JP2016079154A - Skin cosmetic, hair cosmetic, and food and drink - Google Patents

Abstract

PROBLEM TO BE SOLVED: To discover an ingredient having an anti-aging action, a skin-whitening action, a hair-growing action, an anti-androgen action, an anti-inflammatory action, an anti-obesity action, a cyclic AMP phosphodiesterase activity inhibitory action, a suppressing action on glutathione decrease, or a dipeptidyl peptidase IV activity inhibitory action; and to provide an anti-aging agent, a skin-whitening agent, a hair-growing agent, anti-androgen agent, an anti-inflammatory agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, an agent for suppressing glutathione decrease, or a dipeptidyl peptidase IV activity inhibitor, which contains the ingredient as an active ingredient.SOLUTION: An anti-aging agent, a skin-whitening agent, a hair-growing agent, an anti-androgen agent, an anti-inflammatory agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, an agent for suppressing glutathione decrease, and a dipeptidyl peptidase IV activity inhibitor contain 4-vinyl catechol as an active ingredient. The 4-vinyl catechol is blended in the skin cosmetic, the hair cosmetic and the food and drink of the present invention.SELECTED DRAWING: None

Description

  The present invention relates to an anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor, dipeptidyl peptidase IV activity inhibitor, skin cosmetics Food, hair cosmetics, and food and drink.

  Glutathione is a tripeptide consisting of three amino acids, glutamic acid, cysteine and glycine, and is a compound having a major cysteine residue in the cell. Intracellular glutathione functions as a radical donor, regulation of cell function by redox, foreign body metabolism, SH donor of various enzymes, and is also known as an antioxidant component against active oxygen and the like. Its action expression is thought to be derived from cysteine residues. However, it has been reported that the amount of glutathione in cells is deficient or decreased due to excessive oxidative stress, addition of foreign substances, aging, etc., which reduces the protective ability of cells against oxidative stress, and the cellular DNA It is also considered to be a cause of damaging components such as proteins.

  Diseases that are known to be associated with pathological conditions such as a decrease or deficiency in intracellular glutathione levels include diseases caused by oxidative stress, as well as liver damage (drinking alcohol, Or it is caused by ingestion of foreign substances such as heavy metals and chemical substances). That is, it is considered that promoting the production of glutathione can enhance the ability of cells to protect against oxidative stress, and can prevent or treat the above-mentioned disease group caused by the decrease or lack of intracellular glutathione level. . As a substance having a glutathione production promoting action, liquiritigenin (see Patent Document 1) and the like are known.

  A basement membrane is present at the boundary between the epidermis and the dermis constituting the skin. The basement membrane not only connects the epidermis and dermis, but also plays an important role in maintaining skin function (see Non-Patent Document 1). The main skeleton of the basement membrane has a network structure composed of type IV collagen. Various glycoproteins mainly composed of laminin 5 are present at the boundary between the basement membrane and the epidermis and connect the basement membrane and the epidermis. Such laminin 5 is produced from epidermal keratinocytes present in the epidermis. Is done. In young skin, the basement membrane works to maintain the homeostatic interaction of the epidermis and dermis, ensuring moisture retention, flexibility, elasticity, and the like. Maintained.

  However, laminin 5 which is a main component of the basement membrane degrades and deteriorates when it is affected by certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc., or when aging progresses. And the basement membrane structure is destroyed (see Non-Patent Document 2). As a result, the skin's moisturizing function and elasticity are lowered, and the keratin begins to exfoliate abnormally, so the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. As described above, changes associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with a decrease in basement membrane components and structural changes in basement membrane, and production of laminin 5 It is considered that aging symptoms of the skin can be prevented or improved by promoting the above.

  Laminin consists of various combinations of α chain, β chain and γ chain, and 15 types (laminin 1 to laminin 15) are known at present. Of these, laminin 5 (α3β3γ2) is present in large amounts in the basement membrane of epithelial tissues such as skin, digestive organs, kidneys and lungs. In genetic diseases caused by congenital abnormalities of the genes encoding each chain of laminin 5 (lethal congenital epidermolysis bullosa, Herlitz junctional epidermolysis bullosa), the epidermis of the whole body may show fatal symptoms Are known. Laminin 5 is known to strongly adhere cells (high cell adhesion activity) and strongly promote cell motility (high cell motility activity) compared to other extracellular matrix molecules.

  Thus, since laminin 5 has high cell motility activity, it is known to promote cell migration in damaged skin and promote damage healing (see Patent Document 2). That is, promoting the production of laminin 5 is important in promoting healing of skin damage that destroys the structure of the basement membrane.

  In recent years, the involvement of matrix metalloproteinases (MMPs) has been pointed out as factors that induce changes associated with skin aging. Among these MMPs, matrix metalloproteinase-1 (MMP-1), which is an enzyme belonging to the collagenase group, is known as an enzyme that degrades collagen, which is the main component of the dermal extracellular matrix. The expression of MMP-1 is greatly increased by irradiation with ultraviolet rays, which contributes to the decrease / denaturation of collagen, and is considered to be a major factor such as the formation of wrinkles on the skin and the decrease in elasticity. Further, when the activity of MMP-1 is enhanced, the extracellular matrix is destroyed. It is known that destruction of extracellular matrix is associated with various diseases such as cancer invasion / metastasis, rheumatoid arthritis, knee osteoarthritis, periodontal disease, age-related macular degeneration, and the like.

  Therefore, it is considered that inhibiting the activity of MMP-1 can prevent, treat or ameliorate skin aging symptoms and treat or prevent diseases associated with the destruction of extracellular matrix. . As what has MMP-1 activity inhibitory action, corosolic acid (refer patent document 3) etc. are known, for example.

  On the other hand, matrix metalloproteinase-2 (MMP-2), which is an enzyme belonging to the gelatinase group, is known as an enzyme that degrades type IV collagen and laminin 5 which are main components of the basement membrane. It is clear that the expression and activity of MMP-2 are greatly increased by irradiation with ultraviolet rays, causing a decrease in basement membrane components and structural changes of the basement membrane due to ultraviolet rays, and a major factor in the formation of wrinkles and sagging in the skin. (See Non-Patent Document 3). In addition, MMP-2 degrades type IV collagen and the like constituting the basement membrane existing under the vascular endothelial cells, and the decomposed vascular endothelial cells migrate to the stroma, proliferate in the stroma, and tube A cavity is formed and new blood vessels are constructed. Then, it is known that the newly formed blood vessels reach the tumor cells, supply nutrient sources and oxygen to the tumor cells, and the tumor grows (see Non-Patent Document 4).

  Therefore, by inhibiting the activity of MMP-2, it is possible to suppress the decrease in basement membrane components and structural changes of the basement membrane, improve the skin function, suppress angiogenesis, and suppress the growth of tumor cells. I think it can be done. As what has a MMP-2 inhibitory effect, the extract from a black-leafed azuki bean (refer patent document 4) etc. are known, for example.

  Carbohydrates are very important as energy sources in living organisms including humans. However, carbohydrates are known to cause glycation reactions with proteins. In the saccharification reaction, a non-enzymatic reaction between a carbonyl group of a carbohydrate and an amino group such as a protein is a first step, and finally a final glycation product (hereinafter referred to as “AGEs”) from a Schiff base through an Amadori compound. Is a series of reactions to form. Due to the saccharification reaction, the protein is modified non-enzymatically by sugar, and as a result, denaturation of the protein, cross-linking between proteins, and the like occur, resulting in a decrease in protein function.

  The glycation reaction not only causes direct damage by modifying and changing the structure of extracellular matrix constituent proteins such as collagen, but also affects cellular responses by being recognized by receptors with glycated proteins as ligands. Bring. In particular, the effect of the saccharification reaction is serious for diabetic patients whose glucose concentration in the blood is high. It is known that protein glycation contributes to diabetic complications such as diabetic neuropathy, diabetic retinopathy, and diabetic nephropathy. In addition, it is known that the glycation reaction in the blood vessel wall leads to the progression of arteriosclerosis due to the damage of endothelial cells and the accumulation of denatured proteins. Further, extracellular matrix components such as collagen occupy a majority of the dry weight in tissues such as bone and skin. Therefore, for example, when collagen is glycated and abnormally crosslinked, osteoporosis, osteoarthritis, etc. occur in bone and cartilage tissue, and elasticity in skin, dullness due to yellowing, etc. occur. . Furthermore, since abnormally cross-linked collagen or the like is less susceptible to degradation by collagenase or the like, the expression of collagenase or the like is induced, causing problems such as degradation to normal collagen.

  For this reason, if the glycation reaction can be suppressed in some way, for example, the formation of AGEs can be suppressed, or the degradation of AGEs can be promoted, the aforementioned diseases, that is, diabetic complications, arteriosclerosis, osteoporosis, deformation It is expected to be useful for the prevention or treatment of osteoarthritis. Furthermore, it is expected to be effective in preventing or improving skin elasticity reduction and dullness.

  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, fibroblasts are actively proliferating, and the interaction between skin tissues such as fibroblasts and collagen keeps their homeostasis, ensuring moisture retention, flexibility, elasticity, etc. It is also kept fresh with tension and gloss.

  However, when there is an influence of certain external factors such as irradiation of ultraviolet rays, significant drying of air, excessive skin washing, etc., or when aging progresses, the production amount of collagen, which is a main component of the extracellular matrix, is increased. Decreases and causes degradation and alteration. As a result, the moisturizing function and elasticity of the skin are lowered and abnormal exfoliation of the keratin occurs, so that the skin loses its tension and gloss, and exhibits aging symptoms such as rough skin and wrinkles. As described above, changes associated with skin aging, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with reduction / denaturation of matrix components such as collagen. Therefore, promoting the production of collagen is important in preventing, treating or improving skin aging.

  Here, collagen is a fibrous protein that contributes to the maintenance of the structure and mechanical strength of skin tissue. If collagen production can be promoted, wrinkles and sagging are less likely to occur, and it is considered that skin aging symptoms such as loss of texture and tension and reduced elasticity can be prevented, treated, or improved.

  Collagen is also present in a large amount in bones, tendons, ligaments, corneas, blood vessels and the like, and it is known that a decrease in collagen production due to aging or the like causes osteoporosis and the like. Furthermore, it is known that in the wound healing process, the amount of collagen produced is increased and becomes a scaffold for fibroblasts to promote wound healing. Therefore, promoting the production of collagen is important from the viewpoint of preventing or treating osteoporosis and the like, and promoting wound healing. As what has a collagen production promotion effect | action, the extract (patent document 5) etc. from Kusunohagashi are known, for example.

  Ceramide is produced by the action of an enzyme such as serine palmitoyltransferase (SPT) known as a rate-limiting enzyme for ceramide synthesis based on serine and palmitoyl-CoA in the process of keratinization of epidermal cells. Ceramide exists specifically as the main component of the keratinocyte lipid covering the outermost layer of the skin, and plays an important role in maintaining the function of the skin itself as a barrier film between the living body and the outside world.

  The stratum corneum structure is compared to brick and mortar, and a strong barrier membrane is formed in such a way that intercellular lipids connect the stratum corneum stacked in about 15 layers. Keratinocytes retain water by containing natural moisturizing factors mainly composed of amino acids in the cells, while stratum corneum lipids are composed of approximately 50% ceramide as a main component, and parents such as cholesterol and fatty acids. It is composed of an amphiphilic lipid, and is characterized by a lamellar structure in which hydrophobic portions and hydrophilic portions are alternately repeated, so-called lamellar structure.

  The decrease in the skin barrier function due to various internal and external factors increases the transepidermal water transpiration, causing skin roughness, desquamation, pruritus, etc., resulting in so-called dry skin. Moreover, the decrease in the barrier function of the skin increases the inflammation of the skin and falls into a vicious circle in which the protective function against various stimuli from the outside world is reduced. In recent studies, keratin ceramide components (so-called intercellular lipids) have been decreased and compositional changes have been reported in patients with atopic dermatitis known as aging or barrier disorders (see Non-Patent Document 5). It is widely known that ceramide is important for maintaining and improving the barrier function of ceramide. As a method for improving the skin barrier function, a method of supplementing ceramide from the outside (see Non-Patent Document 6), a method of enhancing the ability of ceramide production in the skin (see Non-Patent Document 7), and the like are known.

