JP2018080206A - Skin cosmetic, hair cosmetic and food or drink - Google Patents

Abstract

PROBLEM TO BE SOLVED: To discover an ingredient having anti-obesity action, a cyclic AMP phosphodiesterase activity inhibitory action, a dipeptidyl peptidase IV activity inhibitory action, anti-aging action, hair-growing action, antiandrogen action, skin-whitening action or anti-inflammatory action from compounds derived from a natural product and to provide an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a dipeptidyl peptidase IV activity inhibitor, an anti-aging agent, a hair-growing agent, an antiandrogen agent, a skin whitening agent or an anti-inflammatory agent, which uses the ingredient as an active ingredient.SOLUTION: Dihydroferulic acid is used as an active ingredient of an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a dipeptidyl peptidase IV activity inhibitor, an anti-aging agent, a hair-growing agent, an anti-androgen agent, a skin-whitening agent and an anti-inflammatory agent. Further, dihydroferulic acid is blended in a skin cosmetic, a hair cosmetic and a food or drink.SELECTED DRAWING: None

Description

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

  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 lifestyle-related diseases such as hyperlipidemia and arteriosclerosis, so it 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. Tubamoside I (see Patent Document 1) and the like are known as those having an inhibitory action on cAMP phosphodiesterase activity.

  Obesity causes abnormalities in lipid metabolism and causes lifestyle-related diseases such as dyslipidemia as described above. Here, dyslipidemia is a disease in which one or more components among serum lipids, that is, cholesterol, triglycerides, phospholipids, free fatty acids and the like are abnormally increased to cause various disorders. Cholesterol is a component of animal biological membranes and a starting material such as steroid hormones and is a very important substance for animals. However, if blood cholesterol is excessive, cholesterol or the like accumulates inside the blood vessels and causes arteriosclerosis, which may lead to fatal diseases such as ischemic heart disease and cerebral infarction. On the other hand, free fatty acids are released from fat cells into the blood and used as an energy source in the body, but the surplus is taken into the liver and re-synthesized into neutral fat. If the blood free fatty acid is excessive, the neutral fat is chronically excessive and the abnormal lipid state continues, increasing the risk of arteriosclerosis. In addition, if the blood free fatty acid is excessive, insulin resistance may increase and the pathological condition of diabetes may progress.

  Therefore, if too high blood lipids (such as blood cholesterol and blood free fatty acids) due to obesity can be reduced, the symptoms of arteriosclerosis and diabetes that progress with obesity can be prevented, alleviated or improved. I think it can be done. As a component having a blood cholesterol reducing action and a blood free fatty acid reducing action, a roasted yellow tuna leaf extract (Patent Document 2) is known.

  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 the epithelial and endothelial cells of tissues such as kidney, liver, intestine, 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 1). 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.

  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 2). 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 3). 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 3). 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-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 4). 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 5).

  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.

  On the other hand, among 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 5) etc. are known, for example.

  The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrices such as collagen, elastin, and hyaluronic acid that are outside these cells and support 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, if there is an influence of certain external factors such as ultraviolet irradiation, drastic air drying, excessive skin washing, etc., or aging progresses, elastin and hyaluronic acid, which are the main components of the extracellular matrix, As production decreases, it 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 accompanying aging of the skin, that is, wrinkles, dullness, disappearance of texture, decrease in elasticity, and the like are associated with reduction / denaturation of matrix components such as elastin and hyaluronic acid. Therefore, promoting the production of elastin or hyaluronic acid is important in preventing, treating or improving skin aging.

  Here, among these extracellular matrix components, hyaluronic acid is a kind of mucopolysaccharide, which has a function of retaining cells by being filled in the intercellular spaces, and also retains moisture in the intercellular spaces. It has many functions such as lubrication and flexibility to the tissue, resistance to external forces such as mechanical failure. If the production of hyaluronic acid can be promoted, it is considered that skin aging symptoms such as rough skin, wrinkles, dullness, loss of texture, reduced elasticity and reduced moisturizing function can be prevented, treated or improved.

  In addition to skin tissue, hyaluronic acid is present in cartilage, joint fluid, umbilical cord, ophthalmic vitreous, and other connective tissues. Among these, the hyaluronic acid contained in the joint fluid covers the surface of the articular cartilage and is useful for the smooth operation of the joint due to the lubricating function of the hyaluronic acid, the cartilage covering / protecting function, and the like. On the other hand, in arthritis such as rheumatoid arthritis, it is known that the concentration of hyaluronic acid in joint fluid is reduced. Therefore, it is considered that by promoting the production of hyaluronic acid, arthritis such as rheumatoid arthritis, osteoarthritis, septic arthritis, gouty arthritis, traumatic arthritis, or osteoarthritis can be prevented or treated. Furthermore, in the healing process of wounds or burns, granulation (tissue) is formed, but it is known that hyaluronic acid is significantly increased in the granulation. Therefore, it is considered that healing of wounds or burns can be promoted by promoting production of hyaluronic acid. As what has a hyaluronic acid production promotion effect | action, the extract (refer patent document 6) etc. from Kusunohagashi are known.

  On the other hand, among the aforementioned extracellular matrix components, elastin is a fiber that gives elasticity to skin tissue. If the production of elastin can be promoted, wrinkles and sagging are less likely to occur, and skin aging symptoms such as loss of tension and reduced elasticity can be prevented, treated or improved.

  In addition to skin tissue, elastin is widely expressed in tissues that require elasticity in the body, such as lungs and blood vessels. It is known that when normal elastin decreases from these tissues with aging, elasticity in the lungs and blood vessels decreases, causing pulmonary diseases such as emphysema and vascular diseases such as hypertension and aneurysms. ing. Therefore, if the production of elastin can be promoted, the elasticity in the lungs and blood vessels is unlikely to decrease, and it is considered that lung diseases such as emphysema and vascular diseases such as hypertension and aneurysms can be prevented or treated. . For example, an extract from a plant belonging to the genus Hippophae is known as one having an elastin production promoting action (see Patent Document 7).

  The aforementioned extracellular matrix components such as collagen are produced by fibroblasts. In young skin, the proliferation of fibroblasts is active, and the retention of moisture, flexibility, elasticity, etc. is ensured by maintaining the homeostasis of the interaction of skin tissues such as fibroblasts and collagen. It is maintained in a fresh state with tension and gloss. However, when there is an influence of certain external factors such as ultraviolet rays, significant drying of air, excessive skin washing, etc., or when aging progresses, the growth of fibroblasts slows down and the skin's moisturizing function and elasticity are descend. And the skin loses its tension and gloss, and exhibits aging symptoms such as roughness and wrinkles. Therefore, promoting the proliferation of fibroblasts is considered to be very important in preventing, treating or improving skin aging.

  Fibroblasts play an important role in the healing process of wounds such as trauma and burns, and are also important in the healing process of skin diseases (eg, skin ulcers such as pressure sores, burn ulcers, diabetic ulcers, etc.). is there. Therefore, it is considered that wounds and skin diseases can be treated by promoting the proliferation of fibroblasts. Furthermore, in the field of regenerative medicine, a method for culturing and proliferating skin cells from a patient's own skin fragment and transplanting it to a patient as a treatment method for a patient suffering from a wound that is difficult to self-heal (such as a severe burn) However, the use of a cell growth promoter is expected to shorten the culture period of skin cells. Conventionally, an extract from Kusunohagashi (see Patent Document 6 described above) or the like is known as one having a fibroblast proliferation promoting action.

