JP5419259B2 - Cosmetics - Google Patents

Description

  The present invention relates to an antioxidant, an anti-inflammatory agent, an anti-aging agent, a hair-restoring agent, an anti-androgen agent, an anti-obesity agent and a cyclic AMP phosphodiesterase activity inhibitor, and a skin cosmetic, a hair cosmetic, and a food and drink.

In recent years, active oxygen has attracted attention as a factor that particularly oxidizes biological components, and its adverse effect on living organisms has become a problem. Active oxygen is generated in the process of energy metabolism in living cells. Examples of active oxygen include superoxide (that is, superoxide anion generated by one-electron reduction of oxygen molecules: .O ₂ ⁻ ), hydrogen peroxide (H ₂ O ₂ ), hydroxy radical (.OH), singlet oxygen ( ¹ O ₂ ) and the like. These active oxygens are essential for the phagocytic sterilization mechanism and play an important role in removing viruses and cancer cells.

  However, excessive production of active oxygen attacks in vivo molecules constituting membranes and tissues in the living body and induces various diseases. Normally, superoxide produced in vivo and used as a starting material for other active oxygen is sequentially eliminated by the catalytic action of superoxide dismutase (SOD) contained in the cells. Is excessive or when the SOD action is reduced, superoxide elimination is insufficient, and the superoxide concentration increases, which may cause tissue damage such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke Cause cataracts, wrinkles, diabetes, arteriosclerosis, stiff shoulders, coldness.

  In particular, since the skin is directly affected by environmental factors such as ultraviolet rays, it is an organ that easily generates superoxide, and therefore, by increasing the concentration of superoxide, for example, the biological tissue such as collagen is decomposed and denatured or It is thought to crosslink and oxidize fats and oils to produce lipid peroxides that damage cells, and the damage caused by active oxygen is the cause of aging such as skin wrinkle formation and skin elasticity reduction (See Non-Patent Document 1). Therefore, by inhibiting and suppressing the generation of active oxygen and in vivo radicals, skin aging such as wrinkle formation and reduced elasticity, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataracts, diabetes, arteries It is considered that various disorders involving active oxygen such as hardening, stiff shoulders, and coldness can be prevented, treated or improved.

  Therefore, attempts have been made to obtain active oxygen scavenging substances and radical scavenging substances from natural products advantageous in terms of safety, and extracts from Brassica family Brassica (Patent Document 1) have such actions. Reference), an extract from a plant belonging to the genus Ryukyubenkei (see Patent Document 2), an extract from an octopus butterfly (see Patent Document 3), an extract from sulfur (see Patent Document 4), and the like. .

  Nitric oxide (NO) is a nitrogen oxide that causes air pollution, acid rain, etc. In recent years, this nitric oxide has become a vascular endothelium-derived relaxing factor (EDRF), a neurotransmitter, and a microorganism in biological defense. -It has been found to be a physiologically active substance exhibiting various functions in vivo, such as tumor cell injury factors. In biological defense, in particular, nitric oxide produced from macrophages protects against bacterial and viral infections.

  However, when nitric oxide is biosynthesized in large quantities, it is not non-toxic to the living body but causes destruction of the self tissue, which causes pathological conditions such as worsening of inflammation, rheumatism, and diabetes. It is also known that large amounts of biosynthesized nitric oxide cause vascular smooth muscle relaxation and excessive permeability increase, and cause endotoxin shock by markedly lowering blood pressure.

  Therefore, it is important to suppress excessive production of nitric oxide in inflammatory diseases. As those having an action of inhibiting the production of nitric oxide, for example, rosemary extract, carnosol, carnosic acid, coffee bean extract, primrose extract, auren extract, buckwheat extract, licorice extract, Inno rose extract, Senkyu extract, Tonin extract, Peonies extract, Yakuinin extract, Red grape extract (see Patent Document 5); , Flower buds, flower buds (see Non-Patent Document 2), and the like are known.

  There are various causes and onset mechanisms of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and various other skin diseases with rough skin. One of the causes is hyaluronidase. Activation, histamine release, platelet aggregation and the like are known. The activation of hyaluronidase promotes histamine degranulation from mast cells, resulting in inflammation. Therefore, prevention, treatment or improvement of inflammatory diseases can be expected by inhibiting the activation of hyaluronidase. As a plant extract having hyaluronidase inhibitory activity, an extract from Japanese maple (see Patent Document 6) and the like are known.

  Further, since hexosaminidase is released at the same time as histamine is released, the histamine release inhibitory action can be evaluated using the release of hexosaminidase as an index. Therefore, it is considered that by suppressing the release of hexosaminidase, the release of histamine can also be suppressed at the same time, which is effective for preventing, treating or improving inflammatory diseases. As a plant extract having a hexosaminidase release inhibitory action, an extract from Fuji tea (see Patent Document 7) and the like are known.

  Platelet aggregates and activates to cause hemostasis physiologically and pathologically thrombus formation, and platelet aggregation is involved in the progression of arteriosclerosis, cancer metastasis, inflammation, etc. It is considered. Therefore, prevention, treatment or improvement of the above diseases can be expected by suppressing platelet aggregation. As a plant extract having a platelet aggregation inhibitory action, an extract from lichen snow tea (see Patent Document 8) and the like are known.

  The dermis and epidermis of the skin is composed of epidermal cells, fibroblasts, and dermal extracellular matrix such as elastin and collagen that are outside these cells and support the skin structure. In young skin, these skin tissues By maintaining the homeostasis, moisture retention, flexibility, elasticity and the like are ensured, and the skin is maintained in a fresh state with a tension and gloss. However, elastin, which is a major component of the dermal extracellular matrix, due to the influence of certain external factors such as ultraviolet irradiation, significant air drying, excessive skin washing, etc. The production amount of collagen and the like is reduced, and degeneration and decomposition are caused. As a result, the stratum corneum begins to exfoliate abnormally, the skin loses its tension and gloss, and exhibits aging symptoms such as rough skin and wrinkles. As described above, changes accompanying aging of the skin, that is, formation of wrinkles, disappearance of tension, decrease in elasticity, and the like are associated with reduction and degeneration of dermal extracellular matrix components such as elastin and collagen.

  Elastin is a fiber that gives elasticity to the skin tissue. When elastin is decomposed with aging or the like, the elasticity of the skin is lost and the elasticity is lowered. Elastase, an enzyme that degrades elastin, is activated by irradiation with ultraviolet rays, thereby accelerating the degradation of elastin, losing skin tension and lowering elasticity. Therefore, it is considered that by inhibiting the activity of elastase, the degradation of elastin can be suppressed, and skin aging symptoms such as loss of tension and reduced elasticity can be prevented and improved. Conventionally, a star fruit fruit extract has been known as a herbal medicine having an elastase inhibitory effect (see Patent Document 9).

  In recent years, the involvement of matrix proteases has been pointed out as a factor that induces changes associated with skin aging. Among matrix proteases, matrix metalloproteinase-1 (MMP-1) is known as an enzyme that degrades collagen, which is a major component of the dermal extracellular matrix of the skin, but its expression is greatly increased by irradiation with ultraviolet rays. It is believed to increase, contribute to the decrease / denaturation of collagen, and to become a major factor such as the formation of wrinkles on the skin and the decrease in elasticity. Therefore, inhibiting MMP-1 activity is important in preventing and improving skin aging symptoms. Conventionally, the extract of the bark of the pine pine genus Futabamatsu is known as a crude drug which has MMP-1 inhibitory action (refer patent document 10).