  In skin cells, it is known that aquaporins, known as water channels, are expressed on the cell membrane and play a role of taking in low-molecular substances such as interstitial water into cells. In humans, the presence of 13 types of aquaporins (AQP0 to AQP12) is known. In epidermal cells, AQP3 is mainly present, and it is considered to play a role of taking in low-molecular compounds such as glycerol and urea involved in water retention action in addition to water.

  However, AQP3 decreases with aging, and it has been suggested that this contributes to a decrease in water retention function. Therefore, by promoting the expression of AQP3, the water retention capacity and barrier function due to aging, etc. Can be controlled (see Non-Patent Document 8). As what has an AQP3 expression promotion effect, the extract from the leaf part of a star fruit (refer patent document 6) etc. are known, for example.

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

  Conventionally, for the prevention, treatment, or improvement of skin pigmentation, stains, buckwheat, etc., a treatment using 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, development of a whitening agent containing a highly safe natural raw material as an active ingredient is desired, and as a substance having an inhibitory action on tyrosinase activity, for example, willow extract (see Patent Document 7) is known. . Moreover, as what has a melanin production inhibitory effect, the extract (refer patent document 8) etc. from a South urea (Saussurea) genus plant are known, for example.

  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 methods for suppressing the generation of active 5α-DHT. So far, for example, an extract from Higashi Shiso (see Patent Document 9) or the like is known as having testosterone 5α-reductase inhibitory action.

  Hair repeats growth and loss according to a periodic hair cycle (hair cycle) consisting of a growth phase, a regression phase, and a resting phase. Of these hair cycles, the stage at which new hair follicles are formed from the resting stage to the growing stage is considered to be the most important for hair growth, and the proliferation and differentiation of hair follicle epithelial cells at this stage It is believed that the dermal papilla cells play an important role. The dermal papilla cell is a cell located inside the hair follicle epithelial cell composed of outer root sheath cells and matrix cells in the vicinity of the hair root, and located in the root portion of the hair root wrapped in the basement membrane. It plays an important role in the growth / differentiation of hair follicle epithelial cells and the formation of hair, such as by acting on epithelial cells to promote their proliferation (see Non-Patent Document 9).

  Thus, dermal papilla cells play an important role in the proliferation and differentiation of hair follicle epithelial cells and the formation of hair. By promoting the proliferation of dermal papilla cells, hair loss can be prevented or improved. It is considered possible. So far, for example, a wild time extract (see Patent Document 10) or the like has been known as one having a dermal papilla cell proliferation promoting action.

  Causes and onset of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, other skin inflammatory diseases with rough skin, rheumatoid arthritis, osteoarthritis, asthma, etc. The mechanism is diverse. As the cause thereof, those mainly due to the enhanced activity of hyaluronidase, those due to the release of histamine, those due to the enhanced activity of cyclooxygenase-2 (COX-2) are known.

  Hyaluronidase is a hydrolase of hyaluronic acid. Hyaluronate that maintains affinity for body tissues is decomposed by ultraviolet rays, oxygen, and the like in a water-containing system, and the water retention effect decreases with a decrease in molecular weight. In addition, hyaluronic acid exists as an intercellular tissue in vivo and is also involved in blood vessel permeability. Furthermore, although hyaluronidase exists in mast cells, it 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 or reduce inflammation. As what has a hyaluronidase activity inhibitory effect, the extract (refer patent document 11) etc. from an Osbecchia plant are known, for example.

  Histamine release is a phenomenon in which histamine in mast cells is released extracellularly, and the released histamine causes an inflammatory reaction. Therefore, attempts have been made to prevent or treat allergic diseases and inflammatory diseases with substances that inhibit or suppress histamine release. However, it is difficult to directly evaluate the release of histamine, and the release of histamine can be evaluated using as an index the release of hexosaminidase that has been confirmed to be released simultaneously with the release of histamine. Therefore, by suppressing the release of hexosaminidase, the release of histamine can also be suppressed at the same time, which is considered to be effective for the prevention, treatment or improvement of inflammatory diseases.

  It is also known that histamine mediates information transmission between cells as a local transmitter, enhances gastric acid secretion in the digestive tract, functions as a neurotransmitter in the central nervous system, and contributes to the maintenance of wakefulness. It has been. Here, excessive release of histamine causes ulcers due to excessive gastric acidity in the digestive tract, and contributes to sleep disorders in the central nervous system. As described above, by inhibiting the release of hexosaminidase, it is also possible to inhibit the release of histamine at the same time, which is considered to prevent, treat, or improve gastric ulcers, sleep disorders, etc. caused by hyperacidity. . As what has a hexosaminidase release inhibitory effect, the extract from Fuji tea (refer patent document 12) etc. are known, for example.

  Inflammation is a complex reaction that exhibits symptoms such as redness, edema, fever, pain, and dysfunction. Microscopically, inflammation consists of common reactions such as vascular reactions that cause plasma leakage, leukocyte infiltration, tissue destruction by inflammatory cells, and the central nervous system such as fever and hyperalgesia is also involved. It may also cause a reaction. It is clear that prostaglandins play an important role in each reaction of such inflammation, and that cyclosogenase-2, which is an inducible cyclooxygenase, is mainly involved in the production of prostaglandins during this inflammation. It has become. For this reason, many cyclooxygenase inhibitors represented by aspirin are used for the purpose of preventing and preventing inflammatory reactions (see Non-Patent Document 10).

  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.

  Here, it is known that cyclic AMP (cAMP) is involved in lipolysis in vivo. cAMP activates lipase present in the living body, and fat is broken down into fatty acid and glycerol by the activated lipase. However, when cAMP phosphodiesterase is activated, cAMP degradation is induced and lipase activation is inhibited. Therefore, it is considered that by inhibiting the activity of cAMP phosphodiesterase, the amount of cAMP in the cell is increased and the decomposition of fat can be promoted.

  In addition, platelet aggregation that causes an inflammatory reaction is related to the concentration of cyclic AMP (cAMP) in the platelet, and it is known that cAMP is degraded by cAMP phosphodiesterase and the concentration of cAMP decreases, so that platelets tend to aggregate. It has been. Therefore, if the action of cAMP phosphodiesterase is suppressed to prevent a decrease in cAMP concentration, platelet aggregation can be prevented, thereby preventing, treating or improving allergic diseases and inflammatory diseases. Tubeamoside I (see Patent Document 13) and the like are known as having cAMP phosphodiesterase activity inhibitory action.

  The liver metabolizes and stores various nutrients absorbed in the intestine, and also performs many vital functions such as bile production and secretion, detoxification and excretion, and is an extremely important organ for humans. It is. However, the liver is prone to acute or chronic damage due to various factors such as irregular life, stress, viruses, drugs, alcohol, malnutrition, hepatic circulatory system disorders, acute hepatitis, chronic hepatitis, fat May cause diseases such as liver, jaundice, cirrhosis (liver dysfunction).

  Here, in the liver, glutathione protects hepatocytes from various oxidative stresses, forms a conjugate with harmful substances such as drugs and reactive compounds, and excretes them outside the cell. It contributes directly to the development of liver function. However, it is known that glutathione is exerted at the same time that glutathione is consumed, and that when liver damage is actually induced in rats by galactosamine or the like, the amount of glutathione in hepatocytes decreases. . Therefore, if it is possible to suppress a decrease in the amount of glutathione due to various stresses, drugs, and the like in the liver, it is considered that liver damage can be suppressed and eventually liver function can be improved.

  Dipeptidyl peptidase IV (hereinafter also referred to as “DPPIV”) is one of serine proteases and has an enzyme activity of recognizing the second proline or alanine from the N-terminus and cleaving the C-terminal side. DPPIV is expressed on epithelial and endothelial cells of tissues such as kidney, liver, intestinal tract, and placenta, and on the cell surface of T cells, and is considered to be involved in various physiological phenomena through its enzyme activity and the like.

  As a substrate for DPPIV, there is a hormone called incretin. Incretin is a general term for hormones secreted from the intestine by stimulation of nutrients and promoting insulin secretion from pancreatic β cells in a blood glucose-dependent manner, and GLP-1 and GIP are known. These incretins not only promote blood glucose-dependent insulin secretion, but also suppress glucagon secretion from pancreatic alpha cells, lower blood pressure, suppress gastric emptying, and suppress feeding by acting on the hypothalamus. It has an effect | action (refer nonpatent literature 11). However, since incretin is degraded by DPPIV, it is known that, for example, the half-life of GLP-1 in vivo is about 1.5 minutes. Therefore, if the enzyme activity of DPPIV can be inhibited, the half-life of incretin in the living body can be extended. As a result, type 2 diabetes, obesity, hypertension, insulin resistance can be achieved through the action of incretin described above. It is expected to be useful for such treatment.

  DPPIV is identical to CD26, which is one of T cell activation markers, and is known to control its activity using many immunoregulatory peptides as substrates. Therefore, it is considered that by controlling the activity of DPPIV, an autoimmune disease such as rheumatoid arthritis and an immune reaction such as rejection due to transplantation can be controlled. Furthermore, DPPIV is known to be involved in metabolism of several neuropeptides and growth hormones; invasion in cancer, metastasis, angiogenesis, etc .; HIV infection of lymphocytes, and the like. Therefore, by inhibiting the activity of DPPIV, neurological disorders such as pain, neurodegenerative diseases and neuropsychiatric diseases (eg sciatica, Alzheimer's disease, depression, etc.); growth hormone deficiency and growth hormone are used for treatment Diseases; cancers (eg, T-cell lymphoma, acute lymphoblastic leukemia, thyroid cancer, basal cell cancer, breast cancer, etc.); diseases such as HIV infection (AIDS) may be treated.

  As for 4-vinylcatechol, it is known that 4-vinylcatechol is detected when certain lactic acid bacteria are cultured in a medium containing caffeic acid (Non-patent Document 12).

JP 2009-269889 A JP 2006-063033 A JP 2006-265232 A JP 2007-217352 A JP 2003-146837 A JP 2009-191039 A JP 2004-083488 A JP 2002-201222 A JP 2010-184915 A JP 2006-219407 A Japanese Patent Laid-Open No. 2003-055242 JP 2003-012532 A JP 2006-056855 A

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  The present invention includes an anti-aging action, whitening action, hair-growth action, anti-androgen action, anti-inflammatory action, anti-obesity action, cyclic AMP phosphodiesterase activity inhibitory action, glutathione reduction inhibitory action or dipeptidyl among natural compounds. An anti-aging agent, a whitening agent, a hair restorer, an antiandrogen, an anti-inflammatory agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a glutathione lowering agent that has a peptidase IV activity inhibitory activity An object is to provide an inhibitor, a dipeptidyl peptidase IV activity inhibitor, and skin cosmetics, hair cosmetics and foods and drinks containing the compound.

  In order to solve the above problems, the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and dipeptidyl peptidase of the present invention The IV activity inhibitor is characterized by containing 4-vinylcatechol as an active ingredient. Moreover, the skin cosmetics, hair cosmetics and food and drink of the present invention are characterized by blending 4-vinylcatechol.

  According to the present invention, by containing 4-vinylcatechol as an active ingredient, an anti-aging agent, a whitening agent, a hair-growth agent, an anti-androgen agent, an anti-inflammatory agent, an anti-obesity agent, a cyclic AMP excellent in action and effect. A phosphodiesterase activity inhibitor, a glutathione reduction inhibitor, and a dipeptidyl peptidase IV activity inhibitor can be provided. Furthermore, the skin cosmetics, hair cosmetics, and food-drinks excellent in the said effect | action can be provided by mix | blending 4-vinyl catechol.

Embodiments of the present invention will be described below.
The anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and dipeptidyl peptidase IV activity inhibitor of this embodiment are 4 -It contains vinyl catechol as an active ingredient. Moreover, the skin cosmetics, hair cosmetics, and foods and beverages of this embodiment are blended with 4-vinylcatechol.

  4-vinylcatechol is a compound having a chemical structure represented by the following formula.

