the kawarayomogi seed koji was changed to rice seed koji ( Aspergillus oryzae "Kawarayomogi fermentation broth 2" was obtained in the same manner as Production Example 3, except that it was changed to (manufactured by Akita Konno Shoten Co., Ltd.).

[0225] <Comparative Production Example 2: Preparation of Kawarayomogi Extract> In Comparative Production Example 1, mugwort was changed to kawarayomogi ( Artemisia capillaris "Kawarayomogi extract" was obtained in the same manner as Comparative Production Example 1, except that it was changed to (manufactured by Albion Co., Ltd.).

[0226] (Test Example A-2: Measurement of Contact Angle) In Test Example A-1, the contact angle was measured in the same manner as Test Example A-1, except that the test sample was changed to the kawarayomogi fermentation broth 1 obtained in Production Example 3, the kawarayomogi fermentation broth 2 obtained in Production Example 2, and the kawarayomogi extract obtained in Comparative Production Example 2. The results are shown in Table 7 below. Also, an example of the liquid droplet at the time of measuring the contact angle of each test sample is shown in FIGS. 2A to 2C.

[0227]

Table 7

[0228] With respect to the mugwort extract obtained in Comparative Production Example 2, both the mugwort fermentation broth 1 obtained in Production Example 3 and the mugwort fermentation broth 2 obtained in Production Example 4 had small contact angles, 81° or less, and were excellent in skin affinity. Furthermore, the mugwort fermentation broth 1 obtained in Production Example 3 had a contact angle of 78° or less and was even more excellent in skin affinity.

[0229] (Test Example 2-1: Test for the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) In Test Example 1-2, the test sample was changed to the mugwort fermentation broth 1 obtained in Production Example 3, the mugwort fermentation broth 2 obtained in Production Example 4, and the mugwort extract obtained in Comparative Production Example 2, and the final concentration of the test sample was changed to the concentrations shown in Table 8 below. The effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA was tested in the same manner as in Test Example 1-2. The results are shown in Table 8 below.

[0230]

Table 8

[0231] (Test Example 2-2: Test for the effect of promoting the production of type I collagen) The mugwort fermentation broth 1 obtained in Production Example 3, the mugwort fermentation broth 2 obtained in Production Example 4, and the mugwort extract obtained in Comparative Production Example 2 were used as test samples, and the effect of promoting the production of type I collagen was tested by the following test method.

[0232] Each test sample was dissolved in Dulbecco's MEM (manufactured by Nissui Pharmaceutical Co., Ltd.) containing 0.25% fetal bovine serum (FBS, manufactured by Biosera) so that the final concentration was the concentration shown in Table 9 below, and a test sample-added medium was prepared. Normal human fibroblasts (NB1RGB, purchased from RIKEN BRC) were cultured using DMEM containing 10% FBS until confluent under the conditions of 37°C and 5% CO2, and then the cells were recovered by trypsin treatment. This was adjusted to 1.6 × 10 5Adjusted to cells / mL. Next, the NB1RGB (1.6×10 5 cells / mL) was seeded at 100 μL per well in a 96-well microplate and cultured overnight under the conditions of 37 °C and 5% CO2. After the culture was completed, the medium was replaced with 100 μL of the test sample-added medium, and the cells were cultured for 3 days under the conditions of 37 °C and 5% CO2. After the culture was completed, the amount of type I collagen in the medium of each well was measured by the ELISA method.

[0233] Specifically, 90 μL of the culture supernatant was transferred to an ELISA plate and adsorbed onto the plate overnight at 4 °C. Then, the solution was discarded, and washing was performed with phosphate-buffered saline containing 0.05% Tween-20 (PBS-T). Thereafter, blocking was performed with phosphate-buffered saline containing 1% FBS. The solution was discarded, and washing was performed with phosphate-buffered saline containing 0.05% Tween-20 (PBS-T), and then reacted with an anti-human collagen type I antibody (rabbit IgG, manufactured by Chemicon). The solution was discarded, and washing was performed with phosphate-buffered saline containing 0.05% Tween-20 (PBS-T), and then reacted with an HRP-labeled anti-rabbit IgG antibody. After that, the same washing operation was performed, and a color reaction was carried out. The promotion rate of type I collagen production was calculated by performing the above ELISA using a standard product to create a calibration curve.

[0234] In addition, as a control, the test sample solution was changed to 0.25% FBS-containing DMEM without the test sample, and the same operations as above and measurement by the ELISA method were performed.

[0235] Based on the obtained measured values, the promotion rate of type I collagen production was calculated according to the following formula 6. The results are shown in Table 9 below. <Formula 6> Promotion rate of type I collagen production (%) = A / B × 100 In Formula 6 above, A and B represent the following, respectively. A: Amount of type I collagen when the test sample is added B: Amount of type I collagen when the test sample is not added

[0236]

Table 9

[0237] (Test Example 2-3: Claudin-1 mRNA Expression Promotion Test) Using the mugwort fermentation broth 1 obtained in Production Example 3, the mugwort fermentation broth 2 obtained in Production Example 4, and the mugwort extract obtained in Comparative Production Example 2 as test samples, the promoting effect on Claudin-1 mRNA expression was tested by the following test method.

[0238] Each test sample was dissolved in normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was the concentration shown in Table 10 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kuraray Co., Ltd.) were cultured using normal human epidermal keratinocyte growth medium (HuMedia-KG2, manufactured by Kuraray Co., Ltd.) at 37°C under 5% CO2 until confluent, and then the cells were collected by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with normal human epidermal keratinocyte growth medium (HuMedia-KG2). Next, the NHEK (1.5×10 5 cells / mL) was seeded at 2 mL in a 35 mm petri dish and cultured overnight at 37°C under 5% CO2. After the culture was completed, the medium was replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2), and further cultured for 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium and cultured at 37°C under 5% CO2 for 24 hours. After the culture was completed, the culture solution 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, and the total RNA was prepared to be 200 ng / μL using purified water.