  The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and an extracellular matrix such as collagen that is outside these cells and supports the skin structure. In young skin, the proliferation of fibroblasts is active, and moisture is retained in the skin by the interaction of skin tissues such as fibroblasts and collagen, and the flexibility and elasticity of the skin are secured. It is maintained in a fresh state with its tension and gloss.

  However, the growth of fibroblasts slows down due to the influence of certain external factors such as ultraviolet rays, drastic drying of air, excessive skin washing, etc., and aging progresses, and the moisture retention function and elasticity of the skin decrease. To do. And the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. Therefore, it is considered that aging of the skin can be prevented or improved by promoting the proliferation of fibroblasts. Based on such an idea, a Kusunohagashi extract (see Patent Document 11) and the like have been proposed as having a fibroblast proliferation promoting action.

  Many steroid hormones are expressed in the form of molecules secreted from production organs and bind to receptors to exert their effects. In the case of male hormones collectively called androgens, for example, testosterone enters the cells of target organs and becomes testosterone. After being reduced to 5α-dihydrotestosterone (5α-DHT) by 5α-reductase, it binds to the receptor and develops an action as an androgen.

  Androgen is an important hormone, but when it works excessively, it is variously preferred, such as androgenetic alopecia, hirsutism, seborrhea, acne (such as acne), benign prostatic hyperplasia, prostate tumor, premature boyhood Not triggering symptoms. Therefore, conventionally, in order to improve these various symptoms, a method for suppressing the action of excess androgen, specifically, by inhibiting the action of testosterone 5α-reductase which reduces testosterone to active 5α-DHT. There are known a method for suppressing the generation of active 5α-DHT, a method for inhibiting the binding of 5α-DHT generated from testosterone to the receptor and not expressing the androgenic activity, and the like.

  Examples of compounds having testosterone 5α-reductase inhibitory action include extracts from plants belonging to the genus Choerospondias (see Patent Document 12), extracts from majito and / or cuture (see Patent Document 13). ), An extract from five lions (see Patent Document 14), an extract from one or more plants selected from the group consisting of red bean cedar, bird cage, hooded umbrella and centripetal lotus (Patent Document 15) For example).

  Moreover, as what has an androgen receptor binding inhibitory action, for example, the extract of a leaf of a yellow skin (refer patent document 16), the extract of a majito and / or a cuttlefish (refer patent document 13), a goat egg Extracts (see Patent Document 14), extracts from wisteria tea branches and leaves (see Patent Document 17), and the like are known.

  In recent years, body fat has increased due to lifestyle habits such as satiety and lack of exercise, and obesity has increased. Such an increase in obesity is seen not only in humans but also in pets and livestock. Obesity is a cause of adult diseases such as hyperlipidemia and arteriosclerosis, which is not only a cosmetic problem but also a major problem in health.

  In order to break down fat in the living body, the role of cyclic AMP is important. Cyclic AMP activates lipase present in the living body, and fat is decomposed into fatty acid and glycerol by the activated lipase. However, when cyclic AMP phosphodiesterase is activated, degradation of cyclic AMP is induced and lipase activation is inhibited. Therefore, it is considered that by inhibiting the activity of cyclic AMP phosphodiesterase, the amount of cyclic AMP in the cell can be increased and the decomposition of fat can be promoted. A peanut astringent skin extract (see Patent Document 18) or the like is known as having such a cyclic AMP phosphodiesterase activity inhibitory action.

JP 2003-81848 A JP 2005-29483 A JP 2006-321730 A JP 2007-8902 A JP 2002-87975 A JP 2003-1113068 A JP 2003-12532 A JP 2005-82532 A Japanese Patent Laid-Open No. 2003-300893 JP 2003-277223 A JP 2003-146837 A JP 2003-55162 A JP 2002-241297 A JP 2002-241296 A JP 2002-87976 A JP 2001-226278 A JP 2002-308790 A JP 2004-26719 A

"Fragrance Journal Extra Number", 1995, No.14, p.156 "Wakan Journal of Pharmaceutical Sciences", 1998, Vol.15, p.302-303

  The present invention relates to an antioxidant, an anti-inflammatory agent, an anti-aging agent, a hair-restoring agent, an anti-androgen agent, an anti-obesity agent and a cyclic, which comprise a highly safe natural extract and / or a natural extract-derived component as an active ingredient. It is an object of the present invention to provide skin cosmetics, hair cosmetics, and foods and drinks that contain an AMP phosphodiesterase activity inhibitor and the natural extract and / or components derived from the natural extract.

In order to achieve the above object, the antioxidant, anti-inflammatory agent, anti-aging agent, hair restorer, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention is extracted from Higashi Shiso. Is contained as an active ingredient.
Moreover, the skin cosmetics, hair cosmetics or foods and beverages of the present invention are characterized by blending an extract from Tosho.

  In the antioxidant of the present invention, it is preferable that the extract from Higashi Shiso has an active oxygen scavenging action and / or a radical scavenging action, and in the anti-inflammatory drug of the present invention, the extract from Higashi Shiso. Preferably has one or more actions selected from the group consisting of a nitric oxide production inhibitory action, a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, and a platelet aggregation inhibitory action. In the aging agent, the extract from Tosho is one or two selected from the group consisting of an elastase activity inhibitory action, a matrix metalloproteinase-1 (MMP-1) activity inhibitory action and a skin fibroblast proliferation promoting action. It is preferable to have more than a kind of action, and in the hair-restoring agent or anti-androgen agent of the present invention, the extract from Toshoso is testosterone. Preferably it has a α- reductase inhibitory activity and / or androgen receptor binding inhibitory action.

  Here, “androgen receptor binding inhibition” in the present invention means inhibition of binding between 5α-DHT and androgen receptor, and the inhibition mode is not particularly limited. For example, competitive antagonists Inhibition as an antagonist, such as a non-competitive antagonist, is contemplated.

  According to the present invention, an extract from Toshiso, which is a natural product, and / or an ingredient derived from the extract as an active ingredient, a highly safe antioxidant, anti-inflammatory agent, anti-aging agent, hair restorer, An anti-androgen agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a skin cosmetic, a hair cosmetic, and a food and drink can be provided.

The present invention will be described below.
[Antioxidants, anti-inflammatory agents, anti-aging agents, hair restorers, anti-androgen agents, anti-obesity agents, cyclic AMP phosphodiesterase activity inhibitors]
The antioxidant, anti-inflammatory agent, anti-aging agent, hair-restoring agent, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention contains an extract from Tosho as an active ingredient.

  Here, in the present invention, the “extract” includes an extract obtained by using Tosoh (scientific name: Elsholtzia bodinieri) as an extraction raw material, a diluted or concentrated solution of the extract, and a dry product obtained by drying the extract. Or any of these crudely purified products or purified products.

  Elsholtzia bodinieri is a plant belonging to the family Lamiaceae and is widely distributed in the southwestern region of China, the southern region of China, etc., and can be easily obtained from these regions. There are no particular limitations on the constituent parts of Higashi Shiso used as the raw material for extraction, and examples include flower parts, leaf parts, branch parts, whole grasses, root parts, etc. Of these, whole grasses are used in particular. Is preferred. Higashi Shiso's whole herb is known as herbal medicine (Houchacha, also known as: Noyama tea, Oyama tea, Kumomatsu tea, Koka tea, Komatsuge tea, Hiyama tea, Fangbushi) , Headache, sore throat, hepatitis, eye pain, indigestion, etc.

  The extract from Higashi Shiso can be obtained by drying the extraction raw material, pulverizing it as it is or using a crusher, and subjecting it to extraction with an extraction solvent. At this time, the extraction raw material may be dried in the sun or by a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing a pretreatment such as degreasing, the extraction raw material can be efficiently extracted with a polar solvent.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as an active ingredient of an antioxidant, anti-inflammatory agent, anti-aging agent, hair restorer, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor as it is, Concentrated liquid or dried product is easier to use.