  4-vinylcatechol is obtained by, for example, fermenting caffeic acid or a composition containing caffeic acid (for example, a plant crushed or extract) with a microorganism having phenolic acid decarboxylase, and converting caffeic acid into 4-vinyl catechol. After conversion into catechol, the obtained fermentation product can be extracted, isolated and purified. Examples of the composition containing caffeic acid include crushed materials and extracts of plants such as coffee, burdock, sweet potato, cacao, tomato, mate, and mugwort. Examples of microorganisms having phenolic acid decarboxylase include lactic acid bacteria such as Lactobacillus plantarum, Lactobacillus brevis and Pediococcus pentosaceus; and yeasts such as Saccharomyces cerevisiae.

  The method for extracting, isolating and purifying 4-vinylcatechol from the plant or fermented product is not particularly limited, and can be carried out according to a conventional method. For example, the extraction treatment may be performed by drying the plant or fermentation product as an extraction raw material, pulverizing it as it is or using a crusher, and subjecting it to extraction with an extraction solvent. Drying may be performed in the sun or using 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 pretreatment such as degreasing, extraction with a polar solvent can be performed efficiently.

  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 this embodiment includes purified water, hot water, ion exchange 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 a mixed liquid of water and lower aliphatic alcohol is used as an extraction solvent, the mixing ratio of water and lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 7: 3 to 2: 8 (capacity ratio). In addition, when a mixed liquid of water and a lower aliphatic ketone is used, the mixing ratio of water and the lower aliphatic ketone is preferably 9: 1 to 2: 8 (volume ratio). When using the liquid mixture with a monohydric alcohol, it is preferable that the mixing ratio of water and a polyhydric alcohol is 5: 5 to 1: 9 (volume ratio).

  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. When the solvent is distilled off from the obtained extract, a paste-like concentrate is obtained, and when this concentrate is further dried, a dried product is obtained.

  The method for isolating and purifying 4-vinylcatechol from the extract obtained as described above, the concentrate of the extract or the dried product of the extract is not particularly limited, and is performed by a conventional method. be able to. For example, the extract is dissolved in a developing solvent and subjected to column chromatography using a porous material such as silica gel or alumina, a porous resin such as styrene-divinylbenzene copolymer or polymethacrylate, and the like. Examples thereof include a method for collecting a fraction containing catechol. In this case, the developing solvent may be appropriately selected according to the stationary phase to be used. For example, when the extract is separated by normal phase chromatography using silica gel as the stationary phase, the developing solvent is chloroform: methanol = 95: 5 etc. are mentioned. Further, the fraction containing 4-vinylcatechol obtained by column chromatography was subjected to reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using ion exchange resin, etc. You may refine | purify using arbitrary organic compound refinement | purification means.

[Anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor, dipeptidyl peptidase IV activity inhibitor]
The 4-vinylcatechol obtained as described above has an excellent anti-aging action, whitening action, hair growth action, anti-androgenic action, anti-inflammatory action, anti-obesity action, cyclic AMP (cAMP) phosphodiesterase activity inhibitory action, glutathione. Anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor because it has a lowering suppressive action and dipeptidyl peptidase IV (DPPIV) activity inhibitory action, It can be used as an active ingredient of a glutathione lowering inhibitor and a DPPIV activity inhibitor. The anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and DPPIV activity inhibitor of this embodiment are pharmaceuticals, quasi drugs It can be used for a wide range of applications such as cosmetics.

  As an active ingredient of the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor and DPPIV activity inhibitor according to this embodiment, Instead of the isolated 4-vinyl catechol, a composition containing 4-vinyl catechol may be used. Here, in the “composition containing 4-vinyl catechol” in the present embodiment, an extract obtained by using a plant containing 4-vinyl catechol as an extraction raw material, a fermented product containing 4-vinyl catechol, and the concerned An extract obtained by using a fermentation product as an extraction raw material is included. The “extract” includes an extract obtained by an extraction process, a diluted or concentrated solution of the extract, or a dried product obtained by drying the extract.

  As an anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and DPPIV activity inhibitor according to this embodiment, When a composition containing vinyl catechol is used, it is preferable to use a composition that has been purified to increase the purity of 4-vinyl catechol. Anti-aging agent, whitening agent, hair-restoring agent, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP, which are more effective by using 4-vinylcatechol purity increased as an active ingredient A phosphodiesterase activity inhibitor, a glutathione reduction inhibitor and a DPPIV activity inhibitor can be obtained.

  The anti-aging action of 4-vinylcatechol includes, for example, glutathione production promoting action, laminin 5 production promoting action, matrix metalloproteinase-1 (MMP-1) activity inhibiting action, final glycation product (AGEs) degradation promoting action, final glycation Product (AGEs) formation inhibitory action, type I collagen production promoting action, matrix metalloproteinase-2 (MMP-2) activity inhibiting action, serine palmitoyltransferase (SPT) mRNA expression promoting action and aquaporin 3 (AQP3) mRNA expression promoting action It is exhibited based on one or more actions selected from the group consisting of: However, the anti-aging action which 4-vinylcatechol has is not limited to the anti-aging action exhibited based on the said action.

  Moreover, 4-vinylcatechol has its glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen production promoting action, MMP-2 activity inhibiting action , SPT mRNA expression promoting action or AQP3 mRNA expression promoting action, glutathione production promoter, laminin 5 production promoter, MMP-1 activity inhibitor, AGEs degradation promoter, AGEs formation inhibitor, type I collagen production promoter, respectively , MMP-2 activity inhibitor, SPT mRNA expression promoter or AQP3 mRNA expression promoter may be used as an active ingredient.

  The whitening action of 4-vinylcatechol is exhibited based on, for example, a tyrosinase activity inhibitory action and / or a melanin production inhibitory action. However, the whitening action of 4-vinylcatechol is not limited to the whitening action exhibited based on the above action. Moreover, 4-vinylcatechol may be used as an active ingredient of a tyrosinase activity inhibitor or a melanin production inhibitor, respectively, utilizing its tyrosinase activity inhibitory action or melanin production inhibitory action.

  The hair growth effect of 4-vinylcatechol is exhibited based on, for example, a testosterone 5α-reductase activity inhibitory effect and / or a hair papillary cell proliferation promoting effect. However, the hair growth action which 4-vinyl catechol has is not limited to the hair growth action exhibited based on the said action. Further, 4-vinylcatechol is used as an active ingredient of a testosterone 5α-reductase activity inhibitor or a hair papilla cell proliferation promoter, utilizing its testosterone 5α-reductase activity inhibitory effect or hair papilla cell proliferation promoting effect, respectively. Also good.

  The anti-androgenic action of 4-vinylcatechol is exhibited, for example, based on the testosterone 5α-reductase activity inhibitory action. However, the anti-androgenic action which 4-vinylcatechol has is not limited to the anti-androgenic action exhibited based on the said action.

  The anti-inflammatory action of 4-vinylcatechol is, for example, one or more actions selected from the group consisting of hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, and cyclooxygenase-2 (COX-2) activity inhibitory action Based on. However, the anti-inflammatory action which 4-vinylcatechol has is not limited to the anti-inflammatory action exhibited based on the said action. Further, 4-vinylcatechol utilizes its hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, or COX-2 activity inhibitory action, respectively, to thereby produce a hyaluronidase activity inhibitor, hexosaminidase release inhibitor, or COX-2, respectively. You may use as an active ingredient of an activity inhibitor.

  The anti-obesity action of 4-vinylcatechol is exhibited based on, for example, the cAMP phosphodiesterase activity inhibitory action. However, the anti-obesity action which 4-vinylcatechol has is not limited to the anti-obesity action exhibited based on the said action.

  The anti-aging agent, whitening agent, hair-restoring agent, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor or DPPIV activity inhibitor of this embodiment is 4-vinylcatechol or 4 -It may consist only of a composition containing vinyl catechol, or may be a formulation of 4-vinyl catechol or a composition containing 4-vinyl catechol.

  The anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and DPPIV activity inhibitor of this embodiment are dextrin, cyclodextrin, etc. Using a pharmaceutically acceptable carrier or any other auxiliary agent, it can be formulated into an arbitrary dosage form such as powder, granule, tablet, and liquid according to a conventional method. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / flavoring agent, and the like can be used. Anti-aging agents, whitening agents, hair restorers, anti-androgen agents, anti-inflammatory agents, anti-obesity agents, cAMP phosphodiesterase activity inhibitors, glutathione reduction inhibitors and DPPIV activity inhibitors may be used in other compositions (eg, skin cosmetics). , Hair cosmetics, etc.) and can also be used as an ointment, a liquid for external use, a patch and the like.

  When the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor or DPPIV activity inhibitor of this embodiment is formulated, 4- The content of the composition containing vinyl catechol or 4-vinyl catechol is not particularly limited, and can be appropriately set according to the purpose.

  In addition, the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor of the present embodiment is used as necessary. Anti-aging action, whitening action, hair-growth action, anti-androgenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione reduction inhibitory action or DPPIV activity inhibitory action, etc. -It can mix | blend with the composition containing vinyl catechol or 4-vinyl catechol, and can be used as an active ingredient.

  As an administration method for patients of the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor of this embodiment, Examples include transdermal administration, oral administration, and the like, and a suitable method for the prevention or treatment of the disease may be appropriately selected depending on the type of the disease. In addition, the dosage of the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor of this embodiment is also a disease. May be appropriately increased or decreased depending on the type, severity, individual differences among patients, administration method, administration period, and the like.

  The anti-aging agent of this embodiment includes glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen, which 4-vinylcatechol as an active ingredient has Through one or more actions selected from the group consisting of production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action and AQP3 mRNA expression promoting action, formation of skin wrinkles, reduced elasticity, reduced moisturizing function Can prevent, treat or ameliorate skin aging symptoms such as However, in addition to these uses, the anti-aging agent of the present embodiment is glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen production promoting It can be used for all purposes that are meaningful for exerting the action, the MMP-2 activity inhibitory action, the SPT mRNA expression promoting action or the AQP3 mRNA expression promoting action.

  For example, the anti-aging agent of the present embodiment or the aforementioned glutathione production promoter is caused by liver injury (drinking of alcohol or ingestion of foreign substances such as heavy metals and chemical substances) through the glutathione production promotion action of 4-vinylcatechol. It is possible to prevent, treat or ameliorate a disease or the like in which a decrease or deficiency in the amount of intracellular glutathione is known to be associated with a disease state.

  In addition to the use described above, the anti-aging agent of the present embodiment or the laminin 5 production promoter described above induces the remodeling of the basement membrane structure through the laminin 5 production promotion action of 4-vinylcatechol, thereby causing wounds in the skin. Can be treated and improved. Moreover, the anti-aging agent of this embodiment or the laminin 5 production promoter described above can be used as a preventive or therapeutic agent for diseases (such as epidermolysis bullosa) caused by laminin 5 deficiency (deficiency).

  In addition to the above-mentioned uses, the anti-aging agent of the present embodiment or the above-mentioned MMP-1 activity inhibitor or MMP-2 activity inhibitor is an MMP-1 activity inhibitory action or an MMP-2 activity inhibitory action of 4-vinylcatechol. Diseases that are known to be associated with pathological conditions through the destruction of extracellular matrix (eg, cancer invasion / metastasis, rheumatoid arthritis, osteoarthritis of the knee, periodontal disease, age-related macular degeneration, etc.) ) Can be used for therapeutic / preventive purposes. Moreover, the anti-aging agent or MMP-2 activity inhibitor of this embodiment can suppress angiogenesis and tumor cell growth by its MMP-2 activity inhibitory action.

  In addition to the above-described uses, the anti-aging agent of the present embodiment, or the above-described AGEs degradation accelerator or AGEs formation inhibitor is capable of diabetic neuropathy, diabetes through the AGEs degradation promoting action or AGEs formation inhibiting action of 4-vinylcatechol. Diabetic complications such as retinopathy and diabetic nephropathy; arteriosclerosis caused by glycation reaction of protein; osteoporosis and osteoarthritis caused by glycation reaction of protein, etc. can be prevented or treated. In addition, the anti-aging agent of the present embodiment or the AGEs formation inhibitor described above can prevent or suppress hair damage due to protein saccharification reaction through its AGEs formation inhibitory action, thereby causing hair irritation. Can be prevented or suppressed, the elasticity and suppleness of the hair can be recovered, and the hair can be given firmness and firmness.