[0239] As a control, the same operations and absorbance measurements as above were performed, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL in the same manner as above.

[0240] Using each of the total RNAs as a template, the expression levels of claudin-1 mRNA and GAPDH mRNA, which is an internal standard, were measured. 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of claudin-1 mRNA without the test sample and with the test sample added were corrected by the expression level of GAPDH mRNA. From this corrected value, the promotion rate of claudin-1 mRNA expression was calculated based on the following formula 7. The results are shown in Table 10 below. <Formula 7> Promotion rate of claudin-1 mRNA expression (%) = A / B × 100 In Formula 7 above, A and B represent the following, respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0241]

Table 10

[0242] (Test Example 2-4: Test for promoting claudin-4 mRNA expression) Using the mugwort fermentation broth 1 obtained in Production Example 3, the mugwort fermentation broth 2 obtained in Production Example 4, and the mugwort extract obtained in Comparative Production Example 2 as test samples, the effect of promoting claudin-4 mRNA expression was tested by the following test method.

[0243] Each test sample was dissolved in normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was as shown in Table 11 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kuraray Co., Ltd.) were cultured using normal human epidermal keratinocyte growth medium (HuMedia-KG2, manufactured by Kuraray Co., Ltd.) at 37 °C under 5% CO2 until confluent, and then the cells were collected by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with normal human epidermal keratinocyte growth medium (HuMedia-KG2). Next, 2 mL of the NHEK (1.5×10 5 cells / mL) was seeded in a 35 mm petri dish and cultured overnight at 37 °C under 5% CO2. After the culture was completed, the medium was replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2), and further cultured for 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium and cultured at 37 °C under 5% CO2 for 24 hours. After the culture was completed, the culture solution was removed, total RNA was extracted with a reagent for RNA extraction (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), the amount of each RNA was measured with a spectrophotometer, and total RNA was prepared to be 200 ng / μL using purified water.

[0244] Also, as a control, the same operations and absorbance measurements as above were performed except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL in the same manner as above.

[0245] Using each total RNA as a template, the expression levels of claudin-4 mRNA and GAPDH mRNA, which is an internal standard, were measured. 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of claudin-4 mRNA without addition of the test sample and with addition of the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the promotion rate of claudin-4 mRNA expression was calculated based on the following formula 8. The results are shown in Table 11 below. <Formula 8> Promotion rate of claudin-4 mRNA expression (%) = A / B × 100 In the above formula 8, A and B represent the following respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0246]

Table 11

[0247] (Test Example 2-5: Occludin mRNA Expression Promotion Test) Using the mugwort fermentation broth 1 obtained in Production Example 3, the mugwort fermentation broth 2 obtained in Production Example 4, and the mugwort extract obtained in Comparative Production Example 2 as test samples, the promotion effect on occludin mRNA expression was tested by the following test method.

[0248] Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was the concentration shown in Table 12 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurabo Industries Ltd.) were cultured using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurabo Industries Ltd.) at 37°C under 5% CO2 until confluent, and then the cells were recovered by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2). Next, 2 mL of the above NHEK (1.5×10 5 cells / mL) was seeded in a 35-mm petri dish and cultured overnight at 37°C under 5% CO2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2), and further cultured for 24 hours. After completion of the culture, the culture solution was removed, and total RNA was extracted with a reagent for RNA extraction (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.). The amount of each RNA was measured with a spectrophotometer, and total RNA was prepared to be 200 ng / μL using purified water.

[0249] Also, as a control, the same operations and absorbance measurements as above were performed, except that 2 mL of the test sample-added medium was changed to 2 mL of a normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL by the same method as above.

[0250] Using each of the above total RNAs as a template, the expression levels of occludin mRNA and GAPDH mRNA, which is an internal standard, were measured. 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of occludin mRNA without addition of the test sample and with addition of the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the promotion rate of occludin mRNA expression was calculated based on the following formula (9). The results are shown in Table 12 below. <Formula 9> Occludin mRNA expression promotion rate (%) = A / B × 100 In the above formula (9), A and B represent the following respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0251]

Table 12

[0252] <Production Example 5: Preparation of Fermentation Liquid 1 of Polygonum bistorta L. In the koji preparation step of Production Example 1 above, except that Artemisia princeps Pamp. was changed to Polygonum bistorta L. Thymus vulgaris (manufactured by Albion Co., Ltd.), "koji of Polygonum bistorta L." was prepared in the same manner as the koji preparation step of Production Example 1 above. Also, in the fermentation step of Production Example 1 above, except that Artemisia princeps Pamp. was changed to Polygonum bistorta L. Thymus vulgaris (manufactured by Albion Co., Ltd.), "fermentation liquid 1 of Polygonum bistorta L." was obtained in the same manner as the fermentation step of Production Example 1 above.

[0253] <Production Example 6: Preparation of Fermentation Liquid 2 of Polygonum bistorta L. In Production Example 5 above, except that the koji of Polygonum bistorta L. was changed to koji made from rice Aspergillus oryzae (manufactured by Akita Konno Shoten Co., Ltd.), "fermentation liquid 2 of Polygonum bistorta L." was obtained in the same manner as Production Example 5 above.

[0254] <Comparative Production Example 3: Preparation of Extract of Polygonum bistorta L. In Comparative Production Example 1 above, except that Artemisia princeps Pamp. was changed to Polygonum bistorta L. Thymus vulgaris"Tachijakousousou extract" was obtained in the same manner as in Comparative Production Example 1, except that it was changed to Albion Co., Ltd.

