JP7253263B2 - Skin cosmetics, hair cosmetics, food and drink - Google Patents

Description

The present invention provides an anti-aging agent, a whitening agent, a hair restorer, an antiandrogenic agent, an anti-inflammatory agent, an anti-obesity agent, a cyclic AMP phosphodiesterase activity inhibitor, a glutathione decrease inhibitor, a dipeptidyl peptidase IV activity inhibitor, and a skin cosmetic. products, hair cosmetics, and food and drink.

Glutathione, a tripeptide consisting of three amino acids, glutamic acid, cysteine and glycine, is the compound with the major cysteine residue in cells. Glutathione in cells functions as a radical scavenger, regulates cell functions by oxidation-reduction, metabolizes foreign substances, and acts as an SH donor for various enzymes, and is also known as an antioxidant component against active oxygen and the like. Its action is believed to be derived from cysteine residues. However, it has been reported that the amount of intracellular glutathione is depleted or decreased due to excessive oxidative stress, addition of foreign substances, aging, etc. This reduces the defense ability of cells against oxidative stress, and increases the DNA content of cells. and is thought to be one of the causes of damage to constituents such as proteins.

Diseases in which a decrease or deficiency in the amount of intracellular glutathione is known to be associated with pathology include, in addition to a group of diseases induced by oxidative stress, liver disorders (excessive alcohol consumption, or caused by ingestion of foreign substances such as heavy metals and chemical substances). That is, promoting the production of glutathione is thought to increase the ability of cells to defend against oxidative stress and prevent or treat the above-mentioned groups of diseases caused by a decrease or deficiency in the amount of intracellular glutathione. . Liquiritigenin (see Patent Literature 1) and the like are known to have a glutathione production promoting effect.

The epidermis has the function of relieving external stimuli and controlling the loss of body components such as water. It has a four-layer structure starting from the basal layer, which is the lowest layer, followed by the stratum spinosum, the stratum granulosum, and the stratum corneum. consists of Most of the cells present in each layer are keratinocytes differentiated from the basal layer. The keratinocytes, which divide and proliferate in the basal layer, differentiate while passing through the stratum spinosum and the stratum granulosum to become keratinocytes. Eventually, it falls off from the stratum corneum as dirt.

The stratum corneum is the outermost layer of the skin and serves as a physical barrier against external stimuli. In order for the skin to have this barrier function, the cycle (keratinization) from the production of keratinocytes in the basal layer to the exfoliation of the skin as dirt (keratinization) is repeated in a cycle of 4 weeks, and the metabolism of the epidermis is carried out. . However, this stratum corneum also loses its metabolic function with aging, and causes skin troubles such as wrinkles, dullness, pigmentation, and rough skin. Therefore, it is thought 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, earth shell mother extract (see Patent Literature 2) and the like are known as substances having epidermal keratinocyte proliferation-promoting action.

In recent years, involvement of matrix metalloproteinases (MMPs) has been pointed out as a factor that induces skin aging-related changes. Among these MMPs, matrix metalloprotease-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 major constituents of the basement membrane. It is clear that the expression and activity of MMP-2 are significantly increased by UV irradiation, causing a decrease in basement membrane components and structural changes in the basement membrane due to UV irradiation, and a major factor in the formation of wrinkles and sagging in the skin. (see Non-Patent Document 1). Increased activity of MMP-2 destroys the extracellular matrix. Destruction of extracellular matrix is known to be associated with various diseases such as cancer invasion/metastasis, rheumatoid arthritis, knee osteoarthritis, periodontal disease, and age-related macular degeneration. Furthermore, MMP-2 degrades type IV collagen, etc., which constitutes the basement membrane present under vascular endothelial cells, and the degraded vascular endothelial cells migrate to the interstitium, proliferate in the interstitium, It forms cavities and constructs new blood vessels. It is known that these newly generated blood vessels reach tumor cells, supply nutrients and oxygen to the tumor cells, and cause the tumor to grow (see Non-Patent Document 2).

Therefore, by inhibiting the activity of MMP-2, it is possible to suppress the reduction of basement membrane components and structural changes in the basement membrane, improve skin function, suppress angiogenesis, and suppress the proliferation of tumor cells. It is considered possible. As a substance having an MMP-2 inhibitory action, for example, an extract from black bean paste (see Patent Document 3) is known.

Ceramide is produced from serine and palmitoyl-CoA in the process of keratinization of epidermal cells through the actions of enzymes including serine palmitoyltransferase (SPT) known as the rate-limiting enzyme for ceramide synthesis. Ceramide exists specifically as a major component of intercorneocyte lipids that cover the outermost layer of the skin, and plays an important role in maintaining the skin's original function as a barrier film between the living body and the outside world.

The structure of the stratum corneum is likened to a brick and a mortar, and forms a strong barrier film in the form of intercellular lipids connecting corneocytes, which are piled up in about 15 layers. Corneocytes retain moisture by containing natural moisturizing factors whose main components are amino acids, while intercorneocyte lipids are mainly composed of about 50% ceramide, and contain cholesterol, fatty acids, and other parent substances. It is composed of lipophilic lipids and is characterized by a so-called lamellar structure in which hydrophobic and hydrophilic portions are alternately repeated.

Deterioration of the skin barrier function due to various internal and external factors increases transepidermal water loss, causing dry skin, desquamation, itching, and the like, resulting in so-called dry skin. In addition, the deterioration of the skin barrier function leads to a vicious cycle in which skin inflammation increases and the defense function against various external stimuli decreases. In recent studies, it has been reported that keratin ceramide components (so-called intercellular lipids) decrease or change in composition due to aging or in patients with atopic dermatitis known as barrier disorder (see Non-Patent Document 3). It has become widely known that ceramide is important for maintaining and improving the barrier function of the skin. Known methods for improving the barrier function of the skin include a method of supplementing ceramide from the outside (see Non-Patent Document 4) and a method of increasing ceramide-producing ability inside the skin (see Non-Patent Document 5).

It is known that in skin cells, aquaporins, known as water channels, are expressed on the cell membrane and play a role in the uptake of low-molecular-weight substances, including intercellular water, into the cells. In humans, 13 kinds of aquaporins (AQP0 to AQP12) are known to exist. Epidermal cells mainly contain AQP3, which is thought to play a role in taking up not only water but also low-molecular-weight compounds such as glycerol and urea that are involved in water retention.

However, AQP3 decreases with age, and it has been suggested that this is one of the reasons for the decline in water retention function. can be controlled (see Non-Patent Document 6). For example, an extract from star fruit leaves (see Patent Document 4) is known to have an AQP3 expression promoting effect.

In the skin, melanin also plays a role in protecting the body from ultraviolet rays, but excessive production 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 tyrosinase, an enzyme biosynthesized in pigment cells, and is then formed via intermediates such as 5,6-dihydroxyindophenol. ing. Therefore, in order to prevent, treat, or improve skin darkening (dermal pigmentation), age spots, freckles, etc., it is necessary to inhibit the activity of tyrosinase involved in melanin production, or to suppress melanin production. can be considered.