  In addition to the use described above, the anti-aging agent of the present embodiment or the collagen production promoter described above is used to prevent diseases caused by a decrease in collagen production such as osteoporosis through the action of promoting the production of type I collagen by 4-vinylcatechol. It can be used for applications such as treatment or improvement; promotion of regeneration of damaged tendons and ligaments; promotion of healing of wounds or burns;

  In addition to the use described above, the anti-aging agent of the present embodiment or the SPT mRNA expression promoter described above enhances the barrier function of the skin through the SPT mRNA expression promoting action of 4-vinylcatechol, in addition to rough skin, dry skin, etc. Dry skin diseases (eg, atopic dermatitis, psoriasis, ichthyosis, etc.) can be prevented, treated or ameliorated. In addition, the anti-aging agent of the present embodiment or the above-described AQP3 mRNA expression promoter can improve water retention ability, barrier function, and the like due to aging through the AQP3 mRNA expression promoting action of 4-vinylcatechol.

  The whitening agent of this embodiment can prevent or improve pigmentation such as skin darkening, spots, and freckles through the tyrosinase activity inhibiting action and / or the melanin production inhibiting action of 4-vinylcatechol, which is an active ingredient. it can. However, the whitening agent of the present embodiment can be used for all uses other than these uses that are meaningful for exhibiting a tyrosinase activity inhibitory action or a melanin production inhibitory action.

  The hair-restoring agent of the present embodiment is effective for male pattern alopecia, alopecia areata, tricotylonia, etc. through the testosterone 5α-reductase activity inhibiting action and / or the papillary cell proliferation promoting action of 4-vinylcatechol as an active ingredient. Alopecia and the like can be prevented, treated or ameliorated, and is particularly suitable for preventing, treating or improving male pattern alopecia. However, the hair-restoring agent of the present invention can be used for all uses other than these uses that are meaningful for exerting a testosterone 5α-reductase activity inhibitory action or a hair papillary cell proliferation promoting action.

  For example, the hair growth agent of the present embodiment or the above-described hair papilla cell proliferation promoter activates the hair papilla cells through the proliferation promoting effect of 4-vinylcatechol on the hair papilla cells, and the growth and differentiation of hair follicle epithelial cells and In addition to promoting hair formation, the hair cycle can be prevented from transitioning from the growth phase to the regression phase and the rest phase, and the growth phase can be extended. Thereby, alopecia can be prevented or improved. In addition, the hair growth agent of the present embodiment or the aforementioned hair dermal papilla cell proliferation promoter is applied to the regenerative medicine field such as hair regeneration using hair nipple cells through the dermal papilla cell proliferation promotion action of 4-vinylcatechol. It can also be used.

  The anti-androgen agent of the present embodiment or the aforementioned testosterone 5α-reductase activity inhibitor is a disease involving male hormones, for example, male type, through the testosterone 5α-reductase activity inhibitory action of 4-vinylcatechol as an active ingredient. Alopecia, hirsutism, 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 uses other than these uses that are meaningful for exerting testosterone 5α-reductase activity inhibitory action.

  The anti-inflammatory agent of this embodiment is one or more selected from the group consisting of hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action and COX-2 activity inhibitory action of 4-vinylcatechol as an active ingredient Through the action, it is possible to prevent, treat or ameliorate contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, and various other skin inflammatory diseases associated with rough skin. However, in addition to these uses, the anti-inflammatory agent of the present embodiment should be used for all purposes that are meaningful for exerting a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, or a COX-2 activity inhibitory action. Can do.

  For example, the anti-inflammatory agent of the present embodiment, the hyaluronidase activity inhibitor, the hexosaminidase release inhibitor or the COX-2 activity inhibitor described above are the hyaluronidase activity inhibitory action and hexosaminidase release inhibition of 4-vinylcatechol. Rheumatoid arthritis, osteoarthritis, asthma and the like can be prevented, treated or ameliorated through the action or the COX-2 activity inhibitory action. Moreover, the anti-inflammatory agent of the present embodiment or the hexosaminidase release inhibitor described above prevents gastric ulcers, sleep disorders, etc. caused by gastric hyperacidity through the hexosaminidase release inhibitory action of 4-vinylcatechol. Can be treated or improved.

  The anti-obesity agent of this embodiment can promote cAMP production and adipocyte degradation through the cAMP phosphodiesterase activity inhibitory action of 4-vinylcatechol, which is an active ingredient. As a result, obesity, Various diseases such as accompanying arteriosclerosis, diabetes, and metabolic syndrome can be prevented or ameliorated.

  The cAMP phosphodiesterase activity inhibitor of the present embodiment promotes cAMP production through the cAMP phosphodiesterase activity inhibitory action of 4-vinylcatechol, which is an active ingredient, and thus can suppress the aggregation of platelets. And various inflammatory diseases can be prevented, treated or ameliorated. In addition, the cAMP phosphodiesterase activity inhibitor of the present embodiment can promote the degradation of adipocytes through the cAMP phosphodiesterase activity inhibitory action of 4-vinylcatechol. As a result, obesity and accompanying arteriosclerosis, Various lifestyle-related diseases such as diabetes and metabolic syndrome can be prevented or improved. However, the cAMP phosphodiesterase activity inhibitor of the present embodiment can be used for all purposes that are meaningful in exhibiting the cAMP phosphodiesterase activity inhibitory action in addition to these uses.

  The glutathione reduction inhibitor of this embodiment has 4-vinylcatechol, which is an active ingredient, when hepatocytes are exposed to various stresses caused by excessive drinking of alcohol, ingestion of foreign substances such as heavy metals and chemical substances, etc. Through the glutathione lowering-inhibiting action, metabolism of harmful substances (for example, extracellular excretion due to the formation of conjugates with glutathione) can be promoted, and liver damage caused by harmful substances can be prevented or reduced. Thereby, diseases (liver dysfunction) caused by liver disorders such as acute hepatitis, chronic hepatitis, fatty liver, jaundice, and cirrhosis can be prevented, treated or ameliorated. However, the glutathione reduction inhibitor of the present embodiment can be used for all uses other than these uses that are meaningful for exhibiting a glutathione reduction suppressing action.

  For example, the glutathione decrease inhibitor of the present embodiment prevents or improves hangover through the glutathione decrease inhibitory action of 4-vinylcatechol; prevention, treatment or symptoms of symptoms (rough skin, fatigue, etc.) caused by accumulation of harmful substances, or It can be used for applications such as improvement;

  The DPPIV inhibitor of this embodiment can prevent or treat type 2 diabetes, obesity, hypertension, insulin resistance, and the like through DPPIV inhibitory action of 4-vinylcatechol, and autoimmune diseases such as rheumatoid arthritis. And transplant rejection can be prevented or treated. However, the DPPIV inhibitor of the present embodiment can be used for all uses that are meaningful for exerting a DPPIV inhibitory action in addition to these uses. For example, the DPPIV inhibitor of the present embodiment has a DPPIV inhibitory action of 4-vinylcatechol, through neuropathy such as pain, neurodegenerative disease, and neuropsychiatric disease (for example, sciatica, Alzheimer's disease, depression, etc.); growth Diseases in which hormone deficiency and growth hormone are used for treatment; cancer (eg T-cell lymphoma, acute lymphoblastic leukemia, thyroid cancer, basal cell cancer, breast cancer, etc.); diseases such as HIV infection (AIDS) Can be prevented or treated.

  In addition, the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor of the present embodiment is an excellent anti-aging agent. Since it has an action, whitening action, hair growth action, anti-androgenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione lowering inhibitory action or DPPIV activity inhibitory action, for example, it is added to skin external preparations Is preferred. In this case, 4-vinylcatechol or a composition containing 4-vinylcatechol may be blended as it is, or an anti-aging agent, whitening agent formulated from 4-vinylcatechol or a composition containing 4-vinylcatechol. An agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor or DPPIV activity inhibitor may be added.

  Here, the external preparation for skin is not limited in its category, and includes a wide range of quasi-drugs and pharmaceuticals used transdermally, in addition to the skin cosmetics described below.

  In addition, the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor of the present embodiment is an excellent anti-aging agent. Reagents for research on the mechanism of action, since it has action, whitening action, hair growth action, anti-androgenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione lowering inhibitory action or DPPIV activity inhibitory action Also, it can be suitably used.

[Skin cosmetics, hair cosmetics]
4-vinylcatechol has an excellent anti-aging effect, whitening effect, hair-restoring effect, anti-androgenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione reduction inhibitory action and DPPIV activity inhibitory action Therefore, it is suitable for blending into skin cosmetics or hair cosmetics. In this case, 4-vinylcatechol or a composition containing 4-vinylcatechol may be blended as it is, or an anti-aging agent and a whitening agent formulated from a composition containing 4-vinylcatechol or 4-vinylcatechol. , Hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor or DPPIV activity inhibitor.

  By blending 4-vinylcatechol or the above-mentioned anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor, Anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen production promoting action, MMP-2 on skin cosmetics or hair cosmetics Activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action, melanin production inhibiting action, hair growth action, testosterone 5α-reductase activity inhibiting action, hair papilla cell proliferation promoting action, anti-androgenic hormone action, Anti-inflammatory action, hyaluronidase activity inhibition Action, hexosaminidase release inhibitory effect, COX-2 inhibitory activity, anti-obesity effect, cAMP phosphodiesterase inhibitory activity, it is possible to impart glutathione reduction-inhibiting or DPPIV inhibitory activity.

  All of these effects exert favorable actions when applied to skin cosmetics or hair cosmetics. Among them, anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 Activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action, melanin production inhibiting action , Anti-inflammatory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, testosterone 5α-reductase activity inhibitory action, anti-androgen action or cAMP phosphodiesterase activity inhibitory action is imparted to skin cosmetics Then, those effects are demonstrated Cheap Therefore, it is particularly suitable.

  Among the actions described above, hair growth action, testosterone 5α-reductase activity inhibition action, hair papillary cell proliferation promotion action, antiandrogen action, anti-inflammatory action, hyaluronidase activity inhibition action, hexosaminidase release inhibition action, COX- 2) Inhibiting action, cAMP phosphodiesterase activity inhibiting action, laminin 5 production promoting action, AGEs formation inhibiting action or type I collagen production promoting action are particularly suitable because they are easily exerted on hair cosmetics. is there.

  Skin in which 4-vinylcatechol or the above-mentioned anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor can be blended The type of cosmetic or hair cosmetic is not particularly limited, and examples of skin cosmetics include ointments, creams, emulsions, lotions, packs, foundations, etc., and hair cosmetics include, for example, , Hair tonic, hair cream, hair liquid, shampoo, pomade, rinse and the like.

  4-vinylcatechol or the above-mentioned anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity inhibitor When blended into hair cosmetics, the blending amount can be appropriately adjusted according to the type of skin cosmetics or hair cosmetics, suitable blending rate is about 0.0001 to 10 mass%, A particularly suitable blending ratio is about 0.001 to 1% by mass in terms of a standard extract.

  The skin cosmetic or hair cosmetic of the present embodiment has an anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action possessed by 4-vinylcatechol. , Type I collagen production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action, melanin production inhibiting action, hair growth action, testosterone 5α-reductase activity inhibiting action, hair Nipple cell proliferation promoting action, anti-androgenic action, anti-inflammatory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action or DPPIV activity inhibition Main agents, auxiliaries or other ingredients used in the production of normal skin cosmetics or hair cosmetics, such as astringents, bactericides / antibacterial agents, whitening agents, UV absorbers, moisturizers, cells, etc. 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. When used in combination, it becomes a more general product, and a synergistic effect with other active ingredients used in combination may lead to an excellent effect that is more than normally expected.