[0255] (Test Example A-3: Measurement of Contact Angle) In Test Example A-1, the contact angle was measured in the same manner as in Test Example A-1, except that the test sample was changed to the Tachijakousousou fermentation broth 1 obtained in Production Example 5, the Tachijakousousou fermentation broth 2 obtained in Production Example 6, and the Tachijakousousou extract obtained in Comparative Production Example 3. The results are shown in Table 13 below. Also, an example of the liquid droplet at the time of measuring the contact angle of each test sample is shown in FIGS. 3A to 3C.

[0256]

Table 13

[0257] With respect to the Tachijakousousou extract obtained in Comparative Production Example 3, both the Tachijakousousou fermentation broth 1 obtained in Production Example 5 and the Tachijakousousou fermentation broth 2 obtained in Production Example 6 had a small contact angle of 87° or less and were excellent in skin compatibility. Furthermore, the Tachijakousousou fermentation broth 1 obtained in Production Example 5 had a contact angle of 81° or less and was more excellent in skin compatibility.

[0258] (Test Example 3-1: Transglutaminase-1 (TGM-1) mRNA Expression Promotion Effect Test) Using the Tachijakousousou fermentation broth 1 obtained in Production Example 5, the Tachijakousousou fermentation broth 2 obtained in Production Example 6, and the Tachijakousousou extract obtained in Comparative Production Example 3 as test samples, the transglutaminase-1 (TGM-1) mRNA expression promotion effect was tested by the following test method.

[0259] Each test sample was dissolved in normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was the concentration shown in Table 14 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurabo Industries Ltd.) were cultured in a medium for normal human epidermal keratinocyte growth (HuMedia-KG2, manufactured by Kurabo Industries Ltd.) at 37°C under 5% CO2 until confluent, and then the cells were recovered by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with a medium for normal human epidermal keratinocyte growth (HuMedia-KG2). Next, 2 mL of the above NHEK (1.5×10 5 cells / mL) was seeded in a 35 mm petri dish and cultured overnight at 37°C under 5% CO2. After the culture was completed, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2), and further cultured for 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium, and cultured at 37°C under 5% CO2 for 24 hours. After the culture was completed, the culture solution was removed, and total RNA was extracted with a reagent for RNA extraction (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA was measured with a spectrophotometer, and the total RNA was prepared to be 200 ng / μL using purified water.

[0260] Also, as a control, the same operations and absorbance measurements as above were performed except that 2 mL of the test sample-added medium was changed to 2 mL of a normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL in the same manner as above.

[0261] Using each of the above total RNAs as a template, the expression levels of transglutaminase-1 (TGM-1) mRNA and GAPDH mRNA, which is an internal standard, were measured. 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of TGM-1 mRNA without addition of the test sample and with addition of the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the promotion rate of TGM-1 mRNA expression was calculated based on the following formula 10. The results are shown in Table 14 below. <Formula 10> Promotion rate of TGM-1 mRNA expression (%) = A / B × 100 In the above formula 10, A and B represent the following respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0262]

Table 14

[0263] (Test Example 3-2: Aquaporin 3 (AQP3) mRNA expression promotion effect test) Using the Tacca chantrieri fermentation broth 1 obtained in Production Example 5, the Tacca chantrieri fermentation broth 2 obtained in Production Example 6, and the Tacca chantrieri extract obtained in Comparative Production Example 3 as test samples, the aquaporin 3 (AQP3) mRNA expression promotion effect was tested by the following test method.

[0264] Each test sample was dissolved in normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kuraray Co., Ltd.) so that the final concentration was the concentration shown in Table 15 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kuraray Co., Ltd.) were cultured using normal human epidermal keratinocyte growth medium (HuMedia-KG2, manufactured by Kuraray Co., Ltd.) at 37°C and 5% CO2 until confluent, and then the cells were collected by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with normal human epidermal keratinocyte growth medium (HuMedia-KG2). Next, the above NHEK (1.5×10 52 mL of cells / mL were seeded and cultured overnight at 37°C under 5% CO2 conditions. After the culture was completed, the medium was replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2), and incubation was continued for an additional 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured for 24 hours at 37°C under 5% CO2 conditions. After the culture was completed, the culture supernatant was removed, and total RNA was extracted using a reagent for RNA extraction (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.). The amount of each RNA was measured using a spectrophotometer, and total RNA was prepared to a concentration of 200 ng / μL using purified water.

[0265] Also, as a control, the same operations and absorbance measurements as above were performed, except that 2 mL of the test sample-added medium was replaced with 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to a concentration of 200 ng / μL in the same manner as above.

[0266] Using each of the total RNAs as a template, the expression levels of aquaporin 3 (AQP3) mRNA and GAPDH mRNA, which is an internal standard, were measured. Detection of mRNA was performed by a two-step real-time PCR reaction using a real-time PCR apparatus (Thermal Cycler DIce (registered trademark) Real Time System III, manufactured by Takara Bio Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (Catalog No.: RR063A), manufactured by Takara Bio Inc.). The expression levels of AQP3 mRNA without and with the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the AQP3 mRNA expression promotion rate was calculated based on the following formula 11. The results are shown in Table 15 below. <Formula 11> AQP3 mRNA expression promotion rate (%) = A / B × 100 In Formula 11 above, A and B represent the following, respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0267]

Table 15

[0268] (Test Example 3-3: Test for promoting Occludin mRNA expression) In Test Example 2-5, the test sample was changed to the fermented liquid 1 of Artemisia princeps Pamp. obtained in Production Example 5, the fermented liquid 2 of Artemisia princeps Pamp. obtained in Production Example 6, and the extract of Artemisia princeps Pamp. obtained in Comparative Production Example 3, and the final concentration of the test sample was changed to the concentration shown in Table 16 below. Except for this, the test for promoting Occludin mRNA expression was carried out in the same manner as in Test Example 2-5. The results are shown in Table 16 below.