Conventionally, skin pigmentation, age spots, freckles, etc. have been prevented, treated or improved by externally applying a whitening agent containing a chemically synthesized product such as hydroquinone as an active ingredient. However, chemically synthesized products such as hydroquinone may cause side effects such as skin irritation and allergies. Therefore, the development of a whitening agent containing highly safe natural raw materials as active ingredients has been desired, and as an agent having a tyrosinase activity inhibitory effect, for example, the extract of Polygonum japonicum (see Patent Document 5) and the like are known. . In addition, for example, extracts from plants of the genus Saussurea (see Patent Document 6) are known as substances having melanin production inhibitory action.

Most steroid hormones are molecules that are secreted from the organ where they are produced and bind to receptors to exert their effects. After being reduced to 5α-dihydrotestosterone (5α-DHT) by 5α-reductase, it binds to receptors and exerts an androgenic action.

Androgens are important hormones, but when they act excessively, they can cause a variety of negative effects, such as male pattern baldness, hirsutism, seborrhea, acne (pimples, etc.), benign prostatic hyperplasia, prostate tumors, and precocious male puberty. induce no symptoms. Therefore, conventional methods for suppressing the action of excessive androgen in order to improve these various symptoms, specifically, by inhibiting the action of testosterone 5α-reductase that reduces testosterone to active 5α-DHT. , a method for inhibiting the production of active 5α-DHT, and a method for inhibiting the expression of androgenic activity by inhibiting the binding of 5α-DHT produced from testosterone to receptors. So far, for example, an extract from Higashi Shiso (see Patent Document 7) is known to have testosterone 5α-reductase inhibitory activity and androgen receptor binding inhibitory activity.

Hair repeats growth and shedding according to a periodic hair cycle (hair cycle) consisting of a growth phase, a regression phase and a resting phase. Within this hair cycle, the stage from the resting phase to the anagen phase, in which new hair follicles are formed, is considered to be the most important for hair growth. Dermal papilla cells are thought to play an important role. Dermal papilla cells are cells located inside hair follicle epithelial cells consisting of outer root sheath cells and matrix cells in the vicinity of the hair root, and located at the root portion of the hair root surrounded by the basement membrane. It plays an important role in proliferation/differentiation of hair follicle epithelial cells and hair formation, such as 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 hair formation, and promoting the proliferation of dermal papilla cells can prevent or improve alopecia. It is possible. So far, for example, wild thyme extract (see Patent Document 8) and the like have been known to have an effect of promoting dermal papilla cell proliferation.

Causes and development of inflammatory diseases such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, various skin inflammatory diseases accompanied by rough skin, rheumatoid arthritis, osteoarthritis, asthma, etc. Mechanisms vary widely. The causes are mainly tumor necrosis factor (TNF-α) produced from macrophages, enhancement of hyaluronidase activity, release of histamine, and enhancement of cyclooxygenase-2 (COX-2) activity. etc. are known.

TNF-α was found as a factor causing tumor necrosis, but recently it is considered to be a cytokine that plays a mediator role in regulating not only tumor but also normal cell functions. TNF-α plays an important role in the process from the onset to the end of inflammation, but its persistent and excessive production causes damage to tissues including the skin, and systemically causes fever and chalkia. resulting in exacerbation of inflammation. Therefore, suppression of excessive production of TNF-α is important in pathological inflammation. For example, extracts from mother earthen shells (see Patent Document 2 mentioned above) and the like are known to have a TNF-α production inhibitory effect.

Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid. Hyaluronate, which maintains affinity for body tissue, is decomposed by ultraviolet light, oxygen, etc. in a water-containing system, and its water-retaining effect decreases as its molecular weight decreases. In addition, hyaluronic acid exists as intercellular tissue in vivo and is also involved in vascular permeability. Furthermore, hyaluronidase, which is present in mast cells, is released by degranulation caused by its activation and acts as an inflammatory chemical mediator. Therefore, inhibition of hyaluronidase activity is expected to enhance moisturization and prevent or reduce inflammation. As substances having hyaluronidase activity inhibitory activity, for example, extracts from plants belonging to the genus Osbechia (see Patent Document 9) are known.

Histamine release is a phenomenon in which histamine in mast cells is released outside the cells, and the released histamine induces an inflammatory reaction. Therefore, attempts have been made to prevent or treat allergic diseases and inflammatory diseases using substances that inhibit or suppress histamine release. However, it is difficult to directly evaluate the release of histamine, and the release of hexosaminidase, which has been confirmed to be released simultaneously with the release of histamine, can be used as an index to evaluate the release of histamine. Therefore, by inhibiting the release of hexosaminidase, it is possible to inhibit the release of histamine at the same time, which is considered to be effective in preventing, treating or improving inflammatory diseases and the like.

In addition, histamine mediates communication between cells as a local transmitter, enhances gastric acid secretion in the digestive system, functions as a neurotransmitter in the central nervous system, and is known to contribute to the maintenance of wakefulness. It is Here, excessive histamine release causes ulcers due to hyperacidity in the digestive organs, and contributes to sleep disorders in the central nervous system. As described above, by suppressing the release of hexosaminidase, the release of histamine can also be suppressed at the same time. Therefore, it is thought that gastric ulcers, sleep disorders, etc. caused by hyperacidity can be prevented, treated or improved. . For example, an extract from wisteria tea (see Patent Document 10) is known as a substance having a hexosaminidase release inhibitory action.

Inflammation is a complex reaction with symptoms such as redness, edema, fever, pain and functional impairment. Microscopically, inflammation consists of common reactions such as vascular reactions that cause plasma leakage, leukocyte infiltration, and tissue destruction by inflammatory cells. It may also cause reactions. Prostaglandins play an important role in each of these inflammatory reactions, and it is clear that cyclooxygenase-2, an inducible cyclooxygenase, is mainly involved in prostaglandin production during inflammation. It has become. Therefore, many cyclooxygenase inhibitors typified by aspirin are used for the purpose of preventing and preventing inflammatory reactions (see Non-Patent Document 8).

In recent years, lifestyle habits such as overeating and lack of exercise have led to an increase in body fat and obesity. Such an increase in obesity is observed not only in humans, but also in pets and livestock. Since obesity causes adult diseases such as hyperlipidemia and arteriosclerosis, it poses not only a cosmetic problem but also a serious health problem.

Here, cyclic AMP (cAMP) is known to be involved in lipolysis in vivo. cAMP activates lipase present in the body, and the activated lipase decomposes fat into fatty acids and glycerol. However, activation of cAMP phosphodiesterase induces cAMP degradation and inhibits lipase activation. Therefore, it is considered that inhibition of cAMP phosphodiesterase activity increases intracellular cAMP and promotes the decomposition of fat.

It is also known that platelet aggregation, which causes an inflammatory reaction, is related to the concentration of cyclic AMP (cAMP) in platelets. It is Therefore, by inhibiting the action of cAMP phosphodiesterase to prevent a decrease in cAMP concentration, it is possible to prevent platelet aggregation, thereby preventing, treating, or ameliorating allergic diseases, inflammatory diseases, and the like. Tsubeimoside I (see Patent Document 11) and the like are known to have cAMP phosphodiesterase activity inhibitory action.

