JP2020164486A - Anti-aging agents, antioxidants, anti-inflammatory agents, and whitening agents, as well as cosmetics - Google Patents

Abstract

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

Description

The present invention comprises an anti-aging agent, an antioxidant, an anti-inflammatory agent, and a whitening agent containing a fermented solution of plant aspergillus as an active ingredient, and the anti-inflammatory agent, the antioxidant, the anti-inflammatory agent, and the anti-inflammatory agent. The present invention relates to a cosmetic containing at least one of the whitening agents.

In recent years, active oxygen has been attracting attention as a factor for oxidizing biological components, and its adverse effect on the living body has become a problem. Active oxygen, which occur in energy metabolism processes within living cells, superoxide [i.e., superoxide anion generated by one-electron reduction of molecular oxygen (· O 2 ^_-), hydrogen peroxide (H 2 _{O 2),} There are singlet oxygen ( ¹ O ₂ ), hydroxyl radical (.OH)] and the like. Such active oxygen is essential for the bactericidal mechanism of phagocytic cells and plays an important role in the removal of viruses and cancer cells.

However, the excessive production of active oxygen attacks in vivo molecules constituting membranes and tissues in the living body and induces various diseases. Superoxide, which is produced in vivo and is also a starting material for other active 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 becomes high. This is one of the causes of tissue disorders such as rheumatoid arthritis and Bechette's disease, myocardial infarction, stroke, cataracts, stains, freckles, wrinkles, diabetes, arteriosclerosis, stiff shoulders, poor circulation, and skin aging. It is considered.

Among these, the skin is an organ in which superoxide is easily generated because it is directly stimulated by environmental factors such as ultraviolet rays. Therefore, as the superoxide concentration increases, for example, biological tissues such as collagen are decomposed and denatured. Or it bridges or oxidizes fats and oils to produce lipid peroxide that damages cells, forming wrinkles on the skin, causing aging such as decreased elasticity of the skin, inflammation, and pigmentation of the skin. (See Non-Patent Document 1). Therefore, by inhibiting or suppressing the production of active oxygen and radicals in the body, skin aging such as wrinkle formation and decreased elasticity, tissue disorders such as rheumatoid arthritis and Bechet's disease, myocardial infarction, stroke, cataracts, diabetes and arteriosclerosis It is considered that various disorders related to active oxygen such as hardening, stiff shoulders, and coldness can be prevented, treated, or ameliorated.

Therefore, attempts have been made to obtain active oxygen scavengers, radical scavengers, hydrogen peroxide scavengers, etc. from natural products that are advantageous in terms of safety, and extracts of cruciferous plants of the genus Brasca (see Patent Document 1). Effectiveness has been confirmed in extracts of plants belonging to the genus Crassulaceae, Crassulaceae (see Patent Document 2), extracts of tamakocho (see Patent Document 3), and extracts of Suiou (see Patent Document 4).

In addition, there are various causes and onset mechanisms of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other various skin diseases accompanied by rough skin. It is known that the cause is mainly due to the increased activity of hyaluronidase.

Hyaluronidase is a hydrolase of hyaluronic acid. Hyaluronic acid, which maintains its affinity for body tissues, is decomposed by ultraviolet rays, oxygen, etc. in a water-containing system, and the water retention effect decreases as the molecular weight decreases. Hyaluronic acid also exists as an intercellular tissue in the living body and is involved in vascular permeability. In addition, hyaluronidase, which is present in mast cells, is released by degranulation caused by its activation and acts as an inflammatory chemical mediator. Therefore, by inhibiting the activity of hyaluronidase, it is expected to enhance moisturization and prevent / reduce inflammation.
As those having such a hyaluronidase activity inhibitory action, for example, an extract of a plant of the genus Osvecchia (see Patent Document 5), a wisteria tea extract (see Patent Document 6) and the like are known.

In addition, melanin also plays a role of protecting the living body from ultraviolet rays in the skin, but overproduction and uneven accumulation cause darkening of the skin and spots. In general, melanin is changed from tyrosine to dopa and from dopa to dopaquinone by the action of the enzyme tyrosinase biosynthesized in pigment cells, and then formed via intermediates such as 5,6-dihydroxyindophenol. ing. Therefore, in order to prevent, treat or improve skin darkening (skin pigmentation), age spots, freckles, etc., it is necessary to inhibit the activity of tyrosinase involved in melanin production or suppress melanin production. Can be considered.

Conventionally, for the prevention, treatment or improvement of skin pigmentation, age spots, freckles and the like, a treatment of externally applying a whitening agent containing a chemically synthesized product such as hydroquinone as an active ingredient has been performed. However, chemically synthesized products such as hydroquinone may cause side effects such as skin irritation and allergies.
Therefore, it is desired to develop a whitening agent containing a highly safe natural raw material as an active ingredient. Examples of the whitening agent having a tyrosinase activity inhibitory action include wisteria tea extract (see Patent Document 7) and water pepper extract (see Patent Document 7). (Refer to Patent Document 8) and the like are known. Further, as those having a melanin production inhibitory action, for example, an extract from the root of castor bean (see Patent Document 9), an extract from a plant belonging to the genus Saussurea (see Patent Document 10), and the like are known. There is.

The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrix such as collagen that is outside these cells and supports the skin structure. In young skin, the interaction of these skin tissues maintains homeostasis to ensure water retention, flexibility, elasticity, etc., and the skin is maintained in a fresh state with appearance of tension and luster.
However, under the influence of certain external factors such as ultraviolet (UV-A, UV-B) irradiation, significant dryness of air, excessive skin cleaning, and aging, the extracellular matrix is major. The amount of collagen produced, which is a constituent, decreases and the elasticity decreases due to cross-linking. As a result, the moisturizing function and elasticity of the skin are reduced, and the keratin begins to exfoliate abnormally, so that the skin loses tension and luster, and exhibits aging symptoms such as roughness and wrinkles.
As described above, various factors are mentioned for the changes associated with skin aging, that is, wrinkles, dullness, loss of texture, decrease in elasticity, etc., but extracellular matrix components such as collagen, hyaluronic acid, and elastin are listed. Reduction and denaturation are involved. Therefore, it is considered that skin aging can be prevented and improved by promoting the production of collagen, hyaluronic acid and the like.

Therefore, an attempt has been made to obtain a substance having a collagen production promoting action from a natural product which is advantageous in terms of safety. As those having a collagen production promoting action, for example, a star fruit leaf extract (see Patent Document 11), a Mallotus philipponica extract (see Patent Document 12), and the like have been confirmed.

In addition, the amino acid which is the main component of the natural moisturizing factor (Natural Moisturizing Factors) is produced by decomposing filaggrin derived from keratohyalin granules in the stratum corneum. This filaggrin is expressed as profilaggrin in the epidermal keratinocytes present in the stratum granulosum just below the stratum corneum. After that, it is immediately phosphorylated, accumulated in keratohyalin granules, decomposed into filaggrin through dephosphorylation and hydrolysis, transferred to the stratum corneum, increases the aggregation efficiency of keratin filaments, and is involved in the internal construction of keratinocytes. It has been reported (see Non-Patent Document 2).
In recent years, it has been reported that this filaggrin is very important and indispensable for water retention of the skin, and that the synthetic ability of filaggrin decreases due to conditions such as dryness, and the amount of amino acids in the stratum corneum decreases (non-). See Patent Document 3).
Therefore, it is expected that the amount of amino acids in the stratum corneum can be increased by promoting the synthesis of filaggrin through the promotion of the expression of profilaggrin mRNA in epidermal keratinocytes, and the water environment of the stratum corneum can be essentially improved.

As a natural product-derived filaggrin synthesis promoter, for example, citrus extract (see Patent Document 13), liquiritin known as a flavanone glycoside contained in natural plants (see Patent Document 14), and a natural product-derived pro. As at least one of filaggrin and filaggrin protein production promoter, a plant extract belonging to the genus Citrus or a yeast extract (see Patent Document 15) has been proposed.

The epidermis has a protective function that protects the skin from various stimuli from the outside world by constantly producing new keratinocytes by division of keratinocytes and subsequent differentiation. In particular, in the process of differentiation of keratinocytes, proteins such as involucrin are expressed from the spinous layer to the granular layer, cross-linked by the action of the enzyme transglutaminase-1, and are insoluble cell membrane-like structures that enclose the keratinocytes. It forms a fide envelope (hereinafter abbreviated as "CE") and contributes to the stability of the cytoskeleton and structure of keratinocytes.
However, when the amount of transglutaminase-1 produced in the epidermis decreases due to various factors, CE formation becomes incomplete and keratinization does not occur normally. As a result, it is considered that the keratin barrier function and the moisturizing function of the skin are deteriorated, and skin symptoms such as rough skin and dry skin are exhibited.
Therefore, by increasing the production of transglutaminase-1 in the epidermis of keratinocytes, promoting the formation of CE and normalizing keratinization, the skin barrier function associated with external stimuli such as dryness and ultraviolet rays can be achieved. It is thought that it can suppress the decrease and prevent / improve various skin symptoms such as dry skin and rough skin.

Noni extract (see Patent Document 16), royal jelly extract (see Patent Document 17), and the like have been proposed as transglutaminase-1 production promoters derived from natural products.

Further, in skin cells, it is known that aquaporin known as a water channel is expressed on the cell membrane and plays a role of taking up low molecular weight substances such as water in the intercellular space into the cell. In humans, the existence of 13 types of aquaporins (AQP0 to AQP12) is known. In epidermal cells, aquaporin 3 (AQP3) is mainly present, and it is considered that it plays a role of taking up low molecular weight compounds such as glycerol and urea involved in water retention in addition to water. ing.

However, AQP3 decreases with aging, and it is suggested that this is one of the causes of the decrease in water retention function. Therefore, by promoting the expression of AQP3, the water retention function and barrier function due to aging, etc. It is considered that it is possible to control (see Non-Patent Document 4). As a substance having an AQP3 expression promoting action, for example, an extract from the leaf part of star fruit (see Patent Document 18) and the like are known.

Conventionally, it was thought that the barrier function of the skin is borne only by the stratum corneum, but when the constituent proteins of tight junctions (hereinafter abbreviated as "TJ") existing in the stratum granulosum of the epidermis are deleted at the gene level, In recent years, TJs are also considered to play an important role in the barrier function of the skin because the barrier function of the skin is disrupted (see Non-Patent Document 5). The TJ is a binding device that not only brings adjacent cells into close contact with each other, but also controls the permeation of a substance by sealing the gap between cells. TJ is composed of cell membrane proteins claudin and occludin, and these proteins are considered to constitute the skeleton of TJ strands and control the barrier function of TJ (see Non-Patent Document 6). From the above, when the expression of claudin and occludin is reduced for some reason, structural destruction of TJ occurs and it does not function as a permeation barrier for substances, resulting in dry skin, rough skin, atopic dermatitis and various types. It is expected to cause skin symptoms such as infectious diseases.

Therefore, by promoting the formation of TJ of epidermal keratinocytes by promoting the production of claudin and occludin in the epidermis, the barrier function and water retention function of the skin can be enhanced, and the skin symptoms can be prevented or ameliorated. Conceivable. Based on this idea, natural product-derived Coptis chinensis extract (see Patent Document 19), spruce extract (see Patent Document 20), and the like have been proposed as those for improving the skin barrier function through the TJ formation promoting action. ing.

In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as a factor that induces changes associated with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1) is known as an enzyme that decomposes collagen, which is a major component of the extracellular matrix of the dermis of the skin, but its expression is greatly increased by irradiation with ultraviolet rays. However, it is thought to contribute to the decrease and degeneration of collagen, and to be a major factor in the formation of wrinkles on the skin and the decrease in elasticity. Therefore, inhibiting the activity of MMP-1 is important for preventing and ameliorating the aging symptoms of the skin.

Examples of those having such an MMP-1 inhibitory effect include an extract from Himalayan cherry tree (see Patent Document 21), an extract from Zingiberaceae Zingiba cassumner or Ficus umbellata of Moraceae (see Patent Document 22). Etc. are known.

On the other hand, although fermented liquids obtained by fermenting Moxa plants with aspergillus (see Patent Document 23) and fermented liquids obtained by fermenting Tachijakousou with aspergillus (see Patent Document 24) are known, these plant fermented liquids are used. It is not known to have anti-aging, anti-oxidant, anti-inflammatory, whitening and other effects.