  Since the extract from Higashi Shiso has a characteristic odor, it can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity. Since it is not used in large quantities when blended into hair cosmetics or foods and drinks, there is no practical problem even if it is not purified.

  The extract from Higashi Shiso obtained as described above has an antioxidant action, an anti-inflammatory action, an anti-aging action, a hair growth action, an anti-androgenic action, an anti-obesity action or a cyclic AMP phosphodiesterase activity inhibition action. Therefore, each action can be used as an active ingredient of an antioxidant, an anti-inflammatory agent, an anti-aging agent, a hair restorer, an anti-androgen agent, an anti-obesity agent or a cyclic AMP phosphodiesterase inhibitor.

  Antioxidant action of the extract from Higashi Shiso is exhibited based on, for example, active oxygen scavenging action and / or radical scavenging action. However, the antioxidant effect which the extract from Dongshio has is not limited to the antioxidant effect exhibited based on these effects.

  The anti-inflammatory action of the extract from Higashi Shiso is, for example, one or two selected from the group consisting of a nitric oxide production inhibitory action, a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, and a platelet aggregation inhibitory action. It is demonstrated based on the above actions. However, the anti-inflammatory action which the Tosho extract has is not limited to the anti-inflammatory action exhibited based on these actions.

  The anti-aging action of the extract from Higashi Shiso is based on, for example, one or more actions selected from the group consisting of an elastase activity inhibitory action, an MMP-1 activity inhibitory action, and a fibroblast proliferation promoting action. Demonstrated. However, the anti-aging action of the Tosho extract is not limited to the anti-aging action exhibited based on these actions.

  The hair-growth action or anti-androgen action of the extract from Dongshio is exhibited based on, for example, testosterone 5α-reductase inhibitory action and / or androgen receptor binding inhibitory action. However, the hair-growth action or anti-androgen action that the extract from Toshiso has is not limited to the hair-growth action or anti-androgen action that is exhibited based on these actions.

  In addition, the extract from Higashi Shiso has an active oxygen scavenging action, radical scavenging action, nitric oxide production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, elastase activity inhibitory action, MMP-1 activity inhibitory action , Fibroblast proliferation promoting action, testosterone 5α-reductase inhibitory action or androgen receptor binding inhibitory action, utilizing these actions, active oxygen scavenger, radical scavenger, nitric oxide production inhibitor, hyaluronidase Used as an active ingredient of an activity inhibitor, hexosaminidase release inhibitor, elastase activity inhibitor, MMP-1 activity inhibitor, fibroblast proliferation promoter, testosterone 5α-reductase inhibitor or androgen receptor binding inhibitor May be.

  The antioxidant, anti-inflammatory agent, anti-aging agent, hair-restoring agent, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase inhibitor of the present invention may consist only of an extract from Tosho. However, it may be a formulation of an extract from Higashi Shiso.

  Extract from Dongseo is any dosage form such as powder, granule, tablet, liquid, etc., using pharmaceutically acceptable carriers such as dextrin, cyclodextrin and other optional auxiliaries according to conventional methods. Can be formulated. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent and the like can be used. In addition, the extract from Dongdo can be used by blending with other compositions (for example, skin cosmetics, hair cosmetics, foods and drinks, etc.).

  The antioxidant, anti-inflammatory agent, anti-aging agent, hair-restoring agent, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention may have an antioxidant effect or an anti-inflammatory effect, if necessary. , Other natural extracts having anti-aging action, hair growth action, anti-androgenic action, anti-obesity action or cyclic AMP phosphodiesterase activity inhibiting action can be used as active ingredients.

  Examples of the method of administering the antioxidant, anti-inflammatory agent, anti-aging agent, hair restorer, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention generally include transdermal administration and oral administration. However, a suitable method for the prevention / treatment or the like may be appropriately selected depending on the type of the disease.

  In addition, the dosage of the antioxidant, anti-inflammatory agent, anti-aging agent, hair-restoring agent, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention is also the type of disease, severity, patient What is necessary is just to increase / decrease suitably according to an individual difference, an administration method, an administration period, etc.

  Antioxidant of the present invention, through the active oxygen scavenging action and / or radical scavenging action of the extract from Tosho, skin aging such as wrinkle formation and reduced elasticity, tissue damage such as rheumatoid arthritis and Behcet's disease, Various disorders involving active oxygen such as myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, shoulder stiffness, and coldness can be prevented and improved. However, the antioxidant of the present invention can be used for all purposes that are meaningful for exhibiting an active oxygen scavenging action and / or a radical scavenging action in addition to these uses.

  The anti-inflammatory agent of the present invention is one or two selected from the group consisting of an inhibitory action on nitric oxide production, an inhibitory action on hyaluronidase activity, an inhibitory action on hexosaminidase release, and an inhibitory action on platelet aggregation. Through the action of more than species, contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other various inflammatory skin diseases associated with rough skin can be prevented and improved. However, in addition to these uses, the anti-inflammatory agent of the present invention is one or two selected from the group consisting of a nitric oxide production inhibitory action, a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, and a platelet aggregation inhibitory action. It can be used for all purposes that are meaningful for exerting more than a kind of action.

  The anti-aging agent of the present invention has one or more actions selected from the group consisting of an elastase activity inhibitory action, an MMP-1 activity inhibitory action, and a fibroblast proliferation promoting action possessed by an extract from Tosho, It can prevent and improve skin aging symptoms and the like. However, the anti-aging agent of the present invention exhibits one or more actions selected from the group consisting of an elastase activity inhibitory action, an MMP-1 activity inhibitory action and a fibroblast proliferation promoting action in addition to these uses. It can be used for all purposes that make sense.

  The hair-growth agent of the present invention is effective for male pattern baldness, hirsutism, seborrhea, acne (acne etc.) through the testosterone 5α-reductase activity inhibitory action and / or androgen receptor binding inhibitory action of the extract from Tosho. ), Benign prostatic hyperplasia, prostate tumor, premature boyhood, etc., and is particularly suitable for the prevention, treatment or improvement of male pattern baldness. However, the hair-restoring agent of the present invention can be used for all purposes that are meaningful for exerting testosterone 5α-reductase activity inhibitory activity and / or androgen receptor binding inhibitory activity in addition to these uses.

  The anti-androgen agent of the present invention can be used for diseases involving male hormones, such as androgenetic alopecia, hirsutism, through the testosterone 5α-reductase activity inhibitory action and / or androgen receptor binding inhibitory action of the extract from Tosho. Symptom, seborrhea, acne (acne etc.), benign prostatic hyperplasia, prostate tumor, premature boyhood, etc. can be prevented, treated or improved. However, the anti-androgenic hormone agent of the present invention can be used for all purposes that are significant in exhibiting testosterone 5α-reductase activity inhibitory activity and / or androgen receptor binding inhibitory activity in addition to these uses.

  The anti-obesity agent of the present invention can promote cyclic AMP production and decompose adipocytes through the cyclic AMP phosphodiesterase inhibitory action of the extract from Tosho, resulting in obesity, Various diseases such as arteriosclerosis, diabetes, and metabolic syndrome can be prevented and improved. However, the anti-obesity agent of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.