[0269]

Table 16

[0270] (Test Example 3-4: Test for DPPH radical scavenging activity) In Test Example 1-3, the test sample was changed to the fermented liquid 1 of Artemisia princeps Pamp. obtained in Production Example 5, the fermented liquid 2 of Artemisia princeps Pamp. obtained in Production Example 6, and the extract of Artemisia princeps Pamp. obtained in Comparative Production Example 3, and the final concentration of the test sample was changed to the concentration shown in Table 17 below. Except for this, the test for DPPH radical scavenging activity was carried out in the same manner as in Test Example 1-3. The results are shown in Table 17 below.

[0271]

Table 17

[0272] <Production Example 7: Preparation of fermented liquid 1 of Mentha canadensis L.> In the koji preparation step of Production Example 1, except that yomogi was changed to Mentha canadensis L. Melissa officinalis (manufactured by Albion Co., Ltd.), "Mentha canadensis L. koji" was prepared in the same manner as the koji preparation step of Production Example 1. Also, in the fermentation step of Production Example 1, except that mugwort was changed to Artemisia princeps Pamp. Melissa officinalis (manufactured by Albion Co., Ltd.), "Artemisia princeps Pamp. fermentation broth 1" was obtained in the same manner as the fermentation step of Production Example 1.

[0273] <Production Example 8: Preparation of Artemisia princeps Pamp. fermentation broth 2> In Production Example 7, except that the Artemisia princeps Pamp. seed koji was changed to rice seed koji Aspergillus oryzae (Shirakami white koji, manufactured by Akita Konno Shoten Co., Ltd.), "Artemisia princeps Pamp. fermentation broth 2" was obtained in the same manner as Production Example 7.

[0274] <Comparative Production Example 4: Preparation of Artemisia princeps Pamp. extract> In Comparative Production Example 1, except that mugwort was changed to Artemisia princeps Pamp. Melissa officinalis (manufactured by Albion Co., Ltd.), "Artemisia princeps Pamp. extract" was obtained in the same manner as Comparative Production Example 1.

[0275] (Test Example A-4: Measurement of contact angle) In Test Example A-1, except that the test sample was changed to the Artemisia princeps Pamp. fermentation broth 1 obtained in Production Example 7, the Artemisia princeps Pamp. fermentation broth 2 obtained in Production Example 8, and the Artemisia princeps Pamp. extract obtained in Comparative Production Example 4, the contact angle was measured in the same manner as Test Example A-1. The results are shown in Table 18 below. Also, an example of the liquid droplet at the time of measuring the contact angle of each test sample is shown in FIGS. 4A to 4C.

[0276]

Table 18

[0277] With respect to the Artemisia princeps Pamp. extract obtained in Comparative Production Example 4, both the Artemisia princeps Pamp. fermentation broth 1 obtained in Production Example 7 and the Artemisia princeps Pamp. fermentation broth 2 obtained in Production Example 8 had small contact angles, which were 85° or less, and were excellent in skin affinity. Furthermore, the Artemisia princeps Pamp. fermentation broth 1 obtained in Production Example 7 had a contact angle of 79° or less and was even more excellent in skin affinity.

[0278] (Test Example 4-1: Test for promoting type I collagen production) In Test Example 2-2, the test sample was changed to the Artemisia princeps Pamp. fermentation broth 1 obtained in Production Example 7, the Artemisia princeps Pamp. fermentation broth 2 obtained in Production Example 8, and the Artemisia princeps Pamp. extract obtained in Comparative Production Example 4, and the final concentration of the test sample was changed to the concentration shown in Table 19 below. The promoting effect on type I collagen production was tested in the same manner as in Test Example 2-2 except for the above changes. The results are shown in Table 19 below.

[0279]

Table 19

[0280] (Test Example 4-2: Test for promoting aquaporin 3 (AQP3) mRNA expression) In Test Example 3-2, the test sample was changed to the Artemisia princeps Pamp. fermentation broth 1 obtained in Production Example 7, the Artemisia princeps Pamp. fermentation broth 2 obtained in Production Example 8, and the Artemisia princeps Pamp. extract obtained in Comparative Production Example 4, and the final concentration of the test sample was changed to the concentration shown in Table 20 below. The promoting effect on aquaporin 3 (AQP3) mRNA expression was tested in the same manner as in Test Example 3-2 except for the above changes. The results are shown in Table 20 below.

[0281]

Table 20

[0282] (Test Example 4-3: Test for DPPH radical scavenging activity) In Test Examples 1-3, the test sample was changed to the Kosui Hakka fermentation broth 1 obtained in Production Example 7, the Kosui Hakka fermentation broth 2 obtained in Production Example 8, and the Kosui Hakka extract obtained in Comparative Production Example 4, and the final concentration of the test sample was changed to the concentration shown in Table 21 below. The DPPH radical scavenging activity was tested in the same manner as in Test Examples 1-3 except for the above changes. The results are shown in Table 21 below.

[0283]

Table 21

[0284] (Test Example 4-4: Hyaluronidase Activity Inhibition Test) In Test Examples 1-4, the test sample was changed to the Kosui Hakka fermentation broth 1 obtained in Production Example 7, the Kosui Hakka fermentation broth 2 obtained in Production Example 8, and the Kosui Hakka extract obtained in Comparative Production Example 4, and the final concentration of the test sample was changed to the concentration shown in Table 22 below. The hyaluronidase activity inhibition was tested in the same manner as in Test Examples 1-4 except for the above changes. The results are shown in Table 22 below.

[0285]

Table 22

[0286] (Test Example 4-5: Tyrosinase Activity Inhibition Test) In Test Examples 1-5, the test sample was changed to the Kosui Hakka fermentation broth 1 obtained in Production Example 7, the Kosui Hakka fermentation broth 2 obtained in Production Example 8, and the Kosui Hakka extract obtained in Comparative Production Example 4, and the final concentration of the test sample was changed to the concentration shown in Table 22 below. The tyrosinase activity inhibition was tested in the same manner as in Test Examples 1-5 except for the above changes. The results are shown in Table 23 below.