Japanese Unexamined Patent Publication No. 2003-81848 Japanese Unexamined Patent Publication No. 2005-29483 Japanese Unexamined Patent Publication No. 2006-321730 JP-A-2007-8902 Japanese Unexamined Patent Publication No. 2003-55242 Japanese Unexamined Patent Publication No. 2003-12532 JP-A-2002-370962 Japanese Unexamined Patent Publication No. 2004-83488 Japanese Unexamined Patent Publication No. 2001-213757 JP-A-2002-20122 JP-A-2002-226323 Japanese Unexamined Patent Publication No. 2003-1486837 JP-A-2002-363504 Japanese Unexamined Patent Publication No. 2003-146886 Japanese Unexamined Patent Publication No. 2001-261568 JP-A-2010-09093 Japanese Unexamined Patent Publication No. 2009-184955 JP-A-2009-191039 JP-A-2007-176830 JP-A-2007-176835 Japanese Unexamined Patent Publication No. 2003-176232 Japanese Unexamined Patent Publication No. 2003-176230 JP 2011-140453 Japanese Unexamined Patent Publication No. 2011-130689

"Fragrance Journal" Extra Edition No. 14, p156, 1995 Fragrance Journal Extra Edition, vol. 17, pp. 14-19 (2000) Arch. Dermatol. Res. , Vol. 288, pp. 442-446 (1996) "Fragrance Journal", 2006, Vol. 34, No. 10, p. 19-23 J. Cell Biol. , Vol. 156, pp. 1099-1111 (2002) Journal of the Japanese Society of Cosmetic Science, vol. 31, pp. 296-301 (2007)

An object of the present invention is to solve the above-mentioned problems in the past and to achieve the following object.
That is, the present invention has an excellent anti-aging effect and a highly safe anti-aging agent, an excellent antioxidant effect and a highly safe antioxidant, and an excellent anti-inflammatory effect. It is an object of the present invention to provide a highly safe anti-inflammatory agent and a highly safe whitening agent having an excellent whitening effect.
The present invention also provides 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. With the goal.

The means for solving the above-mentioned problems are as follows. That is,
<1> Containing at least one of the fermented liquor of the genus Jiuqu plant, the fermented liquor of Tachijakousou aspergillus, the fermented liquor of Kousuihakka aspergillus, and the fermented liquor of Yagurumagiku as the active ingredient. It is a characteristic anti-aging agent.
<2> Containing at least one of the fermented liquor of the genus Yomogi plant, the fermented liquor of Tachijakousou, the fermented liquor of Kousuihakka, and the fermented liquor of Yagurumagiku as the active ingredient. It is a characteristic antioxidant.
<3> An anti-inflammatory agent characterized by containing at least one of a fermented liquor produced by Aspergillus oryzae of a plant of the genus Moxa and a fermented liquor produced by Aspergillus oryzae of Kousuihakka as an active ingredient.
<4> A whitening agent characterized by containing at least one of a fermented liquid of Jiuqu of a plant of the genus Moxa, a fermented liquid of Jiuqu of Kousuihakka, and a fermented liquid of Jiuqu of cornflower as an active ingredient.
<5> From the group consisting of the anti-aging agent according to <1>, the antioxidant according to <2>, the anti-inflammatory agent according to <3>, and the whitening agent according to <4>. It is a cosmetic characterized by containing at least one selected.

According to the present invention, it is possible to solve the above-mentioned problems in the past, achieve the above-mentioned object, have an excellent anti-aging effect, and have a highly safe anti-aging agent and an excellent antioxidant effect. It is possible to provide a highly safe antioxidant, an anti-inflammatory agent having an excellent anti-inflammatory effect and a high safety, and a whitening agent having an excellent whitening effect and a high safety.
The present invention also provides 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. Can be done.

FIG. 1A is a photograph showing an example of droplets at the time of measuring the contact angle of the Yamayomogi fermented liquid 1 of Production Example 1. FIG. 1B is a photograph showing an example of droplets at the time of measuring the contact angle of the Yamayomogi fermented liquid 2 of Production Example 2. FIG. 1C is a photograph showing an example of droplets at the time of measuring the contact angle of the Yamayomogi extract of Comparative Production Example 1. FIG. 2A is a photograph showing an example of droplets at the time of measuring the contact angle of the capillary wormwood fermented liquid 1 of Production Example 3. FIG. 2B is a photograph showing an example of droplets at the time of measuring the contact angle of the capillary wormwood fermented liquid 2 of Production Example 4. FIG. 2C is a photograph showing an example of droplets at the time of measuring the contact angle of the Capillary wormwood extract of Comparative Production Example 2. FIG. 3A is a photograph showing an example of droplets at the time of measuring the contact angle of the Tachijakousou fermented liquid 1 of Production Example 5. FIG. 3B is a photograph showing an example of droplets at the time of measuring the contact angle of the Tachijakousou fermented liquid 2 of Production Example 6. FIG. 3C is a photograph showing an example of droplets at the time of measuring the contact angle of the Tachijakousou extract of Comparative Production Example 3. FIG. 4A is a photograph showing an example of droplets at the time of measuring the contact angle of the Kousuihakka fermented liquid 1 of Production Example 7. FIG. 4B is a photograph showing an example of droplets at the time of measuring the contact angle of the Kousuihakka fermented liquid 2 of Production Example 8. FIG. 4C is a photograph showing an example of droplets at the time of measuring the contact angle of the Kousuihakka extract of Comparative Production Example 4. FIG. 5A is a photograph showing an example of droplets at the time of measuring the contact angle of the cornflower fermented liquid 1 of Production Example 9. FIG. 5B is a photograph showing an example of droplets at the time of measuring the contact angle of the cornflower fermented liquid 2 of Production Example 10. FIG. 5C is a photograph showing an example of droplets at the time of measuring the contact angle of the cornflower extract of Comparative Production Example 5.

(Anti-aging agents, antioxidants, anti-inflammatory agents, and whitening agents)
<Anti-aging agent>
The anti-aging agent of the present invention is a fermented liquor produced by aspergillus of a plant of the genus Yomogi (hereinafter, may be referred to as "fermented liquor of a plant of the genus Yomogi") and a fermented liquor produced by aspergillus of Tachijakousou (hereinafter, "fermented liquor of Tachijakousou"). (Sometimes referred to as), fermented liquid with aspergillus of Kousuihakka (hereinafter sometimes referred to as "fermented liquid of Kousuihakka"), and fermented liquid with aspergillus of Yagurumagiku (hereinafter sometimes referred to as "fermented liquid of Yagurumagiku") The fermented liquor of at least one of the above is contained as an active ingredient, and if necessary, other ingredients are further contained.

The fermented liquid of the genus Claudin, the fermented liquid of Tachijakousou, the fermented liquid of Kousuihakka, and the fermented liquid of Yagurumagiku have a matrix metalloproteinase-1 (MMP-1) activity inhibitory action, hyaluronan synthase 3 (Hyaluronan synthesis 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 (transglutaminase-) 1; TGM-1) has at least one action selected from the group consisting of mRNA expression promoting action, aquaporin 3 (AQP3) mRNA expression promoting action, and phyllagulin mRNA expression promoting action, and these Utilizing its action, it can be used as an active ingredient of an antiaging agent.

Therefore, the anti-aging agent has an MMP-1 activity inhibitory action, 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, It has at least one action selected from the group consisting of an occludin mRNA expression promoting action, a transglutaminase-1 mRNA expression promoting action, an aquaporin 3 mRNA expression promoting action, and a phyllagulin mRNA expression promoting action.

The fermented liquid of the genus Yomogi, the fermented liquid of Tachijakousou, the fermented liquid of Kousuihakka, and the fermented liquid of Yagurumagiku have an MMP-1 activity inhibitory action, a hyaluronic acid synthase 3 mRNA expression promoting action, and a 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 phyllagulin mRNA expression promoting action. Although the details of the substance exhibiting the above are unknown, the fermented liquid of the genus Yomogi, the fermented liquid of Tachijakousou, the fermented liquid of Kousuihakka, and the fermented liquid of Yagurumagiku have such excellent effects and are anti-fermenting. The usefulness as an aging agent has not been known at all in the past, and is a new finding by the present inventors.

<Antioxidant>
The antioxidant of the present invention is a fermented liquor of a plant of the genus Yomogi (fermented liquor of a genus Yomogi), a fermented liquor of fermented liquor of Tachijakousou (fermented liquor of Tachijakousou), and a fermented liquor of fermented liquor of Kousuihakka ), And at least one of the fermented liquids of Yagurumagiku fermented liquid (Yagurumagiku fermented liquid) as an active ingredient, and if necessary, other ingredients are contained.

The fermented liquid of the genus Lemon balm, the fermented liquid of Tachijakousou, the fermented liquid of Kousuihakka, and the fermented liquid of Yagurumagiku have a diphenyl-p-picrylhydrazil (DPPH) radical scavenging action, and this action is utilized. Then, it can be used as an active ingredient of an antioxidant.
Therefore, the antioxidant has a DPPH radical scavenging effect.

The details of the substance exhibiting the DPPH radical scavenging action of the fermented liquid of the genus Lemon balm, the fermented liquid of Tachijakousou, the fermented liquid of Kousuihakka, and the fermented liquid of Yagurumagiku are unknown, but the fermented liquid of the genus Lemon balm. , The Tachijakousou fermented liquid, the Kousuihakka fermented liquid, and the Yagurumagiku fermented liquid have such an excellent action and are useful as an antioxidant, and it has not been known at all in the present invention. This is a new finding by the people.

<Anti-inflammatory agent>
The anti-inflammatory agent of the present invention contains at least one of a fermented liquid of Moxa plant (Moxa plant fermented liquid) and a fermented liquid of Kousuihakka aspergillus (Moxa fermented liquid) as an active ingredient. However, if necessary, it contains other components.

The fermented liquid of the genus Moxa and the fermented liquid of Kousuihakka have a hyaluronidase activity inhibitory action, and this action can be utilized as an active ingredient of an anti-inflammatory agent.
Therefore, the anti-inflammatory agent has a hyaluronidase activity inhibitory action.

The details of the substance that exerts the hyaluronidase activity inhibitory action of the fermented moxa plant and the fermented Kousuihakka are unknown, but the fermented moxa plant and the fermented Kousuihakka have such excellent effects. It has not been known at all in the past that it is useful as an anti-inflammatory agent, and it is a new finding by the present inventors.

<Whitening agent>
The whitening agent of the present invention is a fermented liquor of Jiuqu of plants of the genus Yomogi (fermented liquor of plants of the genus Yomogi), a fermented liquor of fermented liquor of Jiuqu of Kousuihakka (fermented liquor of Jiuqu), and a fermented liquor of fermented liquor of Jiuqu of Yagurumagiku (fermented liquid of Yagurumagiku). It contains at least one of the fermentation broths as an active ingredient, and further contains other ingredients as needed.

The fermented liquid of the genus Moxa, the fermented liquid of Kousuihakka, and the fermented liquid of cornflower have at least one action of inhibiting tyrosinase activity and suppressing melanin production, and utilizing this action, a whitening agent. It can be used as an active ingredient.
Therefore, the whitening agent has at least one action of inhibiting tyrosinase activity and suppressing melanin production.

The details of the substance exhibiting at least one of the tyrosinase activity inhibitory action and the melanin production inhibitory action of the moxa genus fermented liquor, the moxa fermented liquor, and the yagurumagiku fermented liquor are unknown, but the moxa plant fermentation It has not been known at all conventionally that the liquid and the fermented mugwort have such an excellent action and are useful as a whitening agent, and it is a new finding by the present inventors.

<< Mugwort plant fermented liquid >>
The moxa plant fermented liquor is a fermented liquor produced by aspergillus of a plant belonging to the genus Artemisia (hereinafter, may be referred to as "Mugwort plant").

-Mugwort plants-
The moxa plant used as the fermentation raw material is a perennial herb belonging to the Asteraceae ( Compositae ) Moxa genus ( Artemisia ), and has been used as a raw material for foods and medicinal products since ancient times. It grows naturally or is widely cultivated in Japan such as Hokkaido, Honshu, Shikoku, and Kyushu, and is easily available from these areas.

The type of the plant of the genus Artemisia is not particularly limited and may be appropriately selected depending on the intended purpose. For example, Artemisia montana (Nakai) Pamp. , Artemisia capillaris Thunbergii, Artemisia princeps Pampan ., Artemisia japonica Thunb., Artemisia absinthium L. , Artemisia lactiflora Wall., Artemisia maritima L., Artemisia scoparia Waldst.et kit. And the like. These may be used alone or in combination of two or more.
The method for obtaining the plant of the genus Artemisia is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used.

The site of use of the moxa plant used as the fermentation raw material is not particularly limited and may be appropriately selected depending on the intended purpose. For example, flowers, buds, fruits, fruit skins, seeds, seed coats, stems, leaves, etc. Above-ground parts such as branches and 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 preferable as the site of use of the plant of the genus Artemisia.

The size of the Artemisia genus plant used as the fermentation raw material is not particularly limited as long as the size is such that the aspergillus can be cultivated, and can be appropriately selected according to the purpose. The size of the sagebrush, the desired size of the cut, the size of the fine powder (powder), and the like.

The state of the Artemisia genus plant used as the fermentation raw material is not particularly limited as long as the aspergillus can be cultivated, and can be appropriately selected depending on the purpose. For example, the state as it is collected. , Dry state, crushed state, squeezed state, extract state and the like. Among these, the as-collected state, the crushed state, the squeezed state, and the extracted state are preferable, and the collected as-is state and the crushed state are more preferable, because the aspergillus is likely to act.

The method for drying the mugwort plant is not particularly limited and may be appropriately selected depending on the intended purpose. For example, it is dried in the sun or using a commonly used dryer. The method etc. can be mentioned.

The method for crushing the wormwood plant is not particularly limited and may be appropriately selected depending on the intended purpose. For example, it is crushed by a mixer, a sugar mill, a power mill, a jet mill, an impact crusher or the like. How to do it.