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

In the liver, glutathione protects hepatocytes from various oxidative stresses, forms conjugates with harmful substances such as drugs and reactive compounds, and excretes them out of the cells. It directly contributes to the expression of liver function. However, when the action of glutathione is exerted, glutathione is also consumed at the same time. In fact, it is known that the amount of glutathione in hepatocytes decreases when liver damage is induced in rats with galactosamine or the like. . Therefore, if it is possible to suppress the decrease in the amount of glutathione in the liver caused by various stresses, drugs, etc., it is thought that liver damage will be suppressed, leading to an improvement in liver function.

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

Substrates of DPPIV include hormones called incretins. Incretin is a general term for hormones that are secreted from the intestinal tract upon nutrient stimulation and promote insulin secretion from pancreatic β cells in a blood sugar-dependent manner, and GLP-1, GIP, and the like are known. These incretins not only promote blood sugar-dependent insulin secretion, but also suppress glucagon secretion from pancreatic α-cells, lower blood pressure, suppress gastric emptying, and act on the hypothalamus to suppress food intake. It has an effect (see Non-Patent Document 9). However, since incretins are degraded by DPPIV, it is known that the in vivo half-life of GLP-1, for example, is about 1.5 minutes. Therefore, if the enzymatic activity of DPPIV can be inhibited, the half-life of the incretin in the body can be extended, thereby improving the effects of type 2 diabetes, obesity, hypertension, and insulin resistance through the action of the incretin described above. It is expected to be useful for the treatment of such as.

In addition, DPPIV is the same as CD26, which is one of the T cell activation markers, and is known to use many immunoregulatory peptides as substrates and regulate their activity. Therefore, by controlling the activity of DPPIV, it is believed that autoimmune diseases such as rheumatoid arthritis and immune reactions such as transplant rejection can be controlled. Furthermore, DPPIV is known to be involved in the metabolism of several neuropeptides and growth hormone; invasion, metastasis, angiogenesis in cancer, etc.; HIV infection of lymphocytes, and the like. Therefore, by inhibiting the activity of DPPIV, neurological disorders such as pain, neurodegenerative and neuropsychiatric disorders (e.g. 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 carcinoma, breast cancer, etc.); HIV infection (AIDS) and other diseases could be treated.

As for 4-vinylguaiacol, it is known that 4-vinylguaiacol is detected when certain lactic acid bacteria are cultured in a medium containing ferulic acid (Non-Patent Document 10).

JP 2009-269889 A JP 2006-056854 A JP 2007-217352 A JP 2009-191039 A Japanese Patent Application Laid-Open No. 2004-083488 Japanese Unexamined Patent Application Publication No. 2002-201122 JP 2010-184915 A JP 2006-219407 A JP 2003-055242 A Japanese Patent Application Laid-Open No. 2003-012532 JP 2006-056855 A

Nature, 1996, Vol.379, No.6563, p.335-339 "Angiogenesis and Matrix Metalloprotease", Proceedings of the 120th Symposium of the Japanese Association of Medical Sciences, Basics and Clinical Angiogenesis, December 13, 2001, pp.43-49 J. Dermatol., 1993, Vol.20, No.1, p.1-6 "Fragrance Journal", 2004, Vol.32, No.11, p.23-32 Br. J. Dermatol., 2000, Vol.143, Issue 3, p.524-531 "Fragrance Journal", 2006, Vol.34, No.10, p.19-23 Trends Genet., 1992, Vol.8, Issue 2, p.55-61 "Pharmacological Atlas", translated by Takehiko Fukuhara, Bunkodo, 1995, p.184 "Journal of Japanese Pharmacology", 2005, Vol.125, No.6, p.379-384 J. Agric. Food Chem., 2009, Vol.57, No.2, pp.490-494

The present invention provides anti-aging action, whitening action, hair growth action, anti-male hormone action, anti-inflammatory action, anti-obesity action, cyclic AMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action, or dipeptidyl among compounds derived from natural products. Anti-aging agent, whitening agent, hair growth agent, anti-androgenic agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione-lowering agent, found to have peptidase IV activity inhibitory action An object of the present invention is to provide inhibitors and dipeptidyl peptidase IV activity inhibitors, as well as skin cosmetics, hair cosmetics, and food and drink containing the compounds.

In order to solve the above problems, the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and dipeptidyl peptidase of the present invention are provided. The IV activity inhibitor is characterized by containing 4-vinylguaiacol as an active ingredient. Further, the skin cosmetic, hair cosmetic and food and drink of the present invention are characterized by containing 4-vinylguaiacol.

According to the present invention, by using 4-vinylguaiacol as an active ingredient, anti-aging agent, whitening agent, hair growth agent, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase, which have excellent effects and effects. Activity inhibitors, glutathione depletion inhibitors and dipeptidyl peptidase IV activity inhibitors can be provided. Furthermore, by blending 4-vinylguaiacol, it is possible to provide skin cosmetics, hair cosmetics, and foods and drinks that are excellent in the above effects.

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below.
4 - containing vinyl guaiacol as an active ingredient; Further, the skin cosmetic, hair cosmetic and food and drink of the present embodiment contain 4-vinylguaiacol.

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

4-vinyl guaiacol is, for example, a compound containing ferulic acid or a ferulic acid derivative such as ethyl ferulate, or a composition containing ferulic acid (for example, a crushed product or an extract of a plant, etc.) and a microorganism having phenolic acid decarboxylase. It can be produced by fermenting with and converting ferulic acid to 4-vinylguaiacol, and then extracting, isolating and purifying the resulting fermented product. Compositions containing ferulic acid include, for example, crushed products and extracts of plants such as coffee, wheat, corn, tomato, mate, mugwort, and burdock. Furthermore, since ferulic acid is a constituent 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 phenolic acid decarboxylase include, for example, lactic acid bacteria such as Lactobacillus plantarum, Lactobacillus brevis, and Pediococcus pentosaceus; yeasts such as Saccharomyces cerevisiae;

The method for extracting, isolating and purifying 4-vinylguaiacol from the plant or fermented product is not particularly limited, and can be carried out according to a conventional method. For example, the extraction treatment may be carried out by drying the plant or fermented product as the raw material for extraction, and then subjecting it to extraction with an extraction solvent, either as it is or pulverized using a coarse crusher. Drying may be performed in the sun or using a commonly used dryer. In addition, it may be used as an extraction raw material after being subjected to pretreatment such as degreasing with a non-polar solvent such as hexane. By performing pretreatment such as degreasing, the extraction treatment with a polar solvent can be efficiently performed.

As the extraction solvent, it is preferable to use a polar solvent, for example, water, hydrophilic organic solvents, etc., which are used alone or in combination of two or more at room temperature or at a temperature below the boiling point of the solvent. is preferred.