  The epidermis functions to alleviate external stimuli and control the loss of body components such as moisture, and starts with the basal layer, the lowest layer, and continues to the spiny layer, granule layer, and stratum corneum. It is composed of Most cells present in each layer are keratinocytes differentiated from the basal layer. The keratinocytes that divide and proliferate in the basal layer differentiate into keratinocytes while passing through the spinous layer and the granule layer, forming a stratum corneum composed of keratin protein fibers with strong cross-linking, Eventually it will fall off the stratum corneum as plaque.

  The stratum corneum is present in the outermost shell of the skin and serves as a physical barrier against irritation from the outside world. In order to have this barrier function in the skin, the cycle (keratinization) from the production of keratinocytes in the basal layer to the peeling off of the keratinocytes (keratinization) is usually repeated at a cycle of 4 weeks to perform epidermal metabolism. . However, the metabolic function of this stratum corneum also deteriorates with aging, and skin troubles such as wrinkles, dullness, pigmentation, and rough skin occur. Therefore, it is considered that skin aging such as wrinkles, dullness, and pigmentation can be improved by promoting the proliferation of keratinocytes and restoring the metabolic function of the skin. Conventionally, as a thing which has an epidermal keratinocyte proliferation promotion effect, a shellfish mother extract (refer patent document 8) etc. 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 6). As what has an AQP3 expression promotion effect, the extract from the leaf part of a star fruit (refer patent document 9) 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.

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

  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 10) and the like have been 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 and 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 7).

  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 11) or the like has been known as one having an action to promote proliferation of dermal papilla cells.

  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, in order to prevent, treat or improve skin darkness (skin pigmentation), spots, buckwheat, etc., it is conceivable to suppress the production of melanin.

  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. Examples of those having a melanin production inhibitory action include extracts from plants belonging to the genus Southurea (see Patent Document 12). ) Etc. are known.

  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 13) 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 14) 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 8).

  As for dihydroferulic acid, it is known that dihydroferulic acid is detected when certain lactic acid bacteria are cultured in a medium containing ferulic acid or ethyl ferulate (Non-patent Document 9, Patent Document 15). reference).

JP 2006-056855 A Japanese Patent Laid-Open No. 7-274832 JP 2006-063033 A JP 2007-217352 A JP 2006-265232 A JP 2003-146837 A JP 2005-022993 A JP 2006-056854 A JP 2009-191039 A JP 2010-184915 A JP 2006-219407 A JP 2002-201222 A Japanese Patent Laid-Open No. 2003-055242 JP 2003-012532 A JP 2014-003929 A

“The Journal of Japanese Pharmacology”, 2005, Vol.125, p.379-384 J. Cell. Biol., 1992, Vol.199, p.695-703 J. Invest. Dermatol., 1979, Vol. 73, p.59-66 Nature, 1996, Vol.379, p.335-339 “Angiogenesis and Matrix Metalloproteinases”, 120th Symposium of the Japan Medical Association Symposium Basics and Clinic of Angiogenesis, December 13, 2001, p.43-49 "Fragrance Journal", 2006, Vol.34, p.19-23 Trends Genet., 1992, Vol. 8, p.55-61 "Pharmacology Atlas", translated by Takehiko Fukuhara, Bunkodo, 1995, p.184 J. Sci. Food Agric., 2012, Vol.92, pp.2291-2296

  The present invention includes anti-obesity action, cyclic AMP phosphodiesterase activity inhibition action, dipeptidyl peptidase IV activity inhibition action, anti-aging action, hair growth action, anti-androgen action, whitening action, or anti-inflammatory among compounds derived from natural products. An anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a dipeptidyl peptidase IV activity inhibitor, an anti-aging agent, a hair restorer, an anti-androgen agent, a whitening agent and an anti-obesity agent An object of the present invention is to provide an inflammatory agent and skin cosmetics, hair cosmetics and foods and drinks containing the compound.

  In order to solve the above problems, the anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, dipeptidyl peptidase IV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent of the present invention Is characterized by containing dihydroferulic acid as an active ingredient. Moreover, the skin cosmetics, hair cosmetics, and foods and beverages of the present invention are characterized by blending dihydroferulic acid.

  According to the present invention, by using dihydroferulic acid as an active ingredient, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a dipeptidyl peptidase IV activity inhibitor, an anti-aging agent, a hair-growth agent, an anti-antibacterial agent having excellent action and effect. Male hormone agents, whitening agents, and anti-inflammatory agents can be provided. Furthermore, the skin cosmetics, hair cosmetics, and food-drinks excellent in the said effect | action can be provided by mix | blending dihydro ferulic acid.

Embodiments of the present invention will be described below.
The anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, dipeptidyl peptidase IV activity inhibitor, anti-aging agent, hair restorer, anti-androgen, whitening agent, and anti-inflammatory agent of this embodiment are effective dihydroferulic acid Ingredients. Moreover, the skin cosmetics, hair cosmetics, and foods and beverages of this embodiment contain dihydroferulic acid.

  Dihydroferulic acid is a cinnamic acid derivative having a chemical structure represented by the following formula.

  For example, dihydroferulic acid is obtained by fermenting a ferulic acid derivative such as ferulic acid or ethyl ferulate, or a composition containing these (for example, a plant fragment or extract) by a microorganism having a phenolic acid reductase. It can also be produced by converting ferulic acid to dihydroferulic acid and then extracting, isolating and purifying the obtained fermentation product. Examples of the composition containing ferulic acid include crushed and extract of plants such as coffee, wheat, corn, tomato, mate, mugwort, and burdock. Furthermore, since ferulic acid is a constituent component of lignin in woody plants, lignin or a composition containing the same may be used as the composition containing ferulic acid. On the other hand, microorganisms having a phenolic acid reductase include, for example, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus delbrueckina, Lactobacillus Lactic acid bacteria.

  The method for extracting, isolating and purifying dihydroferulic acid from the plant or fermented product is not particularly limited, and can be performed 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 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 dihydroferulic acid from the extract obtained as described above, the concentrate of the extract or the dried product of the extract is not particularly limited, and should be performed by a conventional method. Can do. 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. And a method for recovering a fraction containing. 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. Furthermore, the fraction containing dihydroferulic acid obtained by column chromatography can be used for any of the reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using ion exchange resin, etc. The organic compound purification means may be used for purification.

[Anti-obesity agents, cyclic AMP phosphodiesterase activity inhibitors, dipeptidyl peptidase IV activity inhibitors, anti-aging agents, hair restorers, anti-androgen agents, whitening agents, anti-inflammatory agents]
The dihydroferulic acid obtained as described above has excellent anti-obesity action, cyclic AMP (cAMP) phosphodiesterase activity inhibitory action, dipeptidyl peptidase IV (DPPIV) activity inhibitory action, anti-aging action, hair growth action, anti-androgen hormone Anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent due to its action, whitening action, and anti-inflammatory action It can be used as an active ingredient. The anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent of this embodiment are pharmaceuticals, quasi drugs, cosmetics, etc. Can be used for a wide range of applications.