  The skin cosmetic according to the present embodiment has an anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGEs formation inhibiting action, type I collagen, which 4-vinylcatechol has. Production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action, melanin production inhibiting action, anti-inflammatory action, hyaluronidase activity inhibiting action, hexosaminidase release inhibiting action , COX-2 activity inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, hair papillary cell proliferation promoting action, anti-androgen action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action and DPPIV activity inhibitory action Group of Prevention, treatment or amelioration of skin aging symptoms such as skin wrinkle formation, reduced elasticity, reduced moisturizing function, etc. through one or more selected actions; accelerated healing of wounds or burns; Prevention, treatment or amelioration of dry skin as well as dry skin diseases (for example, atopic dermatitis, psoriasis, ichthyosis, etc.); prevention, treatment or amelioration of pigmentation such as skin darkening, spots and freckles; Prevention, treatment or amelioration of contact dermatitis (rash), psoriasis, pemphigus vulgaris and other skin inflammatory diseases associated with rough skin; prevention, treatment or amelioration of seborrhea, acne (acne etc.); obesity And the prevention, treatment or improvement of lifestyle-related diseases such as arteriosclerosis, diabetes, and metabolic syndrome associated therewith.

  Further, the hair cosmetic composition of the present embodiment has a hair growth action, testosterone 5α-reductase activity inhibition action, hair papillae cell proliferation promotion action, antiandrogen action, anti-inflammatory action, hyaluronidase activity inhibition action, Sosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action , AGEs formation inhibitory action, type I collagen production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action, melanin production inhibiting action, glutathione decrease inhibiting action and DPPIV activity From inhibitory action Prevention, treatment or amelioration of alopecia such as androgenetic alopecia, alopecia areata, trichotiromania; contact dermatitis (rash), psoriasis, vulgaris, through one or more actions selected from the group Prevention, treatment or improvement of various skin inflammatory diseases associated with pimples and other rough skin; treatment of dry skin diseases (for example, atopic dermatitis, psoriasis, ichthyosis, etc.) in addition to rough skin and dry skin; It is possible to prevent or suppress sticking, restore the elasticity and suppleness of the hair, and give the hair firmness and firmness.

[Food and Drink]
4-vinylcatechol has an excellent anti-aging effect, whitening effect, hair-restoring effect, anti-androgenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione reduction inhibitory action and DPPIV activity inhibitory action Therefore, it is suitable for blending into food and drink. In this case, 4-vinylcatechol may be blended as it is, or an anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity formulated from 4-vinylcatechol You may mix | blend an inhibitor, a glutathione fall inhibitor, or a DPPIV activity inhibitor.

  Here, food and drink are those that are less likely to harm human health and that are taken by oral or gastrointestinal administration in normal social life. It is not limited to such categories. Therefore, “food and drink” in the present embodiment includes general foods that are taken orally, health foods (functional foods and drinks), health functional foods (special health foods, nutritional functional foods), quasi drugs, and pharmaceuticals. And so on.

  4-Vinylcatechol or the above-mentioned anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen, anti-inflammatory agent or whitening agent is added to food and drink Anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action, DPPIV activity inhibitory action, anti-aging action, glutathione production promoting action, laminin 5 production promoting action, MMP-1 activity inhibiting action, AGEs degradation promoting action, AGE formation inhibitory action, type I collagen production promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, hair growth action, testosterone 5α-reductase activity inhibiting action, hair papilla cell proliferation promoting action, anti-androgen Action, anti-inflammatory action , Hyaluronidase inhibitory effect, hexosaminidase release inhibitory effect, COX-2 inhibitory activity, can be imparted to the whitening effect, tyrosinase inhibitory activity or melanogenesis inhibiting food or drink action. These actions are suitable because they are easily exerted by being applied to food and drink.

  Anti-aging agent, whitening agent, hair-restoring agent, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor or DPPIV activity formulated from 4-vinyl catechol or 4-vinyl catechol When blending inhibitors into foods and drinks, the amount of active ingredients in them can be appropriately changed in consideration of the purpose of use, symptoms, sex, etc., but the general intake of foods and drinks to be added In consideration of the above, it is preferable that the daily intake of the extract is about 1 to 1000 mg. In addition, when the food or drink to be added is a granular, tablet or capsule food or drink, 4-antigen catechol, or an anti-aging agent formulated from 4-vinyl catechol, a whitening agent, a hair restorer, an antiandrogen, The addition amount of the anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor or DPPIV activity inhibitor is usually 0.1 to 100% by mass, preferably 5 to 5% with respect to the food or drink to be added. 100% by mass.

  The food / beverage products of this embodiment may be blended with any food / beverage product that does not interfere with the activity of 4-vinylcatechol, and is a dietary supplement mainly composed of 4-vinylcatechol. Also good.

  When producing the food and drink of this embodiment, for example, sugars such as dextrin and starch; proteins such as gelatin, soybean protein and corn protein; amino acids such as alanine, glutamine and isoleucine; Arbitrary auxiliary agents, such as saccharides; oils and fats, such as soybean oil and medium chain fatty acid triglyceride, can be added, and it can be set as food / beverage products of arbitrary shapes.

  Although the food and drink which can mix | blend 4-vinyl catechol is not specifically limited, As the specific example, Beverages, such as a soft drink, a carbonated drink, a nutritive drink, a fruit drink, and a lactic acid drink (Concentrated undiluted solution and adjustment powder of these drinks Ice cream, ice sherbet, shaved ice and other frozen desserts; buckwheat noodles, udon, harusame, gyoza skin, cucumber skin, Chinese noodles, instant noodles and other noodles; salmon, chewing gum, candy, gum, chocolate, tablet confectionery Confectionery such as snacks, biscuits, jelly, jam, cream, baked confectionery; fishery and livestock processed foods such as kamaboko, ham and sausage; dairy products such as processed milk and fermented milk; salad oil, tempura oil, margarine, mayonnaise, Fats and processed foods such as shortening, whipped cream and dressing; seasonings such as sauces and sauces Soups, stews, salads, prepared dishes, pickles; various other forms of health / nutritional supplements; tablets, capsules, drinks, etc., etc. Usually used when blending 4-vinylcatechol into these foods and drinks Supplementary raw materials and additives to be used can be used in combination.

  In addition, the anti-aging agent, whitening agent, hair restorer, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor, DPPIV activity inhibitor, skin cosmetic, hair of this embodiment Cosmetics and foods and drinks are suitably applied to humans, but as long as their respective effects are exhibited, animals other than humans (eg, mice, rats, hamsters, dogs, cats, cows, It can also be applied to pigs, monkeys, etc.).

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In this test example, 4-vinylcatechol (manufactured by Toronto Research Chemicals, sample 1) was used as a test sample.

[Test Example 1] Glutathione production promoting action test For 4-vinylcatechol (sample 1), the glutathione production promoting action was tested as follows.

Normal human skin fibroblasts (NB1RGB) were cultured using α-MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with 10% FBS-containing α-MEM medium to a cell density of 2.0 × 10 ⁵ cells / mL, then seeded at 200 μL per well in a 48-well plate and cultured overnight.

  After culturing, 200 μL of a test sample (sample 1, refer to Table 1 below for sample concentration) dissolved in 1% FBS-containing Dulbecco's MEM medium was added to each well and cultured for 24 hours. As a control, the culture was similarly performed using Dulbecco's MEM medium containing 1% FBS without addition of the test sample. After completion of the culture, the medium was removed from each well, washed with 400 μL of PBS (−) buffer, and then the cells were lysed using 150 μL of M-PER (PIERCE).

  Of these, 100 μL was used to quantify total glutathione. That is, 100 μL of lysed cell extract, 50 μL of 0.1 mol / L phosphate buffer, 2 μL of NADPH, and 25 μL of glutathione reductase (final concentration: 17.5 unit / mL) were added to a 96-well plate at 37 ° C. for 10 minutes. After heating, 25 mmol of 10 mmol / L 5,5′-dithiobis (2-nitrobenzoic acid) was added, and the absorbance at a wavelength of 412 nm until 5 minutes later was measured to obtain ΔOD / min. The total glutathione concentration was calculated based on a calibration curve prepared using oxidized glutathione (manufactured by Wako Pure Chemical Industries, Ltd.). After correcting the obtained value to the amount of glutathione per total protein amount, the glutathione production promotion rate (%) was calculated by the following formula.

Glutathione production promotion rate (%) = B / A × 100
Each term in the formula represents the following.
A: Glutathione amount per total protein amount when no sample is added B: Glutathione amount per total protein amount when test sample is added Table 1 shows the results.

  As shown in Table 1, it was recognized that 4-vinylcatechol (Sample 1) has an excellent glutathione production promoting action.

[Test Example 2] Laminin 5 production promoting action test With respect to 4-vinylcatechol (sample 1), laminin 5 production promoting action was tested as follows.

Normal human neonatal epidermal keratinocytes (NHEK) are precultured in an 80 cm ² flask using a growth medium for normal human epidermal keratinocytes (KGM) at 37 ° C. and 5% CO ₂ -95% air. The cells were collected by trypsin treatment. The collected cells were diluted with a medium (KGM-BPE) obtained by removing BPE from KGM so that the cell density was 1.0 × 10 ⁵ cells / mL, and then seeded at 500 μL per well in a 24-well plate. and cultured for two days under the conditions of _{℃ · 5% CO 2 -95%} air.

After completion of the culture, the medium was removed and 500 μL of a test sample dissolved in KGM-BPE medium (sample 1, sample concentration as shown in Table 2 below) was added to each well at 37 ° C., 5% CO ₂ -95%. The cells were cultured for 48 hours under air conditions. In addition, it culture | cultivated similarly using the KGM-BPE culture medium without a sample as control. After completion of the culture, 100 μL of the supernatant was transferred to an ELISA plate and adsorbed on the plate at 37 ° C. for 2 hours, and then the amount of adsorbed laminin 5 was determined using a monoclonal anti-human laminin 5 antibody (mouse IgG) (manufactured by Chemicon). It was measured by the ELISA method used. From the obtained measurement results, the laminin 5 production promotion rate (%) was calculated by the following formula.

Laminin 5 production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Laminin 5 amount with addition of test sample B: Laminin 5 amount without addition of sample Table 2 shows the results.

  As shown in Table 2, it was confirmed that 4-vinylcatechol (sample 1) has an excellent laminin 5 production promoting action.

Test Example 3 MMP-1 Activity Inhibitory Action Test With respect to 4-vinylcatechol (sample 1), the MMP-1 activity inhibitory action was tested as follows.

  Test sample dissolved in 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 3 below for sample concentration) 50 μL MMP-1 solution (Sigma, COLLAGENASE Type IV from Clostridium histolyticum) 50 μL and Pz peptide solution (BACHEM Feinchemikalien AG, Pz-Pro-Leu-Gly-Pro-D-Arg-OH) 400 μL were mixed. The mixture was 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 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. Similarly, a blank test was performed and corrected. From the obtained results, the MMP-1 activity inhibition rate (%) was calculated by the following formula.

MMP-1 activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
Each term in the formula represents the following.
A: Absorbance at a wavelength of 320 nm with no sample added / enzyme added B: Absorbance at a wavelength of 320 nm with no sample added / enzyme added C: Absorbance at a wavelength of 320 nm with test sample added / enzyme added D: Test sample added / enzyme Absorbance at a wavelength of 320 nm without addition is shown in Table 3.

  As shown in Table 3, it was recognized that 4-vinylcatechol (sample 1) has an excellent MMP-1 activity inhibitory action.

[Test Example 4] AGEs degradation promoting action test With respect to 4-vinylcatechol (sample 1), the degradation promoting action of final glycation products (AGEs) was tested as follows.

  To a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.), add 100 μL of 0.2MD (−)-ribose solution prepared in PBS (−) buffer, and leave it at 37 ° C. for 2 weeks. Formed. In addition, what added only PBS (-) buffer as a negative control was left still similarly. Two weeks later, 100 μL of each test sample (sample 1, refer to Table 4 below for sample concentration) prepared in PBS (−) buffer was added, and the mixture was further allowed to stand at 37 ° C. for 2 weeks. In addition, only PBS (-) buffer solution as a positive control and PBS (-) buffer solution as a negative control were allowed to stand in the same manner. Two weeks later, the amount of AGEs was measured by an ELISA method using an anti-AGE antibody (manufactured by Transgenic), and the AGEs degradation promoting action was evaluated. From the obtained results, the AGE decomposition rate (%) was calculated by the following formula.

AGE decomposition rate (%) = {(BC) / (BA)} × 100
Each term in the formula represents the following.
A: Absorbance at a wavelength of 405 nm in a negative control B: Absorbance at a wavelength of 405 nm without addition of a sample (positive control) C: Absorbance at a wavelength of 405 nm with addition of a test sample Table 4 shows the results.