[0287]

Table 23

[0288] <Production Example 9: Preparation of Fermentation Broth 1 of Jaguruma Kikku> In the koji preparation step of Production Example 1, except that mugwort was changed to Centaurea Centaurea cyanus Artemisia lactiflora Wall. ex DC. (manufactured by Albion Co., Ltd.), "Artemisia lactiflora Wall. ex DC. koji" was prepared in the same manner as the koji preparation step of Production Example 1. In addition, in the fermentation step of Production Example 1, except that mugwort was changed to Centaurea Centaurea cyanus Artemisia lactiflora Wall. ex DC. (manufactured by Albion Co., Ltd.), "Artemisia lactiflora Wall. ex DC. fermentation broth 1" was obtained in the same manner as the fermentation step of Production Example 1.

[0289] <Production Example 10: Preparation of Artemisia lactiflora Wall. ex DC. fermentation broth 2> In Production Example 9, except that Artemisia lactiflora Wall. ex DC. koji was changed to rice koji Aspergillus oryzae , Shirakami white koji, manufactured by Akita Konno Shoten Co., Ltd.), "Artemisia lactiflora Wall. ex DC. fermentation broth 2" was obtained in the same manner as Production Example 9.

[0290] <Comparative Production Example 5: Preparation of Artemisia lactiflora Wall. ex DC. extract> In Comparative Production Example 1, except that mugwort was changed to Centaurea Centaurea cyanus Artemisia lactiflora Wall. ex DC. (manufactured by Albion Co., Ltd.), "Artemisia lactiflora Wall. ex DC. extract" was obtained in the same manner as Comparative Production Example 1.

[0291] (Test Example A-5: Measurement of contact angle) In Test Example A-1, except that the test sample was changed to the Artemisia lactiflora Wall. ex DC. fermentation broth 1 obtained in Production Example 9, the Artemisia lactiflora Wall. ex DC. fermentation broth 2 obtained in Production Example 10, and the Artemisia lactiflora Wall. ex DC. extract obtained in Comparative Production Example 5, the contact angle was measured in the same manner as Test Example A-1. The results are shown in Table 24 below. In addition, an example of the liquid droplet at the time of measuring the contact angle of each test sample is shown in FIGS. 5A to 5C.

[0292]

Table 24

[0293] With respect to the yaguruma aster extract obtained in Comparative Production Example 5, both the yaguruma aster fermentation broth 1 obtained in Production Example 9 and the yaguruma aster fermentation broth 2 obtained in Production Example 10 had small contact angles, 85° or less, and were excellent in skin affinity. Furthermore, the yaguruma aster fermentation broth 1 obtained in Production Example 9 had a contact angle of 79° or less and was even more excellent in skin affinity.

[0294] (Test Example 5-1: Type I Collagen Production Promotion Test) In Test Example 2-2, the test sample was changed to the yaguruma aster fermentation broth 1 obtained in Production Example 9, the yaguruma aster fermentation broth 2 obtained in Production Example 10, and the yaguruma aster extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentrations shown in Table 25 below. The promotion effect on type I collagen production was tested in the same manner as in Test Example 2-2 except for this. The results are shown in Table 25 below.

[0295]

Table 25

[0296] (Test Example 5-2: Transglutaminase-1 mRNA Expression Promotion Test) In Test Example 3-1, the test sample was changed to the yaguruma aster fermentation broth 1 obtained in Production Example 9, the yaguruma aster fermentation broth 2 obtained in Production Example 10, and the yaguruma aster extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentrations shown in Table 26 below. The promotion effect on transglutaminase-1 mRNA expression was tested in the same manner as in Test Example 3-1 except for this. The results are shown in Table 26 below.

[0297]

Table 26

[0298] (Test Example 5-3: Filaggrin mRNA Expression Promotion Test) Using the sunflower aster fermented liquid 1 obtained in Production Example 9, the sunflower aster fermented liquid 2 obtained in Production Example 10, and the sunflower aster extract obtained in Comparative Production Example 5 as test samples, the promoting effect on filaggrin mRNA expression was tested by the following test method.

[0299] Each test sample was dissolved in normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurabo Industries Ltd.) so that the final concentration was the concentration shown in Table 27 below, and a test sample-added medium was prepared. Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurabo Industries Ltd.) were cultured using normal human epidermal keratinocyte growth medium (HuMedia-KG2, manufactured by Kurabo Industries Ltd.) at 37 °C under 5% CO2 until confluent, and then the cells were recovered by trypsin treatment. This was adjusted to 1.5×10 5 cells / mL with normal human epidermal keratinocyte growth medium (HuMedia-KG2). Next, 2 mL of the NHEK (1.5×10 5 cells / mL) was seeded in a 35 mm petri dish and cultured overnight at 37 °C under 5% CO2. After the culture was completed, the medium was replaced with normal human epidermal keratinocyte basal medium (HuMedia-KB2) and cultured for another 24 hours. After the culture was completed, the medium was replaced with 2 mL of the test sample-added medium and cultured at 37 °C under 5% CO2 for 24 hours. After the culture was completed, the culture solution was removed, and total RNA was extracted with a reagent for RNA extraction (ISOGEN II (Catalog No.: 311-07361), manufactured by Nippon Gene Co., Ltd.). The amount of each RNA was measured with a spectrophotometer, and total RNA was prepared to be 200 ng / μL using purified water.

[0300] Also, as a control, the same operations and absorbance measurements as above were performed except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) not containing the test sample, and total RNA was prepared to be 200 ng / μL in the same manner as above.