  In addition, as an active ingredient of the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent according to this embodiment, isolated dihydro Instead of ferulic acid, a composition containing dihydroferulic acid may be used. Here, the “composition containing dihydroferulic acid” in the present embodiment includes an extract obtained by using a plant containing dihydroferulic acid as an extraction raw material, a fermented product containing dihydroferulic acid, and the fermented product. An extract obtained 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 active ingredient of the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent according to this embodiment, dihydroferulic acid is contained. When using the composition, it is preferable to use a composition that has been purified to increase the purity of dihydroferulic acid. Anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair-restoration agent, anti-androgen, which are more excellent in action and effect by using a compound with increased purity of dihydroferulic acid as an active ingredient Agents, whitening agents, and anti-inflammatory agents can be obtained.

  The anti-obesity action of dihydroferulic acid is, for example, one or more selected from the group consisting of cAMP phosphodiesterase activity inhibitory action, blood cholesterol reducing action, blood free fatty acid reducing action, and DPPIV activity inhibiting action Demonstrated based on action. However, the anti-obesity action of dihydroferulic acid is not limited to the anti-obesity action exhibited based on the above action.

  In addition, dihydroferulic acid uses its blood lipid reducing action (particularly blood cholesterol reducing action and blood free fatty acid reducing action) to reduce blood lipid reducing agent, blood cholesterol reducing agent or blood free fatty acid. You may use as an active ingredient of an agent.

  Examples of the anti-aging action of dihydroferulic acid include laminin 5 production promoting action, matrix metalloproteinase-2 (MMP-2) activity inhibiting action, hyaluronic acid production promoting action, epidermal keratinocyte proliferation promoting action, elastin production promoting action 1 selected from the group consisting of a matrix metalloproteinase-1 (MMP-1) activity inhibitory action, a fibroblast proliferation promoting action, an aquaporin 3 (AQP3) mRNA expression promoting action, and a final glycation product (AGEs) formation inhibitory action It is exhibited based on the action of two or more species. However, the anti-aging action of dihydroferulic acid is not limited to the anti-aging action exhibited based on the above action.

  Further, dihydroferulic acid has its laminin 5 production promoting action, MMP-2 activity inhibiting action, hyaluronic acid production promoting action, epidermal keratinocyte proliferation promoting action, elastin production promoting action, MMP-1 activity inhibiting action, fibroblasts Laminin 5 production promoter, MMP-2 activity inhibitor, hyaluronic acid production promoter, epidermal keratinocyte proliferation promoter, elastin production promotion using growth promoting action, AQP3 mRNA expression promoting action, or AGEs formation inhibitory action, respectively It may be used as an active ingredient of an agent, an MMP-1 activity inhibitor, a fibroblast proliferation promoter, an AQP3 mRNA expression promoter, or an AGEs formation inhibitor.

  The hair growth effect of dihydroferulic acid 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 dihydroferulic acid has is not limited to the hair growth action exhibited based on the said action. Further, dihydroferulic acid may be used as an active ingredient of a testosterone 5α-reductase activity inhibitor or a hair papilla cell proliferation promoter, respectively, utilizing its testosterone 5α-reductase activity inhibitory action or hair papilla cell proliferation promoting action. Good.

  The antiandrogenic action of dihydroferulic acid is exhibited based on, for example, the testosterone 5α-reductase activity inhibitory action. However, the antiandrogenic action of dihydroferulic acid is not limited to the antiandrogenic action exerted based on the above action.

  The whitening action of dihydroferulic acid is exhibited based on, for example, a melanin production inhibitory action. However, the whitening action of dihydroferulic acid is not limited to the whitening action exhibited based on the above action. In addition, dihydroferulic acid may be used as an active ingredient of a melanin production inhibitor using its melanin production inhibitory action.

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

  The anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, antiandrogen, whitening agent, or anti-inflammatory agent of this embodiment contains dihydroferulic acid or dihydroferulic acid It may consist of only the composition, or may be prepared by formulating dihydroferulic acid or a composition containing dihydroferulic acid.

  The anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent of this embodiment are pharmaceutically acceptable such as dextrin and cyclodextrin. It can be formulated into an arbitrary dosage form such as powder, granule, tablet, liquid, etc. according to a conventional method using a carrier and other optional auxiliaries. 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-obesity agents, cAMP phosphodiesterase activity inhibitors, DPPIV activity inhibitors, anti-aging agents, hair restorers, anti-androgen agents, whitening agents, and anti-inflammatory agents may be used in other compositions (eg, skin cosmetics, hair cosmetics). Etc.), and can also be used as an ointment, a solution for external use, a patch and the like.

  When the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, antiandrogen, whitening agent, or anti-inflammatory agent of the present embodiment is formulated, dihydroferulic acid or dihydroferula Content of the composition containing an acid is not specifically limited, According to the objective, it can set suitably.

  In addition, the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen hormone agent, whitening agent, or anti-inflammatory agent of this embodiment may have an anti-obesity action as necessary. , CAMP phosphodiesterase activity inhibitory action, DPPIV activity inhibitory action, anti-aging action, hair growth action, anti-androgenic action, whitening action, or other natural extracts having anti-inflammatory action, containing dihydroferulic acid or dihydroferulic acid It can mix | blend with the composition to perform and can be used as an active ingredient.

  As an administration method to the patient of the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, or anti-inflammatory agent of this embodiment, transdermal administration, Although oral administration etc. are mentioned, What is necessary is just to select suitably the method suitable for the prevention or treatment etc. according to the kind of disease. In addition, the dose of the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen hormone agent, whitening agent, or anti-inflammatory agent of this embodiment is also determined depending on the type of disease, severe It may be appropriately increased or decreased depending on the degree, individual patient differences, administration method, administration period, and the like.

  The anti-obesity agent of the present embodiment is selected from the group consisting of cAMP phosphodiesterase activity inhibitory action, blood cholesterol reduction action, blood free fatty acid reduction action, and DPPIV activity inhibition action possessed by dihydroferulic acid as an active ingredient Through the action of two or more species, it can promote cAMP production, promote fat breakdown in adipocytes, reduce blood lipid levels, and increase in vivo half-life of incretin can do. As a result, the anti-obesity agent of this embodiment can prevent or ameliorate various diseases such as obesity, arteriosclerosis associated with it, diabetes, and metabolic syndrome. However, the anti-obesity agent of the present embodiment has a blood cholesterol reducing action or blood free fatty acid reducing action in addition to the above-described uses, in addition to the use as a cAMP phosphodiesterase activity inhibitor or DPPIV activity inhibitor described later. It can be used for all purposes that are meaningful to exert.

  For example, the anti-obesity agent of the present embodiment or the blood lipid-reducing agent, blood cholesterol-reducing agent, or blood free fatty acid-reducing agent described above is not dyslipidemic due to obesity (for example, blood cholesterol or blood Symptoms with high free fatty acids) can be improved, and various diseases such as arteriosclerosis, diabetes, and metabolic syndrome without obesity can be prevented or improved.