The method for bringing the mugwort plant into the state of squeezing is not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include squeezing.

The method for bringing the Artemisia plant into the state of the extract is not particularly limited, and a method generally used for extracting the plant can be appropriately selected depending on the intended purpose.

The Artemisia plant used as the fermentation raw material is preferably sterilized before inoculation with the Jiuqu. The means for sterilizing the plant of the genus Artemisia is not particularly limited, and can be appropriately selected from known methods.

-Jiuqu-
As the koji mold to ferment the Artemisia plant is not particularly limited and may be appropriately selected depending on the purpose, for example, Aspergillus oryzae (Aspergillus oryzae), Aspergillus sojae (Aspergillus sojae) yellow koji molds such as; Aspergillus Ruchu Black aspergillus such as Ensis ( Aspergillus luchuensis ); White Aspergillus such as Aspergillus kawauchii ; mutant strains thereof and the like can be mentioned. These may be used alone or in combination of two or more. Among them, as the koji mold, Aspergillus oryzae (Aspergillus oryzae) is an anti-aging effect, antioxidant, anti-inflammatory, and preferable from the viewpoint of excellent least one of the effects of whitening effect.
The method for obtaining the aspergillus is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used. Further, as the aspergillus, seed koji made from rice or the like may be used, or the moxa plant seed koji described later may be used, and the aspergillus cultivated in a medium (agar medium, liquid medium, etc.) is used. You may. Among these, it is preferable to use the above-mentioned Artemisia plant species koji because it is excellent in at least one of anti-aging action, antioxidant action, anti-inflammatory action, and whitening action.

The amount of the aspergillus inoculated into the moxa plant used as the fermentation raw material is not particularly limited as long as the amount of the moxa plant can be fermented, and may be appropriately selected depending on the purpose. However, when the fermentation raw material is in a liquid state, 1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL is preferable, and when the fermentation raw material is in a solid state, 1 × 10 ³ pieces / g to 1 × 10 ⁸ pieces / g are preferable.

When inoculating the Mugwort plant with the Jiuqu, it is preferable to add water. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose, but it is preferable to add 500 parts by mass to 5,000 parts by mass with respect to 100 parts by mass of the plant of the genus Moxa. , 1,000 parts by mass to 4,000 parts by mass is more preferable, and 1,500 parts by mass to 3,000 parts by mass is particularly preferable.

The temperature of the fermentation (culture) is not particularly limited as long as it is within the temperature range in which fermentation by the aspergillus can be performed, and can be appropriately selected depending on the intended purpose. However, 20 ° C to 40 ° C is preferable, and 25 ° C. ℃ -35 ℃ is more preferable. If the fermentation temperature is less than 20 ° C., the moxa plant cannot be sufficiently fermented, and at least one of the anti-aging effect, the antioxidant effect, the anti-inflammatory effect, and the whitening effect is ineffective. May be sufficient. If the temperature exceeds 50 ° C., the aspergillus may not grow.

The fermentation (culture) time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 10 hours to 40 hours, more preferably 20 hours to 30 hours. If the fermentation time is less than 10 hours, the plant of the genus Artemisia cannot be sufficiently fermented, and at least one of the anti-aging effect, the antioxidant effect, the anti-inflammatory effect, and the whitening effect is ineffective. May be sufficient.

The method for stopping the fermentation (culture) is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof 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 cannot grow, and can be appropriately selected depending on the intended purpose. However, 50 ° C. or higher is preferable, and 70 ° C. or higher is more preferable. It is 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 anti-aging action, antioxidant action, anti-inflammatory action, and whitening action is ineffective. May be sufficient.
The heating time for stopping the fermentation is not particularly limited as long as the aspergillus can be prevented from growing, and can be appropriately selected depending on the intended purpose, but is preferably 5 minutes or more, preferably 10 minutes to. 20 minutes is more preferable. If the heating time is less than 5 minutes, the fermentation may not be stopped, and if it exceeds 20 minutes, at least one of the anti-aging effect, the antioxidant effect, the anti-inflammatory effect, and the whitening effect is ineffective. May be sufficient.

It is preferable that the fermented liquid of the genus Artemisia after the fermentation is stopped is cooled. The cooling method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of allowing the material to stand at room temperature or in a refrigerator.

The number of times the moxa plant is fermented by the aspergillus is not particularly limited, and can be appropriately selected depending on the intended purpose, and may be once or a plurality of times. You may.

When the fermentation is carried out a plurality of times, the aspergillus may be inoculated only for the first time, may be inoculated only several times, or may be inoculated at all times, but it is preferable to inoculate only at the first time.
When the fermentation is carried out a plurality of times, the fermentation temperature and the fermentation time may be different or the same.

--- Mugwort plant species Jiuqu ---
The moxa plant seed koji is obtained by using the moxa plant as a seed koji raw material and inoculating the seed koji raw material with aspergillus fungi to inoculate the moxa plant with a sufficient amount of spores. By using this in the fermentation, it is advantageous in that a fermented liquid of the genus Artemisia, which is excellent in familiarity with the skin, can be obtained more efficiently and easily.

As the Mugwort plant used as the seed koji raw material, the same plant as described in the above-Mugwort plant-can be used, and the mode, size, state and the like of the Mugwort plant are used. Is the same.

As the aspergillus used in the production of the mugwort plant species aspergillus, the same as those described in the above-aspergillus-can be used.

The inoculation amount of the Moxa bacterium to the Moxa plant used as the seed koji raw material is not particularly limited and may be appropriately selected depending on the purpose, but with respect to 100 parts by mass of the Moxa plant. It is preferable to inoculate 5 parts by mass to 100 parts by mass of aspergillus (1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL) suspended in sterilized water, and it is more preferable to inoculate 10 parts by mass to 50 parts by mass. It is preferable to inoculate 20 parts by mass to 30 parts by mass, and it is particularly preferable. If the inoculation amount of the Aspergillus oryzae to 100 parts by mass of the Moxa plant is less than 5 parts by mass, a sufficient amount of spores may not be formed on the Moxa plant, and if it exceeds 100 parts by mass, the water content is excessive. May overgrow.

It is preferable to add water when inoculating the Mugwort plant used as a raw material for the seed koji. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 10 parts by mass to 250 parts by mass with respect to 100 parts by mass of the plant of the genus Moxa. It is more preferable to add parts by mass to 200 parts by mass, and it is particularly preferable to add parts by mass to 150 parts by mass. If the amount of water added to 100 parts by mass of the plant of the genus Artemisia is less than 10 parts by mass, a sufficient amount of spores may not be formed on the plant of the genus Artemisia.

The temperature of the culture is not particularly limited as long as it is within the temperature range in which the aspergillus can grow, and can be appropriately selected depending on the intended purpose, but is preferably 20 ° C to 40 ° C, preferably 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 plant of the genus Artemisia. If the temperature exceeds 50 ° C., the aspergillus may not grow.

The culture time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the culture time is less than 80 hours, a sufficient amount of spores may not be formed on the Artemisia plant, and if it exceeds 210 hours, the germination rate of spores may decrease.

The fermented sagebrush plant may contain the cells of the aspergillus or the cells of the aspergillus may be removed, but the cells of the aspergillus may be removed. Is preferable.

The state of the moxa plant fermented liquid is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the moxa plant fermented liquid itself may be used, and the moxa plant fermented liquid may be purified. It may be a product, a concentrate of the moxa plant fermented liquid, a diluted product of the moxa plant fermented liquid, or the like. In addition, the moxa plant fermented liquid may be a dried product of the moxa plant fermented liquid mixed or dissolved in a solvent such as water or a hydrophilic solvent again.

The purified product of the fermented wormwood plant is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the solid content in the fermented wormwood plant (for example, the plant body of the wormwood plant). , Moxa cells, mugwort, etc.) have been removed.
The removing means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include filtration.
The filtration method is not particularly limited, and can be appropriately selected from known methods according to the purpose.

The dilution of the fermented moxa plant and the concentrate of the fermented moxa plant are not particularly limited and may be appropriately selected depending on the intended purpose. For example, the fermented moxa plant has a desired concentration. Examples include prepared products.
The diluting means is not particularly limited, and can be appropriately selected from known methods according to the purpose.
The means for concentration is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include vacuum concentration.

The dried product of the fermented mugwort plant is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a dried product of the fermented mugwort plant.
The drying means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include freeze-drying.

The fermented moxa plant is not particularly limited as long as it is a fermented moxa plant fermented with aspergillus, and can be appropriately selected depending on the purpose. However, the contact angle is 81 ° or less because it is familiar to the skin. The fermented moxa plant is preferable, and the fermented moxa plant having a contact angle of 78 ° or less is more preferable.

In the present specification, the contact angle is defined as a contact angle / surface tension measuring device (FTA1000 Falcon, manufactured by First Ten Angstroms), and 3 μL of the measurement sample is dropped onto the sample stage of the device, and the temperature is 22 ° C. The measurement is carried out by the sessile drop method under the condition of a relative humidity of 20%, and the contact angle θ (°) of 1,000 ms is represented by the value obtained by the θ / 2 method.
The contact angle is used as an index to indicate "wetting", and "the angle between the liquid surface and the solid surface (takes the angle inside the liquid) at the place where the free surface of the stationary liquid touches the solid wall" is used. It is defined as "Iwanami Shoten Co., Ltd., 4th edition of the Physics and Chemistry Dictionary). The contact angle is determined by the magnitude relationship between the cohesive force between the liquid molecules and the adhesive force between the solid walls, and is an acute angle when the liquid wets the solid (the adhesive force is large) and an obtuse angle when the liquid does not wet. Therefore, the smaller the contact angle, the easier it is to get wet, that is, the better the skin fits. Therefore, the lower limit of the contact angle of the fermented mugwort plant is not particularly limited and may be appropriately selected according to the purpose. it can.

<< Tachijakousou Fermented Liquid >>
The fermented liquid of Tachijakousou is a fermented liquid of Jiuqu of Tachijakousou.

-Tachijakousou-
Thymus vulgaris to be used as the raw material for fermentation (Thymus vulgaris Linne) is a Labiatae (Labiatae) Ibuki thyme (Thymus) perennial woody belonging to the genus, is a kind of herb, is used since ancient times as a raw material for edible and medicinal ing. Another name is common time. The place of origin is the Mediterranean coast, but it is also native or cultivated in Japan and is easily available from these areas.
The method for obtaining the garden thyme is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used.

The site of use of the Tachijakousou used as the fermentation raw material is not particularly limited and may be appropriately selected depending on the intended purpose. For example, flowers, buds, fruits, peels, seeds, seed coats, stems, leaves and branches. , Bark, trunk, branches and leaves, etc. above ground; roots, rhizomes, etc. underground. These may be used alone or in combination of two or more. Among these, the above-ground portion is preferable as the site where the garden thyme is used.

The size of the Tachijakousou used as the fermentation raw material is not particularly limited as long as it is a size capable of culturing the aspergillus, and can be appropriately selected according to the purpose. Examples thereof include a size, a desired size cut, and a finely powdered size.

The state of the Tachijakousou used as the fermentation raw material is not particularly limited as long as the aspergillus can be cultivated, and can be appropriately selected depending on the purpose. For example, the state as it is collected, Examples include a dry state, a crushed state, a squeezed state, and an extract state. Among these, the as-collected state, the crushed state, the squeezed state, and the extracted state are preferable, and the collected as-is state and the crushed state are more preferable, because the aspergillus is likely to act.

The method for drying the garden thyme is not particularly limited and may be appropriately selected depending on the intended purpose. For example, a method of drying in the sun or a method of drying using a commonly used dryer. And so on.

The method for bringing the thyme into the crushed state is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the thyme is crushed by a mixer, a sugar mill, a power mill, a jet mill, an impact crusher or the like. The method etc. can be mentioned.

The method for bringing the Tachijakousou into the state of squeezing is not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include squeezing.

The method for bringing the Thyme into the state of the extract is not particularly limited, and a method generally used for extracting plants can be appropriately selected depending on the intended purpose.

The Thyme japonicum used as the fermentation raw material is preferably sterilized before inoculation with the Jiuqu. The means for sterilizing the garden thyme is not particularly limited, and can be appropriately selected from known methods.

-Jiuqu-
The aspergillus that ferments the Tachijakousou is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include those described in << Mugwort plant fermented liquid >>. These may be used alone or in combination of two or more. Among them, as the koji mold, Aspergillus oryzae (Aspergillus oryzae) is preferable from the viewpoint of excellent least one of the effects of anti-aging effect and antioxidant activity.
The method for obtaining the aspergillus is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used. Further, as the aspergillus, seed koji made from rice or the like may be used, or Tachijakousou seed koji, which will be described later, may be used, and aspergillus cultivated in a medium (agar medium, liquid medium, etc.) may be used. You may. Among these, it is preferable to use the Tachijakousou seed koji because it is excellent in at least one of the anti-aging action and the antioxidant action.

The amount of the aspergillus inoculated into the Tachijakousou used as the fermentation raw material is not particularly limited as long as the amount of the Tachijakousou can be fermented, and can be appropriately selected depending on the intended purpose. When the fermentation raw material is in a liquid state, 1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL is preferable, and when the fermentation raw material is in a solid state, 1 × 10 ³ pieces / mL. / G to 1 × 10 ⁸ pieces / g is preferable.