[0301] Using each of the total RNAs as a template, the expression levels of filaggrin mRNA and GAPDH mRNA, which is an internal standard, were measured. Detection of the 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 Inc.) and SYBR (registered trademark) PrimeScript (registered trademark) RT-PCR Kit (Perfect Real Time (catalog number: RR063A), manufactured by Takara Bio Inc.). The expression levels of filaggrin mRNA without addition of the test sample and with addition of the test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the filaggrin mRNA expression promotion rate was calculated based on the following formula 12. The results are shown in Table 27 below. <Formula 12> Filaggrin mRNA expression promotion rate (%) = A / B × 100 In the above formula 12, A and B represent the following respectively. A: Corrected value when the test sample is added B: Corrected value when the test sample is not added

[0302]

Table 27

[0303] (Test Example 5-4: Aquaporin 3 (AQP3) mRNA expression promotion test) In Test Example 3-2, the test sample was changed to the Hieracium pilosella fermentation broth 1 obtained in Production Example 9, the Hieracium pilosella fermentation broth 2 obtained in Production Example 10, and the Hieracium pilosella extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentrations shown in Table 28 below. An aquaporin 3 (AQP3) mRNA expression promotion test was performed in the same manner as in Test Example 3-2 except for the above changes. The results are shown in Table 28 below.

[0304]

Table 28

[0305] (Test Example 5-5: Test for the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA) In Test Example 1-2, the test sample was changed to the fermented liquid 1 of Hieracium pilosella obtained in Production Example 9, the fermented liquid 2 of Hieracium pilosella obtained in Production Example 10, and the Hieracium pilosella extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 29 below. Except for this, the effect of promoting the expression of hyaluronic acid synthase 3 (HAS3) mRNA was tested in the same manner as in Test Example 1-2. The results are shown in Table 29 below.

[0306]

Table 29

[0307] (Test Example 5-6: Test for the effect of promoting the expression of claudin-1 mRNA) In Test Example 2-3, the test sample was changed to the fermented liquid 1 of Hieracium pilosella obtained in Production Example 9, the fermented liquid 2 of Hieracium pilosella obtained in Production Example 10, and the Hieracium pilosella extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 30 below. Except for this, the effect of promoting the expression of claudin-1 mRNA was tested in the same manner as in Test Example 2-3. The results are shown in Table 30 below.

[0308]

Table 30

[0309] (Test Example 5-7: Test for the effect of promoting the expression of claudin-4 mRNA) In Test Example 2-4, the test sample was changed to the fermented liquid 1 of Hieracium pilosella obtained in Production Example 9, the fermented liquid 2 of Hieracium pilosella obtained in Production Example 10, and the Hieracium pilosella extract obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 31 below. Except for this, the effect of promoting the expression of claudin-4 mRNA was tested in the same manner as in Test Example 2-4. The results are shown in Table 31 below.

[0310]

Table 31

[0311] (Test Example 5-8: Test for the effect of promoting Occludin mRNA expression) In Test Examples 2-5, the test sample was changed to the Fermented Liquid of Young Marigold 1 obtained in Production Example 9, the Fermented Liquid of Young Marigold 2 obtained in Production Example 10, and the Extract of Young Marigold obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 32 below. The effect of promoting Occludin mRNA expression was tested in the same manner as in Test Examples 2-5. The results are shown in Table 32 below.

[0312]

Table 32

[0313] (Test Example 5-9: Test for DPPH radical scavenging effect) In Test Examples 1-3, the test sample was changed to the Fermented Liquid of Young Marigold 1 obtained in Production Example 9, the Fermented Liquid of Young Marigold 2 obtained in Production Example 10, and the Extract of Young Marigold obtained in Comparative Production Example 5, and the final concentration of the test sample was changed to the concentration shown in Table 33 below. The DPPH radical scavenging effect was tested in the same manner as in Test Examples 1-3. The results are shown in Table 33 below.

[0314]

Table 33

[0315] (Test Example 5-10: Test for the effect of suppressing melanin production on B16 melanoma cells) The Fermented Liquid of Young Marigold 1 obtained in Production Example 9, the Fermented Liquid of Young Marigold 2 obtained in Production Example 10, and the Extract of Young Marigold obtained in Comparative Production Example 5 were used as test samples, and the effect of suppressing melanin production on B16 melanoma cells was tested by the following test method.

[0316] Each test sample was dissolved in DMEM (manufactured by Nissui Pharmaceutical Co., Ltd.) containing 10% FBS (manufactured by Biosera) and 1 mmol / L theophylline (manufactured by Fujifilm Wako Pure Chemical Corporation) to prepare a test sample-added medium. B16 melanoma cells were cultured in DMEM containing 10% FBS at 37 °C under 5% CO2 until confluent, and then the cells were collected by trypsin treatment. This was adjusted to 2.4×10 5 cells / mL with DMEM containing 10% FBS and 1 mmol / L theophylline. Next, the B16 melanoma cells (2.4×10 5 cells / mL) were seeded at 300 μL per well in a 48-well plate and cultured at 37 °C under 5% CO2 for 6 hours. After the culture, the medium was replaced with 100 μL of the test sample-added medium and cultured at 37 °C under 5% CO2 for 3 days. After the culture, 300 μL of the test sample-added medium was added per well and cultured at 37 °C under 5% CO2 for 4 days. The final concentration of the test sample at this time was the concentration shown in Table 34 below.

[0317] After the culture, the medium was removed from each well, 200 μL of 2 mol / L sodium hydroxide solution was added, and the cells were disrupted with an ultrasonic crusher, and the absorbance at a wavelength of 475 nm was measured. Based on the calibration curve prepared using synthetic melanin (manufactured by SIGMA), the amount of melanin was calculated from the measured absorbance value.