  The cAMP phosphodiesterase activity inhibitor of the present embodiment promotes cAMP production through the cAMP phosphodiesterase activity inhibitory action of dihydroferulic acid, which is an active ingredient, and thus can suppress aggregation of platelets. Various inflammatory diseases and the like can be prevented, treated or improved. In addition, the cAMP phosphodiesterase activity inhibitor of the present embodiment can promote fat degradation in adipocytes through the cAMP phosphodiesterase activity inhibitory action of dihydroferulic acid, resulting in obesity and arteriosclerosis associated therewith. In addition, 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 DPPIV activity inhibitor of this embodiment can prevent or treat type 2 diabetes, obesity, hypertension, insulin resistance, etc. through the DPPIV activity inhibitory action of dihydroferulic acid, and autoimmunity such as rheumatoid arthritis. Diseases and transplant rejection can be prevented or treated. However, the DPPIV activity inhibitor of the present embodiment can be used for all purposes that are meaningful in exhibiting the DPPIV activity inhibitory action in addition to these uses. For example, the DPPIV activity inhibitor of the present embodiment allows neuropathy (for example, sciatica, Alzheimer's disease, depression, etc.) such as pain, neurodegenerative disease, and neuropsychiatric disorder through the DPPIV activity inhibitory action of dihydroferulic acid; Growth hormone deficiency and diseases in which growth hormone is 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.

  The anti-aging agent of the present embodiment is a laminin 5 production promoting action, an MMP-2 activity inhibiting action, a hyaluronic acid production promoting action, an epidermal keratinocyte proliferation promoting action, an elastin production promoting action, which dihydroferulic acid as an active ingredient has, Through the action of one or more selected from the group consisting of MMP-1 activity inhibitory action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, and AGEs formation inhibitory action, formation of skin wrinkles, elasticity It is possible to prevent, treat or ameliorate skin aging symptoms such as a decrease or a decrease in moisture retention function. However, in addition to these uses, the anti-aging agent of the present embodiment is capable of promoting laminin 5 production, inhibiting MMP-2 activity, promoting hyaluronic acid production, promoting epidermal keratinocyte proliferation, promoting elastin production, MMP, -1 activity inhibiting action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action or AGEs formation inhibiting action can be used for all purposes that are meaningful.

  For example, the anti-aging agent of this embodiment or the laminin 5 production promoter described above can promote the production of laminin 5 by the action of dihydroferulic acid. Thereby, reconstruction of a basement membrane structure can be induced | guided | derived and the wound in 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 use described above, the anti-aging agent of the present embodiment, the MMP-2 activity inhibitor, or the MMP-1 activity inhibitor described above is an MMP-2 activity inhibitory action or an MMP-1 activity inhibitory action of dihydroferulic acid. 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-mentioned uses, the anti-aging agent of the present embodiment or the hyaluronic acid production promoter described above has rheumatoid arthritis, osteoarthritis, purulent arthritis, gouty arthritis, traumatic arthritis due to its hyaluronic acid production promoting action. , Prevention, treatment or amelioration of arthritis such as osteoarthritis; promotion of wound or burn healing; In addition, the anti-aging agent of the present embodiment or the above-described epidermal keratinocyte proliferation promoter restores skin metabolism through the epidermal keratinocyte proliferation promoting action of dihydroferulic acid, causing wrinkles, dullness, pigmentation, etc. Can be used for prevention, treatment or amelioration; regenerative medicine;

  In addition to the above-mentioned uses, the anti-aging agent of the present embodiment or the above-mentioned elastin production promoting agent prevents or treats pulmonary diseases such as emphysema; vascular diseases such as hypertension and aneurysms; Or it can use for the use of improvement. In addition, the anti-aging agent of the present embodiment or the above-mentioned fibroblast proliferation promoter promotes healing of wounds such as trauma and burn through the fibroblast proliferation promoting action of dihydroferulic acid; skin diseases (for example, pressure sores, It can be used for applications such as treatment or improvement of burn ulcers, skin ulcers such as diabetic ulcers, and regenerative medicine.

  In addition to the above-described uses, the anti-aging agent of the present embodiment or the above-described AQP3 mRNA expression promoter can improve water retention ability and barrier function due to aging through the AQP3 mRNA expression promoting action of dihydroferulic acid.

  In addition to the above-mentioned uses, the anti-aging agent of the present embodiment or the above-described AGEs formation inhibitor is capable of diabetic neuropathy, diabetic retinopathy, diabetic nephropathy and other diabetes through the AGEs formation inhibitory action of dihydroferulic acid. Complications; arteriosclerosis caused by protein glycation reaction; osteoporosis, osteoarthritis, etc. caused by protein glycation reaction 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.

  The hair-restoring agent of the present embodiment is a hair loss such as male pattern alopecia, alopecia areata, tricotylonia, etc. through a testosterone 5α-reductase activity inhibiting action and / or a hair papillary cell proliferation promoting action possessed by dihydroferulic acid as an active ingredient. 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 aforementioned hair dermal papilla cell proliferation promoter activates the hair dermal papilla cells through the hair dermal papilla cell proliferation promotion action of dihydroferulic acid, and the growth and differentiation of hair follicle epithelial cells and hair Can be promoted, the transition from the growth phase to the regression phase and the resting phase can be prevented in the hair cycle, and the growth phase can be extended. Thereby, alopecia can be prevented or improved. Further, the hair growth agent of the present embodiment or the hair dermal papilla cell proliferation promoter described above is used for application to the regenerative medicine field such as hair regeneration using hair dermal papilla cells through the dermal papilla cell proliferation promotion action of dihydroferulic acid. You can also

  The anti-androgen agent of the present embodiment or the testosterone 5α-reductase activity inhibitor described above is a disease involving male hormones, such as male pattern hair loss, through the testosterone 5α-reductase activity inhibitory action of dihydroferulic acid as an active ingredient. Can prevent, treat or ameliorate symptom, hirsutism, seborrhea, acne (acne etc.), benign prostatic hyperplasia, prostate tumor, premature boyhood. 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 whitening agent of this embodiment can prevent or improve pigmentation such as skin darkening, spots, and freckles through the melanin production inhibitory action of dihydroferulic acid, which is an active ingredient. However, the whitening agent of this embodiment can be used for all uses other than these uses, which are meaningful for exhibiting a melanin production inhibitory action.

  The anti-inflammatory agent of this embodiment is one or two selected from the group consisting of a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, and a COX-2 activity inhibitory action of dihydroferulic acid as an active ingredient Through the above actions, contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, and other various skin inflammatory diseases associated with rough skin can be prevented, treated or improved. However, the anti-inflammatory agent of the present embodiment is used for all uses that are meaningful for exerting a hyaluronidase activity inhibitory action, a hexosaminidase release inhibitory action, or a COX-2 activity inhibitory action in addition to these uses. be able to.

  For example, the anti-inflammatory agent of the present embodiment or the hyaluronidase activity inhibitor, hexosaminidase release inhibitor, or COX-2 activity inhibitor described above is an inhibitory action of dihydroferulic acid on hyaluronidase activity, inhibition of hexosaminidase release. Rheumatoid arthritis, osteoarthritis, asthma and the like can be prevented, treated or ameliorated through the action or the COX-2 activity inhibitory action. Further, the anti-inflammatory agent of the present embodiment or the aforementioned hexosaminidase release inhibitor prevents or treats gastric ulcers, sleep disorders, etc. caused by excessive gastric acid through the action of inhibiting hexosaminidase release of dihydroferulic acid. Or can be improved.