When inoculating the Aspergillus oryzae with the Tachijakousou, it is preferable to add water. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 500 parts by mass to 5,000 parts by mass with respect to 100 parts by mass of the Tachijakousou. It is more preferable to add 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.

The temperature of the fermentation (culture) is not particularly limited as long as it is within the temperature range in which fermentation by the aspergillus can be performed, and can be appropriately selected depending on the intended purpose. However, 20 ° C to 40 ° C is preferable, and 25 ° C. ℃ -35 ℃ is more preferable. If the fermentation temperature is less than 20 ° C., the garden thyme may not be sufficiently fermented, and at least one of the anti-aging action and the antioxidant action may be insufficient.

The fermentation (culture) time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 10 hours to 40 hours, more preferably 20 hours to 30 hours. If the fermentation time is less than 10 hours, the garden thyme may not be sufficiently fermented, and at least one of the anti-aging action and the antioxidant action may be insufficient.

The method for stopping the fermentation (culture) is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof 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 cannot grow, and can be appropriately selected depending on the intended purpose. However, 50 ° C. or higher is preferable, and 70 ° C. or higher is more preferable. It is 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 action and the antioxidant action may be insufficient.
The heating time for stopping the fermentation is not particularly limited as long as the aspergillus can be prevented from growing, and can be appropriately selected depending on the intended purpose, but is preferably 5 minutes or more, preferably 10 minutes to. 20 minutes is more preferable. If the heating time is less than 5 minutes, the fermentation may not be stopped, and if it exceeds 20 minutes, at least one of the anti-aging action and the antioxidant action may be insufficient.

It is preferable that the fermented thyme broth after the fermentation is stopped is cooled. The cooling method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of allowing the material to stand at room temperature or in a refrigerator.

The number of times the Tachijakousou is fermented by the Jiuqu is not particularly limited, and can be appropriately selected depending on the purpose, and may be once or a plurality of times. May be good.

When the fermentation is carried out a plurality of times, the aspergillus may be inoculated only for the first time, may be inoculated only several times, or may be inoculated at all times, but it is preferable to inoculate only at the first time.
When the fermentation is carried out a plurality of times, the fermentation temperature and the fermentation time may be different or the same.

--Tachijakousou Seed Jiuqu ---
The Tachijakousou seed koji is obtained by using the Tachijakousou as a seed koji raw material, inoculating the seed koji raw material with aspergillus, and causing a sufficient amount of spores to grow on the Tachijakousou. By using this in the fermentation, it is advantageous in that a fermented Tachijakousou liquid having excellent skin compatibility can be obtained more efficiently and easily.

As the Tachijakousou used as the raw material for the seed koji, the same ones as those described in the above-Tachijakousou can be used, and the same applies to the usage site, size, state and the like of the Tachijakousou. is there.

As the aspergillus used in the production of the Tachijakousou seed koji, the same as those described in the above-aspergillus-can be used.

The amount of the aspergillus inoculated into the Tachijakousou used as the raw material for the seed koji is not particularly limited and may be appropriately selected depending on the intended purpose. However, sterilized water is added to 100 parts by mass of the Tachijakousou. It is preferable to inoculate 5 parts by mass to 100 parts by mass of aspergillus (1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL) suspended in, and more preferably 10 parts by mass to 50 parts by mass. It is particularly preferable to inoculate 20 parts by mass to 30 parts by mass. If the inoculation amount of the aspergillus to 100 parts by mass of the Tachijakousou is less than 5 parts by mass, a sufficient amount of spores may not be formed on the Tachijakousou, and if it exceeds 100 parts by mass, it is abnormal due to excessive water content. May breed.

It is preferable to add water when inoculating the Aspergillus oryzae to the Tachijakousou used as the seed koji raw material. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 10 parts by mass to 250 parts by mass with respect to 100 parts by mass of the garden thyme, and 20 parts by mass. It is more preferable to add ~ 200 parts by mass, and it is particularly preferable to add 30 parts by mass to 150 parts by mass. If the amount of water added to 100 parts by mass of the garden thyme is less than 10 parts by mass, a sufficient amount of spores may not be formed on the garden thyme.

The temperature of the culture is not particularly limited as long as it is within the temperature range in which the aspergillus can grow, and can be appropriately selected depending on the intended purpose, but is preferably 20 ° C to 40 ° C, preferably 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 garden thyme. If the temperature exceeds 50 ° C., the aspergillus may not grow.

The culture time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the culture time is less than 80 hours, a sufficient amount of spores may not be formed on the garden thyme, and if it exceeds 210 hours, the germination rate of spores may decrease.

The fermented thyme liquor may contain the cells of the aspergillus or the cells of the aspergillus may be removed, but the fermented solution of the aspergillus may be removed. Is preferable.

The state of the fermented thyme is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the fermented thyme may be the fermented thyme itself, or the purified product of the fermented thyme. It may be a concentrate of the fermented thyme, a diluted product of the fermented thyme, or the like. Further, the fermented Tachijakousou liquid may be a dried product of the fermented Tachijakousou liquid mixed or dissolved in a solvent such as water or a hydrophilic solvent again.

The purified product of the Tachijakousou fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the solid content in the Tachijakousou fermented liquor (for example, the plant body of the Tachijakousou and the aspergillus) Examples include those from which bacterial cells, sediments, etc.) have been removed.
The removing means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include filtration.
The filtration method is not particularly limited, and can be appropriately selected from known methods according to the purpose.

The dilution of the Tachijakousou fermented liquor and the concentrate of the Tachijakousou fermented liquor are not particularly limited and may be appropriately selected depending on the intended purpose. For example, the Tachijakousou fermented liquor was prepared to a desired concentration. Things can be mentioned.
The diluting means is not particularly limited, and can be appropriately selected from known methods according to the purpose.
The means for concentration is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include vacuum concentration.

The dried product of the fermented Tachijakousou is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a dried product of the fermented Tachijakousou.
The drying means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include freeze-drying.

The fermented thyme broth is not particularly limited as long as it is a fermented thyme broth made from aspergillus of thyme, and can be appropriately selected depending on the intended purpose, but the contact angle is 87 ° or less in terms of familiarity with the skin. The fermented thyme liquor is preferable, and the fermented thyme liquor having a contact angle of 81 ° or less is more preferable.

<< Kousuihakka Fermented Liquid >>
The fermented balm of Lemon balm is a fermented balm of Lemon balm.

-Kousui Hacker-
The perfume peppermint used as a fermentation feedstock (Melissa officinalis Linne) is a Labiatae (Labiatae) Perfume mint (Melissa) perennial herb belonging to the genus. It is a kind of herb and has been used as an edible and medicinal ingredient since ancient times. Other names include lemon balm and Seiyoyamahakka. The place of origin is Southern Europe, but it is also native or cultivated in Japan and is easily available from these areas.
The method for obtaining the lemon balm is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used.

The site of use of the Kousuihakka used as the fermentation raw material is not particularly limited and may be appropriately selected depending on the intended purpose. For example, flowers, buds, fruits, pericarps, seeds, seed coats, stems, leaves and branches. , Above-ground parts such as branches and leaves; Underground parts such as roots and rhizomes. These may be used alone or in combination of two or more. Among these, the above-ground part is preferable as the part where the Kousuihakka is used.

The size of the lemon balm used as the fermentation raw material is not particularly limited as long as it is large enough to cultivate the aspergillus, and can be appropriately selected according to the purpose. For example, as it is collected. Examples thereof include a size, a desired size cut, and a finely powdered size.

The state of the lemon balm used as the fermentation raw material is not particularly limited as long as the aspergillus can be cultivated, and can be appropriately selected depending on the purpose. For example, the state as it is collected, Examples include a dry state, a crushed state, a squeezed state, and an extract state. Among these, the as-collected state, the crushed state, the squeezed state, and the extracted state are preferable, and the collected as-is state and the crushed state are more preferable, because the aspergillus is likely to act.

The method for drying the lemon balm is not particularly limited and may be appropriately selected depending on the intended purpose. For example, a method of drying in the sun or a method of drying using a commonly used dryer. And so on.

The method for bringing the Kousui hacker into the crushed state is not particularly limited and may be appropriately selected depending on the intended purpose. For example, it is crushed by a mixer, a sugar mill, a power mill, a jet mill, an impact crusher or the like. The method etc. can be mentioned.

The method for bringing the lemon balm into the state of juice is not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include pressing.

The method for bringing the lemon balm into the state of the extract is not particularly limited, and a method generally used for extracting plants can be appropriately selected depending on the intended purpose.

The lemon balm used as the fermentation raw material is preferably sterilized before inoculation with the aspergillus. The means for sterilizing the lemon balm is not particularly limited, and can be appropriately selected from known methods.

-Jiuqu-
The aspergillus that ferments the mugwort is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include those described in << Mugwort plant fermented liquid >>. These may be used alone or in combination of two or more. Among them, as the koji mold, Aspergillus oryzae (Aspergillus oryzae) is an anti-aging effect, antioxidant, anti-inflammatory, and preferable from the viewpoint of excellent least one of the effects of whitening effect.
The method for obtaining the aspergillus is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used. Further, as the aspergillus, seed koji made from rice or the like may be used, Kousuihakka seed koji described later may be used, and aspergillus cultivated in a medium (agar medium, liquid medium, etc.) is used. You may. Among these, it is preferable to use the Kousuihakka seed koji because it is excellent in at least one of an anti-aging effect, an antioxidant effect, an anti-inflammatory effect, and a whitening effect.

The amount of the aspergillus inoculated into the Kousuihakka used as the fermentation raw material is not particularly limited as long as the amount of the Kousuihakka can be fermented, and can be appropriately selected depending on the intended purpose. When the fermentation raw material is in a liquid state, 1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL is preferable, and when the fermentation raw material is in a solid state, 1 × 10 ³ pieces / mL. / G to 1 × 10 ⁸ pieces / g is preferable.

It is preferable to add water when inoculating the aspergillus oryzae to the lemon balm. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 500 parts by mass to 5,000 parts by mass with respect to 100 parts by mass of the Kousuihakka. It is more preferable to add 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.

The temperature of the fermentation (culture) is not particularly limited as long as it is within the temperature range in which fermentation by the aspergillus can be performed, and can be appropriately selected depending on the intended purpose. However, 20 ° C to 40 ° C is preferable, and 25 ° C. ℃ -35 ℃ is more preferable. If the temperature of the fermentation is less than 20 ° C., the lemon balm cannot be sufficiently fermented, and at least one of the anti-aging effect, the antioxidant effect, the anti-inflammatory effect, and the whitening effect is insufficient. May become.

The fermentation (culture) time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 10 hours to 40 hours, more preferably 20 hours to 30 hours. If the fermentation time is less than 10 hours, the lemon balm cannot be sufficiently fermented, and at least one of anti-aging, antioxidant, anti-inflammatory, and whitening effects is insufficient. May become.

The method for stopping the fermentation (culture) is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof 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 cannot grow, and can be appropriately selected depending on the intended purpose. However, 50 ° C. or higher is preferable, and 70 ° C. or higher is more preferable. It is 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 anti-aging action, antioxidant action, anti-inflammatory action, and whitening action is ineffective. May be sufficient.
The heating time for stopping the fermentation is not particularly limited as long as the aspergillus can be prevented from growing, and can be appropriately selected depending on the intended purpose, but is preferably 5 minutes or more, preferably 10 minutes to. 20 minutes is more preferable. If the heating time is less than 5 minutes, the fermentation may not be stopped, and if it exceeds 20 minutes, at least one of the anti-aging effect, the antioxidant effect, the anti-inflammatory effect, and the whitening effect is ineffective. May be sufficient.

It is preferable that the Kousuihakka fermented liquid after the fermentation is stopped is cooled. The cooling method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of allowing the material to stand at room temperature or in a refrigerator.

The number of times of fermentation of the lemon balm by the aspergillus is not particularly limited, and can be appropriately selected depending on the purpose, and may be once or a plurality of times. May be good.

When the fermentation is carried out a plurality of times, the aspergillus may be inoculated only for the first time, may be inoculated only several times, or may be inoculated at all times, but it is preferable to inoculate only at the first time.
When the fermentation is carried out a plurality of times, the fermentation temperature and the fermentation time may be different or the same.

--Kousuihakka Seed Jiuqu ---
The Kousuihakka seed koji is obtained by using the Kousuihakka as a seed koji raw material, inoculating the seed koji raw material with aspergillus, and causing a sufficient amount of spores to grow on the Kousuihakka. By using this in the fermentation, it is advantageous in that a Kousuihakka fermented liquid having excellent skin compatibility can be obtained more efficiently and easily.

As the Kousui hacker used as the raw material for the seed koji, the same one as described in the above-Kousui hacker can be used, and the same applies to the usage site, size, state and the like of the Kousui hacker. is there.

As the aspergillus used in the production of the Kousuihakka seed koji, the same as those described in the above-aspergillus-can be used.