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

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 include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

When using a mixture of two or more polar solvents as the extraction solvent, the mixture ratio can be adjusted as appropriate. For example, when a mixture of water and a lower aliphatic alcohol is used as the extraction solvent, the mixing ratio of water and the lower aliphatic alcohol is preferably 9: 1 to 1: 9 (volume ratio), More preferably, it is 7:3 to 2:8 (volume ratio). When using a mixture of water and lower aliphatic ketone, the mixing ratio of water and lower aliphatic ketone is preferably 9:1 to 2:8 (volume ratio). When using a mixed solution with a hydric alcohol, the mixing ratio of water and polyhydric alcohol is preferably 5:5 to 1:9 (volume ratio).

The extraction treatment is not particularly limited as long as the soluble components contained in the raw material for extraction can be eluted into the extraction solvent, and can be carried out according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent of 5 to 15 times the amount (mass ratio) of the extraction raw material, the soluble component is extracted at room temperature or under reflux heating, and then filtered to remove the extraction residue. You can get the liquid. A paste-like concentrate is obtained by distilling off the solvent from the obtained extract, and a dried product is obtained by further drying the concentrate.

The method for isolating and purifying 4-vinylguaiacol from the extract obtained as described above, the concentrate of the extract, or the dried product of the extract is not particularly limited, and is carried out by a conventional method. be able to. For example, the extract is dissolved in a developing solvent and subjected to column chromatography using a porous material such as silica gel or alumina, or a porous resin such as styrene-divinylbenzene copolymer or polymethacrylate to obtain 4-vinyl Examples thereof include a method for recovering a fraction containing guaiacol. In this case, the developing solvent may be appropriately selected according to the stationary phase 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 and the like. Furthermore, the fraction containing 4-vinylguaiacol obtained by column chromatography is subjected to reverse phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, column chromatography using an ion exchange resin, etc. Any organic compound purification means may be used for purification.

[Anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cyclic AMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor, dipeptidyl peptidase IV activity inhibitor]
The 4-vinylguaiacol obtained as described above has excellent anti-aging, whitening, hair-growth, anti-male hormone, anti-inflammatory, anti-obesity, cyclic AMP (cAMP) phosphodiesterase activity inhibitory effects, and glutathione. Since it has a dipeptidyl peptidase IV (DPPIV) activity inhibitory action and a dipeptidyl peptidase IV (DPPIV) activity inhibitory action, it can be used as an anti-aging agent, a whitening agent, a hair restorer, an anti-androgenic agent, an anti-inflammatory agent, an anti-obesity agent, a cAMP phosphodiesterase activity inhibitor, It can be used as an active ingredient of glutathione depletion inhibitors and DPPIV activity inhibitors. The anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and DPPIV activity inhibitor of the present embodiment are pharmaceuticals and quasi-drugs. , cosmetics, etc.

In addition, as an active ingredient of the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and DPPIV activity inhibitor according to the present embodiment, Compositions containing 4-vinylguaiacol may be used in place of isolated 4-vinylguaiacol. Here, the "composition containing 4-vinylguaiacol" in the present embodiment includes an extract obtained from a plant containing 4-vinylguaiacol as an extraction raw material, a fermented product containing 4-vinylguaiacol, and the Extracts obtained by using fermented products as extraction raw materials are included. In addition, the "extract" includes an extract obtained by an extraction process, a diluted or concentrated liquid of the extract, or a dried product obtained by drying the extract.

As an active ingredient of the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and DPPIV activity inhibitor according to the present embodiment, 4- When using a composition containing vinyl guaiacol, it is preferable to use 4-vinyl guaiacol that has been purified to increase its purity. Anti-aging agent, whitening agent, hair restorer, anti-androgenic agent, anti-inflammatory agent, anti-obesity agent, cAMP, which have further excellent effects by using 4-vinylguaiacol with increased purity as an active ingredient. Phosphodiesterase activity inhibitors, glutathione depletion inhibitors and DPPIV activity inhibitors can be obtained.

The anti-aging action of 4-vinylguaiacol is, for example, glutathione production promoting action, epidermal keratinocyte proliferation promoting action, matrix metalloprotease-2 (MMP-2) activity inhibitory action, and serine palmitoyltransferase (SPT) mRNA expression promoting action. and aquaporin 3 (AQP3) mRNA expression promoting action. However, the anti-aging action possessed by 4-vinylguaiacol is not limited to the anti-aging action exhibited based on the above action.

In addition, 4-vinyl guaiacol utilizes its glutathione production promoting action, epidermal keratinocyte proliferation promoting action, MMP-2 activity inhibitory action, SPT mRNA expression promoting action or AQP3 mRNA expression promoting action to promote glutathione production, epidermis, respectively. It may be used as an active ingredient of a keratinocyte proliferation promoter, MMP-2 activity inhibitor, SPT mRNA expression promoter or AQP3 mRNA expression promoter.

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

The hair growth action of 4-vinylguaiacol is exhibited based on one or more actions selected from the group consisting of, for example, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action and dermal papilla cell proliferation promoting action. be done. However, the hair growth action of 4-vinylguaiacol is not limited to the hair growth action exhibited based on the above action. In addition, 4-vinylguaiacol is a testosterone 5α-reductase activity inhibitor and androgen receptor binding inhibitor by utilizing its testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action or dermal papilla cell growth promoting action, respectively. Alternatively, it may be used as an active ingredient of a dermal papilla cell proliferation-promoting agent.

The antiandrogen action of 4-vinylguaiacol is exhibited based on, for example, testosterone 5α-reductase activity inhibitory action and/or androgen receptor binding inhibitory action. However, the antiandrogen action of 4-vinylguaiacol is not limited to the antiandrogen action exhibited based on the above action.

The anti-inflammatory action of 4-vinylguaiacol is, for example, tumor necrosis factor (TNF-α) production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action and cyclooxygenase-2 (COX-2) activity inhibitory action. It is exerted based on one or two or more actions selected from the group consisting of: However, the anti-inflammatory action possessed by 4-vinylguaiacol is not limited to the anti-inflammatory action exhibited based on the above action. In addition, 4-vinyl guaiacol utilizes its TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action or COX-2 activity inhibitory action to act as a TNF-α production inhibitor and a hyaluronidase activity inhibitor, respectively. It may be used as an active ingredient of inhibitors, hexosaminidase release inhibitors or COX-2 activity inhibitors.

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

The anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor or DPPIV activity inhibitor of the present embodiment is 4-vinylguaiacol or 4 - A composition containing vinyl guaiacol may be used alone, or a composition containing 4-vinyl guaiacol or 4-vinyl guaiacol may be formulated.

The anti-aging agent, whitening agent, hair restorer, anti-androgenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and DPPIV activity inhibitor of the present embodiment are dextrin, cyclodextrin and the like. Using a pharmaceutically acceptable carrier and any other adjuvant, it can be formulated into any dosage form such as powder, granule, tablet, liquid and the like according to a conventional method. At this time, as auxiliary agents, for example, excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like can be used. Anti-aging agents, whitening agents, hair restorers, antiandrogen agents, anti-inflammatory agents, anti-obesity agents, cAMP phosphodiesterase activity inhibitors, glutathione lowering inhibitors and DPPIV activity inhibitors are used in other compositions (e.g., skin cosmetics , hair cosmetics, etc.), and can also be used as ointments, liquids for external use, patches, and the like.