  In addition, the anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, or anti-inflammatory agent of the present embodiment has an excellent anti-obesity action, cAMP phosphodiesterase Since it has activity inhibitory action, DPPIV activity inhibitory action, anti-aging action, hair growth action, anti-androgenic action, whitening action, or anti-inflammatory action, it is suitable for blending into, for example, a skin external preparation. In this case, a dihydroferulic acid or a composition containing dihydroferulic acid may be blended as it is, or an antiobesity agent and a cAMP phosphodiesterase activity inhibitor formulated from the composition containing dihydroferulic acid or dihydroferulic acid. , DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen hormone agent, whitening agent, or anti-inflammatory agent 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-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, or anti-inflammatory agent of the present embodiment has an excellent anti-obesity action, cAMP phosphodiesterase Since it has activity inhibitory action, DPPIV activity inhibitory action, anti-aging action, hair growth action, anti-androgenic action, whitening action, or anti-inflammatory action, it can be suitably used as a reagent for research on these action mechanisms it can.

[Skin cosmetics, hair cosmetics]
Dihydroferulic acid has an excellent anti-aging effect, hair-growth effect, anti-androgen action, whitening effect, anti-inflammatory action, cAMP phosphodiesterase activity inhibitory action, DPPIV activity inhibitory action, and anti-obesity action. It is suitable for blending into cosmetics or hair cosmetics. In this case, a dihydroferulic acid or a composition containing dihydroferulic acid may be blended as it is, or an anti-aging agent, hair restorer, anti-androgen prepared from the composition containing dihydroferulic acid or dihydroferulic acid An agent, whitening agent, anti-inflammatory agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, or anti-obesity agent may be added.

  Skin cosmetics or hair by blending dihydroferulic acid or the above-mentioned anti-aging agent, hair restorer, anti-androgen agent, whitening agent, anti-inflammatory agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, or anti-obesity agent Anti-aging effect on cosmetics, laminin 5 production promotion effect, MMP-2 activity inhibition effect, hyaluronic acid production promotion effect, epidermal keratinocyte proliferation promotion effect, elastin production promotion effect, MMP-1 activity inhibition effect, fibroblast proliferation Promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, hair papillary cell proliferation promoting action, anti-androgen action, whitening action, melanin production inhibitory action, anti-inflammatory action, hyaluronidase activity Inhibitory action, hexosaminidase release inhibitory action, COX-2 activity Adverse effects, cAMP phosphodiesterase inhibitory activity, DPPIV inhibitory activity, anti-obesity effect can be imparted to blood cholesterol lowering action, or blood free fatty acid lowering action.

  All of these effects exert favorable actions when applied to skin cosmetics or hair cosmetics. Among them, anti-aging action, laminin 5 production promoting action, MMP-2 activity inhibiting action, hyaluron, among others Acid production promoting action, epidermal keratinocyte proliferation promoting action, elastin production promoting action, MMP-1 activity inhibiting action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibiting action, whitening action, melanin production inhibiting action, Anti-inflammatory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, testosterone 5α-reductase activity inhibitory action, or antiandrogen action is imparted to skin cosmetics Then, since those actions are easily exhibited, 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 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, laminin 5 production promoting action, hyaluronic acid production promoting action, elastin production promoting action, fibroblast proliferation promoting action, or AGEs formation inhibiting action is imparted to hair cosmetics, Since these functions are easily exhibited, it is particularly suitable.

  Skin cosmetics or hair cosmetics that may contain dihydroferulic acid, or the above-mentioned anti-aging agent, hair restorer, anti-androgen agent, whitening agent, anti-inflammatory agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, or anti-obesity agent The type of the preparation is not particularly limited, and examples of the skin cosmetics include ointments, creams, emulsions, lotions, packs, foundations, and the like, and examples of hair cosmetics include, for example, hair tonics, hair Examples include cream, hair liquid, shampoo, pomade, rinse and the like.

  Combination of dihydroferulic acid or the above-mentioned anti-aging agent, hair restorer, antiandrogen, whitening agent, anti-inflammatory agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor or anti-obesity agent in skin cosmetics or hair cosmetics In this case, the blending amount can be appropriately adjusted according to the type of skin cosmetic or hair cosmetic, but the preferred blending ratio is about 0.0001 to 10% by mass, and the particularly preferred blending ratio. Is about 0.001 to 1% by mass in terms of standard extract.

  The skin cosmetic or hair cosmetic of the present embodiment includes an anti-aging action, laminin 5 production promoting action, MMP-2 activity inhibiting action, hyaluronic acid production promoting action, epidermal keratinocyte proliferation promoting action, elastin, which dihydroferulic acid has. Production promoting action, MMP-1 activity inhibiting action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibiting action, hair growth action, testosterone 5α-reductase activity inhibiting action, hair papillary cell proliferation promoting action, anti-androgenic hormone action Whitening action, melanin production inhibitory action, anti-inflammatory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, DPPIV activity inhibitory action, anti-obesity action, blood Cholesterol-reducing action or blood free fatty acid reduction Main ingredients, 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, cell activation Agents, 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 of this embodiment has an anti-aging action, laminin 5 production promoting action, MMP-2 activity inhibiting action, hyaluronic acid production promoting action, epidermal keratinocyte proliferation promoting action, elastin production promoting action, which dihydroferulic acid has, MMP-1 activity inhibitory action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibitory action, whitening action, melanin production inhibitory action, anti-inflammatory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX -2 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, hair papilla cell proliferation promoting action, anti-androgen action, DPPIV activity inhibitory action, anti-obesity action, blood cholesterol reducing action, And from the group consisting of blood free fatty acid reducing action Prevention, treatment or amelioration of skin aging symptoms such as skin wrinkle formation, reduced elasticity, reduced moisturizing function through one or more selected actions; accelerated healing of wounds or burns; rough skin Prevention, treatment or amelioration of dry skin diseases (eg, atopic dermatitis, psoriasis, ichthyosis, etc.); prevention, treatment or improvement of skin darkening, pigmentation such as spots, buckwheat Prevention, treatment or improvement of contact dermatitis (rash), psoriasis, pemphigus vulgaris and other skin inflammatory diseases associated with rough skin; prevention, treatment or improvement of seborrhea, acne (acne etc.); Prevention, treatment or amelioration of obesity and associated lifestyle-related diseases such as arteriosclerosis, diabetes, and metabolic syndrome.

  In addition, the hair cosmetic composition of the present embodiment has a hair growth action, a testosterone 5α-reductase activity inhibitory action, a hair papillary cell proliferation promoting action, an antiandrogen action, an anti-inflammatory action, a hyaluronidase activity inhibitory action, hexosa, which dihydroferulic acid has. Minidase release inhibitory action, COX-2 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, anti-aging action, laminin 5 production promoting action, MMP-2 activity inhibiting action, hyaluronic acid production promoting action, epidermal keratinocyte proliferation promoting action, Elastin production promoting action, MMP-1 activity inhibiting action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibiting action, whitening action, melanin production inhibiting action, DPPIV activity inhibiting action, anti-obesity action, blood cholesterol reduction Action and blood free fatty acid reducing action Prevention, treatment or amelioration of alopecia such as androgenetic alopecia, alopecia areata, trichotillomania, etc. through one or more selected actions; rough skin, dry skin, etc., dry skin disease ( For example, treatment of atopic dermatitis, psoriasis, ichthyosis, etc .; prevention, treatment or amelioration of contact dermatitis (rash), psoriasis, pemphigus vulgaris and other skin inflammatory diseases associated with rough 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]
Dihydroferulic acid has excellent anti-obesity action, cAMP phosphodiesterase activity inhibition action, DPPIV activity inhibition action, anti-aging action, hair growth action, anti-androgen action, whitening action, and anti-inflammatory action. It is suitable for blending. In this case, dihydroferulic acid may be blended as it is, or antiobesity agent formulated from dihydroferulic acid, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen, whitening An agent and an anti-inflammatory agent may be added.