The amount of the aspergillus inoculated into the Kousui hacker used as the raw material for the seed koji is not particularly limited and may be appropriately selected depending on the intended purpose. However, sterilized water is added to 100 parts by mass of the Kousui hacker. It is preferable to inoculate 5 parts by mass to 100 parts by mass of aspergillus (1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL) suspended in, and more preferably 10 parts by mass to 50 parts by mass. It is particularly preferable to inoculate 20 parts by mass to 30 parts by mass. If the inoculation amount of the aspergillus to 100 parts by mass of the lemon balm is less than 5 parts by mass, a sufficient amount of spores may not be formed on the lemon balm, and if it exceeds 100 parts by mass, it is abnormal due to excessive water content. May breed.

It is preferable to add water when inoculating the aspergillus fungus into the Kousuihakka used as the seed koji raw material. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 10 parts by mass to 250 parts by mass with respect to 100 parts by mass of the Kousuihakka, and 20 parts by mass. It is more preferable to add parts to 200 parts by mass, and it is particularly preferable to add 30 parts to 150 parts by mass. If the amount of water added to 100 parts by mass of the lemon balm is less than 10 parts by mass, a sufficient amount of spores may not be formed on the lemon balm.

The temperature of the culture is not particularly limited as long as it is within the temperature range in which the aspergillus can grow, and can be appropriately selected depending on the intended purpose, but is preferably 20 ° C to 40 ° C, preferably 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 lemon balm. If the temperature exceeds 50 ° C., the aspergillus may not grow.

The culture time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the culture time is less than 80 hours, a sufficient amount of spores may not be formed on the lemon balm, and if it exceeds 210 hours, the germination rate of spores may decrease.

The fermented balm of Kousuihakka may contain the cells of the aspergillus or the cells of the aspergillus may be removed, but the fermented liquid of the aspergillus may be removed. Is preferable.

The state of the Kousui hacker fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the Kousui hacker fermented liquor itself may be used, and the purified product of the Kousui hacker fermented liquor, said. It may be a concentrate of the fermented balm of Kousuihakka, a diluted product of the fermented balm of Kousuihakka, or the like. Further, the fermented balm may be a dried product of the fermented lemon balm mixed or dissolved in a solvent such as water or a hydrophilic solvent.

The purified product of the Kousuihakka fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the solid content in the Kousuihakka fermented liquor (for example, the plant body of the Kousuihakka, aspergillus) Examples include those from which bacterial cells, ori, etc. have been removed.
The removing means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include filtration.
The filtration method is not particularly limited, and can be appropriately selected from known methods according to the purpose.

The dilution of the Kousui hacker fermented liquor and the concentrate of the Kousui hacker fermented liquor are not particularly limited and may be appropriately selected depending on the intended purpose. For example, the Kousui hacker fermented liquor was prepared to a desired concentration. Things can be mentioned.
The diluting means is not particularly limited, and can be appropriately selected from known methods according to the purpose.
The means for concentration is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include vacuum concentration.

The dried product of the fermented lemon balm is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a dried product of the fermented lemon balm.
The drying means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include freeze-drying.

The fermented balm of Kousuihakka is not particularly limited as long as it is a fermented balm of lemon balm of Kousuihakka, and can be appropriately selected depending on the intended purpose. The Kousuihakka fermented liquid is preferable, and the Kousuihakka fermented liquid having a contact angle of 79 ° or less is more preferable.

<< Cornflower fermented liquid >>
The fermented cornflower is a fermented cornflower fermented with aspergillus.

-Cornflower-
Cornflower to be used as the raw material for fermentation (Centaurea cyanus Linne) is an annual herb belonging to the Asteraceae (Compositae) Centaurea (Centaurea), has been used since ancient times as a raw material for edible and medicinal. Other names include Knapweed, Kentaurea, and Centurea. The place of origin is Europe, but it is also native or cultivated in Japan and is easily available from these areas.
The method for obtaining the cornflower is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used.

The site of use of the yagrumagiku used as the fermentation raw material is not particularly limited and may be appropriately selected depending on the intended purpose. For example, flowers, buds, fruits, pericarps, seeds, seed coats, stems, leaves, branches, etc. Above-ground parts such as branches and leaves; underground parts such as roots and rhizomes. These may be used alone or in combination of two or more. Among these, the above-ground portion is preferable as the site where the cornflower is used.

The size of the cornflower used as the fermentation raw material is not particularly limited as long as it is a size capable of culturing the aspergillus, and can be appropriately selected according to the purpose. For example, the size as it is collected. The desired size of the cut, the size of the fine powder (powder), and the like can be mentioned.

The state of the cornflower used as the fermentation raw material is not particularly limited as long as the aspergillus can be cultivated, and can be appropriately selected depending on the purpose. For example, the collected state as it is or dried. Examples include fermented state, crushed state, squeezed state, and extract state. Among these, the as-collected state, the crushed state, the squeezed state, and the extracted state are preferable, and the collected as-is state and the crushed state are more preferable, because the aspergillus is likely to act.

The method for drying the cornflower is not particularly limited and may be appropriately selected depending on the intended purpose. For example, a method of drying in the sun, a method of drying using a commonly used dryer, or the like. Can be mentioned.

The method for crushing the cornflower is not particularly limited and may be appropriately selected depending on the intended purpose. For example, a method of crushing the cornflower with a mixer, a sugar mill, a power mill, a jet mill, an impact crusher or the like. And so on.

The method for bringing the cornflower into the state of squeezing is not particularly limited and may be appropriately selected depending on the intended purpose, and examples thereof include squeezing.

The method for bringing the cornflower into the state of the extract is not particularly limited, and a method generally used for extracting plants can be appropriately selected depending on the intended purpose.

The cornflower used as the fermentation raw material is preferably sterilized before inoculation with the aspergillus. The means for sterilizing the cornflower is not particularly limited, and can be appropriately selected from known methods.

-Jiuqu-
The aspergillus that ferments the cornflower is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include those described in << Mugwort plant fermented liquid >>. These may be used alone or in combination of two or more. Among them, as the koji mold, Aspergillus oryzae (Aspergillus oryzae) is an anti-aging effect, antioxidant effect, and preferred from the viewpoint of excellent least one of the effects of whitening effect.
The method for obtaining the aspergillus is not particularly limited and may be appropriately selected depending on the intended purpose, and may be collected from the natural world or a commercially available product may be used. Further, as the aspergillus, seed koji made from rice or the like may be used, yagurumagiku seed koji described later may be used, or aspergillus cultivated in a medium (agar medium, liquid medium, etc.) may be used. May be good. Among these, it is preferable to use the cornflower seed koji because it is excellent in at least one of an anti-aging effect, an antioxidant effect, and a whitening effect.

The amount of the aspergillus inoculated into the yagrumagi used as the fermentation raw material is not particularly limited as long as the amount of the aspergillus can be fermented, and can be appropriately selected depending on the intended purpose. When the fermentation raw material is in a liquid state, 1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL is preferable, and when the fermentation raw material is in a solid state, 1 × 10 ³ pieces / g. ~ 1 × 10 ⁸ pieces / g is preferable.

When inoculating the cornflower with the aspergillus, it is preferable to add water. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 500 parts by mass to 5,000 parts by mass with respect to 100 parts by mass of the cornflower. It is more preferable to add 3,000 parts by mass to 4,000 parts by mass, and particularly preferably 1,500 parts by mass to 3,000 parts by mass.

The temperature of the fermentation (culture) is not particularly limited as long as it is within the temperature range in which fermentation by the aspergillus can be performed, and can be appropriately selected depending on the intended purpose. However, 20 ° C to 40 ° C is preferable, and 25 ° C. ℃ -35 ℃ is more preferable. If the fermentation temperature is less than 20 ° C., the cornflower may not be sufficiently fermented, and at least one of the anti-aging effect, the antioxidant effect, and the whitening effect may be insufficient. ..

The fermentation (culture) time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 10 hours to 40 hours, more preferably 20 hours to 30 hours. If the fermentation time is less than 10 hours, the cornflower may not be sufficiently fermented, and at least one of the anti-aging effect, the antioxidant effect, and the whitening effect may be insufficient. ..

The method for stopping the fermentation (culture) is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof 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 cannot grow, and can be appropriately selected depending on the intended purpose. However, 50 ° C. or higher is preferable, and 70 ° C. or higher is more preferable. It is 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, the antioxidant effect, and the whitening effect becomes insufficient. There is.
The heating time for stopping the fermentation is not particularly limited as long as the aspergillus can be prevented from growing, and can be appropriately selected depending on the intended purpose, but is preferably 5 minutes or more, preferably 10 minutes to. 20 minutes is more preferable. If the heating time is less than 5 minutes, the fermentation may not be stopped, and if it exceeds 20 minutes, at least one of the anti-aging effect, the antioxidant effect, and the whitening effect becomes insufficient. There is.

It is preferable that the cornflower fermented liquid after the fermentation is stopped is cooled. The cooling method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a method of allowing the material to stand at room temperature or in a refrigerator.

The number of times the cornflower is fermented by the aspergillus is not particularly limited, and may be appropriately selected depending on the intended purpose, and may be once or a plurality of times. Good.

When the fermentation is carried out a plurality of times, the aspergillus may be inoculated only for the first time, may be inoculated only several times, or may be inoculated at all times, but it is preferable to inoculate only at the first time.
When the fermentation is carried out a plurality of times, the fermentation temperature and the fermentation time may be different or the same.

--- Cornflower Jiuqu ---
The cornflower seed koji is obtained by using the cornflower as a seed koji raw material and inoculating the seed koji raw material with aspergillus to inoculate the cornflower with a sufficient amount of spores. By using this in the fermentation, it is advantageous in that a cornflower fermented liquid having excellent skin compatibility can be obtained more efficiently and easily.

As the cornflower used as the raw material for the seed koji, the same cornflower as described in the above-Yagurumagiku-can be used, and the same applies to the mode of use, size, state and the like of the cornflower.

As the aspergillus used in the production of the cornflower seed koji, the same as those described in the above-aspergillus may be used.

The amount of the aspergillus inoculated into the yagurumagi used as the raw material for the seed koji is not particularly limited and may be appropriately selected depending on the intended purpose. However, 100 parts by mass of the yagurumagi is submerged in sterilized water. It is preferable to inoculate 5 parts by mass to 100 parts by mass of turbid aspergillus (1 × 10 ³ pieces / mL to 1 × 10 ⁸ pieces / mL), more preferably 10 parts by mass to 50 parts by mass, and 20 parts by mass. It is particularly preferable to inoculate parts to 30 parts by mass. If the inoculation amount of the aspergillus to 100 parts by mass of the cornflower is less than 5 parts by mass, a sufficient amount of spores may not be formed on the cornflower, and if it exceeds 100 parts by mass, abnormal growth occurs due to excessive water content. Sometimes.

When inoculating the cornflower used as the seed koji raw material with the aspergillus, it is preferable to add water. The amount of water added is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is preferable to add 10 parts by mass to 250 parts by mass with respect to 100 parts by mass of the cornflower, and 20 parts by mass. It is more preferable to add ~ 200 parts by mass, and it is particularly preferable to add 30 parts by mass to 150 parts by mass. If the amount of water added to 100 parts by mass of the cornflower is less than 10 parts by mass, a sufficient amount of spores may not be formed on the cornflower.

The temperature of the culture is not particularly limited as long as it is within the temperature range in which the aspergillus can grow, and can be appropriately selected depending on the intended purpose, but is preferably 20 ° C to 40 ° C, preferably 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 cornflower. If the temperature exceeds 50 ° C., the aspergillus may not grow.

The culture time is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 80 hours to 210 hours, more preferably 100 hours to 190 hours, and particularly preferably 120 hours to 170 hours. If the culture time is less than 80 hours, a sufficient amount of spores may not be formed on the cornflower, and if it exceeds 210 hours, the germination rate of spores may decrease.

The fermented cornflower liquid may contain the cells of the aspergillus or the cells of the aspergillus may be removed, but the fermented liquid of the aspergillus may be the one from which the cells of the aspergillus have been removed. preferable.

The state of the cornflower fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the cornflower fermented liquor itself may be used, a purified product of the cornflower fermented liquor, or the cornflower fermented liquor. It may be a concentrate of the above, a diluted product of the fermented cornflower, or the like. Further, the cornflower fermented liquor may be a dried product of the cornflower fermented liquor mixed or dissolved in a solvent such as water or a hydrophilic solvent again.

The purified product of the cornflower fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. For example, the solid content in the cornflower fermented liquor (for example, the cornflower plant, the aspergillus bacterium, etc. Examples include those from which sediment has been removed.
The removing means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include filtration.
The filtration method is not particularly limited, and can be appropriately selected from known methods according to the purpose.

The dilution of the cornflower fermented liquor and the concentrate of the cornflower fermented liquor are not particularly limited and may be appropriately selected depending on the intended purpose. For example, a cornflower fermented liquor prepared to a desired concentration may be used. Can be mentioned.
The diluting means is not particularly limited, and can be appropriately selected from known methods according to the purpose.
The means for concentration is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include vacuum concentration.

The dried cornflower fermented liquor is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a dried cornflower fermented liquor.
The drying means is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include freeze-drying.