[0318] Also, to measure cell viability, B16 melanoma cells were cultured using the test sample-added medium in the same manner as the above method. After the culture, the medium was removed and the cells were washed with 400 μL of PBS buffer. Next, a solution prepared by dissolving neutral red at a final concentration of 0.05 mg / mL in DMEM containing 10% FBS was added to each well at 200 μL and cultured for 2.5 hours. After the culture, 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 dye. After the extraction, the absorbance at a wavelength of 540 nm was measured.

[0319] As a control, the same operations and absorbance measurements as above were performed, except that the test sample solution was changed to DMEM containing 10% FBS and 1 mmol / L theophylline without the test sample.

[0320] From the obtained measured values, 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 Table 34 below. <Formula 13> Cell survival rate (%) = (C / D) × 100 In the above formula 13, C and D represent the following respectively. C: Absorbance at a wavelength of 540 nm without the test sample D: Absorbance at a wavelength of 540 nm with the test sample added <Formula 14> Melanin production inhibition rate (%) = {1 - (B / D) / (A / C)} × 100 In the above formula 14, A to D represent the following respectively. A: Amount of melanin without the test sample added B: Amount of melanin with the test sample added C: Absorbance at a wavelength of 540 nm without the test sample D: Absorbance at a wavelength of 540 nm with the test sample added

[0321]

Table 34

Industrial Applicability

[0322] The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention have at least one of excellent anti-aging action, antioxidant action, anti-inflammatory action, and whitening action, and are natural product-based with high safety. Therefore, they can be widely used regardless of fields such as cosmetics, foods, and pharmaceuticals. Since the cosmetic of the present invention contains at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent of the present invention, it can be suitably used in skin cosmetics such as lotion, emulsion, cream, ointment, beauty liquid, lotion, pack, jelly, lip cream, lipstick, foundation, bath agent, soap, body shampoo, etc.; scalp and hair cosmetics such as astringent, hair tonic, hair cream, hair liquid, pomade, shampoo, rinse, etc.

Claims

Claims 1. With respect to 100 parts by mass of Artemisia princeps Pamp., 5 to 100 parts by mass of Aspergillus oryzae at 1×10³ cells / mL to 1×10⁸ cells / mL are inoculated and cultured at 20°C to 40°C for 80 to 210 hours to obtain a koji preparation step for Artemisia princeps Pamp. koji, and after adding 500 to 5,000 parts by mass of water to 100 parts by mass of Artemisia princeps Pamp., the above Artemisia princeps Pamp. koji at 1×10³ cells / mL to 1×10⁸ cells / mL is inoculated and fermented at 20°C to 40°C for 10 to 40 hours, and the Artemisia princeps Pamp. fermentation broth obtained by this is contained as an active ingredient, An anti-aging agent characterized by having at least one action selected from the group consisting of a matrix metalloprotease-1 activity inhibitory action and a hyaluronic acid synthase 3 mRNA expression promoting action. Claims 2. With respect to 100 parts by mass of Artemisia lavandulaefolia DC., 5 to 100 parts by mass of Aspergillus oryzae at 1×10³ cells / mL to 1×10⁸ cells / mL are inoculated and cultured at 20°C to 40°C for 80 to 210 hours to obtain a koji preparation step for Artemisia lavandulaefolia DC. koji, and after adding 500 to 5,000 parts by mass of water to 100 parts by mass of Artemisia lavandulaefolia DC., the above Artemisia lavandulaefolia DC. koji at 1×10³ cells / mL to 1×10⁸ cells / mL is inoculated and fermented at 20°C to 40°C for 10 to 40 hours, and the Artemisia lavandulaefolia DC. fermentation broth obtained by this is contained as an active ingredient, An anti-aging agent characterized by having at least one action selected from the group consisting of a hyaluronic acid synthase 3 mRNA expression promoting action, a type I collagen production promoting action, a claudin-1 mRNA expression promoting action, a claudin-4 mRNA expression promoting action, and an occludin mRNA expression promoting action. **Claim 3**: A koji preparation step of inoculating 5 to 100 parts by mass of Aspergillus oryzae at 1 × 10³ cells / mL to 1 × 10⁸ cells / mL with respect to 100 parts by mass of Rhizopus chinensis, and culturing at 20°C to 40°C for 80 to 210 hours to obtain a Rhizopus chinensis koji; and a fermentation step of adding 500 to 5,000 parts by mass of water with respect to 100 parts by mass of Rhizopus chinensis, then inoculating 1 × 10³ cells / mL to 1 × 10⁸ cells / mL of the above-mentioned Rhizopus chinensis koji, and fermenting at 20°C to 40°C for 10 to 40 hours, and containing the Rhizopus chinensis fermentation broth obtained thereby as an active ingredient, An anti-aging agent characterized by having at least one action selected from the group consisting of an action of promoting occludin mRNA expression, an action of promoting transglutaminase-1 mRNA expression, and an action of promoting aquaporin 3 mRNA expression. **Claim 4**: A koji preparation step of inoculating 5 to 100 parts by mass of Aspergillus oryzae at 1 × 10³ cells / mL to 1 × 10⁸ cells / mL with respect to 100 parts by mass of Mentha canadensis, and culturing at 20°C to 40°C for 80 to 210 hours to obtain a Mentha canadensis koji; and a fermentation step of adding 500 to 5,000 parts by mass of water with respect to 100 parts by mass of Mentha canadensis, then inoculating 1 × 10³ cells / mL to 1 × 10⁸ cells / mL of the above-mentioned Mentha canadensis koji, and fermenting at 20°C to 40°C for 10 to 40 hours, and containing the Mentha canadensis fermentation broth obtained thereby as an active ingredient, An anti-aging agent characterized by having at least one action selected from the group consisting of an action of promoting type I collagen production and an action of promoting transglutaminase-1 mRNA expression. **Claim 5**: A koji preparation step of inoculating 5 to 100 parts by mass of Aspergillus oryzae at 1×10³ to 1×10⁸ cells / mL with respect to 100 parts by mass of Taraxacum officinale, culturing at 20°C to 40°C for 80 to 210 hours to obtain a koji of Taraxacum officinale, and after adding 500 to 5,000 parts by mass of water with respect to 100 parts by mass of Taraxacum officinale, inoculating the koji of Taraxacum officinale at 1×10³ to 1×10⁸ cells / mL, and fermenting at 20°C to 40°C for 10 to 40 hours. The fermented liquid of Taraxacum officinale obtained by the fermentation step contains an active ingredient, An anti-aging agent having at least one action selected from the group consisting of hyaluronic acid synthase 3 mRNA expression promoting action, type I collagen production promoting action, claudin-1 mRNA expression promoting action, claudin-4 mRNA expression promoting action, occludin mRNA expression promoting action, transglutaminase-1 mRNA expression promoting action, aquaporin 3 mRNA expression promoting action, and filaggrin mRNA expression promoting action. **Claim 6**: A koji preparation step of inoculating 5 to 100 parts by mass of Aspergillus oryzae at 1×10³ to 1×10⁸ cells / mL with respect to 100 parts by mass of Artemisia princeps, culturing at 20°C to 40°C for 80 to 210 hours to obtain a koji of Artemisia princeps, and after adding 500 to 5,000 parts by mass of water with respect to 100 parts by mass of Artemisia princeps, inoculating the koji of Artemisia princeps at 1×10³ to 1×10⁸ cells / mL, and fermenting at 20°C to 40°C for 10 to 40 hours. The fermented liquid of Artemisia princeps obtained by the fermentation step, A koji preparation step of inoculating 5 to 100 parts by mass of Aspergillus oryzae at 1×10³ to 1×10⁸ cells / mL with respect to 100 parts by mass of Ligularia dentata, culturing at 20°C to 40°C for 80 to 210 hours to obtain a koji of Ligularia dentata, and after adding 500 to 5,000 parts by mass of water with respect to 100 parts by mass of Ligularia dentata, inoculating the koji of Ligularia dentata at 1×10³ to 1×10⁸ cells / mL, and fermenting at 20°C to 40°C for 10 to 40 hours. The fermented liquid of Ligularia dentata obtained by the fermentation step, To 100 parts by mass of *Artemisia princeps* Pamp., 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL is inoculated and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Artemisia princeps* Pamp. koji. And then, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Artemisia princeps* Pamp., 1×10³ cells / mL to 1×10⁸ cells / mL of the obtained *Artemisia princeps* Pamp. koji is inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Artemisia princeps* Pamp. fermentation broth obtained by the above steps, and To 100 parts by mass of *Taraxacum officinale* Weber var. *latifolium* DC., 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL is inoculated and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Taraxacum officinale* Weber var. *latifolium* DC. koji. And then, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Taraxacum officinale* Weber var. *latifolium* DC., 1×10³ cells / mL to 1×10⁸ cells / mL of the obtained *Taraxacum officinale* Weber var. *latifolium* DC. koji is inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Taraxacum officinale* Weber var. *latifolium* DC. fermentation broth obtained by the above steps, contains at least one fermentation broth selected from the group consisting of the above as an active ingredient, and is characterized by having a diphenyl-p-picrylhydrazyl radical scavenging action, an antioxidant.