  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.

  By incorporating dihydroferulic acid or the above-mentioned anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, anti-inflammatory agent, or whitening agent into food and drink, anti-obesity Action, blood cholesterol reduction action, blood free fatty acid reduction action, cAMP phosphodiesterase activity inhibition action, DPPIV activity inhibition action, anti-aging action, laminin 5 production promotion action, MMP-2 activity inhibition action, hyaluronic acid production promotion action, epidermis Keratinocyte growth promoting action, elastin production promoting action, MMP-1 activity inhibiting action, fibroblast proliferation promoting action, AQP3 mRNA expression promoting action, AGEs formation inhibiting action, hair growth action, testosterone 5α-reductase activity inhibiting action, hair papillary cell Growth-promoting action, antiandrogen action, anti-inflammatory action Hyaluronidase inhibitory effect, hexosaminidase release inhibitory effect, COX-2 inhibitory activity, can impart whitening effect, or the melanin production inhibitory effect on food products. These actions are suitable because they are easily exerted by being applied to food and drink.

  Dihydroferulic acid, or anti-obesity agent formulated from dihydroferulic acid, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, antiandrogen, whitening agent, or anti-inflammatory agent in food and drink When blended, the amount of the active ingredient in them can be appropriately changed in consideration of the purpose of use, symptoms, sex, etc., but in consideration of the general intake of food and drink to be added, adults 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, dihydroferulic acid or an anti-obesity agent formulated from dihydroferulic acid, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging The addition amount of the agent, hair restorer, anti-androgen hormone agent, whitening agent, or anti-inflammatory agent is usually 0.1 to 100% by mass, preferably 5 to 100% by mass, with respect to the food to be added.

  The food / beverage products of this embodiment may be blended with any food / beverage product that does not hinder its activity, or may be a dietary supplement based on dihydroferulic acid. .

  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.

  Foods and drinks that can be mixed with dihydroferulic acid are not particularly limited, but specific examples thereof include beverages such as soft drinks, carbonated drinks, nutrition drinks, fruit drinks, and lactic acid drinks (concentrated concentrates and powders for adjustment of these drinks). Ice cream, ice sherbet, shaved ice, etc .; noodles such as buckwheat, udon, harusame, gyoza skin, shumai skin, chinese noodles, instant noodles; rice cake, chewing gum, candy, gum, chocolate, tablet confectionery, Sweets such as snacks, biscuits, jelly, jam, cream, baked goods; processed fishery and livestock products such as kamaboko, ham, sausage; dairy products such as processed milk, fermented milk; salad oil, tempura oil, margarine, mayonnaise, shortening , Whipped cream, oils and fats such as dressings and processed foods; seasonings such as sauces and sauces; Various types of health and nutritional supplements; tablets, capsules, drinks, etc., usually used when blending dihydroferulic acid with these foods and drinks Supplementary raw materials and additives to be used can be used in combination.

  The anti-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, anti-inflammatory agent, skin cosmetics, hair cosmetics, and food and drink of this embodiment The product is preferably applied to humans, but as long as each effect is exhibited, animals other than humans (for example, mice, rats, hamsters, dogs, cats, cows, 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, dihydroferulic acid (manufactured by Tokyo Chemical Industry Co., Ltd., sample 1) was used as a test sample.

[Test Example 1] Cyclic AMP phosphodiesterase activity inhibitory action test Dihydroferulic acid (Sample 1) was tested for cyclic AMP phosphodiesterase activity inhibitory action 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 a test sample solution (sample 1, sample concentration: see Table 1 below) 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 condition 3. Further, as a control, the same operation was performed by adding only the solvent without the sample.

<High performance liquid chromatography condition 3>
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 1.

  As shown in Table 1, it was confirmed that dihydroferulic acid (sample 1) has an excellent cyclic AMP phosphodiesterase activity inhibitory action.

[Test Example 2] Blood lipid reducing action test Dihydroferulic acid (Sample 1) was tested for blood lipid reducing action as follows.

  After 4 weeks old TSOD mice (purchased from Shimizu Experimental Materials Co., Ltd.) were fully quarantined, the general condition was observed and the body weight was measured, and animals with good health were selected and used at 5 weeks of age. Animals are housed in plastic cages under conditions of temperature: 22 ± 3 ° C, humidity: 55 ± 15%, ventilation: always fresh, 12 hours lighting / day (6am to 6pm). And raised. The feed was freely inoculated with laboratory animal solid feed lab MR stock (manufactured by Nippon Agricultural Industry Co., Ltd.), and drinking water was ingested freely with an automatic water supply device.

Five-week-old mice were divided into a dihydroferulic acid 50 mg / kg / day administration group and a control group (each group consisted of 7 mice, 14 mice in total). Intragastric administration once a day using a sonde And kept for 7 weeks. Fasting was performed for 16 hours from the day before the end of the test, and blood was collected under isoflurane anesthesia at the end of the test. Plasma was prepared from the collected blood by centrifugation (3,000 rpm, 10 minutes), and the amounts of total cholesterol and free fatty acids were measured. In addition, DRI-CHEM3030 (made by Fujifilm) was used for the measurement of total cholesterol (T-CHO), and NEFA-C test Wako (made by Wako Pure Chemical Industries) was used for the measurement of free fatty acid (NEFA). .
The results are shown in Table 2.

  As shown in Table 2, it was confirmed that dihydroferulic acid (sample 1) is excellent in blood cholesterol reducing action and blood free fatty acid reducing action.

Test Example 3 DPPIV Activity Inhibitory Action Test Dihydroferulic acid (Sample 1) was tested for dipeptidyl peptidase IV (DPPIV) inhibitory action as follows.

  In a 96-well plate, 25 μL of a test sample prepared with 25 mM Tris-HCl buffer (pH 8.0) (sample 1, refer to Table 3 below for final concentration) and 0.4 μg / ml prepared with the above buffer 25 mL of mL DPPIV (rhCD26, manufactured by R & D System) 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 without addition of sample / addition of enzyme B: Absorbance at a wavelength of 415 nm without addition of sample / addition of enzyme C: Absorbance at wavelength of 415 nm with addition of test sample / addition of enzyme D: Addition of test sample / enzyme Absorbance at a wavelength of 415 nm without addition is shown in Table 3.

  As shown in Table 3, dihydroferulic acid (Sample 1) showed an excellent DPPIV inhibitory action.

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

Human normal neonatal epidermal keratinocytes (NHEK) were precultured in an 80 cm ² flask using human normal epidermal keratinocyte medium (KGM) under conditions of 37 ° C. and 5% CO ₂ -95% air. Cells were collected by trypsinization. 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, see Table 4 below for each sample concentration) was added to each well, and 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 used for monoclonal anti-human laminin 5 antibody (mouse IgG, manufactured by Chemicon). It was measured by the ELISA method. 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: Absorbance at a wavelength of 405 nm when a test sample is added B: Absorbance at a wavelength of 405 nm when no sample is added Table 4 shows the results.