The fermented cornflower is not particularly limited as long as it is a fermented cornflower fermented with aspergillus, and can be appropriately selected depending on the purpose. However, the cornflower fermentation having a contact angle of 85 ° or less is good for the skin. The liquid is preferable, and the fermented cornflower liquid having a contact angle of 79 ° or less is more preferable.

<< Other ingredients >>
The other components in the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent are not particularly limited and may be appropriately selected depending on the intended purpose. For example, excipients, moisture-proofing agents, and antiseptic agents. Agents, strengtheners, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, colorants, fragrances, whitening agents, moisturizers, oily ingredients, UV absorbers, surfactants, thickeners, Examples include alcohols, powder components, coloring agents, aqueous components, water, and skin nutrients. These may be used alone or in combination of two or more.
The content of the other components is not particularly limited and may be appropriately selected depending on the intended purpose.

-Use-
Since the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention have at least one of an excellent anti-aging effect, antioxidant effect, anti-inflammatory effect, and whitening effect, for example, a pharmaceutical product. , It can be suitably used as a non-pharmaceutical product, cosmetics, food and drink, etc., and the blending amount, usage, and dosage form thereof can be appropriately selected according to the purpose of use.

The blending amount can be appropriately adjusted depending on the physiological activity of the fermentation broth and the like. Further, the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent may be the fermented liquid itself.

The usage is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include oral, parenteral, and external use. Of these, external use is preferable.

The dosage form is not particularly limited and may be appropriately selected depending on the intended purpose. For example, oral administration agents such as tablets, powders, capsules, granules, extracts and syrups; injections and instillations. Parenteral agents such as preparations and suppositories; lotions, emulsions, creams, ointments, beauty liquids, lotions, packs, jellies, lip balms, lipsticks, foundations, bathing agents, soaps, body shampoos, astringents, hair tonics, Examples include external preparations such as hair cream, hair liquid, pomade, shampoo, and rinse.

In addition, the anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention can also be used as a reagent for research on the action mechanism of anti-aging action, antioxidant action, anti-inflammatory action, or whitening action. ..

The anti-aging agent, antioxidant, anti-inflammatory agent, and whitening agent of the present invention are preferably applied to humans, but as long as their respective actions and effects are exhibited, animals other than humans (for example, , Mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.).

(Cosmetics)
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, and further contains other components as necessary. To do.

<Anti-aging agents, antioxidants, anti-inflammatory agents, whitening agents>
The content of at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent in the cosmetic is not particularly limited and may be appropriately selected depending on the intended 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. When the content of at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent is less than 5% by volume, the anti-aging effect, the antioxidant effect, At least one of the anti-inflammatory and whitening effects may be inadequate. The content of at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent is preferably as high as possible, and the upper limit thereof is not particularly limited and is intended. It can be appropriately selected according to the above. 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.

<Other ingredients>
If necessary, various main agents, auxiliaries, and other ingredients usually used in the production of cosmetics can be added to the cosmetics as long as the object and effects of the present invention are not impaired.
The other components are not particularly limited and may be appropriately selected depending on the intended purpose. For example, an astringent agent, a bactericidal agent, an antibacterial agent, an ultraviolet absorber, a cell activator, oils and fats, waxes and hydrocarbons. Classes, fatty acids, alcohols, esters, surfactants, fragrances and the like. These may 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, the antioxidant, the anti-inflammatory agent, and the whitening agent, they act synergistically and are usually expected. It may bring about the above-mentioned excellent action and effect.
The content of the other components in the cosmetics is not particularly limited as long as the effects of the present invention are not impaired, and can be appropriately selected depending on the intended purpose.

<Use>
The use of the cosmetic is not particularly limited and may be appropriately selected from general cosmetics, for example, lotion, milky lotion, cream, ointment, beauty liquid, lotion, pack, jelly, lip balm. , Lipsticks, foundations, bathing agents, soaps, body shampoos and other skin cosmetics; scalp hair cosmetics such as astringents, hair tonics, hair creams, hair liquids, pomades, shampoos, rinses and the like.

As the cosmetic, at least one selected from the group consisting of the anti-aging agent, the antioxidant, the anti-inflammatory agent, and the whitening agent was blended into any cosmetic so as not to interfere with its activity. It may be a cosmetic containing 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.

The cosmetics of the present invention are suitably applied to humans, but animals other than humans (for example, mice, rats, hamsters, dogs, cats, cows, etc. It can also be applied to pigs, monkeys, etc.).

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 excellent in anti-aging when used on the skin. It is useful in that it exerts at least one of an aging effect, an antioxidant effect, an anti-inflammatory effect, and a whitening effect.

The present invention will be specifically described below with reference to production examples and test examples, but the present invention is not limited to these test examples.

<Production Example 1: Preparation of Yamayomogi Fermented Liquid 1>
-Seed koji preparation process-
Koji mold (Aspergillus oryzae, strain name: AOK1714, Ltd. Akita Konno made shopping) taken in the platinum ear, to prepare a koji mold solution was suspended in sterile water 50mL. The number of bacteria of the Aspergillus solution, was calculated using Thoma hemocytometer (manufactured EKDS), it was 1.0 × 10 ⁵ cells / mL.
Next, 10 g of Yamayomogi (manufactured by Albion Co., Ltd.) cut into 0.5 cm to 5 cm was placed in an Erlenmeyer flask, sterilized under pressure, inoculated with 2 mL of the Jiuqu solution, and statically cultured at 30 ° C. for 168 hours. After completion of the culture, the mixture was dried at 45 ° C. for 24 hours to obtain "Yamayomogi Jiuqu".

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

<Production Example 2: Preparation of Yamayomogi Fermented Liquid 2>
In Production Example 1, the Yamayomogi seed koji was changed to a rice seed koji ( Aspergillus oryzae , Shirakoji Shirakami, manufactured by Akita Konno Shoten Co., Ltd.) (hereinafter, may be referred to as "rice seed koji"). "Yamayomogi fermented liquid 2" was obtained by the same method as in Production Example 1 except for the above.

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

(Test Example A-1: Measurement of contact angle)
The contact angle was measured by the following method using the Yamayomogi fermented liquid 1 obtained in Production Example 1, the Yamayomogi fermented liquid 2 obtained in Production Example 2, and the Yamayomogi extract obtained in Comparative Production Example 1 as test samples. ..
Specifically, using a dynamic contact angle / 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 temperature was 22 ° C. The measurement was carried out by the sessile drop method under the condition of 20% relative humidity. The contact angle θ (°) of 1,000 ms was determined by the θ / 2 method. The contact angle was measured three times, and the average value was calculated. The results are shown in Table 1 below. In addition, examples of droplets at the time of measuring the contact angle of each test sample are shown in FIGS. 1A to 1C.

Compared to the Yamayomogi extract obtained in Comparative Production Example 1, the Yamayomogi fermented liquid 1 obtained in Production Example 1 and the Yamayomogi fermented liquid 2 obtained in Production Example 2 both have a small contact angle and are 81 ° or less. It was excellent in familiarity with the skin. Further, the fermented yamayomogi solution 1 obtained in Production Example 1 had a contact angle of 78 ° or less, and was more familiar to the skin.

(Test Example 1-1: Matrix metalloproteinase-1 (MMP-1) activity inhibitory action test)
The Wunch and Heidrich method was partially modified using the Yamayomogi fermented liquid 1 obtained in Production Example 1, the Yamayomogi fermented liquid 2 obtained in Production Example 2, and the Yamayomogi extract obtained in Comparative Production Example 1 as test samples. The effect of inhibiting matrix metalloproteinase-1 (MMP-1) activity was tested by the following test method.

In a test tube with a lid, each test sample was dissolved in 0.1 mol / L Tris-HCl buffer (pH 7.1) containing 20 mmol / L calcium chloride. Next, 50 μL of the solution of the test sample, 50 μL of a solution of MMP-1 (COLLAGENASE Type IV from Clostridium history, manufactured by Sigma), Pz-peptide (Pz-Pro-Leu-Gly-Pro-D-Arg-OH), 400 μL of a solution (manufactured by BACHEEM Feinchemikalien AG) was 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 of the ethyl acetate layer at a wavelength of 320 nm was measured.

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

Further, as a control, the same operation as above and the same operation as described above except that the solution of the test sample was changed to a 20 mmol / L calcium chloride-containing 0.1 mol / L Tris-HCl buffer (pH 7.1) containing no test sample. Absorbance was measured.

From the obtained measured absorbance, the MMP-1 activity inhibition rate was calculated based on the following formula 1. The results are shown in Table 2 below.
<Equation 1>
MMP-1 activity inhibition rate (%) = {1- (CD) / (AB)} × 100
In the above formula 1, A to D represent the following, respectively.
A: Absorbance at wavelength 320 nm with no test sample added, B: Absorbance at wavelength 320 nm without test sample added, without enzyme added C: Absorbance at wavelength 320 nm with test sample added, enzyme added D: Test sample added Absorbance at 320 nm without enzyme addition

(Test Example 1-2: Hyaluronic acid synthase 3 (HAS3) mRNA expression promoting action test)
Hyaluronic acid was used as a test sample using the Yamayomogi fermented liquid 1 obtained in Production Example 1, the Yamayomogi fermented liquid 2 obtained in Production Example 2, and the Yamayomogi extract obtained in Comparative Production Example 1 as test samples. The synthase 3 (HAS3) mRNA expression promoting action was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 3 below, and a test sample-added medium was prepared.
Normal human epidermal keratinocytes (Normal Human Epidermal Keratinocytes; NHEK, manufactured by Kurashiki Spinning Co., Ltd.) were used in a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C., 5 ° C. After culturing until confluent under the condition of% CO ₂ , cells were collected by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

Using each of the above total RNAs as a template, the expression levels of hyaluronan synthase 3 (HAS3) mRNA and the internal standard glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA were measured. For the detection of mRNA, 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.) by 2-step RT-PCR reaction.
The expression level of HAS3 mRNA without the addition of the test sample and the addition of the test sample was 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.
<Equation 2>
HAS3 mRNA expression promotion rate (%) = A / B × 100
In the above formula 2, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

(Test Example 1-3: DPPH radical scavenging action test)
Using the fermented yamayomogi liquor 1 obtained in Production Example 1, the fermented yamayomogi liquor 2 obtained in Production Example 2, and the yamayomogi extract obtained in Comparative Production Example 1 as test samples, the DPPH radicals were subjected to the following test method. The erasing effect was tested.

Each test sample was dissolved in an ethanol solution (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.) to prepare a test sample solution.
To 3 mL of a 150 μmol / L DPPH (diphenyl-p-picrylhydrazyl) ethanol solution, 3 mL of the test sample solution was added, the container was immediately sealed, shaken, allowed to stand for 30 minutes, and then 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.

Further, as a blank, the same operation and absorbance measurement as described above were performed except that the DPPH ethanol solution was changed to an ethanol solution containing no DPPH.

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

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

(Test Example 1-4: Hyaluronidase activity inhibitory action test)
Using the Yamayomogi fermented liquid 1 obtained in Production Example 1, the Yamayomogi fermented liquid 2 obtained in Production Example 2, and the Yamayomogi extract obtained in Comparative Production Example 1 as test samples, hyaluronidase activity was carried out by the following test method. The inhibitory effect was tested.

Each test sample was dissolved in 0.1 mol / L acetate buffer (pH 3.5) to prepare a test sample solution.
To 0.2 mL of the test sample solution, 0.1 mL of a hyaluronidase solution (Type IV-S (from testis), 400 NF units / mL, manufactured by SIGMA) was added, and the mixture was reacted at 37 ° C. for 20 minutes. Then, 0.2 mL of 2.5 mmol / L calcium chloride was added as an activator, and the reaction was further carried out at 37 ° C. for 20 minutes. To this was added 0.5 mL of a 0.8 mg / mL sodium hyaluronate solution (from rooster comb) (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.), 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, and after cooling, 0.2 mL of a boric acid solution was added to each reaction solution and boiled for 3 minutes. After ice cooling, 6 mL of p-DABA reagent (10 g of p-dimethylaminobenzaldehyde dissolved in a mixture of 12.5 mL of 10N hydrochloric acid and 87.5 mL of acetic acid and diluted 10-fold with acetic acid) was added, and 20 at 37 ° C. Reacted for minutes. Then, the absorbance at a wavelength of 585 nm was measured.

Further, as a blank, the same operation and absorbance measurement as described above were carried out except that the hyaluronidase solution (enzyme solution) was changed to 0.1 mol / L acetate buffer (pH 3.5).

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

From the obtained measured absorbance, the hyaluronidase activity inhibition rate was calculated based on the following formula 4. The results are shown in Table 5 below.
<Equation 4>
Hyaluronidase activity inhibition rate (%) = {1- (CD) / (AB)} × 100
In the above formula 4, A to D represent the following, respectively.
A: Absorbance at wavelength 585 nm without test sample added, B: Absorbance at wavelength 585 nm without test sample added, C: Absorbance at 585 nm with test sample added, enzyme added D: Test sample added, Absorbance at a wavelength of 585 nm without the addition of enzymes

(Test Example 1-5: Tyrosinase activity inhibitory action test)
Using the Yamayomogi fermented liquid 1 obtained in Production Example 1, the Yamayomogi fermented liquid 2 obtained in Production Example 2, and the Yamayomogi extract obtained in Comparative Production Example 1 as test samples, the tyrosinase activity was carried out by the following test method. The inhibitory effect was tested.