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

In addition, the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, or DPPIV activity inhibitor of the present embodiment may be used if necessary. 4. other natural extracts having anti-aging, whitening, hair-growth, anti-androgenic, anti-androgenic, anti-inflammatory, anti-obesity, cAMP phosphodiesterase activity inhibitory, glutathione lowering inhibitory or DPPIV activity inhibitory effects; -It can be used as an active ingredient in combination with a composition containing vinyl guaiacol or 4-vinyl guaiacol.

The anti-aging agent, whitening agent, hair restorer, anti-androgenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, or DPPIV activity inhibitor of the present embodiment can be administered to patients as follows: Transdermal administration, oral administration and the like can be mentioned, and a suitable method for prevention or treatment may be appropriately selected according to the type of disease. In addition, the dosage of the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, or DPPIV activity inhibitor of the present embodiment also affects the disease. The dose may be increased or decreased as appropriate depending on the type, severity, individual patient differences, administration method, administration period, etc.

The anti-aging agent of this embodiment consists of glutathione production-promoting action, epidermal keratinocyte proliferation-promoting action, MMP-2 activity inhibiting action, SPT mRNA expression-promoting action and AQP3 mRNA expression-promoting action possessed by the active ingredient 4-vinylguaiacol. Through one or more actions selected from the group, it is possible to prevent, treat or improve skin aging symptoms such as formation of wrinkles, loss of skin elasticity and loss of moisturizing function. However, in addition to these uses, the anti-aging agent of the present embodiment exhibits glutathione production promoting action, epidermal keratinocyte proliferation promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, or AQP3 mRNA expression promoting action. It can be used for all uses that make sense for

For example, in addition to the above-described uses, the anti-aging agent of the present embodiment or the above-described glutathione production enhancer has liver damage (drinking alcohol, heavy metals, chemicals, etc.) through the glutathione production-enhancing action of 4-vinylguaiacol. It is possible to prevent, treat, or ameliorate diseases in which a decrease or deficiency of intracellular glutathione is known to be associated with pathology, such as those caused by ingestion of foreign substances such as glutathione.

In addition to the uses described above, the anti-aging agent of the present embodiment or the MMP-2 activity inhibitor described above is related to the pathology by the destruction of the extracellular matrix through the MMP-2 activity inhibitory action of 4-vinylguaiacol. It can be used for the treatment and prevention of diseases known to cause cancer (eg, cancer invasion/metastasis, rheumatoid arthritis, knee osteoarthritis, periodontal disease, age-related macular degeneration, etc.). In addition, the anti-aging agent or MMP-2 activity inhibitor of the present embodiment can suppress angiogenesis and tumor cell proliferation by virtue of its MMP-2 activity inhibitory action.

In addition to the uses described above, the anti-aging agent of the present embodiment or the SPTmRNA expression promoter described above enhances the skin barrier function through the SPTmRNA expression-enhancing action of 4-vinylguaiacol, and treats rough skin, dry skin, etc. It can prevent, treat or improve dry skin diseases (eg, atopic dermatitis, psoriasis, ichthyosis, etc.). In addition, the anti-aging agent of the present embodiment or the AQP3 mRNA expression promoter described above can improve age-related water retention capacity, barrier function, etc. through the AQP3 mRNA expression promoting action of 4-vinylguaiacol.

The whitening agent of the present embodiment can prevent or improve pigmentation such as skin darkening, age spots, and freckles through the tyrosinase activity inhibitory action and/or melanin production inhibitory action of the active ingredient 4-vinylguaiacol. can. However, the whitening agent of the present embodiment can be used for all uses other than these uses where it is significant to exhibit tyrosinase activity inhibitory action or melanin production inhibitory action.

The hair restorer of the present embodiment is one or two selected from the group consisting of testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, and dermal papilla cell proliferation-promoting action possessed by the active ingredient 4-vinylguaiacol. It can prevent, treat, or ameliorate male pattern baldness, alopecia areata, alopecia such as trichotillomania, etc., and is particularly suitable for the prevention, treatment, or amelioration of male pattern baldness. However, the hair restorer of the present invention may be used for all other uses that are significant in exhibiting testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, or dermal papilla cell proliferation-promoting action, in addition to these uses. can be done.

For example, the hair restorer of the present embodiment or the aforementioned dermal papilla cell proliferation-promoting agent activates dermal papilla cells through the dermal papilla cell proliferation-promoting action possessed by 4-vinylguaiacol, resulting in proliferation and differentiation of hair follicle epithelial cells and It can promote the formation of hair, prevent the growth phase from shifting to the regression phase and the telogen phase in the hair cycle, and prolong the growth phase. Thereby, alopecia can be prevented or improved. In addition, the hair restorer of the present embodiment or the dermal papilla cell growth promoting agent described above can be applied to the field of regenerative medicine such as hair regeneration using dermal papilla cells through the dermal papilla cell proliferation-promoting action of 4-vinylguaiacol. can also be used.

The antiandrogen agent of the present embodiment or the above-described testosterone 5α-reductase activity inhibitor or androgen receptor binding inhibitor has testosterone 5α-reductase activity inhibitory action and/or androgen receptor activity possessed by 4-vinylguaiacol as an active ingredient. Prevents, treats or improves male hormone-related diseases such as male pattern baldness, hirsutism, seborrhea, acne (pimples, etc.), prostatic hyperplasia, prostatic tumor, and precocious male puberty, etc. can do. However, the anti-androgen agent of the present invention can be used for all purposes other than these uses in which it is meaningful to exhibit testosterone 5α-reductase activity inhibitory action or androgen receptor binding inhibitory action.

The anti-inflammatory agent of the present embodiment is selected from the group consisting of TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action and COX-2 activity inhibitory action possessed by 4-vinylguaiacol as an active ingredient. It is possible to prevent, treat or ameliorate contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, and various other skin inflammatory diseases associated with rough skin through one or more of the actions. However, the anti-inflammatory agent of the present embodiment is significant in exhibiting TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, or COX-2 activity inhibitory action in addition to these uses. It can be used for any purpose.

For example, the anti-inflammatory agent of the present embodiment or the aforementioned TNF-α production inhibitor, hyaluronidase activity inhibitor, hexosaminidase release inhibitor or COX-2 activity inhibitor is Rheumatoid arthritis, osteoarthritis, asthma, etc. can be prevented, treated or ameliorated through inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action or COX-2 activity inhibitory action. In addition, the anti-inflammatory agent of the present embodiment or the hexosaminidase release inhibitor described above prevents gastric ulcers, sleep disorders, etc. caused by hyperacidity through the hexosaminidase release inhibitory action of 4-vinylguaiacol. can be treated or ameliorated.