  As shown in Table 4, 4-vinylcatechol (Sample 1) showed an excellent AGE decomposition degradation action.

[Test Example 5] AGEs formation inhibitory action test 4-vinylcatechol (sample 1) was tested for the formation inhibitory action of final glycation products (AGEs) as follows.

  In a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.), 0.2M D (−)-ribose prepared with PBS (−) buffer and a test sample (Sample 1, see Table 5 below for sample concentration) After adding 100 μL of the mixture, the mixture was allowed to stand at 37 ° C. for 2 weeks to form AGEs. In addition, the PBS (-) buffer only as a negative control and a 0.2M D (-)-ribose solution prepared with a PBS (-) buffer as a positive control were allowed to stand in the same manner. Two weeks later, the amount of AGEs was measured by an ELISA method using an anti-AGE antibody (manufactured by Transgenic) to evaluate the AGEs formation inhibitory action. From the obtained results, the AGE formation inhibition rate (%) was calculated by the following formula.

AGE formation inhibition rate (%) = {(BC) / (BA)} × 100
Each term in the formula represents the following.
A: Absorbance at a wavelength of 405 nm in a negative control B: Absorbance at a wavelength of 405 nm without addition of a sample (positive control) C: Absorbance at a wavelength of 405 nm with addition of a test sample Table 5 shows the results.

  As shown in Table 5, 4-vinylcatechol (sample 1) exhibited an excellent AGE formation inhibitory action.

[Test Example 6] Type I collagen production promoting action test 4-vinylcatechol (Sample 1) was tested for type I collagen production promoting action as follows.

Normal human skin fibroblasts (NB1RGB) were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with the above medium to a cell density of 1.6 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well microplate, and cultured overnight.

  After completion of the culture, the medium was removed, and 150 μL of a test sample dissolved in Dulbecco's MEM medium containing 0.25% FBS (Sample 1, see Table 6 below for each sample concentration) was added to each well and cultured for 3 days. As a control, the culture was similarly performed using Dulbecco's MEM medium containing 0.25% FBS without any sample. After culture, the amount of type I collagen in the medium of each well was measured by ELISA. From the measurement results, the type I collagen production promotion rate (%) was calculated by the following formula.

Type I collagen production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Type I collagen amount with addition of test sample B: Type I collagen amount without addition of sample Table 6 shows the results.

  As shown in Table 6, it was confirmed that 4-vinylcatechol (sample 1) has an excellent type I collagen production promoting effect.

[Test Example 7] MMP-2 activity inhibitory action test MMP-2 prepared by the method described in JP-A-2007-217352 was used. That is, the MMP-2 protein was expressed in a large amount using a Escherichia coli gene expression system as a recombinant proMMP protein having 6 histidine residues at the C-terminus, purified and refolded using Ni-NTA resin, and then activated. Moved to mold. This was used as an enzyme preparation and appropriately diluted to prepare an enzyme solution.
Using the obtained enzyme solution, 4-vinylcatechol (sample 1) was tested for MMP-2 activity inhibitory action as follows.

Using a 96-well plate for fluorescence intensity measurement, 20 μL of test sample (Sample 1, see Table 7 below for sample concentration), 40 μL of enzyme solution and buffer (0.05 mol / L Tris, 150 mmol / L NaCl, 10 mmol / L CaCl) ₂ , 50 μmol / L ZnSO ₄ , 0.02% NaN ₃ , 0.05% Brij 35 (pH 7.5)) 20 μL were mixed and allowed to stand at 37 ° C. for 15 minutes. Thereafter, 120 μL of a fluorescent substrate peptide (MOCAc / DNP peptide) prepared to 4.16 μmol / L was added, and the fluorescence intensity was immediately measured at an excitation wavelength of 340 nm and a fluorescence wavelength of 420 nm, and this was taken as the fluorescence intensity immediately after the addition of the substrate. Immediately after the measurement, the reaction was carried out at 37 ° C. for 120 minutes. During this time, the fluorescence intensity at an excitation wavelength of 340 nm and a fluorescence wavelength of 420 nm was measured every 15 minutes, and these were taken as the fluorescence intensity after addition of the substrate. A blank test was performed in the same manner and corrected. From the obtained results, the MMP-2 activity inhibition rate (%) was calculated by the following formula.

MMP-2 activity inhibition rate (%) = {1− (C−D) / (A−B)} × 100
Each term in the formula represents the following.
A: Fluorescence intensity at a wavelength of 420 nm 120 minutes after no sample addition / substrate addition B: Fluorescence intensity at a wavelength 420 nm immediately after no sample addition / substrate addition C: At 420 nm wavelength 120 minutes after addition of a test sample / substrate addition Fluorescence intensity D: Fluorescence intensity at a wavelength of 420 nm immediately after the addition of the test sample and the substrate is shown in Table 7.

  As shown in Table 7, it was confirmed that 4-vinylcatechol (sample 1) has an excellent MMP-2 activity inhibitory action.

[Test Example 8] Serine palmitoyltransferase (SPT) mRNA expression promoting action test 4-vinylcatechol (sample 1) was tested for SPT mRNA expression promoting action as follows.

Normal human neonatal epidermal keratinocytes (NHEK) were cultured in a culture medium for long-term culture of normal human epidermal keratinocytes (EpiLife-KG2) in an 80 cm ² flask under conditions of 37 ° C. and 5% CO ₂ -95% air. And the cells were recovered by trypsin treatment. The collected cells are diluted with Epilife-KG2 medium to a cell density of 20 × 10 ⁴ cells / mL, and then seeded at 2 mL each in a 35 mm Petri dish (FALCON) (40 × 10 ⁴ cells / Petri dish), 37 They were cultured for 24 hours under the conditions of _{℃ · 5% CO 2 -95%} air.

After culturing, the medium was removed, and 2 mL each of the test samples dissolved in Epilife-KG2 medium (sample 1, sample concentration as shown in Table 8 below) was added to each dish, and the mixture was incubated at 37 ° C., 5% CO ₂ -95% air. The culture was performed under the conditions of 24 hours. In addition, it culture | cultivated similarly as control using Epilife-KG2 culture medium without a sample. After culturing, the medium is removed, and total RNA is extracted with ISOGEN (Nippon Gene, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression level of mRNA was measured for SPT and GAPDH as an internal standard. The detection was performed by real-time 2 Step RT-PCR reaction using a real-time PCR device Smart Cycler (Cepheid) and a TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A). As for the expression level of SPT, a correction value was obtained from the value of GAPDH based on the total RNA preparation prepared from the cells cultured in “addition of test sample” and “no addition of sample”, respectively. From the obtained value, the SPT mRNA expression promotion rate (%) was calculated by the following formula.

SPT mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value without addition of sample Table 8 shows the results.

  As shown in Table 8, it was confirmed that 4-vinylcatechol (sample 1) has an excellent SPT mRNA expression promoting action.

[Test Example 9] Aquaporin 3 (AQP3) mRNA expression promoting action test 4-vinylcatechol (sample 1) was tested for AQP3 mRNA expression promoting action as follows.

Normal human neonatal epidermal keratinocytes (NHEK) were precultured in an 80 cm ² flask using growth medium for normal human epidermal keratinocytes (KGM) under conditions of 37 ° C. and 5% CO ₂ -95% air. The cells were collected by trypsin treatment. The collected cells are diluted with KGM medium to a cell density of 20 × 10 ⁴ cells / mL, and then seeded at 2 mL each in a 35 mm petri dish (FALCON) (40 × 10 ⁴ cells / petri dish) at 37 ° C. They were cultured for 24 hours under the conditions of 5% CO ₂ -95% air.

After culturing, the medium was removed, and 2 mL of the KGM medium of the test sample dissolved in KGM medium (Sample 1, see Table 9 below for sample concentration) was added to each petri dish at 37 ° C., 5% CO ₂ -95% air. The culture was performed under the conditions of 24 hours. As a control, the same culture was performed using a sample-free KGM medium. After culturing, the medium is removed, and total RNA is extracted with ISOGEN (Nippon Gene, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression level of mRNA was measured for AQP3 and GAPDH as an internal standard. Detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). . The expression level of AQP3 mRNA was determined as a GAPDH value based on the total RNA preparation prepared from the cells cultured in “addition of test sample” and “no addition of sample”, respectively. From the obtained value, the AQP3 mRNA expression promotion rate (%) was calculated by the following formula.

AQP3 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value without addition of sample Table 9 shows the results.

  As shown in Table 9, it was confirmed that 4-vinylcatechol (sample 1) has an excellent AQP3 mRNA expression promoting action.

[Test Example 10] Tyrosinase activity inhibitory action test 4-vinylcatechol (sample 1) was tested for tyrosinase activity inhibitory action as follows.

  Test sample dissolved in 0.2 mL of Mclvaine buffer (pH 6.8), 0.06 mL of 0.3 mg / mL tyrosine solution, 25% DMSO solution in 48-well plate (Sample 1, see Table 10 below for sample concentration) 0.18 mL was added and allowed to stand at 37 ° C. for 10 minutes. To this, 0.02 mL of 800 units / mL tyrosinase solution was added, followed by reaction at 37 ° C. for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 475 nm was measured.

  Further, as a blank, the same operation and absorbance measurement were performed when no enzyme solution was added. Further, as a control, the same measurement was performed when distilled water was added without adding the sample solution. From the obtained measurement results, the tyrosinase activity inhibition rate (%) was calculated by the following formula.

Tyrosinase activity inhibition rate (%) = {1− (St−Sb) / (Ct−Cb)} × 100
Each term in the formula represents the following.
St: Absorbance at a wavelength of 475 nm when a test sample is added and an enzyme is added Sb: Absorbance at a wavelength of 475 nm when a test sample is added and an enzyme is not added Ct: Absorbance at a wavelength of 475 nm when no sample is added and an enzyme is added Cb: No sample is added and an enzyme Table 10 shows the results of absorbance at a wavelength of 475 nm without addition.

  As shown in Table 10, it was recognized that 4-vinylcatechol (Sample 1) has an excellent tyrosinase activity inhibitory action.

[Test Example 11] Test for inhibiting melanin production on B16 melanoma cells With respect to 4-vinylcatechol (sample 1), the melanin production inhibiting action on B16 melanoma cells was tested as follows.

B16 melanoma cells were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS and 1 mmol / L theophylline to a cell density of 24.0 × 10 ⁴ cells / mL, and then seeded at 300 μL per well in a 48-well plate. Incubate for hours.

  After completion of the culture, 300 μL of a test sample dissolved in Dulbecco MEM medium containing 10% FBS and 1 mmol / L theophylline (Sample 1, see Table 11 below for sample concentration) was added to each well and cultured for 4 days. In addition, it culture | cultivated similarly using 10% FBS without a sample and 1 mmol / L theophylline containing Dulbecco's MEM culture medium as control. After completion of the culture, the medium was removed, 200 μL of 2 mol / L NaOH solution was added, the cells were broken with 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 a calibration curve prepared using synthetic melanin (manufactured by SIGMA).

  In order to measure the cell viability, after culturing in the same manner as described above, the medium was removed, washed with 400 μL of PBS (−) buffer, and Dulbecco containing 10% FBS at a final concentration of 0.05 mg / mL. 200 μL of neutral red dissolved in MEM was added to each well and cultured for 2.5 hours. After incubation, the neutral red solution was removed, and 200 μL of ethanol / acetic acid solution (ethanol: acetic acid: water = 50: 1: 49) was added to each well to extract the dye. After extraction, the absorbance at a wavelength of 540 nm was measured. From the obtained results, the melanin production inhibition rate (%) corrected by the cell viability was calculated by the following formula.

Melanin production inhibition rate (%) = {1− (B / D) / (A / C)} × 100
Each term in the formula represents the following.
A: Amount of melanin without addition of sample B: Amount of melanin with addition of test sample C: Absorbance at a wavelength of 540 nm without addition of sample D: Absorbance at a wavelength of 540 nm with addition of test sample Table 11 shows the results.

  As shown in Table 11, it was recognized that 4-vinylcatechol (sample 1) has an excellent melanin production inhibitory action.