Each test sample was dissolved in 25% DMSO solution to prepare a test sample solution.
To a 48-well plate, 0.2 mL of Mclvine 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, and the mixture was allowed to stand at 37 ° C. for 10 minutes. To this, 0.02 mL of an 800 unit / mL tyrosinase solution (manufactured by SIGMA) was added, and the mixture was further reacted at 37 ° C. for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 475 nm was measured. The final concentration of the test sample at this time was the concentration shown in Table 6 below.

Further, as a blank, the same operation and absorbance measurement as described above were carried out except that the tyrosinase solution (enzyme solution) was changed to Mclvine buffer (pH 6.8).

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

From the obtained measured absorbance, the tyrosinase activity inhibition rate was calculated based on the following formula 5. The results are shown in Table 6 below.
<Equation 5>
Tyrosinase activity inhibition rate (%) = {1- (CD) / (AB)} × 100
In the above formula 5, A to D represent the following, respectively.
A: Absorbance at wavelength 475 nm without test sample added, B: Absorbance at wavelength 475 nm without test sample added, C: Absorbance at 475 nm with test sample added, enzyme added D: Test sample added, Absorbance at a wavelength of 475 nm without the addition of enzymes

<Production Example 3: Preparation of Capillary Wormwood Fermented Liquid 1>
In the seed koji preparation step of Production Example 1, "Capillary wormwood seeds" was prepared in the same manner as the seed koji preparation step of Production Example 1 except that the capillary wormwood was changed to Capillary wormwood ( Artemisia capillaris Thumbergii) (manufactured by Albion Co., Ltd.). "Koji" was prepared.
Further, in the fermentation step of Production Example 1, "Capillary wormwood fermented liquid 1" is carried out in the same manner as in the fermentation step of Production Example 1 except that the capillary wormwood is changed to Capillary wormwood ( Artemisia capillaris Tumbergii) (manufactured by Albion Co., Ltd.). I got.

<Production Example 4: Preparation of Capillary Wormwood Fermented Liquid 2>
In Production Example 3, "Capillary wormwood fermented liquid" is produced in the same manner as in Production Example 3 except that the capillary wormwood seed koji is changed to rice seed koji ( Aspergillus oryzae , Shirakoji Shirakami, manufactured by Akita Konno Shoten Co., Ltd.). 2 ”was obtained.

<Comparative Production Example 2: Preparation of Capillary Wormwood Extract>
In Comparative Production Example 1, "Capillary wormwood extract" was obtained in the same manner as in Comparative Production Example 1 except that the capillary wormwood was changed to Capillary wormwood ( Artemisia capillaris Thumbergii) (manufactured by Albion Co., Ltd.).

(Test Example A-2: Measurement of contact angle)
In Test Example A-1, the test sample was changed to Capillary wormwood fermented liquid 1 obtained in Production Example 3, Capillary wormwood fermented liquid 2 obtained in Production Example 2, and Capillary wormwood extract obtained in Comparative Production Example 2. The contact angle was measured in the same manner as in Test Example A-1 except for the above. The results are shown in Table 7 below. Further, FIGS. 2A to 2C show examples of droplets when measuring the contact angle of each test sample.

Compared to the Capillary wormwood extract obtained in Comparative Production Example 2, the Capillary wormwood fermented liquid 1 obtained in Production Example 3 and the Capillary wormwood fermented liquid 2 obtained in Production Example 4 both have a small contact angle and are 81 ° or less. It was excellent in familiarity with the skin. Further, the capillary wormwood fermented liquid 1 obtained in Production Example 3 had a contact angle of 78 ° or less, and was more familiar to the skin.

(Test Example 2-1: Hyaluronan synthase 3 (HAS3) mRNA expression promoting action test)
In Test Example 1-2, the test sample was changed to the capillary wormwood fermented liquid 1 obtained in Production Example 3, the capillary wormwood fermented liquid 2 obtained in Production Example 4, and the capillary wormwood extract obtained in Comparative Production Example 2. The hyaluronan synthase 3 (HAS3) mRNA expression promoting action was tested in the same manner as in Test Example 1-2 except that the final concentration of the test sample was changed to the concentration shown in Table 8 below. The results are shown in Table 8 below.

(Test Example 2-2: Type I collagen production promoting action test)
Using the capillary wormwood fermented liquid 1 obtained in Production Example 3, the capillary wormwood fermented liquid 2 obtained in Production Example 4, and the capillary wormwood extract obtained in Comparative Production Example 2 as test samples, type I was used by the following test method. The collagen production promoting effect was tested.

Dissolve each test sample in 0.25% fetal bovine serum (FBS, manufactured by biosera) in Dalbeco MEM, manufactured by Nissui Pharmaceutical Co., Ltd. so that the final concentration is as shown in Table 9 below. Then, a test sample-added medium was prepared.
Normal human fibroblasts (NB1RGB, purchased from RIKEN BRC) were cultured in DMEM containing 10% FBS under the conditions of 37 ° C. and 5% CO ₂ until they became confluent, and then the cells were recovered by trypsin treatment. This was adjusted to 1.6 × 10 ⁵ cells / mL by 10% FBS-containing DMEM.
Then, the in 96well microplate NB1RGB (1.6 × 10 ⁵ cells / mL), seeded by 100μL per one well, 37 ° C., and cultured overnight under conditions of 5% CO _2. After completion of the culture, the medium was replaced with 100 μL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 3 days. After completion of the culture, the amount of type I collagen in the medium of each well was measured by the ELISA method.

Specifically, 90 μL of the culture supernatant was transferred to an ELISA plate, adsorbed on the plate at 4 ° C. overnight, the solution was discarded, and a phosphate buffer solution (PBS-) containing 0.05% Tween-20 was discarded. Washing was performed in T). Then, a blocking operation was performed with a phosphate physiological buffer solution containing 1% FBS. The solution was discarded, washed with phosphate buffered buffer (PBS-T) containing 0.05% Tween-20, and reacted with an anti-human collagen type I antibody (rabbit IgG, manufactured by Chemi-Con). Discard the solution, wash with phosphate buffered buffer (PBS-T) containing 0.05% Tween-20, react with HRP-labeled anti-rabbit IgG antibody, and then perform the same washing operation to develop color. The reaction was carried out.
The type I collagen production promotion rate was calculated by performing the above-mentioned ELISA using a standard product and preparing a calibration curve.

In addition, as a control, the same operation as described above and the measurement by the ELISA method were performed except that the test sample solution was changed to DMEM containing 0.25% FBS containing no test sample.

From the obtained measured values, the type I collagen production promotion rate was calculated based on the following formula 6. The results are shown in Table 9 below.
<Equation 6>
Type I collagen production promotion rate (%) = A / B x 100
In the above formula 6, A and B represent the following, respectively.
A: Amount of type I collagen when test sample is added B: Amount of type I collagen when no test sample is added

(Test Example 2-3: Claudin-1 mRNA expression promoting action test)
Using the capillary wormwood fermented liquor 1 obtained in Production Example 3, the capillary wormwood fermented liquor 2 obtained in Production Example 4, and the capillary wormwood extract obtained in Comparative Production Example 2 as test samples, claudin was used by the following test method. -1 The effect of promoting mRNA expression was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 10 below, and a test sample-added medium was prepared.
Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

Using each of the above total RNAs as a template, the expression levels of claudin-1 mRNA and GAPDH mRNA, which is an internal standard, were measured. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression levels of the test sample-free and test sample-added claudin-1 mRNA were corrected by the expression levels of GAPDH mRNA. From this corrected value, the claudin-1 mRNA expression promotion rate was calculated based on the following formula 7. The results are shown in Table 10 below.
<Equation 7>
Claudin-1 mRNA expression promotion rate (%) = A / B × 100
In the above formula 7, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

(Test Example 2-4: Claudin-4 mRNA expression promoting action test)
Using the capillary wormwood fermented liquid 1 obtained in Production Example 3, the capillary wormwood fermented liquid 2 obtained in Production Example 4, and the capillary wormwood extract obtained in Comparative Production Example 2 as test samples, claudin was used by the following test method. -4 The effect of promoting mRNA expression was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning 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 Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

Using each of the above total RNAs as a template, the expression levels of claudin-4 mRNA and GAPDH mRNA, which is an internal standard, were measured. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression levels of the test sample-free and test sample-added claudin-4 mRNA were corrected by the expression levels of GAPDH mRNA. From this corrected value, the claudin-4 mRNA expression promotion rate was calculated based on the following formula 8. The results are shown in Table 11 below.
<Equation 8>
Claudin-4 mRNA expression promotion rate (%) = A / B × 100
In the above formula 8, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

(Test Example 2-5: Occludin mRNA expression promoting action test)
Using the capillary wormwood fermented liquid 1 obtained in Production Example 3, the capillary wormwood fermented liquid 2 obtained in Production Example 4, and the capillary wormwood extract obtained in Comparative Production Example 2 as test samples, the occludin mRNA was prepared by the following test method. The expression promoting effect was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 12 below, and a test sample-added medium was prepared.
Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

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. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression levels of occludin mRNA without test sample and with test sample were corrected by the expression level of GAPDH mRNA. From this corrected value, the occludin mRNA expression promotion rate was calculated based on the following formula 9. The results are shown in Table 12 below.
<Equation 9>
Occludin mRNA expression promotion rate (%) = A / B × 100
In the above formula 9, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

<Production Example 5: Preparation of Fermented Tachijakousou 1>
In the seed koji preparation step of Production Example 1, "Tachi" is carried out in the same manner as the seed koji preparation step of Production Example 1 except that the yamayomogi is changed to Tachijakousou ( Thymus vulgaris Linne) (manufactured by Albion Co., Ltd.). "Garden Thyme" was prepared.
Further, in the fermentation step of Production Example 1, "Tachijakousou fermentation" was carried out in the same manner as in the fermentation step of Production Example 1 except that the yamayomogi was changed to Thymus vulgaris Linne (manufactured by Albion Co., Ltd.). Liquid 1 ”was obtained.

<Production Example 6: Preparation of Tachijakousou Fermented Liquid 2>
In Production Example 5, "Tachijakousou" is produced in the same manner as in Production Example 5, except that the Tachijakousou seed koji is changed to rice seed koji ( Aspergillus oryzae , Shirakoji Shirakami, manufactured by Akita Konno Shoten Co., Ltd.). Fermented liquid 2 ”was obtained.

<Comparative Production Example 3: Preparation of Tachijakousou Extract>
In Comparative Production Example 1, "Tachijakousou extract" was obtained in the same manner as in Comparative Production Example 1 except that the yamayomogi was changed to Tachijakousou ( Thymus vulgaris Line) (manufactured by Albion Co., Ltd.).

(Test Example A-3: Measurement of contact angle)
In Test Example A-1, the test sample was used as the Tachijakousou fermented liquid 1 obtained in Production Example 5, the Tachijakousou fermented liquid 2 obtained in Production Example 6, and the Tachijakousou fermented liquid obtained in Comparative Production Example 3. The contact angle was measured in the same manner as in Test Example A-1 except that the solution was changed to a liquid. The results are shown in Table 13 below. Further, examples of droplets at the time of measuring the contact angle of each test sample are shown in FIGS. 3A to 3C.

Compared to the Tachijakousou extract obtained in Comparative Production Example 3, the Tachijakousou fermented liquid 1 obtained in Production Example 5 and the Tachijakousou fermented liquid 2 obtained in Production Example 6 both had a small contact angle. It was 87 ° or less, and was excellent in familiarity with the skin. Further, the Tachijakousou fermented liquid 1 obtained in Production Example 5 had a contact angle of 81 ° or less, and was more familiar to the skin.

(Test Example 3-1: Transglutaminase-1 (TGM-1) mRNA expression promoting action test)
Using the Tachijakousou fermented liquid 1 obtained in Production Example 5, the Tachijakousou fermented liquid 2 obtained in Production Example 6, and the Tachijakousou extract obtained in Comparative Production Example 3 as test samples, the following test method was used. , Transglutaminase-1 (TGM-1) mRNA expression promoting action was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 14 below, and a test sample-added medium was prepared.
Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

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. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression level of TGM-1 mRNA without the test sample added and with the test sample added was corrected by the expression level of GAPDH mRNA. From this corrected value, the TGM-1 mRNA expression promotion rate was calculated based on the following formula 10. The results are shown in Table 14 below.
<Equation 10>
TGM-1 mRNA expression promotion rate (%) = A / B × 100
In the above formula 10, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

(Test Example 3-2: Aquaporin 3 (AQP3) mRNA expression promoting action test)
Using the Tachijakousou fermented liquid 1 obtained in Production Example 5, the Tachijakousou fermented liquid 2 obtained in Production Example 6, and the Tachijakousou extract obtained in Comparative Production Example 3 as test samples, the following test method was used. , Aquaporin 3 (AQP3) mRNA expression promoting action was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 15 below, and a test sample-added medium was prepared.
Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

Using each of the above total RNAs as a template, the expression levels of aquaporin 3 (AQP3) mRNA and GAPDH mRNA, which is an internal standard, were measured. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression level of AQP3 mRNA without the addition of the test sample and with the addition of the test sample was 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.
<Equation 11>
AQP3 mRNA expression promotion rate (%) = A / B × 100
In the above formula 11, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

(Test Example 3-3: Occludin mRNA expression promoting action test)
In Test Example 2-5, the test sample was used as the Tachijakousou fermented solution 1 obtained in Production Example 5, the Tachijakousou fermented solution 2 obtained in Production Example 6, and the Tachijakousou extract obtained in Comparative Production Example 3. The occludin mRNA expression promoting action was tested in the same manner as in Test Example 2-5, except that the final concentration of the test sample was changed to the concentration shown in Table 16 below. The results are shown in Table 16 below.