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

[Test Example 5] 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 resulting enzyme solution, dihydroferulic acid (Sample 1) was tested for MMP-2 activity inhibitory action as follows.

Using a 96-well plate for fluorescence intensity measurement, 20 μL of a test sample (sample 1, see the following table 5 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 420 nm 120 minutes after addition of substrate without addition of sample B: Fluorescence intensity at 420 nm immediately after addition of substrate without addition of sample C: Fluorescence intensity at 420 nm 120 minutes after addition of substrate with addition of test sample D : Fluorescence intensity at 420 nm immediately after addition of substrate in addition of test sample The results are shown in Table 5.

  As shown in Table 5, it was confirmed that dihydroferulic acid (sample 1) has an excellent MMP-2 activity inhibitory action.

[Test Example 6] Dermal hyaluronic acid production promoting action test Human normal dermal fibroblasts (NB1RGB) were cultured in an α-MEM medium containing 10% FBS, and the cells were collected by trypsin treatment. The collected cells were diluted with α-MEM medium containing 0.5% FBS to a cell density of 1.6 × 10 ⁵ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured overnight. .

  After completion of the culture, the medium was removed, and 100 μL of a test sample dissolved in 0.5% FBS-containing α-MEM medium (Sample 1, see Table 6 below for sample concentration) was added to each well and cultured for 3 days. In addition, it culture | cultivated similarly using (alpha) -MEM culture medium containing 0.5% FBS without a sample as control. After the culture, the amount of hyaluronic acid in the medium of each well was measured by a sandwich method using hyaluronic acid binding protein (HABP). From the obtained results, the hyaluronic acid production promotion rate (%) was calculated by the following formula.

Dermal hyaluronic acid production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Hyaluronic acid amount when test sample is added B: Hyaluronic acid amount when test sample is not added Table 6 shows the results.

  As shown in Table 6, it was confirmed that dihydroferulic acid (sample 1) has an excellent dermal hyaluronic acid production promoting action.

[Test Example 7] Epidermal keratinocyte proliferation promoting action test Dihydroferulic acid (sample 1) was examined for epidermal keratinocyte proliferation promoting action as follows.

Normal human newborn epidermal keratinocytes (NHEK) were cultured using normal human epidermal keratinocyte growth medium (KGM), and then cells were collected by trypsin treatment. The collected cells were diluted with KGM medium to a cell density of 3.0 × 10 ⁴ cells / mL, and then seeded at 100 μL per well in a collagen-coated 96-well plate and cultured overnight. After completion of the culture, 100 μL of a test sample dissolved in KGM medium (Sample 1, see Table 7 below for sample concentration) was added to each well and cultured for 3 days. As a control, the same culture was performed using a KMG medium without a sample.

  The epidermal keratinocyte proliferation promoting action was measured using the MTT assay. That is, after culturing for 3 days, the medium was removed, and 100 μL of MTT dissolved in PBS (−) buffer at a final concentration of 0.4 mg / mL was added. After culturing for 2 hours, blue formazan produced in the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. From the obtained results, the epidermal keratinocyte proliferation promotion rate (%) was calculated by the following formula.

Epidermal keratinocyte proliferation promotion rate (%) = St / Ct × 100
Each term in the formula represents the following.
St: Blue formazan production amount in cells to which test sample was added Ct: Blue formazan production amount in cells to which no sample was added Table 7 shows the results.

  As shown in Table 7, it was confirmed that dihydroferulic acid (sample 1) has an excellent epidermal keratinocyte proliferation promoting action.

[Test Example 8] Elastin production promoting action test Dihydroferulic acid (sample 1) was tested for elastin production promoting action as follows.

Human normal 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 2.2 × 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 0.25% FBS-containing Dulbecco's MEM medium (Sample 1, see Table 8 below for sample concentration) was added to each well and cultured for 2 days. As a control, the culture was similarly performed using Dulbecco's MEM medium containing 0.25% FBS without any sample. After completion of the culture, the supernatant was collected, and the amount of elastin released into the culture supernatant was measured by ELISA. From the measurement results, the elastin production promotion rate (%) was calculated by the following formula.

Elastin production promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Elastin amount at the time of adding the test sample B: Elastin amount at the time of not adding the sample Table 8 shows the results.

  As shown in Table 8, dihydroferulic acid (Sample 1) showed an excellent elastin production promoting action.

[Test Example 9] MMP-1 activity inhibitory action test Dihydroferulic acid (Sample 1) was tested for MMP-1 activity inhibitory action as follows.

  50 μL of 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 9 below for sample concentration) 50 μL of MMP-1 solution (Sigma, COLLAGENASE Type IV from Clostridium histolyticum) and 400 μL of Pz peptide solution (BACHEM Feinchemikalien AG, Pz-Pro-Leu-Gly-Pro-D-Arg-OH) 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 sample added / enzyme added D: Sample added / no enzyme added Table 9 shows the results of absorbance at a wavelength of 320 nm.

  As shown in Table 9, it was recognized that dihydroferulic acid (Sample 1) has an excellent MMP-1 activity inhibitory action.

[Test Example 10] Skin fibroblast proliferation promoting action test Dihydroferulic acid (sample 1) was tested for skin fibroblast proliferation promoting action 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 recovered cells were diluted with 5% FBS-containing α-MEM medium to a cell density of 7.0 × 10 ⁴ cells / mL, then seeded at 100 μL per well in a 96-well plate, and cultured overnight. After completion of the culture, the medium was removed, and 100 μL of a test sample dissolved in 5% FBS-containing α-MEM medium (sample 1, refer to Table 10 below for sample concentration) was added to each well and cultured for 3 days. In addition, it culture | cultivated similarly as control using the alpha-MEM culture medium containing 5% FBS without a sample.

  Fibroblast proliferation promoting action was measured using MTT assay. That is, after culturing for 3 days, the medium was removed, and 100 μL of MTT dissolved in PBS (−) buffer at a final concentration of 5 mg / mL was added to each well. After culturing for 2 hours, blue formazan produced in the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. From the obtained results, the fibroblast proliferation promotion rate (%) was calculated by the following formula.

Fibroblast proliferation promotion rate (%) = St / Ct × 100
Each term in the formula represents the following.
St: Blue formazan production amount with addition of test sample Ct: Blue formazan production amount without addition of sample Table 10 shows the results.

  As shown in Table 10, it was confirmed that dihydroferulic acid (Sample 1) has an excellent fibroblast proliferation promoting action.

[Test Example 11] Aquaporin 3 (AQP3) mRNA expression promoting action test With respect to dihydroferulic acid (sample 1), the AQP3 mRNA expression promoting action was tested as follows.

Human normal neonatal epidermal keratinocytes (NHEK) were precultured in an 80 cm ² flask using medium for normal human epidermal keratinocytes (KGM) under conditions of 37 ° C. and 5% CO ₂ -95% air. Cells were collected by trypsinization. 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 the culture, the medium was removed, and 2 mL of the KGM medium of the test sample dissolved in the KGM medium (Sample 1, see Table 11 below for each 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 when test sample is added B: Correction value when sample is not added Table 11 shows the results.