[Test Example 12] Testosterone 5α-reductase inhibitory action test With respect to 4-vinylcatechol (sample 1), testosterone 5α-reductase inhibitory action was tested as follows.

  In a V-bottom tube with a lid, 20 μL of a 4.2 mg / mL testosterone (manufactured by Wako Pure Chemical Industries) solution prepared with propylene glycol and 5 mmol / L Tris-HCl (pH 7.13) containing 1 mg / mL NADPH 825 μL of buffer was mixed.

  Furthermore, 80 μL of a test sample solution prepared in 50% ethanol (Sample 1, see Table 12 below for sample concentration) and 75 μL of S-9 (Oriental Yeast Co., Ltd., rat liver homogenate) were added and mixed, Incubated at 37 ° C. for 30 minutes. Thereafter, 1 mL of methylene chloride was added to stop the reaction. This was centrifuged (1600 × g, 10 minutes), the methylene chloride layer was fractionated, and the fractionated methylene chloride layer was subjected to gas chromatography analysis under the following conditions to obtain 3α-androstanediol, 5α. -The concentrations of dihydrotestosterone (5α-DHT) and testosterone were quantified. As a control, the same amount (80 μL) of the sample solvent was used in place of the test sample solution, and the same treatment was performed for gas chromatography analysis.

<Gas chromatography conditions>
Equipment used: Shimadzu GC-2010 (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.
Standards 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 relationship (1) Based on.

A = B × C / D (1)
Each term in the formula represents the following.
A: Concentration of 3α-androstanediol, 5α-DHT or testosterone B: Peak area of 3α-androstanediol, 5α-DHT or testosterone C: Concentration of standard product D: Peak area of standard product

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

Conversion rate = (E + F) / (E + F + G) (2)
Each term in the formula represents the following.
E: concentration of 3α-androstanediol (μg / mL)
F: concentration of 5α-DHT (μg / mL)
G: Testosterone concentration (μg / mL)

Using the conversion rate calculated based on the formula (2), the testosterone 5α-reductase inhibition rate (%) was calculated based on the following formula (3).
Testosterone 5α-reductase inhibition rate (%) = (1−H / I) × 100 (3)
Each term in the formula represents the following.
H: Conversion rate with addition of test sample I: Conversion rate without addition of sample Table 12 shows the results.

  As shown in Table 12, it was confirmed that 4-vinylcatechol (sample 1) has an excellent testosterone 5α-reductase inhibitory action.

[Test Example 13] Hair papilla cell proliferation promoting action test 4-vinylcatechol (sample 1) was tested for hair papilla cell proliferation promoting action as follows.

Normal human hair dermal papilla cells (HFDPC, derived from male parietal region) were cultured using a growth medium for dermal papilla cells (PCGM, manufactured by Toyobo Co., Ltd.) containing 1% FCS and a growth additive, followed by trypsin treatment. Cells were collected. The collected cells were diluted to a cell density of 1.0 × 10 ⁴ cells / mL using DMEM medium containing 10% FBS, and then seeded at 200 μL per well in a collagen-coated 96-well plate. Cultured for days.

  Thereafter, the medium was removed, 200 μL of a test sample dissolved in serum-free DMEM medium (Sample 1, see Table 13 below for sample concentration) was added to each well, and further cultured for 4 days. As a control, the same culture was performed using a serum-free DMEM medium containing no sample. After completion of the culture, the dermal papilla cell proliferation promoting effect was measured by MTT assay. That is, after removing the medium, adding 200 μL of 0.4 mg / mL MTT prepared in serum-free DMEM medium, and further culturing for 2 hours, the blue formazan produced in the cells was extracted with 100 μL of 2-propanol. About this extract, the light absorbency of 570 nm with a blue formazan absorption maximum point 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. From the measurement results, the hair papillary cell proliferation promotion rate (%) was calculated based on the following formula.

Growth rate of dermal papilla cells (%) = A / B × 100
Each term in the formula represents the following.
A: Blue formazan production amount with addition of test sample B: Blue formazan production amount without addition of sample Table 13 shows the results.

  As shown in Table 13, it was recognized that 4-vinylcatechol (Sample 1) has an excellent hair papillary cell proliferation promoting action.

[Test Example 14] Hyaluronidase activity inhibitory action test 0.2 mL of a test sample dissolved in 0.1 mol / L acetate buffer (pH 3.5) (Sample 1, see Table 14 below for sample concentration) was added to a hyaluronidase solution (SIGMA 0.1 mL (Type IV-S, from bovine tests, 400 NF units / mL) was added and allowed to stand at 37 ° C. for 20 minutes. Furthermore, 0.2 mL of 2.5 mmol / L calcium chloride was added as an activator and allowed to stand at 37 ° C. for 20 minutes. To this, 0.5 mL of 0.8 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 cooled, and then 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.

Further, as a blank, the same operation and absorbance measurement were performed when no enzyme solution was added. Further, as a control, the same measurement was performed when distilled water was added without adding the sample solution. From the obtained results, the hyaluronidase activity inhibition rate (%) was calculated by the following formula.
Hyaluronidase activity inhibition rate (%) = {1− (St−Sb) / (Ct−Cb)} × 100
Each term in the formula represents the following.
St: Absorbance at a wavelength of 585 nm when a test sample is added and an enzyme is added Sb: Absorbance at a wavelength of 585 nm when a test sample is added and an enzyme is not added Ct: Absorbance at a wavelength of 585 nm when no sample is added and an enzyme is added Cb: No sample is added and an enzyme Table 14 shows the results of absorbance at a wavelength of 585 nm without addition.

  As shown in Table 14, it was confirmed that 4-vinylcatechol (sample 1) has an excellent hyaluronidase activity inhibitory action. It was also confirmed that the degree of hyaluronidase activity inhibitory action can be adjusted by the concentration of 4-vinylcatechol.

[Test Example 15] Hexosaminidase release inhibitory action test With respect to 4-vinylcatechol (sample 1), hexosaminidase release inhibitory action was tested as follows.

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

  After the culture, the medium was removed, and washing was performed twice with 100 μL of Silaganian buffer. Next, 10 μL of a test sample dissolved in the same buffer (Sample 1, see Table 15 below for sample concentration) and 30 μL of the same buffer were added to each well and allowed to stand at 37 ° C. for 10 minutes. As a control, the same operation was carried out using 40 μL of a sample-free addition of Siraganian buffer. Subsequently, 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 was collected in a new 96-well plate, 10 μL of 1 mmol / L p-NAG (p-nitrophenyl-N-acetyl-β-D-glucosaminide) solution was added to each well, and the temperature was 37 ° C. For 1 hour.

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 wavelengths of 415 nm and 650 nm was measured. The value obtained by subtracting the absorbance at 650 nm from the absorbance at 415 nm was used as a correction value. Further, as a blank, absorbance at wavelengths of 415 nm and 650 nm of a mixed solution of 10 μL of cell supernatant and 250 mol of 0.1 mol / L Na ₂ CO ₃ / NaHCO ₃ was measured, and a correction value was calculated. From the obtained measurement results, the hexosaminidase release inhibition rate (%) was calculated by the following formula.

Inhibition rate of hexosaminidase release (%) = {1− (BC) / A} × 100
Each term in the formula represents the following.
A: Correction value without addition of sample B: Correction value with addition of test sample C: Correction value with addition of test sample / no addition of p-NAG The results are shown in Table 15.

  As shown in Table 15, it was confirmed that 4-vinylcatechol (sample 1) has an excellent hexosaminidase release inhibitory action.

[Test Example 16] COX-2 activity inhibitory action test in mouse macrophages (PGE ₂ production inhibitory action test)
After culturing mouse macrophage cells (RAW264.7) using Dulbecco's MEM medium containing 10% FBS, the cells were collected with a cell scraper. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS to a concentration of 2.0 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured for 18 hours.

After culturing, in order to acetylate and inactivate COX-1 already present and COX-2 expressed in a small amount, the medium was replaced with a medium containing 500 μmol / L aspirin and cultured for 4 hours. Thereafter, the cells were washed three times with PBS (−) buffer and dissolved in Dulbecco's MEM medium containing 10% FBS containing 0.5% final DMSO (Sample 1, see Table 16 below for sample concentration). Was added to each well, and then 100 μL of lipopolysaccharide (LPS) (DIFCO, E. coli 0111; B4) dissolved in Dulbecco MEM containing 10% FBS at a final concentration of 1 μg / mL was added and cultured for 16 hours. did. As a control, the culture was similarly performed using Dulbecco's MEM medium containing 10% FBS containing a final concentration of 0.5% DMSO without addition of the sample. After completion of the culture, the amount of prostaglandin E _{2 in} the culture supernatant of each well was quantified using a PGE ₂ EIA Kit (Cayman Chemical). From the obtained results, the COX-2 activity inhibition rate (%, PGE ₂ production inhibition rate) was calculated by the following formula.

Macrophage COX-2 activity inhibition rate (%) = {1− (A−C) / (B−C)} × 100
Each term in the formula represents the following.
A: Prostaglandin E ₂ amount of test sample added · LPS stimulated B: amount prostaglandin E ₂ in the sample without additives · LPS stimulated C: prostaglandin E ₂ weight results for the samples without additives · LPS non-stimulated Is shown in Table 16.

  As shown in Table 16, it was confirmed that 4-vinylcatechol (sample 1) has an excellent COX-2 activity inhibitory action in macrophages.

[Test Example 17] COX-2 activity inhibitory action test on keratinocytes (PGE ₂ production inhibitory action test)
Normal human neonatal epidermal keratinocytes (NHEK) were cultured using a growth medium for normal human epidermal keratinocytes (KGM), and then cells were collected by trypsin treatment. The collected cells are diluted with KGM medium to a cell density of 12.5 × 10 ⁴ cells / mL, and then seeded at 200 μL per well in a collagen-coated 48-well plate (2.5 × 10 ⁴ cells / well). ) And cultured overnight. After confirming that the cells had settled, the cells were cultured for 24 hours in 200 μL of KGM medium from which hydrocortisone (hydrocortisone) had been removed.

After culturing, in order to acetylate and inactivate COX-1 already present and COX-2 expressed in a small amount, 200 μL of 500 μL / L aspirin-containing KGM medium (without hydrocortisone) was added and cultured for 4 hours. After culturing, the cells were washed three times with PBS (−) buffer, 100 μL of PBS (−) buffer was added, UV-B irradiation (50 mJ / cm ² ) was performed, and then dissolved in KGM medium (without hydrocortisone). 400 μL of the test sample (sample 1, refer to Table 17 below for sample concentration) was added to each well and cultured for 24 hours at 37 ° C. and 5% CO ₂ . In addition, it culture | cultivated similarly as control using the KGM culture medium (without hydrocortisone) without a sample. After completion of the culture, the amount of prostaglandin E _{2 in} the culture supernatant of each well was quantified using a PGE ₂ EIA Kit (Cayman Chemical). From the obtained results, the COX-2 activity inhibition rate (%, PGE ₂ production inhibition rate) was calculated by the following formula.

Keratinocyte COX-2 activity inhibition rate (%) = {1- (A−C) / (B−C)} × 100
Each term in the formula represents the following.
A: Prostaglandin E ₂ amount of test sample added to and UV irradiation B: amount prostaglandin E ₂ in the sample without additives ultraviolet irradiation C: Prostaglandin E ₂ weight results for the samples without additives not irradiated with ultraviolet rays Is shown in Table 17.

  As shown in Table 17, it was confirmed that 4-vinylcatechol (sample 1) has an excellent COX-2 inhibitory action in keratinocytes.

[Test Example 18] Cyclic AMP phosphodiesterase activity inhibitory action test For 4-vinylcatechol (sample 1), the cyclic AMP phosphodiesterase activity inhibitory action was tested as follows.

  To 0.2 mL of 50 mmol / L Tris-HCl buffer (pH 7.5) containing 5 mmol / L magnesium chloride, 0.1 mL of 2.5 mg / mL bovine serum albumin solution, 0.1 mg / mL cyclic AMP phosphodiesterase solution 0.1 mL and 0.05 mL of the test sample solution (Sample 1, see Table 18 below for sample concentration) were added and allowed to stand at 37 ° C. for 5 minutes. Thereafter, 0.05 mL of a 0.5 mg / mL cyclic AMP solution was added and reacted at 37 ° C. for 60 minutes. After completion of the reaction, the reaction was stopped by boiling on a boiling water bath for 3 minutes, and this was centrifuged (2260 × g, 10 minutes, 4 ° C.), and cyclic AMP, which is a reaction substrate in the supernatant, was expressed as follows. Analysis was performed under high performance liquid chromatography conditions. Further, as a control, the same operation was performed by adding only the solvent without the sample.