The anti-obesity agent of the present embodiment can promote cAMP production and decompose adipocytes through cAMP phosphodiesterase activity inhibitory action possessed by 4-vinylguaiacol as an active ingredient, and as a result, obesity, and Various diseases such as arteriosclerosis, diabetes, and metabolic syndrome can be prevented or improved.

The cAMP phosphodiesterase activity inhibitor of the present embodiment promotes the production of cAMP through the cAMP phosphodiesterase activity inhibitory action of the active ingredient 4-vinylguaiacol, so that it can suppress platelet aggregation, thereby suppressing allergic diseases. and various inflammatory diseases can be prevented, treated or improved. In addition, the cAMP phosphodiesterase activity inhibitor of the present embodiment can promote the decomposition of adipocytes through the cAMP phosphodiesterase activity inhibitory action of 4-vinylguaiacol, and as a result, obesity and arteriosclerosis associated therewith, 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 uses other than these uses in which it is significant to exhibit cAMP phosphodiesterase activity inhibitory action.

The glutathione lowering inhibitor of the present embodiment has 4-vinylguaiacol as an active ingredient when hepatocytes are exposed to various stresses caused by excessive alcohol intake, intake of foreign substances such as heavy metals and chemical substances. Through glutathione depletion inhibitory action, metabolism of harmful substances (for example, extracellular excretion by formation of conjugate with glutathione, etc.) can be promoted, and liver injury caused by harmful substances can be prevented or reduced. This makes it possible to prevent, treat, or ameliorate diseases (liver dysfunction) caused by liver disorders such as acute hepatitis, chronic hepatitis, fatty liver, jaundice, and cirrhosis. However, the glutathione decrease inhibitor of the present embodiment can be used for all uses other than these uses that are significant in exhibiting the glutathione decrease inhibitory action.

For example, the glutathione lowering inhibitor of the present embodiment prevents or improves hangovers through the glutathione lowering inhibitory action of 4-vinylguaiacol; can be used for purposes such as improvement;

The DPPIV inhibitor of this embodiment can prevent or treat type 2 diabetes, obesity, hypertension, insulin resistance and the like through the DPPIV inhibitory action of 4-vinylguaiacol, and can also prevent or treat autoimmune diseases such as rheumatoid arthritis. and transplant rejection can be prevented or treated. However, the DPPIV inhibitor of the present embodiment can be used for all uses other than these uses that are significant in exhibiting the DPPIV inhibitory action. For example, the DPPIV inhibitor of the present embodiment can reduce pain, neurodegenerative diseases, and neurological disorders such as neuropsychiatric disorders (eg, sciatica, Alzheimer's disease, depression, etc.) through the DPPIV inhibitory action of 4-vinylguaiacol; diseases such as hormone deficiencies and diseases for which growth hormone is used for treatment; cancers (eg, T-cell lymphoma, acute lymphoblastic leukemia, thyroid cancer, basal cell carcinoma, breast cancer, etc.); diseases such as HIV infection (AIDS); It can be prevented or treated.

In addition, the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, or DPPIV activity inhibitor of the present embodiment is an excellent anti-aging agent. action, whitening action, hair growth action, antiandrogen action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action, or DPPIV activity inhibitory action. preferred. In this case, 4-vinylguaiacol or a composition containing 4-vinylguaiacol may be blended as it is, or an anti-aging agent and whitening agent formulated from 4-vinylguaiacol or a composition containing 4-vinylguaiacol. agents, hair growth agents, anti-androgenic agents, anti-inflammatory agents, anti-obesity agents, cAMP phosphodiesterase activity inhibitors, glutathione lowering inhibitors or DPPIV activity inhibitors may be added.

Here, the category of external preparations for skin is not limited, and includes a wide range of skin cosmetics, quasi-drugs used transdermally, pharmaceuticals, and the like, which will be described later.

In addition, the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, or DPPIV activity inhibitor of the present embodiment is an excellent anti-aging agent. action, whitening action, hair growth action, antiandrogenic action, anti-inflammatory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action, or DPPIV activity inhibitory action. It can also be suitably used as

[Skin cosmetics, hair cosmetics]
4-Vinylguaiacol has excellent anti-aging, whitening, hair-growth, anti-androgenic, anti-inflammatory, anti-obesity, cAMP phosphodiesterase activity inhibitory, glutathione decrease inhibitory and DPPIV activity inhibitory activities. Therefore, it is suitable for blending in skin cosmetics or hair cosmetics. In this case, 4-vinylguaiacol or a composition containing 4-vinylguaiacol may be blended as it is, or an anti-aging agent or whitening agent formulated from 4-vinylguaiacol or a composition containing 4-vinylguaiacol. , a hair restorer, an antiandrogenic agent, an anti-inflammatory agent, an anti-obesity agent, a cAMP phosphodiesterase activity inhibitor, a glutathione lowering inhibitor, or a DPPIV activity inhibitor may be added.

By blending 4-vinylguaiacol or the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor or DPPIV activity inhibitor, Anti-aging action, glutathione production promoting action, epidermal keratinocyte proliferation promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibiting action in skin cosmetics or hair cosmetics, Melanin production inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogenic action, anti-inflammatory action, TNF-α production inhibitory action, hyaluronidase activity inhibitory action , hexosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action or DPPIV activity inhibitory action.

All of these actions exhibit favorable actions when applied to skin cosmetics or hair cosmetics. -2 activity inhibitory action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibitory action, melanin production inhibitory action, anti-inflammatory action, TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release When a skin cosmetic is imparted with inhibitory action, COX-2 activity inhibitory action, cAMP phosphodiesterase activity inhibitory action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action or antiandrogenic action, these actions are exhibited. It is particularly suitable because it is easy to be

In addition, among the above-mentioned actions, hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogenic action, anti-inflammatory action, TNF-α production inhibitory action, hyaluronidase It is particularly preferable to impart activity-inhibiting action, hexosaminidase release-inhibiting action, COX-2 activity-inhibiting action, or cAMP phosphodiesterase activity-inhibiting action to the hair cosmetic, since these actions are likely to be exhibited.

Skin to which 4-vinylguaiacol, or the above anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor or DPPIV activity inhibitor can be formulated The types of cosmetics or hair cosmetics are not particularly limited, and skin cosmetics include, for example, ointments, creams, milky lotions, lotions, packs, and foundations. Hair cosmetics include, for example, , hair tonics, hair creams, hair liquids, shampoos, pomades, rinses and the like.

4-vinyl guaiacol, or the above anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor or DPPIV activity inhibitor in skin cosmetics or When blended in a hair cosmetic, the blending amount can be appropriately adjusted according to the type of skin cosmetic or hair cosmetic. A particularly suitable blending ratio is about 0.001 to 1% by weight in terms of standard extract.