  As shown in Table 11, it was confirmed that dihydroferulic acid (Sample 1) has an excellent AQP3 mRNA expression promoting action.

[Test Example 12] AGEs formation inhibitory action test Dihydroferulic acid (Sample 1) was tested for the formation inhibitory action of final glycation products (AGEs) as follows.

  A 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.) was charged with 0.2M D (−)-ribose prepared in PBS (−) buffer and a test sample (Sample 1, see Table 14 for sample concentration). After adding the mixed solution to 100 μL, 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 in a positive control C: Absorbance at a wavelength of 405 nm when a test sample was added Table 12 shows the results.

  As shown in Table 12, dihydroferulic acid (Sample 1) exhibited an excellent AGE formation inhibitory action.

[Test Example 13] Testosterone 5α-reductase inhibitory action test With respect to dihydroferulic acid (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 (rat liver homogenate, manufactured by Oriental Yeast Co., Ltd.) 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 13 shows the results.

  As shown in Table 13, it was confirmed that dihydroferulic acid (Sample 1) has an excellent testosterone 5α-reductase inhibitory action.

[Test Example 14] Hair papillary cell proliferation promoting action test With respect to dihydroferulic acid (sample 1), hair papillary cell proliferation promoting action was tested as follows.

Normal human hair hair papilla cells (HFDPC, derived from male parietal region) were cultured using hair papilla cell growth medium (PCGM, manufactured by Toyobo Co., Ltd.) containing 1% FCS and growth additives, and then cells were treated with trypsin. Was recovered. 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. As a control, the same culture was performed using a serum-free DMEM medium containing no sample.

  Thereafter, the medium was removed, 200 μL of a test sample dissolved in serum-free DMEM medium (Sample 1, see Table 16 for sample concentration) was added to each well, and further cultured for 4 days. 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 14 shows the results.

  As shown in Table 14, it was recognized that dihydroferulic acid (Sample 1) has an excellent hair papillary cell growth promoting action.

[Test Example 15] Melanin production inhibitory action test on B16 melanoma cells Dihydroferulic acid (Sample 1) was tested for melanin production inhibitory action on B16 melanoma cells 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's MEM medium containing 10% FBS and 1 mmol / L theophylline (Sample 1, refer to Table 13 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 disrupted 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 cell viability, the cells were cultured in the same manner as described above, then the medium was removed, washed with 400 μL of PBS buffer, and dissolved in Dulbecco MEM containing 10% FBS at a final concentration of 0.05 mg / mL. 200 μL of the neutral red 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 540 nm without addition of sample D: Absorbance at 540 nm with addition of test sample Table 15 shows the results.

  As shown in Table 15, it was recognized that dihydroferulic acid (Sample 1) has an excellent melanin production inhibitory action.

[Test Example 16] Hyaluronidase activity inhibitory action test To 0.2 mL of a test sample dissolved in 0.1 mol / L acetate buffer (pH 3.5) (sample 1, sample concentration see Table 14 below), hyaluronidase solution (Type 0.1 mL of IV-S (from bovine testes), 400 NF units / mL, manufactured by SIGMA) was added and the mixture was 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 wavelength 585 nm of test sample solution Sb: Absorbance at wavelength 585 nm of test sample solution blank Ct: Absorbance at wavelength 585 nm of control solution Cb: Absorbance at wavelength 585 nm of control solution blank Table 16 shows the results.

  As shown in Table 16, it was confirmed that dihydroferulic acid (Sample 1) has an excellent hyaluronidase activity inhibitory action.

[Test Example 17] Hexosaminidase release inhibitory action test Dihydroferulic acid (Sample 1) was tested for hexosaminidase release inhibitory action 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 17.

  As shown in Table 17, it was confirmed that dihydroferulic acid (Sample 1) has an excellent hexosaminidase release inhibitory action.

[Test Example 18] 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, E. coli 0111; B4, manufactured by DIFCO) dissolved in Dulbecco MEM containing 10% FBS at a final concentration of 1 μg / mL was added and cultured for 16 hours. . 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 when test sample is added / LPS stimulated B: Prostaglandin E ₂ amount when sample is not added / LPS stimulated C: Prostaglandin E ₂ amount when sample is not added / LPS stimulated Is shown in Table 18.

  As shown in Table 18, it was confirmed that dihydroferulic acid (sample 1) has an excellent COX-2 activity inhibitory action in macrophages.

[Formulation Example 1]
An emulsion having the following composition was produced by a conventional method.
Dihydroferulic acid 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.
Dihydroferulic acid 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.
Dihydroferulic acid 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.
Dihydroferulic acid 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.
Dihydroferulic acid 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.
Dihydroferulic acid 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)>
Dihydroferulic acid 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-obesity agent, cAMP phosphodiesterase activity inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgen agent, whitening agent, and anti-inflammatory agent of the present invention prevent, treat or ameliorate skin aging symptoms; Promotion of wound or burn healing; prevention, treatment or amelioration of dry skin disease; prevention, treatment or improvement of alopecia, especially male pattern baldness; pigmentation of skin darkening, spots, freckles, etc. Prevention or improvement; prevention, treatment or improvement of type 2 diabetes, obesity, hypertension, insulin resistance, etc .; prevention or improvement of various skin inflammatory diseases; prevention, treatment or improvement of obesity;

Claims (12)

  A whitening agent comprising dihydroferulic acid as an active ingredient.   The whitening agent according to claim 1, wherein the dihydroferulic acid has a melanin production inhibitory action.   An anti-inflammatory agent comprising dihydroferulic acid as an active ingredient.   The dihydroferulic acid 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. 3. The anti-inflammatory agent according to 3.   A hair restorer comprising dihydroferulic acid as an active ingredient.   6. The dihydroferulic acid has one or more actions selected from the group consisting of testosterone 5α-reductase activity inhibitory action and / or hair papilla cell proliferation promoting action. Hair restorer.   An antiandrogenic agent comprising dihydroferulic acid as an active ingredient.   An anti-aging agent characterized by comprising dihydroferulic acid as an active ingredient (excluding anti-aging applications based on DPPH radical scavenging action).   The dihydroferulic acid promotes laminin 5 production, inhibits matrix metalloproteinase-2 activity, promotes hyaluronic acid production, promotes epidermal keratinocyte proliferation, promotes elastin production, inhibits matrix metalloproteinase-1 activity, fiber 9. The anti-aging agent according to claim 8, wherein the anti-aging agent has one or more actions selected from the group consisting of a blast cell proliferation promoting action, an aquaporin 3 mRNA expression promoting action, and a final glycation product formation inhibiting action. . A skin cosmetic containing dihydroferulic acid,
It is used for one or more uses selected from the group consisting of whitening use, anti-inflammatory use, anti-androgen use, and anti-aging use (excluding anti-aging use based on DPPH radical scavenging action). Skin cosmetics.   A hair cosmetic characterized by containing dihydroferulic acid. A food or drink containing dihydroferulic acid,
Used for one or more applications selected from the group consisting of whitening applications, anti-inflammatory applications, hair growth applications, anti-androgen applications, and anti-aging applications (excluding anti-aging applications based on DPPH radical scavenging action) Characteristic food and drink.