  The cyclic AMP phosphodiesterase inhibitor of the present invention can promote the production of cyclic AMP and promote the degradation of adipocytes through the cyclic AMP phosphodiesterase inhibitory action of the extract from Higashi Shiso. Various diseases such as obesity, accompanying arteriosclerosis, diabetes, and metabolic syndrome can be prevented and improved. However, the cyclic AMP phosphodiesterase inhibitor of the present invention can be used for all purposes other than these uses that are meaningful for exerting a cyclic AMP phosphodiesterase inhibitory action.

[Skin cosmetics, hair cosmetics]
The extract from Higashi Shiso has antioxidant, anti-inflammatory, anti-aging, hair-growth, anti-androgenic, anti-obesity, or cyclic AMP phosphodiesterase activity inhibitory effects on skin or hair (scalp) It is suitable for blending into skin cosmetics or hair cosmetics because of its excellent usability and safety when applied to (1).

  In this case, the skin cosmetics or hair cosmetics may be blended with an extract from Toshiso as it is, or an antioxidant, an anti-inflammatory agent, an anti-aging agent formulated from an extract from Tosho. , Hair restorer, anti-androgen, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor.

  Antioxidant, anti-inflammatory agent, anti-aging agent, hair restorer, anti-androgen agent, anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor formulated from an extract from Dong Shiso or an extract from Tosho By blending, it is possible to impart an antioxidant action, an anti-inflammatory action, an anti-aging action, a hair growth action, an anti-androgenic action, an anti-obesity action or a cyclic AMP phosphodiesterase activity inhibitory action to skin cosmetics or hair cosmetics. .

  The kind of skin cosmetics or hair cosmetics that can be blended with the extract from Dongdo is not particularly limited, and examples of skin cosmetics include ointments, creams, emulsions, lotions, packs, foundations, etc. Examples of hair cosmetics include hair tonics, hair creams, hair liquids, shampoos, pomades, rinses, and the like.

  When blending the extract from Higashi Shiso into skin cosmetics or hair cosmetics, the blending amount can be appropriately adjusted according to the type of skin cosmetics or hair cosmetics, It is about 0.0001 to 10% by mass in terms of a standard extract, and a particularly suitable blending ratio is about 0.001 to 1% by mass in terms of a standard extract.

  Skin cosmetic or hair cosmetic of the present invention is an antioxidant action, anti-inflammatory action, anti-aging action, hair growth action, anti-androgen action, anti-obesity action or cyclic AMP phosphodiesterase activity inhibition of an extract from Tosho As long as the action is not hindered, main agents, auxiliaries or other components used in the production of normal skin cosmetics or hair cosmetics, such as astringents, bactericides / antibacterial agents, ultraviolet absorbers, humectants, cell activators, Anti-inflammatory / antiallergic agents, antioxidant / active oxygen scavengers, fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances and the like can be used in combination. By using together, it becomes a more general product, and the synergistic action between the above-mentioned components used in combination may bring about a superior use effect than would normally be expected.

  The skin cosmetic of the present invention is highly safe and comprises a group consisting of an antioxidant action, an anti-inflammatory action, an anti-aging action, an antiandrogenic action, an anti-obesity action and a cyclic AMP phosphodiesterase activity inhibiting action. Through one or more selected actions, skin aging such as wrinkle formation and reduced elasticity, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataract, diabetes, arteriosclerosis, stiff shoulders, coldness Various disorders involving active oxygen such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other inflammatory skin diseases with rough skin; skin aging symptoms; seborrhea, acne (acne etc.) , Skin aging symptoms, seborrhea, acne (acne etc.), benign prostatic hyperplasia, prostate tumor, premature boyhood; prevent, cure obesity, arteriosclerosis associated with obesity, diabetes, metabolic syndrome, etc. Or it can be improved. Moreover, the hair cosmetic composition of the present invention has high safety, and can prevent, treat or improve male pattern baldness or the like through a hair growth action and / or an antiandrogenic action.

[Food and Drink]
The extract from Higashi Shiso has antioxidant, anti-inflammatory, anti-aging, hair-growth, anti-androgen, anti-obesity or cyclic AMP phosphodiesterase inhibitory action and is digested in the digestive tract. It is confirmed that it is not such a thing, and since it is excellent also in safety | security, it is suitable for mix | blending with food-drinks.

When blending extracts from Higashi Shiso into foods and drinks, the amount of active ingredients in them can be changed as appropriate in consideration of the purpose of use, symptoms, sex, etc. In consideration of the intake, it is preferable that the daily intake of the extract is about 1-1000 mg.

  The food / beverage product of the present invention may be a blend of any food / beverage product that does not impede its activity with an extract from Higashi Shiso, or a dietary supplement based on an extract from Higashi Shiso. It may be.

  In producing the food and drink of the present invention, for example, sugars such as dextrin and starch; proteins such as gelatin, soy protein and corn protein; amino acids such as alanine, glutamine and isoleucine; polysaccharides such as cellulose and gum arabic Any additive such as oils and fats such as soybean oil and medium chain fatty acid triglycerides can be added to make foods and drinks of any shape.

  The foods and drinks that can be blended with the extract from Higashi Shiso are not particularly limited, but specific examples include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated concentrates and preparations of these drinks) Ice cream, ice sherbet, shaved ice and other frozen desserts; buckwheat noodles, udon, harsame, gyoza skin, cucumber skin, Chinese noodles, instant noodles and other noodles; rice cakes, chewing gum, candy, gum, chocolate, Confectionery such as tablet confectionery, snack confectionery, biscuits, jelly, jam, cream, baked confectionery; processed fishery and livestock products such as kamaboko, ham, sausage; dairy products such as processed milk, fermented milk; salad oil, tempura oil, margarine, Mayonnaise, shortening, whipped cream, dressing and other fats and oils and processed foods; sauces, sauces and other seasonings; , Stew, salad, side dish, pickles; various other forms of health and nutritional supplements; tablets, capsules, drinks, etc. The auxiliary raw materials and additives used can be used in combination.

  The antioxidant, anti-inflammatory agent, anti-aging agent, hair-restoring agent, anti-androgen agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, skin cosmetic, hair cosmetic or food and drink of the present invention are human. However, it can also be applied to animals other than humans as long as each effect is exhibited.

  Hereinafter, although a manufacture example, a test example, and a compounding example are shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all.

[Production Example 1] Manufacture of Toshoso extract To 42.7 g of dried Toshio whole herb, 400 mL of 50 vol% ethanol (volume ratio of water to ethanol = 1: 1) was added, and reflux extractor was added. And heated at 80 ° C. for 2 hours and filtered while hot. The residue was further extracted in the same manner. The obtained extracts were combined, concentrated under reduced pressure, and further dried to obtain 12.6 g of Toshiso extract (Sample 1).

[Test Example 1] Superoxide scavenging action test (NBT method)
The Toshiso extract obtained in Production Example 1 (Sample 1) was tested for superoxide scavenging action as follows.

In a test tube, 2.4 mL of 0.05 M Na ₂ CO ₃ buffer (pH 10.2), and 3 mM xanthine, 3 mM EDTA, bovine serum albumin solution and 0.75 mM NBT (nitroblue tetrazolium) were added. 1 mL each was added, and 0.1 mL of the sample solution (sample 1, sample concentration: see Table 1 below) was added thereto, and the mixture was allowed to stand at 25 ° C. for 10 minutes. After standing, a xanthine oxidase solution as an enzyme solution was added, stirred rapidly, and allowed to stand at 25 ° C. for 20 minutes. Thereafter, 0.1 mL of 6 mM copper chloride was added to stop the reaction, and the absorbance at a wavelength of 560 nm was measured.

Even when the enzyme solution was not added, the same operation and absorbance were measured, and the same measurement was performed when distilled water was added without adding the sample solution. From the obtained results, the superoxide elimination rate (%) was calculated by the following formula, and the sample concentration IC ₅₀ (μg / mL) at which the superoxide elimination rate was 50% was calculated.