7. To 100 parts by mass of *Artemisia lavandulaefolia* DC., 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL is inoculated and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Artemisia lavandulaefolia* DC. koji. And then, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Artemisia lavandulaefolia* DC., 1×10³ cells / mL to 1×10⁸ cells / mL of the obtained *Artemisia lavandulaefolia* DC. koji is inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Artemisia lavandulaefolia* DC. fermentation broth obtained by the above steps, and For 100 parts by mass of *Bacillus subtilis*, 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL are inoculated, and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Bacillus subtilis* seed koji. In the seed koji preparation step, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Bacillus subtilis*, 1×10³ cells / mL to 1×10⁸ cells / mL of the above-mentioned *Bacillus subtilis* seed koji are inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Bacillus subtilis* fermentation broth obtained by the above steps, contains at least one fermentation broth selected from the group consisting of an anti-inflammatory agent characterized by having a hyaluronidase activity inhibitory effect.

8. For 100 parts by mass of *Bacillus subtilis*, 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL are inoculated, and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Bacillus subtilis* seed koji. In the seed koji preparation step, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Bacillus subtilis*, 1×10³ cells / mL to 1×10⁸ cells / mL of the above-mentioned *Bacillus subtilis* seed koji are inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Bacillus subtilis* fermentation broth obtained by the above steps is contained as an active ingredient, a whitening agent characterized by having a tyrosinase activity inhibitory effect.

9. For 100 parts by mass of *Sonchus asper*, 5 to 100 parts by mass of *Aspergillus oryzae* at 1×10³ cells / mL to 1×10⁸ cells / mL are inoculated, and cultured at 20°C to 40°C for 80 to 210 hours to obtain *Sonchus asper* seed koji. In the seed koji preparation step, after adding 500 to 5,000 parts by mass of water to 100 parts by mass of *Sonchus asper*, 1×10³ cells / mL to 1×10⁸ cells / mL of the above-mentioned *Sonchus asper* seed koji are inoculated and fermented at 20°C to 40°C for 10 to 40 hours. The *Sonchus asper* fermentation broth obtained by the above steps is contained as an active ingredient, a whitening agent characterized by having at least one of the effects of suppressing melanin production. Cosmetic product characterized by containing at least one selected from the group consisting of the anti-aging agent according to claim 1, 2 or 4, the antioxidant according to claim 6, the anti-inflammatory agent according to claim 7, and the whitening agent according to claim 8 (however, excluding those containing the fermentation broth of *Tatizakou-sou* or the fermentation broth of *Yagurumagiku*).

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