The skin cosmetic or hair cosmetic of this embodiment has anti-aging action, glutathione production promoting action, epidermal keratinocyte proliferation promoting action, MMP-2 activity inhibiting action, SPT mRNA expression promoting action, and AQP3 mRNA expression possessed by 4-vinylguaiacol. stimulating action, whitening action, tyrosinase activity inhibitory action, melanin production inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogen action, anti-inflammatory action, As long as it does not interfere with TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action or DPPIV activity inhibitory action , Main ingredients, auxiliaries or other ingredients used in the production of ordinary skin cosmetics or hair cosmetics, such as astringents, bactericidal / antibacterial agents, whitening agents, UV absorbers, moisturizers, cell activators, anti-inflammatory / Antiallergic agents, antioxidants/active oxygen removers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, and the like can be used in combination. Such combination may result in a more generalized product and may provide greater benefits than would normally be expected due to synergy between the other active ingredients used in combination.

The skin cosmetic of this embodiment has anti-aging effect, glutathione production promoting effect, epidermal keratinocyte proliferation promoting effect, MMP-2 activity inhibiting effect, SPT mRNA expression promoting effect, AQP3 mRNA expression promoting effect, and whitening effect possessed by 4-vinylguaiacol. action, tyrosinase activity inhibitory action, melanin production inhibitory action, anti-inflammatory action, TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity 1 selected from the group consisting of inhibitory action, hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogenic action, glutathione decrease inhibitory action and DPPIV activity inhibitory action Or through two or more actions, prevention, treatment or improvement of skin aging symptoms such as wrinkle formation, loss of skin elasticity, loss of moisturizing function; acceleration of healing of wounds or burns; rough skin, dry skin, etc. , prevention, treatment or improvement of dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.); prevention, treatment or improvement of pigmentation such as skin darkening, age spots, freckles; contact dermatitis ( rash), psoriasis, pemphigus vulgaris, prevention, treatment or improvement of various skin inflammatory diseases associated with rough skin; prevention, treatment or improvement of seborrhea, acne (pimples, etc.); prevention, treatment, or improvement of lifestyle-related diseases such as arteriosclerosis, diabetes, and metabolic syndrome;

In addition, the hair cosmetic of the present embodiment has a hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogen action, and anti-inflammatory action possessed by 4-vinylguaiacol. , TNF-α production inhibitory action, hyaluronidase activity inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, anti-obesity action, cAMP phosphodiesterase activity inhibitory action, anti-aging action, glutathione production promotion action, epidermal keratinization Selected from the group consisting of cell proliferation promoting action, MMP-2 activity inhibitory action, SPT mRNA expression promoting action, AQP3 mRNA expression promoting action, whitening action, tyrosinase activity inhibitory action, melanin production inhibitory action, glutathione decrease inhibitory action and DPPIV activity inhibitory action. prevention, treatment or improvement of alopecia such as androgenetic alopecia, alopecia areata, and trichotillomania; contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other rough skin Prevention, treatment or improvement of various skin inflammatory diseases associated therewith; treatment of rough skin, dry skin, etc., as well as dry skin diseases (eg, atopic dermatitis, psoriasis, ichthyosis, etc.);

[Food and drink]
4-Vinylguaiacol has excellent anti-aging, whitening, hair-growth, anti-androgenic, anti-inflammatory, anti-obesity, cAMP phosphodiesterase activity inhibitory, glutathione decrease inhibitory and DPPIV activity inhibitory activities. Therefore, it is suitable for blending in foods and drinks. In this case, 4-vinylguaiacol may be blended as it is, or an anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, and cAMP phosphodiesterase activity formulated from 4-vinylguaiacol. An inhibitor, a glutathione lowering inhibitor or a DPPIV activity inhibitor may be added.

Here, “food and drink” refers to those that are less likely to harm human health and are ingested orally or through the gastrointestinal tract in normal social life. It is not limited to categories. Therefore, the "food and drink" in the present embodiment includes a wide range of orally ingested general foods, health foods (functional foods and drinks), foods with health claims (foods for specified health uses, foods with nutrient function claims), and quasi-drugs. includes.

4-vinyl guaiacol or the above anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor, DPPIV activity inhibitor, anti-aging agent, hair restorer, anti-androgenic agent, anti-inflammatory agent or whitening agent is blended in food or drink. Anti-obesity action, cAMP phosphodiesterase activity inhibitory action, glutathione decrease inhibitory action, DPPIV activity inhibitory action, anti-aging action, glutathione production promotion action, epidermal keratinocyte proliferation promotion action, MMP-2 activity inhibition action, SPT mRNA expression promotion action, AQP3 mRNA expression promoting action, hair growth action, testosterone 5α-reductase activity inhibitory action, androgen receptor binding inhibitory action, dermal papilla cell proliferation promoting action, antiandrogenic action, anti-inflammatory action, TNF-α production inhibitory action, hyaluronidase activity Inhibitory action, hexosaminidase release inhibitory action, COX-2 activity inhibitory action, whitening action, tyrosinase activity inhibitory action or melanin production inhibitory action can be imparted to food and drink. These actions are suitable because they are likely to exert their actions and effects when they are added to food and drink.

4-Vinylguaiacol, or an anti-aging agent, a whitening agent, a hair restorer, an antiandrogenic agent, an anti-inflammatory agent, an anti-obesity agent, a cAMP phosphodiesterase activity inhibitor, a glutathione lowering inhibitor, or a DPPIV activity formulated from 4-vinylguaiacol When the inhibitor is added to food and drink, the amount of active ingredient in them can be changed as appropriate in consideration of the purpose of use, symptoms, gender, etc., but the general intake of the food and drink to which the inhibitor is added Considering the above, it is preferable that the daily intake of the extract for an adult is about 1-1000 mg. If the food or drink to be added is a granular, tablet or capsule-shaped food or drink, 4-vinylguaiacol, or an anti-aging agent formulated from 4-vinylguaiacol, a whitening agent, a hair restorer, an antiandrogenic agent, The amount of anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor or DPPIV activity inhibitor added is usually 0.1 to 100% by mass, preferably 5 to 100% by mass.

The food and drink of the present embodiment may be any food or drink containing 4-vinylguaiacol that does not interfere with its activity, or may be a dietary supplement containing 4-vinylguaiacol as a main ingredient. good too.

When producing the food or drink of the present 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; Sugars; soybean oil, oils and fats such as medium-chain fatty acid triglycerides, etc. can be added to make foods and drinks of any shape.

Food and drink in which 4-vinylguaiacol can be blended is not particularly limited, but specific examples thereof include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (concentrated undiluted solutions and adjustment powders of these drinks). frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as buckwheat, udon, vermicelli, gyoza skin, dumpling skin, Chinese noodles, instant noodles; candy, chewing gum, candies, gum, chocolate, tablets , confectionery such as snacks, biscuits, jelly, jam, cream, and baked goods; Processed marine and livestock foods such as fish cakes, hams and sausages; Dairy products such as processed milk and fermented milk; Salad oil, tempura oil, margarine, mayonnaise Fats and fats such as shortening, whipped cream, and dressings, and processed fats and oils; seasonings such as sauces and sauces; soups, stews, salads, side dishes, and pickles; health and nutritional supplements in various forms; tablets, capsules, and drinks and the like, and when 4-vinylguaiacol is blended into these foods and drinks, commonly used auxiliary raw materials and additives can be used together.