Superoxide erasure rate (%) = {1- (AB) / (CD)} × 100
In the formula, A represents “absorbance when enzyme solution is added / sample solution added”, B represents “absorbance when enzyme solution is not added / sample solution is added”, and C is “when enzyme solution is added / sample solution is not added” "D" represents "absorbance when no enzyme solution is added / sample solution is not added".
The results are shown in Table 1.

  As shown in Table 1, it was confirmed that the East Shiso extract (Sample 1) has an excellent active oxygen scavenging action. Moreover, it was confirmed that the degree of the active oxygen scavenging action can be adjusted by the concentration of the Toshio extract.

[Test Example 2] Radical scavenging action test With respect to the East Shiso extract (Sample 1) obtained in Production Example 1, the radical scavenging action was tested as follows.

3 mL of a sample solution (sample 1, see the following table 2 for the sample concentration) was added to 3 mL of 1.5 × 10 ⁻⁴ M DPPH (diphenyl-p-picrylhydrazyl) ethanol solution, sealed, then shaken and allowed to stand for 30 minutes. . After standing, the absorbance at a wavelength of 520 nm was measured. As a control, the same operation was performed using only the solvent in which the sample was dissolved instead of the sample solution, and the absorbance at a wavelength of 520 nm was measured. Further, as a blank, after adding 3 mL of the sample solution to ethanol, the absorbance at a wavelength of 520 nm was measured immediately. From the obtained results, the radical scavenging rate (%) was calculated by the following formula, and the sample concentration IC ₅₀ (μg / mL) at which the radical scavenging rate was 50% was calculated.

Radical scavenging rate (%) = {1- (BC) / A} × 100
In the formula, A represents “absorbance of control”, B represents “absorbance upon addition of sample solution”, and C represents “absorbance of blank”.
The results are shown in Table 2.

  As shown in Table 2, it was confirmed that the East Shiso extract (Sample 1) has an excellent radical scavenging action. In addition, it was confirmed that the degree of radical scavenging action can be adjusted by the concentration of Tosho extract.

[Test Example 3] Nitric Oxide Production Inhibitory Action Test The Toshiso extract (Sample 1) obtained in Production Example 1 was tested for nitric oxide production inhibitory action as follows.

Mouse macrophage cells (RAW264.7 cells, manufactured by Dainippon Pharmaceutical Co., Ltd.) were precultured in DMEM medium (Nissui Pharmaceutical Co., Ltd.) supplemented with 10% FBS, and the cells were collected with a cell scraper. The collected cells are diluted with D-MEM containing 10% FBS and not containing phenol red to a cell density of 3.0 × 10 ⁶ cells / mL, and then seeded at 100 μL per well in a 96-well plate. Cultured for 4 hours.

  After completion of the culture, the medium was removed, and a sample solution (sample 1, sample concentration: 200 μg / mL) dissolved in D-MEM containing 10% FBS containing DMSO at a final concentration of 2% and not containing phenol red was added to each well. Add 100 μL, add 100 μL of lipopolysaccharide (LPS, final concentration 1 μg / mL, E. coli 0111: B4, manufactured by DIFCO) dissolved in D-MEM containing 10% FBS and no phenol red, and culturing for 48 hours did.

Nitric oxide (NO) production was measured using the amount of nitrite ion (NO ₂ ⁻ ) as an index. After completion of the culture, add the same amount of grease reagent (1% by weight sulfanilamide, 0.1% N-1-naphthylethylendiamine dihydrochlpride in 5% phosphoric acid solution) to the culture medium of each well, The reaction was allowed to proceed for 10 minutes at room temperature. After the reaction, absorbance at a wavelength of 540 nm was measured. Nitric oxide (NO) production inhibition rate (%) was calculated by the following formula based on the amount of nitric oxide (NO) produced when no sample was added (control).

NO production inhibition rate (%) = {1- (C−D) / (A−B)} × 100
In the formula, A represents “absorbance without addition of sample”, B represents “absorbance of blank without addition of sample”, C represents “absorbance upon addition of sample”, and D represents “absorbance upon addition of sample”. "Absorbance of blank".

  As a result of the calculation as described above, the nitric oxide inhibition rate of the East Shiso extract (Sample 1) was 15.3%. From this, it was confirmed that the Tosho extract has an excellent nitric oxide production inhibitory action.

[Test Example 4] Hyaluronidase activity inhibitory action test The East Shiso extract obtained in Production Example 1 (Sample 1) was tested for hyaluronidase activity inhibitory action as follows.

  Hyaluronidase solution (Type IV-S (from bovine testis), SIGMA) in 0.2 mL of 0.1 mol / L acetate buffer (pH 3.5) in which the sample was dissolved (Sample 1, see Table 3 below for sample concentration) 0.1 mL (manufactured by 400NF units / mL) was added and reacted at 37 ° C. for 20 minutes. Furthermore, 0.2 mL of 2.5 mmol / L calcium chloride was added as an activator and reacted at 37 ° C. for 20 minutes. To this, 0.5 mL of 0.4 mg / mL sodium hyaluronate solution (from Rooster Comb) was added and reacted at 37 ° C. for 40 minutes.

  Thereafter, 0.2 mL of 0.4 mol / L sodium hydroxide was added to stop the reaction, and after cooling, 0.2 mL of boric acid solution was added to each reaction solution and boiled for 3 minutes. After cooling with ice, 6 mL of p-DABA reagent was added and reacted at 37 ° C. for 20 minutes. Thereafter, the absorbance at a wavelength of 585 nm was measured. A blank test was performed and corrected in the same manner. From the obtained measurement results, the hyaluronidase activity inhibition rate (%) was calculated by the following formula.

Inhibition rate (%) = {1− (St−Sb) / (Ct−Cb)} × 100
In the formula, St represents “absorbance of the sample solution at a wavelength of 585 nm”, Sb represents “absorbance of the sample solution at a wavelength of 585 nm”, Ct represents “absorbance of the control solution at a wavelength of 585 nm”, and Cb represents “control”. The absorbance of the solution blank at a wavelength of 585 nm is expressed.
The results are shown in Table 3.

  As shown in Table 3, it was confirmed that the East Shiso extract (Sample 1) has an excellent hyaluronidase activity inhibitory action. In addition, it was confirmed that the degree of hyaluronidase activity inhibitory action can be adjusted by the concentration of Tosho extract.

[Test Example 5] Hexosaminidase release inhibitory action test The Toshosho extract (Sample 1) obtained in Production Example 1 was tested for hexosaminidase release inhibitory action as follows.

Rat basophil leukemia cells (RBL-2H3) were cultured in S-MEM medium supplemented with 15% FBS, and then cells were collected by trypsin treatment. The collected cells are diluted to a cell density of 4.0 × 10 ⁵ cells / mL, DNP-specific IgE is added to a final concentration of 0.5 μg / mL, and then seeded at 100 μL per well in a 96-well plate. And cultured overnight.

  After completion of the culture, the medium was removed and washing was performed twice with 100 μL of Silligalian buffer. Next, 30 μL of the same buffer solution and 10 μL of the sample solution prepared with the same buffer solution (sample 1, see the following Table 4 for the sample concentration) were added, and the mixture was left at 37 ° C. for 10 minutes. Thereafter, 10 μL of a 100 ng / mL DNP-BSA solution was added, and the mixture was allowed to stand at 37 ° C. for 15 minutes to release hexosaminidase.