(Test Example 3-4: DPPH radical scavenging action test)
In Test Example 1-3, the test samples were used as the Tachijakousou fermented solution 1 obtained in Production Example 5, the Tachijakousou fermented solution 2 obtained in Production Example 6, and the Tachijakousou extract obtained in Comparative Production Example 3. The DPPH radical scavenging action was tested in the same manner as in Test Example 1-3, except that the final concentration of the test sample was changed to the concentration shown in Table 17 below. The results are shown in Table 17 below.

<Production Example 7: Preparation of Lemon Balm 1>
In the above Production Example 1 Seed koji preparation step, the sagebrush, perfume mint (Melissa officinalis Linne) was changed to (Co. Albion), wherein in Production Example 1 Seed koji preparation step and the same method as "hard water "Hacka seed koji" was prepared.
Further, in the above fermentation step of Production Example 1, the sagebrush, except for changing the hard water mint (Melissa officinalis Linne) (Co. Albion), "Perfume peppermint fermented in the same manner as in the processes of fermentation of Preparation Example 1 Liquid 1 ”was obtained.

<Production Example 8: Preparation of Lemon Balm 2>
In Production Example 7, " Aspergillus oryzae , Shirakoji Shirakami, manufactured by Akita Konno Shoten Co., Ltd." is used in the same manner as in Production Example 7, except that the Aspergillus oryzae is changed to Aspergillus oryzae . Fermented liquid 2 ”was obtained.

<Comparative Production Example 4: Preparation of Lemon Balm Extract>
In the Comparative Preparation Example 1, the sagebrush, except for changing the hard water mint (Melissa officinalis Linne) (Co. Albion), to obtain a "hard water mint extract" in the same manner as in Comparative Preparation Example 1.

(Test Example A-4: Measurement of contact angle)
In Test Example A-1, the test sample was used as the Kousuihakka fermented liquid 1 obtained in Production Example 7, the Kousuihakka fermented liquid 2 obtained in Production Example 8, and the Kousuihakka extraction obtained in Comparative Production Example 4. The contact angle was measured in the same manner as in Test Example A-1 except that the solution was changed to liquid. The results are shown in Table 18 below. In addition, examples of droplets when measuring the contact angle of each test sample are shown in FIGS. 4A to 4C.

Compared to the Kousuihakka extract obtained in Comparative Production Example 4, the Kousuihakka fermented liquid 1 obtained in Production Example 7 and the Kousuihakka fermented liquid 2 obtained in Production Example 8 both had a small contact angle. The temperature was 85 ° or less, and it was excellent in familiarity with the skin. Further, the Kousuihakka fermented liquid 1 obtained in Production Example 7 had a contact angle of 79 ° or less, and was more familiar to the skin.

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

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

(Test Example 4-3: DPPH radical scavenging action test)
In Test Example 1-3, the test sample was used as the Kousuihakka fermented solution 1 obtained in Production Example 7, the Kousuihakka fermented solution 2 obtained in Production Example 8, and the Kousuihakka extract obtained in Comparative Production Example 4. The DPPH radical scavenging action was tested in the same manner as in Test Example 1-3, except that the final concentration of the test sample was changed to the concentration shown in Table 21 below. The results are shown in Table 21 below.

(Test Example 4-4: Hyaluronidase activity inhibitory action test)
In Test Examples 1-4, the test samples were used as the Kousuihakka fermented liquid 1 obtained in Production Example 7, the Kousuihakka fermented liquid 2 obtained in Production Example 8, and the Kousuihakka extract obtained in Comparative Production Example 4. The hyaluronidase activity inhibitory effect was tested in the same manner as in Test Example 1-4 except that the final concentration of the test sample was changed to the concentration shown in Table 22 below. The results are shown in Table 22 below.

(Test Example 4-5: Tyrosinase activity inhibitory action test)
In Test Example 1-5, the test sample was used as the Kousuihakka fermented liquid 1 obtained in Production Example 7, the Kousuihakka fermented liquid 2 obtained in Production Example 8, and the Kousuihakka extract obtained in Comparative Production Example 4. The tyrosinase activity inhibitory action was tested in the same manner as in Test Example 1-5, except that the final concentration of the test sample was changed to the concentration shown in Table 22 below. The results are shown in Table 23 below.

<Production Example 9: Preparation of cornflower fermented liquid 1>
In the seed koji preparation step of Production Example 1, "Knapweed seeds" was prepared in the same manner as in the seed koji preparation step of Production Example 1 except that the cornflower was changed to cornflower ( Centurea cyanus Line) (manufactured by Albion Co., Ltd.). "Knapweed" was prepared.
Further, in the fermentation step of Production Example 1, "Knapweed Fermentation Solution 1" was carried out in the same manner as in the fermentation step of Production Example 1 except that the cornflower was changed to cornflower ( Centurea cyanus Line) (manufactured by Albion Co., Ltd.). I got.

<Production Example 10: Preparation of cornflower fermented liquid 2>
In the above-mentioned production example 9, "cornflower fermented liquid" was obtained in the same manner as in the above-mentioned production example 9 except that the cornflower seed koji was changed to rice seed koji ( Aspergillus oryzae , white koji shirakami, manufactured by Akita Konno Shoten Co., Ltd.). 2 ”was obtained.

<Comparative Production Example 5: Preparation of Cornflower Extract>
A " cornflower extract" was obtained in the same manner as in Comparative Production Example 1 except that the cornflower was changed to cornflower ( Centurea cyanus Line) (manufactured by Albion Co., Ltd.).

(Test Example A-5: Measurement of contact angle)
In Test Example A-1, the test sample was changed to the cornflower fermented liquid 1 obtained in Production Example 9, the cornflower fermented liquid 2 obtained in Production Example 10, and the cornflower extract obtained in Comparative Production Example 5. The contact angle was measured in the same manner as in Test Example A-1 except for the above. The results are shown in Table 24 below. Further, an example of a droplet when measuring the contact angle of each test sample is shown in FIGS. 5A to 5C.

Compared to the cornflower extract obtained in Comparative Production Example 5, the cornflower fermented liquor 1 obtained in Production Example 9 and the cornflower fermented liquor 2 obtained in Production Example 10 both had a small contact angle of 85 ° or less. It was excellent in familiarity with the skin. Further, the cornflower fermented liquid 1 obtained in Production Example 9 had a contact angle of 79 ° or less, and was more familiar to the skin.

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

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

(Test Example 5-3: Filaggrin mRNA expression promoting action test)
Using the cornflower fermented liquor 1 obtained in Production Example 9, the cornflower fermented liquor 2 obtained in Production Example 10, and the cornflower extract obtained in Comparative Production Example 5 as test samples, the fillagrin mRNA was prepared by the following test method. The expression promoting effect was tested.

Each test sample was dissolved in a normal human epidermal keratinocyte basal medium (HuMedia-KB2, manufactured by Kurashiki Spinning Co., Ltd.) so that the final concentration was as shown in Table 27 below, and a test sample-added medium was prepared.
Normal human neonatal epidermal keratinocytes (NHEK, manufactured by Kurashiki Spinning Co., Ltd.) using a medium for normal human epidermal keratinocyte proliferation (HuMedia-KG2, manufactured by Kurashiki Spinning Co., Ltd.) at 37 ° C. and 5% CO ₂ conditions. After culturing until confluent underneath, cells were harvested by trypsin treatment. This was adjusted to 1.5 × 10 ⁵ cells / mL by normal human epidermal keratinocytes growth medium (HuMedia-KG2).
Then, the in 35mm petri dish NHEK the (1.5 × 10 ⁵ cells / mL) were 2mL seeded, 37 ° C., and cultured overnight in 5% CO _2. After completion of the culture, the medium was replaced with a normal human epidermal keratinocyte basal medium (HuMedia-KB2) for an additional 24 hours. After completion of the culture, the medium was replaced with 2 mL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 24 hours. After completion of the culture, the culture solution is removed, total RNA is extracted with an RNA extraction reagent (ISOGEN II (catalog number: 311-07361), manufactured by Nippon Gene Co., Ltd.), and the amount of each RNA is measured with a spectrophotometer. Then, purified water was used to prepare total RNA to 200 ng / μL.

In addition, as a control, the same operation and absorbance measurement were performed as described above, except that 2 mL of the test sample-added medium was changed to 2 mL of normal human epidermal keratinocyte basal medium (HuMedia-KB2) containing no test sample. Total RNA was prepared to 200 ng / μL by the same method as described above.

Using each of the above total RNAs as a template, the expression levels of filaggrin mRNA and GAPDH mRNA, which is an internal standard, were measured. For the detection of mRNA, a real-time PCR device (Thermal Cycler DISE (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.) was used for a 2-step real-time PCR reaction.
The expression level of filaggrin mRNA without the addition of the test sample and with the addition of the test sample was 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.
<Equation 12>
Filaggrin mRNA expression promotion rate (%) = A / B × 100
In the formula 12, A and B represent the following, respectively.
A: Correction value when test sample is added B: Correction value when test sample is not added

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

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

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

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

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

(Test Example 5-9: DPPH radical scavenging action test)
In Test Example 1-3, the test sample was changed to the cornflower fermented solution 1 obtained in Production Example 9, the cornflower fermented solution 2 obtained in Production Example 10, and the cornflower extract obtained in Comparative Production Example 5. The DPPH radical scavenging action was tested in the same manner as in Test Example 1-3 except that the final concentration of the test sample was changed to the concentration shown in Table 33 below. The results are shown in Table 33 below.

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

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 Fuji Film Wako Pure Chemical Industries, Ltd.) to prepare a test sample-added medium.
B16 melanoma cells were cultured in DMEM containing 10% FBS under the conditions of 37 ° C. and 5% CO ₂ until they became confluent, and then the cells were recovered by trypsin treatment. This was adjusted to 2.4 × 10 ⁵ cells / mL by 10% FBS and 1 mmol / L theophylline-containing DMEM.
Then, the B16 melanoma cells in 48well plates (2.4 × 10 ⁵ cells / mL), seeded by 300μL per one well, 37 ° C., and 6 hours of culture under the condition of 5% CO _2. After completion of the culture, the medium was replaced with 100 μL of the test sample-added medium, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 3 days. After completion of the culture, 300 μL of each of the test sample-added media was added per well, and the cells were cultured under the conditions of 37 ° C. and 5% CO ₂ for 4 days. The final concentration of the test sample at this time was the concentration shown in Table 34 below.

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

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

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

From the obtained measured values, the melanin production suppression rate (%) corrected by the cell viability calculated based on the following formula 13 was calculated based on the following formula 14. The results are shown in Table 34 below.
<Equation 13>
Cell viability (%) = (C / D) x 100
In the above formula 13, C and D represent the following, respectively.
C: Absorbance at wavelength 540 nm without test sample added D: Absorbance at wavelength 540 nm with test sample added <Equation 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 test sample added B: Amount of melanin with test sample added C: Absorbance at wavelength 540 nm without test sample D: Absorbance at wavelength 540 nm with test sample added

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 safe. Since it is an expensive natural product, it can be widely used in various 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, a lotion, a milky lotion, and the like. Skin cosmetics such as creams, ointments, beauty essences, lotions, packs, jellies, lip creams, lipsticks, foundations, baths, soaps, body shampoos; It can be suitably used for hair styling products and the like.

Claims (5)

It is characterized by containing at least one of the fermented liquor of the genus Lemon balm, the fermented liquor of Tachijakousou, the fermented liquor of Kousuihakka, and the fermented liquor of Yagurumagiku as the active ingredient. Anti-aging agent. It is characterized by containing at least one of the fermented liquor of the genus Lemon balm, the fermented liquor of Tachijakousou, the fermented liquor of Kousuihakka, and the fermented liquor of Yagurumagiku as the active ingredient. Antioxidant. An anti-inflammatory agent comprising at least one of a fermented liquor produced by Aspergillus oryzae of a plant of the genus Moxa and a fermented liquor produced by Aspergillus oryzae of Kousuihakka as an active ingredient. A whitening agent containing at least one of a fermented liquor of a moxa plant, a fermented liquor of Kousuihakka, and a fermented liquor of Yagurumagiku as an active ingredient. At least one selected from the group consisting of the anti-aging agent according to claim 1, the antioxidant agent according to claim 2, the anti-inflammatory agent according to claim 3, and the whitening agent according to claim 4. A cosmetic characterized by containing.