<High performance liquid chromatography conditions>
Product name: Chromatocoder 12 (manufactured by SYSTEM INSTRUMENTS)
Stationary phase: Wakosil C ₁₈ -ODS 5 μm (Wako Pure Chemical Industries, Ltd.)
Column length: 250mm
Mobile phase: 1 mmol / L TBAP in 25 mmol / L KH ₂ PO ₄ : CH ₃ CN = 90: 10
Mobile phase flow rate: 1.0 mL / min
Detection: 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 the cyclic AMP phosphodiesterase when no sample is added, and the cycle time when the test sample is added. The peak area (B2) of the supernatant of the reaction solution of the click AMP standard product and the cyclic AMP phosphodiesterase was determined. From the obtained results, the degradation rate (C) of the cyclic AMP standard product when no sample was added and the degradation rate (D) of the cyclic AMP standard product when the test sample was added were calculated according to the following equations.

Standard product decomposition rate without addition of sample (C,%) = (1−B1 / A) × 100
Degradation rate of standard product with addition of test sample (D,%) = (1−B2 / A) × 100

Thereafter, the cyclic AMP phosphodiesterase activity inhibition rate (%) was calculated by the following formula based on the respective degradation rates (C, D) calculated by the above formula.
cAMP phosphodiesterase activity inhibition rate (%) = (1-D / C) × 100
The results are shown in Table 18.

  As shown in Table 18, it was confirmed that 4-vinylcatechol (sample 1) has an excellent cyclic AMP phosphodiesterase activity inhibitory action.

[Test Example 19] Glutathione reduction inhibitory test using normal human hepatocytes Normal human hepatocytes were cultured using Dulbecco's MEM medium containing 10% FBS, and then cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS to a cell density of 10 × 10 ⁴ cells / mL, then seeded at 200 μL per well in a 48-well plate and cultured overnight.

  After completion of the culture, 200 μL of a test sample dissolved in Dulbecco MEM containing 1% FBS (Sample 1, see Table 19 below for sample concentration) and 200 μL of galactosamine hydrochloride having a final concentration of 5 mM are added to each well, and further 24 hours. Cultured. Similarly, after cell seeding, cells that were not treated with galactosamine hydrochloride and cells that were treated with galactosamine hydrochloride after cell seeding and were not added with the test sample were also measured in the same manner, which were defined as an untreated group and a treated group, respectively. After completion of the culture, the medium was removed from each well, washed with 400 μL of PBS (−) buffer, and the cells were lysed using 150 μL of M-PER (PIERCE).

  The total glutathione was quantified using 50 μL of this. Specifically, 50 μL of dissolved cell extract, 100 μL of 0.1 M phosphate buffer, 2 μM NADPH, and 25 μL glutathione reductase (final concentration: 17.5 unit / mL) were added to a 96-well plate and heated at 37 ° C. for 10 minutes. 25 μL of 10 mM 5,5′-dithiobis (2-nitrobenzoic acid) was added, and the absorbance at a wavelength of 412 nm until 5 minutes later was measured to obtain ΔOD / min. The total glutathione concentration was calculated based on a calibration curve prepared using oxidized glutathione (manufactured by Wako Pure Chemical Industries, Ltd.). After correcting the obtained value to the amount of glutathione per total protein amount, the glutathione decrease suppression rate was calculated according to the following formula.

Glutathione suppression rate (%) = {(Nt-C)-(Nt-Sa)} / (Nt-C) × 100
Each term in the formula represents the following.
Nt: Glutathione amount per total protein in galactosamine hydrochloride untreated (non-treated group) C: No sample added / Glutathione amount per total protein in galactosamine hydrochloride treated (treated group) Sa: Test sample added / galactosamine Table 19 shows the amount of glutathione per total protein in the hydrochloride treatment.

  As shown in Table 19, it was confirmed that 4-vinylcatechol (sample 1) has an excellent glutathione lowering suppressing action.

Test Example 20 DPPIV Activity Inhibitory Action Test For 4-vinylcatechol (sample 1), dipeptidyl peptidase IV (DPPIV) inhibitory action was tested as follows.

  In a 96-well plate, 25 μL of a test sample (sample 1, refer to Table 20 below for the final concentration) prepared with 25 mM Tris-HCl buffer (pH 8.0) and 0.4 μg / ml prepared with the above buffer. 25 μL of mL DPPIV (R & D System, rhCD26) solution was mixed and preincubated at 37 ° C. for 5 minutes. Thereafter, 50 μL of 0.5 mM Gly-Pro-p-NA · Tos (manufactured by Peptide Laboratories) prepared in the above buffer solution was added and reacted at 37 ° C. for 90 minutes. After completion of the reaction, absorbance at a wavelength of 415 nm was measured. From the obtained results, the DPPIV inhibition rate (%) was calculated by the following formula.

DPPIV inhibition rate (%) = {1− (CD) / (A−B)} × 100
Each term in the formula represents the following.
A: Absorbance at a wavelength of 415 nm when no sample is added / enzyme added B: Absorbance at a wavelength of 415 nm when no sample is added / enzyme added C: Absorbance at a wavelength of 415 nm when a test sample is added / enzyme is added D: Test sample is added / enzyme Absorbance at a wavelength of 415 nm without addition is shown in Table 20.

  As shown in Table 20, 4-vinylcatechol (Sample 1) showed an excellent DPPIV inhibitory action.

[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
4-vinyl catechol 0.01g
Jojoba oil 4.00 g
1,3-butylene glycol 3.00 g
Arbutin 3.00g
Polyoxyethylene cetyl ether (20E.O.) 2.50g
2.00 g of olive oil
Squalane 2.00g
Cetanol 2.00g
Glyceryl monostearate 2.00g
Oleic acid polyoxyethylene sorbitan (20E.O.) 2.00g
Methyl paraoxybenzoate 0.15g
Stearyl glycyrrhizinate 0.10g
Twilight extract 0.10g
Dipotassium glycyrrhizinate 0.10g
Ginkgo biloba extract 0.10g
Conchiolin 0.10g
Oat extract 0.10g
Chamomile extract 0.10g
Fragrance 0.05g
Purified water remainder (total amount is 100 g)

[Formulation Example 2]
A cream having the following composition was produced by a conventional method.
4-vinylcatechol 0.05g
Kujin extract 0.1g
Ogon Extract 0.1g
Liquid paraffin 5.0g
Salami beeswax 4.0g
Squalane 10.0g
Cetanol 3.0g
Lanolin 2.0g
Stearic acid 1.0g
Oleic acid polyoxyethylene sorbitan (20E.O.) 1.5g
3.0 g glyceryl monostearate
Oil soluble licorice extract 0.1g
1,3-butylene glycol 6.0 g
1.5 g of methyl paraoxybenzoate
Fragrance 0.1g
Purified water remainder (total amount is 100 g)

[Composition Example 3]
A serum having the following composition was produced by a conventional method.
4-vinyl catechol 0.01g
Chamomile extract 0.1g
Carrot extract 0.1g
Xanthan gum 0.3g
Hydroxyethylcellulose 0.1g
Carboxyvinyl polymer 0.1g
1,3-butylene glycol 4.0 g
0.1g dipotassium glycyrrhizinate
Glycerin 2.0g
Potassium hydroxide 0.25g
Fragrance 0.01g
Preservative (Methyl paraoxybenzoate) 0.15g
Ethanol 2.0g
Purified water remainder (total amount is 100 g)

[Formulation Example 4]
A hair tonic having the following composition was produced by a conventional method.
4-vinyl catechol 0.4g
Tocopherol acetate appropriate amount Cephalatin 0.002g
Isopropylmethylphenol 0.1g
Sodium hyaluronate 0.15g
Glycerin 15.0g
15.0 g of ethanol
Fragrance Appropriate amount Chelating agent (Sodium edetate) Appropriate amount Preservative (hinokitiol) Appropriate amount Solubilizer (Polyoxyethylene cetyl ether) Appropriate amount Purified water The remainder (100% total)

[Formulation Example 5]
A shampoo having the following composition was produced by a conventional method.
4-vinylcatechol 0.5g
Marjoram extract 1.0g
Plum fruit part extract 0.2g
Coconut oil fatty acid methyl taurine sodium 10.0g
Coconut oil fatty acid amidopropyl betaine 10.0g
Sodium polyoxyethylene alkyl ether sulfate 20.0g
Coconut oil fatty acid diethanolamide 4.0g
Propylene glycol 2.0g
Perfume Appropriate amount Purified water The remainder (100g total amount)

[Composition Example 6]
A tablet having the following composition was produced by a conventional method.
4-Vinylcatechol 5.0mg
Dolomite (containing 20% calcium and 10% magnesium) 83.4mg
Casein phosphopeptide 16.7mg
Vitamin C 33.4mg
Maltitol 136.8mg
Collagen 12.7mg
Sucrose fatty acid ester 12.0mg

[Formulation Example 7]
By an ordinary method, an oral liquid preparation having the following composition was produced.
<Composition in one ampoule (one 100 mL)>
4-Vinylcatechol 0.3% by mass
Sorbit 12.0% by mass
Sodium benzoate 0.1% by mass
Fragrance 1.0% by mass
Calcium sulfate 0.5% by mass
Purified water balance (100% by mass)

  The anti-aging agent, whitening agent, hair-restoring agent, anti-androgen agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione reduction inhibitor and DPPIV activity inhibitor of the present invention prevent skin aging symptoms, Treatment or improvement; Promotion of healing of wounds or burns; Prevention, treatment or improvement of dry skin disease; Prevention or improvement of pigmentation of skin darkening, spots, freckles, etc .; Alopecia, especially alopecia of male pattern alopecia Prevention, treatment or improvement of various diseases; Prevention or improvement of various skin inflammatory diseases; Prevention, treatment or improvement of obesity; Improvement or improvement of liver function; Prevention or treatment of type 2 diabetes, obesity, hypertension, insulin resistance, etc. Or it can greatly contribute to improvement.

Claims (16)

  An anti-aging agent comprising 4-vinylcatechol as an active ingredient.   The 4-vinylcatechol promotes glutathione production, laminin 5 production, matrix metalloproteinase-1 activity inhibition, final glycation product degradation promotion, final glycation product formation inhibition, type I collagen production promotion, matrix metallo 2. Anti-aging according to claim 1, having one or more actions selected from the group consisting of protease-2 activity inhibitory action, serine palmitoyltransferase mRNA expression promoting action and aquaporin 3 mRNA expression promoting action Agent.   A whitening agent comprising 4-vinylcatechol as an active ingredient.   The whitening agent according to claim 3, wherein the 4-vinylcatechol has a tyrosinase activity inhibitory action and / or a melanin production inhibitory action.   A hair restorer comprising 4-vinylcatechol as an active ingredient.   6. The hair restorer according to claim 5, wherein the 4-vinylcatechol has a testosterone 5α-reductase activity inhibitory action and / or a dermal papilla cell growth promoting action.   An anti-androgenic agent comprising 4-vinylcatechol as an active ingredient.   An anti-inflammatory agent comprising 4-vinylcatechol as an active ingredient.   The 4-vinylcatechol has one or more actions selected from the group consisting of hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, and cyclooxygenase-2 activity inhibitory action. 8. The anti-inflammatory agent according to 8.   An antiobesity agent comprising 4-vinylcatechol as an active ingredient.   A cyclic AMP phosphodiesterase activity inhibitor comprising 4-vinylcatechol as an active ingredient.   A glutathione reduction inhibitor comprising 4-vinylcatechol as an active ingredient.   A dipeptidyl peptidase IV activity inhibitor comprising 4-vinylcatechol as an active ingredient.   A skin cosmetic comprising 4-vinylcatechol.   A hair cosmetic comprising 4-vinylcatechol.   A food and drink characterized by blending 4-vinylcatechol.