  Thereafter, the release was stopped by allowing the 96-well plate to stand on ice. 10 μL of cell supernatant in each well and 10 μL of 1 mmol / L p-NAG (p-nitrophenyl-N-acetyl-β-D-glucoside) solution were added to a new 96-well plate and incubated at 37 ° C. for 1 hour. Reacted.

After completion of the reaction, 250 μL of 0.1 mol / L Na ₂ CO ₃ / NaHCO ₃ was added to each well, and the absorbance at a wavelength of 415 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured and corrected as turbidity. As a blank test, the absorbance at a wavelength of 415 nm of a mixed solution of 10 μL of cell supernatant and 250 μL of 0.1 mol / L Na ₂ CO ₃ / NaHCO ₃ was measured in the same manner. From the obtained measurement results, the hexosaminidase release inhibition rate (%) was calculated by the following formula.

Inhibition rate of hexosaminidase release (%) = {1− (B−C) / A} × 100
In the formula, A represents “absorbance without addition of sample”, B represents “absorbance without addition of sample”, and C represents “absorbance without addition of sample / p-NAG”.
The results are shown in Table 4.

  As shown in Table 4, it was confirmed that the East Shiso extract (Sample 1) has an excellent hexosaminidase release inhibitory action.

Test Example 6 Platelet Aggregation Inhibitory Action Test The Tohso extract obtained in Production Example 1 (Sample 1) was tested for platelet aggregation inhibitory action as follows.

  1/10 amount of Japanese Pharmacopoeia heparin injection solution is added to the collected blood of the rabbit, centrifuged (180 × g, 10 minutes, room temperature), and a platelet rich liquid (PRP) is obtained. Obtained.

  Next, 1 μL of a 200 mmol / L calcium chloride solution was added to 223 μL of platelet suspension (PRP) and reacted at 37 ° C. for 1 minute. Add 1 μL of the sample solution (sample 1, sample concentration as shown in Table 5 below), react for another 2 minutes, add a stir bar and stir for 1 minute, add 25 μL of the collagen solution, and add at 37 ° C. for 10 minutes. The platelet aggregation rate was measured. Separately, as a control, the platelet aggregation rate was measured in the same manner as above except that the solvent of the test sample solution was added instead of the test sample solution. From the measurement results obtained, the platelet aggregation inhibition rate (%) was calculated by the following formula.

Platelet aggregation inhibition rate (%) = {(A−B) / A} × 100
In the formula, A represents “control platelet aggregation rate”, and B represents “platelet aggregation rate at the time of sample solution addition”.
The results are shown in Table 5.

  From the results shown in Table 5, it was confirmed that the Tosho extract (sample 1) has an excellent platelet aggregation inhibitory action.

[Test Example 7] Elastase Activity Inhibitory Action Test The East Shiso extract (Sample 1) obtained in Production Example 1 was tested for elastase activity inhibitory action as follows.

  Sample solution (sample 1, sample concentration: 400 μg / mL) 50 μL and 20 μg / mL elastase type III solution 50 μL prepared with 0.2 mol / L Tris-HCl buffer (pH 8.0) in a 96-well plate Were mixed. Thereafter, 100 μL of 0.4514 mg / mL N-succinyl-Ala-Ala-Ala-p-nitroanilide prepared in the above buffer solution was added and reacted at 25 ° C. for 15 minutes.

  After completion of the reaction, absorbance at a wavelength of 415 nm was measured. A blank test was performed and corrected in the same manner. From the obtained results, the elastase activity inhibition rate (%) was calculated by the following formula.

Elastase activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
In the formula, A represents “absorbance at a wavelength of 415 nm when no sample is added / enzyme added”, B represents “absorbance at a wavelength of 415 nm when no sample is added / enzyme is added”, and C is “sample added / enzyme added” "Absorbance at a wavelength of 415 nm", and D represents "absorbance at a wavelength of 415 nm when a sample is added and no enzyme is added".

  As a result of the calculation as described above, the elastase activity inhibition rate of the East Shiso extract (Sample 1) was 15.5%. From this, it was confirmed that the Toshiso extract has an excellent elastase activity inhibitory action.

[Test Example 8] MMP-1 Activity Inhibitory Action Test With respect to the East Shiso extract (Sample 1) obtained in Production Example 1, the MMP-1 activity inhibitory action was tested as follows.

  Sample solution prepared with 0.1 mol / L Tris-HCl buffer (pH 7.1) containing 20 mmol / L calcium chloride in a test tube with a lid (Sample 1, see Table 6 below for sample concentration) 50 μL, 0.1 mg / mL of matrix metalloproteinase-1 (MMP-1, COLLAGENASE Type IV from Clostridium histolyticum, manufactured by SIGMA) 50 μL, and 0.5 mol / L of Pz-peptide (Pz-Pro-Leu-Gly -Pro-D-Arg-OH, manufactured by BACHEM Feinchemikalien AG) 400 μL was mixed and reacted at 37 ° C. for 30 minutes, and then 1 mL of a 25 mmol / L citric acid solution was added to stop the reaction.

Thereafter, 5 mL of ethyl acetate was further added and shaken vigorously. This was centrifuged (1600 × g, 10 minutes), and the absorbance of the ethyl acetate layer at a wavelength of 320 nm was measured. A blank test was conducted by the same method. From the obtained results, the MMP-1 activity inhibition rate (%) was calculated by the following formula, and the sample concentration IC ₅₀ (μg / mL) that inhibited MMP-1 activity by 50% was calculated.

MMP-1 activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
In the formula, A represents “absorbance at the time of no sample addition / enzyme addition”, B represents “absorbance at the time of sample addition / no enzyme addition”, and C represents “absorbance at the time of sample addition / enzyme addition”. , D represents “absorbance when sample is added and enzyme is not added”.
The results are shown in Table 6.

  As shown in Table 6, it was confirmed that the East Shiso extract (Sample 1) has an excellent MMP-1 activity inhibitory action. In addition, it was confirmed that the degree of MMP-1 activity inhibitory action can be adjusted by the concentration of Tosho extract.

[Test Example 9] Skin fibroblast proliferation promoting action test The East Shiso extract (Sample 1) obtained in Production Example 1 was tested for skin fibroblast proliferation promoting action as follows.

Human normal skin fibroblasts (NB1RGB) were cultured using α-MEM containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with 5% FBS-containing α-MEM to a cell density of 7.0 × 10 ⁴ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured overnight.

  After completion of the culture, 100 μL of a sample solution dissolved in 5% FBS-containing α-MEM (Sample 1, see Table 7 below for sample concentration) was added to each well and cultured for 3 days. Fibroblast proliferation was measured using the MTT assay.

  After completion of the culture, 100 μL of the medium was removed from each well, and 20 μL of MTT dissolved in PBS (−) at a final concentration of 5 mg / mL was added to each well. After culturing for 4.5 hours, 100 μL of a 0.01 mol / L hydrochloric acid solution in which 10% SDS was dissolved was added to each well and incubated overnight, and then the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. In addition, a blank test was performed and corrected in the same manner. From the obtained results, the fibroblast proliferation promotion rate (%) was calculated by the following formula.

Fibroblast proliferation promotion rate (%) = (St−Sb) / (Ct−Cb) × 100
In the formula, St represents “absorbance in cells to which a sample was added”, Sb represents “absorbance in a blank test to which a sample was added”, Ct represents “absorbance in cells to which no sample was added”, and Cb It represents “absorbance of blank test without adding sample”.
The results are shown in Table 7.

  As shown in Table 7, it was confirmed that the East Shiso extract (Sample 1) has an excellent fibroblast proliferation promoting action.

[Test Example 10] Testosterone 5α-reductase inhibitory action test The Tosho Su extract (Sample 1) obtained in Production Example 1 was tested for testosterone 5α-reductase inhibitory action as follows.

  In a V-bottom test tube with a lid, 20 μL of 4.2 mg testosterone (manufactured by Wako Pure Chemical Industries, Ltd.) dissolved in 1 mL of propylene glycol and 5 mmol / L Tris-HCl buffer (pH 7) containing 1 mg / mL NADPH. .13) 825 μL was mixed.

  Furthermore, 80 μL of ethanol aqueous solution of each sample (Sample 1, see Table 8 below) and 75 μL of S-9 (rat liver homogenate, manufactured by Oriental Yeast Co., Ltd.) were added and mixed, and incubated at 37 ° C. for 30 minutes did. Thereafter, 1 mL of methylene chloride was added to stop the reaction. This was centrifuged (1600 × g, 10 minutes), the methylene chloride layer was separated, and the separated methylene chloride layer was subjected to gas chromatography analysis under the following conditions to obtain 3α-androstanediol, 5α. -The concentration of dihydrotestosterone (5α-DHT) and testosterone (μg / mL) was quantified. As a control, the same amount (80 μL) of the sample solvent was used instead of the sample solution, and the same treatment was performed for gas chromatography analysis.

[Gas chromatography conditions]
Equipment used: Shimadzu GC-7A (manufactured by Shimadzu Corporation)
Column: DB-1701 (inner diameter: 0.53 mm, length: 30 m, film thickness: 1.0 μm, manufactured by J & W Scientific)
Column temperature: 240 ° C
Inlet temperature: 300 ° C
Detector: FID
Sample injection volume: 1 μL
Split ratio: 1: 2
Carrier gas: Nitrogen gas Carrier gas flow rate: 3 mL / min

Quantification of the concentrations of 3α-androstanediol, 5α-DHT and testosterone was performed by the following method.
Standard products of 3α-androstanediol, 5α-DHT and testosterone were dissolved in methylene chloride, and the solution was subjected to gas chromatography analysis. From the concentration (μg / mL) and peak area of these compounds, peak area and compound Correspondence with the concentration of was previously determined. Then, the concentration per peak area of 3α-androstanediol, 5α-DHT and testosterone after the reaction of testosterone and S-9 is calculated using the following formula (1). Based on.

A = B × C / D (1)
In the formula, A represents “3α-androstanediol, 5α-DHT or testosterone concentration (μg / mL)”, B represents “3α-androstanediol, 5α-DHT or testosterone peak area”, and C Represents “concentration of standard product (μg / mL)”, and D represents “peak area of standard product”.

  Using the compound concentration calculated based on the formula (1), based on the following formula (2), the conversion rate (the concentrations of 3α-androstanediol and 5α-DHT produced by reducing testosterone by testosterone 5α-reductase) And the ratio of the initial concentration of testosterone).

Conversion rate = (E + F) / (E + F + G) (2)
In the formula, E represents “3α-androstanediol concentration (μg / mL)”, F represents “5α-DHT concentration (μg / mL)”, and G represents “testosterone concentration (μg / mL)”. ".

Using the conversion rate calculated based on the formula (2), the testosterone 5α-reductase inhibition rate (%) is calculated based on the following formula (3), and the testosterone 5α-reductase inhibition rate is 50%. The concentration IC ₅₀ (μg / mL) was calculated.
Inhibition rate (%) = (1-H / I) × 100 (3)
In the formula, H represents “conversion rate at the time of sample addition”, and I represents “control conversion rate”.
The results are shown in Table 8.

  As shown in Table 8, it was confirmed that the East Shiso extract (Sample 1) has an excellent testosterone 5α-reductase inhibitory action. Moreover, it was confirmed that the degree of testosterone 5α-reductase inhibitory action can be adjusted by the concentration of the Tosho extract.

Test Example 11 Androgen Receptor Binding Inhibitory Action Test With respect to the East Shiso extract (Sample 1) obtained in Production Example 1, the androgen receptor binding inhibitory action was tested as follows.

Androgen-dependent mouse breast cancer cells (SC-3 cells) cloned from mouse spontaneous breast cancer (Shionogi cancer; SC115) were cultured in MEM medium containing 2% DCC-FBS and 10 ⁻⁸ mol / L testosterone (hereinafter, After culturing using MEM-2, the cells were collected by trypsin treatment. The collected cells are diluted with MEM-2 medium so as to have a cell density of 1.0 × 10 ⁵ cells / μL, seeded at 100 μL per well in a 96-well plate, 37 ° C., 5% CO ₂ -95%. The cells were cultured under air conditions.

After 24 hours, the sample (Sample 1, see Table 9 below for sample concentration) and 0.5% BSA-containing HamF12 + MEM medium (hereinafter referred to as HMB medium) supplemented with 1.0 × 10 ⁻⁹ mol / L DHT. The medium was changed and the culture was continued for 48 hours.

  Thereafter, the medium was replaced with MEM-2 medium containing 0.97 mmol / L of MTT, and after culturing for 2 hours, the medium was replaced with isopropanol, and blue formazan produced in the cells was extracted. For the isopropanol containing the eluted blue formazan, the absorbance at 570 nm where the absorption maximum of blue formazan is present was measured.

  In order to correct the influence of adherent cells, the absorbance at 650 nm was also measured at the same time, and the difference between the two absorbances was taken as a value proportional to the amount of blue formazan produced (the absorbance in the formula for calculating the binding inhibition rate is the corrected absorbance below). Is). In parallel with the above, in order to examine the effect of the sample alone on the SC-3 cells, the same culture and measurement were performed by adding only the sample without adding DHT to the HMB medium.

Further, as a control, the same measurement was performed when culturing in an HMB medium without addition of a sample and DHT, and when culturing in an HMB medium without addition of a sample and only DHT. From the measurement results, the androgen receptor binding inhibition rate (%) was calculated according to the following formula, and the sample concentration IC ₅₀ (μg / mL) that inhibits the binding of androgen to the receptor by 50% was calculated.

Androgen receptor binding inhibition rate (%) = {1− (A−B) / (C−D)} × 100
In the formula, A represents “absorbance when DHT is added / sample added”, B represents “absorbance when DHT is not added / sample is added”, and C is “absorbance when DHT is added / sample is not added”. D represents “absorbance when DHT is not added / sample is not added”.
The results are shown in Table 9.

  As shown in Table 9, it was confirmed that the East Shiso extract (Sample 1) has an excellent androgen receptor binding inhibitory action. Moreover, it was confirmed that the degree of the androgen receptor binding inhibitory action can be adjusted by the concentration of the Tosho extract.

[Test Example 12] Cyclic AMP phosphodiesterase activity inhibitory action test The Tosho Su extract (Sample 1) obtained in Production Example 1 was tested for cyclic AMP phosphodiesterase activity inhibitory action as follows.

  0.1 mL of a 2.5 mg / mL bovine serum albumin solution, 0.1 mg / mL cyclic AMP phosphodiesterase solution in 0.2 mL of 50 mM Tris-HCl buffer (pH 7.5) containing 5 mM magnesium chloride 0.05 mL of a sample solution dissolved in 1 mL and 50% DMSO (Sample 1, see Table 10 below for sample concentration) was added, and incubated at 37 ° C. for 5 minutes.

  To this reaction solution, 0.05 mL of a 0.5 mg / mL cyclic AMP solution was added and incubated at 37 ° C. for 60 minutes. The reaction was stopped by boiling on a boiling water bath for 3 minutes, this was centrifuged (2260 × g, 10 minutes, 4 ° C.), and cyclic AMP, which is a reaction substrate in the supernatant, was subjected to a high-speed liquid under the following conditions. Analyzed using chromatography. In addition, a blank test was performed in the same manner without adding the sample solution.

<High performance liquid chromatography conditions>
Product name: Chromatocorder 12 (manufactured by SYSTEM INSTRUMENTS)
Stationary phase: Wakosil 5C ₁₈ (Wako Pure Chemical Industries, Ltd.)
Column diameter: 4.6 mm
Column length: 250mm
Mobile phase: 1 mM TBAP in 25 mM KH ₂ PO ₄ : CH ₃ CN = 90: 10
Mobile phase flow rate: 1.0 mL / min
Detection: UV, 260nm

  Next, the peak area (A) of the cyclic AMP standard product, the peak area (B1) of the supernatant of the reaction solution of the cyclic AMP standard product and cyclic AMP phosphodiesterase when no sample is added, and the cyclic when the sample is added The peak area (B2) of the supernatant of the reaction solution of AMP standard product and cyclic AMP phosphodiesterase was determined. From the obtained results, the decomposition rate (C) of the cyclic AMP standard product when no sample was added and the decomposition rate (D) of the cyclic AMP standard product when the sample was added were calculated according to the following equations.

Standard product decomposition rate when no sample is added (C,%) = (1−B1 / A) × 100
Decomposition rate of standard product at the time of sample addition (D,%) = (1−B2 / A) × 100

Thereafter, a cyclic AMP phosphodiesterase activity inhibition rate (%) is calculated by the following formula based on the respective degradation rates (C, D) calculated by the above formula, and the cyclic AMP phosphodiesterase activity inhibition rate is 50%. The concentration IC ₅₀ (μg / mL) was calculated.
Inhibition rate (%) = (1-D / C) × 100
The results are shown in Table 10.

  As shown in Table 10, it was confirmed that the East Shiso extract (Sample 1) has an excellent cyclic AMP phosphodiesterase activity inhibitory action. In addition, it was confirmed that the degree of the cyclic AMP phosphodiesterase activity inhibitory action can be adjusted by the concentration of the Tosho extract.

[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.1g
Jojoba oil 4.0g
Olive oil 2.0g
Squalane 2.0g
Cetanol 2.0g
Glyceryl monostearate 2.0g
Polyoxyethylene cetyl ether (20E.O.) 2.5g
Oleic acid polyoxyethylene sorbitan (20E.O.) 2.0g
Twilight extract 0.1g
0.1g dipotassium glycyrrhizinate
Ginkgo biloba extract 0.1g
Conchiolin 0.1g
Oat extract 0.1g
Chamomile extract 0.1g
1,3-butylene glycol 3.0 g
Methyl paraoxybenzoate 0.15g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Formulation Example 2]
A lotion having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.1g
Glycerin 3.0g
1,3-butylene glycol 3.0 g
Oleic acid polyoxyethylene sorbitan (20E.O.) 0.5g
Methyl paraoxybenzoate 0.15g
Citric acid 0.1g
Sodium citrate 0.1g
Oil soluble licorice extract 0.1g
Seaweed extract 0.1g
Xylobiose Mixture 0.5g
Kujin extract 0.1g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Composition Example 3]
A cream having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.1g
Liquid paraffin 5.0g
Salami beeswax 4.0g
Cetanol 3.0g
Squalane 10.0g
Lanolin 2.0g
Stearic acid 1.0g
Oleic acid polyoxyethylene sorbitan (20E.O.) 1.5g
3.0 g glyceryl monostearate
1,3-butylene glycol 6.0 g
Yeast extract 0.1g
Perilla extract 0.1g
Linden extract 0.1g
Jiuyu Extract 0.1g
1.5 g of methyl paraoxybenzoate
Fragrance 0.1g
Purified water remainder (total amount is 100 g)

[Formulation Example 4]
A hair nourishing hair tonic having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.2g
0.1 g of pyridoxine hydrochloride
Resorcin 0.01g
D-pantothenyl alcohol 0.1g
0.1g dipotassium glycyrrhizinate
L-Menthol 0.05g
1,3-butylene glycol 4.0 g
Carrot extract 0.5g
Ethanol 25.0g
Fragrance 0.01g
Purified water remainder (total amount is 100 g)

[Formulation Example 5]
A shampoo (cream shampoo) having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.2g
Sodium polyoxyethylene alkyl ether sulfate 30.0g
Polyoxyethylene alkyl ether ammonium sulfate 20.0g
Coconut oil fatty acid amidopropyl betaine 6.0g
Coconut oil fatty acid modiethanolamide 4.0g
2.0g ethylene distearate
Preservative (Methyl paraoxybenzoate) 0.15g
Mukuroji extract 0.2g
Twilight extract 0.5g
Oat extract 0.3g
Rosemary extract 0.5g
1,3-butylene glycol 3.0 g
Fragrance 0.01g
Purified water remainder (total amount is 100 g)

[Composition Example 6]
A rinse having the following composition was produced by a conventional method.
East Shiso Extract (Production Example 1) 0.2g
Stearyltrimethylammonium chloride 1.5g
Polyoxyethylene cetyl ether 1.0g
Cetyl alcohol 2.0g
Octyldodecanol 1.0g
Cationized cellulose 0.5g
Propylene glycol 5.0g
Mukuroji extract 0.2g
Twilight extract 0.5g
Oat extract 0.3g
Rosemary extract 0.5g
Fragrance 3.0g
Purified water remainder (total amount is 100 g)

[Formulation Example 7]
The following mixture was tableted to produce a tablet-shaped dietary supplement.
East Shiso extract (Production Example 1) 50 parts by mass Powdered sugar (sucrose) 188 parts by mass Glycerin fatty acid ester 12 parts by mass

[Formulation Example 8]
The following mixture was granulated to produce a dietary supplement.
East Shiso extract (Production Example 1) 50 parts by weight Beet oligosaccharide 1000 parts by weight Vitamin C 167 parts by weight Stevia extract 10 parts by weight

  The antioxidant of the present invention is used to prevent and improve various disorders caused by reactive oxygen species, the anti-inflammatory agent of the present invention is used to prevent and improve various inflammatory diseases, and the anti-aging agent of the present invention is used to prevent skin For the prevention / improvement of aging, the hair-restoring agent or anti-androgen hormone agent of the present invention can be used for male pattern alopecia, hirsutism, seborrhea, acne (acne etc.), benign prostatic hyperplasia, prostate tumor, premature male sexuality Prevention / improvement, the anti-obesity agent or cyclic AMP phosphodiesterase activity inhibitor of the present invention can greatly contribute to the prevention / improvement of obesity.

Claims (8)

  A hair restorer comprising an extract from Higashi Shiso as an active ingredient. 2. The hair restorer according to claim 1 , wherein the extract from Higashi Shiso has testosterone 5α-reductase activity inhibitory action and / or androgen receptor binding inhibitory action.   An antiandrogenic agent characterized by containing an extract from Higashi Shiso as an active ingredient. 4. The anti-androgen agent according to claim 3 , wherein the extract from Higashi Shiso has testosterone 5α-reductase activity inhibitory action and / or androgen receptor binding inhibitory action.   An anti-obesity agent characterized by containing an extract from East Shiso as an active ingredient.   A cyclic AMP phosphodiesterase activity inhibitor characterized by containing an extract from Higashi Shiso as an active ingredient. A skin cosmetic for the prevention, treatment or improvement of seborrhea or acne, or slimming skin, characterized by containing an extract from Higashi Shiso. A hair growth cosmetic for hair growth characterized by blending an extract from Higashi Shiso.