In addition, the anti-aging agent, whitening agent, hair restorer, antiandrogenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione decrease inhibitor, DPPIV activity inhibitor, skin cosmetics, and hair of the present embodiment. Cosmetics and foods and drinks are preferably applied to humans, but as long as the effects of each are exhibited, animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, etc.) It can also be applied to pigs, monkeys, etc.).

The present invention will be specifically described below with reference to test examples, but the present invention is not limited to the following examples. In this test example, 4-vinylguaiacol (manufactured by Wako Pure Chemical Industries, Ltd., sample 1) was used as a test sample.

[Test Example 1] Glutathione production promoting effect test 4-Vinylguaiacol (Sample 1) was tested for glutathione production promoting effect as follows.

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

After culturing, 200 μL of a test sample (Sample 1, see Table 1 below for sample concentrations) dissolved in Dulbecco's MEM medium containing 1% FBS was added to each well and cultured for 24 hours. As a control, 1% FBS-containing Dulbecco's MEM medium containing no test sample was used and cultured in the same manner. After the culture was completed, the medium was removed from each well, washed with 400 μL of PBS(-) buffer, and then the cells were lysed using 150 μL of M-PER (manufactured by PIERCE).

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

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

As shown in Table 1, 4-vinylguaiacol (Sample 1) was found to have an excellent glutathione production promoting effect.

[Test Example 2] Epidermal Keratinocyte Proliferation Promoting Action Test 4-Vinylguaiacol (Sample 1) was tested for epidermal keratinocyte proliferation promoting action as follows.

After culturing normal human neonatal epidermal keratinocytes (NHEK) using a normal human epidermal keratinocyte growth medium (KGM), the cells were collected by trypsinization. After diluting the collected cells with KGM medium to a cell density of 3.0×10 ⁴ cells/mL, 100 μL of each well was seeded on a collagen-coated 96-well plate and cultured overnight. After completion of the culture, 100 μL of a test sample (Sample 1, see Table 2 below for sample concentrations) dissolved in KGM medium was added to each well and cultured for 3 days. As a control, KMG medium containing no sample was cultured in the same manner.

Epidermal keratinocyte proliferation-promoting activity was measured using the MTT assay method. 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 to each well. After culturing for 2 hours, intracellularly produced blue formazan was extracted with 100 μL of 2-propanol. After extraction, absorbance was measured at a wavelength of 570 nm. At the same time, absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was taken 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 x 100
Each term in the formula represents the following.
St: Amount of blue formazan produced with addition of test sample Ct: Amount of blue formazan produced with no addition of sample Table 2 shows the results.

As shown in Table 2, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent epidermal keratinocyte proliferation-promoting effect.

[Test Example 3] 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 large amounts as a recombinant proMMP protein having 6 histidine residues at the C-terminus using an E. coli gene expression system, purified using Ni-NTA resin, and refolded. transferred to the mold. This was used as an enzyme sample, and diluted appropriately to prepare an enzyme solution.
Using the resulting enzyme solution, 4-vinylguaiacol (Sample 1) was tested for MMP-2 activity inhibitory action as follows.

Using a 96-well plate for fluorescence intensity measurement, 20 μL of test sample (Sample 1, see Table 3 below for sample concentration), 40 μL of enzyme solution and buffer solution (0.05 mol/L Tris, 150 mmol/L NaCl, 10 mmol/L CaCl 20 μL of ₂ , 50 μmol/L ZnSO ₄ , 0.02% NaN ₃ , 0.05% Brij35 (pH 7.5)) was mixed and allowed to stand at 37° C. for 15 minutes. After that, 120 μL of a fluorescent substrate peptide (MOCAc/DNP peptide) adjusted 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 defined as the fluorescence intensity immediately after addition of the substrate. Immediately after the measurement, reaction was carried out at 37° C. for 120 minutes, during which 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. Also, 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 wavelength 420 nm after 120 minutes without sample addition/substrate addition B: Fluorescence intensity at wavelength 420 nm immediately after addition of sample/substrate addition C: Test sample addition/substrate addition 120 minutes after addition at wavelength 420 nm Fluorescence intensity D: Fluorescence intensity at a wavelength of 420 nm immediately after the addition of the test sample and the substrate Table 3 shows the results.

As shown in Table 3, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent MMP-2 activity inhibitory effect.

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

Normal human neonatal epidermal keratinocytes (NHEK) were cultured in an 80 cm ² flask using normal human epidermal keratinocyte long-term culture growth medium (EpiLife-KG2) under conditions of 37° C., 5% CO ₂ and 95% air. and cells were harvested by trypsinization. After diluting the collected cells with Epilife-KG2 medium to a cell density of 20×10 ⁴ cells/mL, 2 mL each was seeded in a 35 mm petri dish (manufactured by FALCON) (40×10 ⁴ cells/dish). C., 5% CO ₂ -95% air, and cultured for 24 hours.

After culturing, the medium was removed, and 2 mL of the test sample dissolved in Epilife-KG2 medium (Sample 1, see Table 4 below for the sample concentration) was added to each petri dish and incubated at 37°C in 5% CO ₂ -95% air. It was cultured for 24 hours under the conditions of As a control, Epilife-KG2 medium containing no sample was used and cultured in the same manner. After culturing, the medium was removed, total RNA was extracted with ISOGEN (manufactured by Nippon Gene, Cat. No. 311-02501), and the amount of each RNA was measured with a spectrophotometer so as to be 200 ng/μL. Total RNA was prepared.

Using this total RNA as a template, the expression level of mRNA was measured for SPT and GAPDH as an internal standard. Detection was performed by real-time 2-Step RT-PCR reaction using a real-time PCR device Smart Cycler (manufactured by Cepheid) and TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (manufactured by Takara Bio, code No. RR063A). The expression level of SPT was corrected with the value of GAPDH based on total RNA preparations prepared from cells cultured with "test sample added" and "sample not added". From the obtained values, the SPT mRNA expression promotion rate (%) was calculated according to the following formula.

SPT mRNA expression promotion rate (%) = A/B x 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value with no addition of sample Table 4 shows the results.

As shown in Table 4, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent SPTmRNA expression promoting effect.

[Test Example 5] Aquaporin 3 (AQP3) mRNA expression promoting action test 4-Vinylguaiacol (Sample 1) was tested for AQP3 mRNA expression promoting action as follows.

Normal human neonatal epidermal keratinocytes (NHEK) were precultured in an 80 cm ² flask using normal human epidermal keratinocyte growth medium (KGM) under the conditions of 37° C., 5% CO ₂ and 95% air. and cells were harvested by trypsinization. After diluting the collected cells with KGM medium to a cell density of 20×10 ⁴ cells/mL, 2 mL each was seeded in a 35 mm petri dish (manufactured by FALCON) (40×10 ⁴ cells/dish). It was cultured for 24 hours under the conditions of 5% CO ₂ -95% air.

After culturing, the medium was removed, and 2 mL of the KGM medium of the test sample (Sample 1, see Table 5 below for the sample concentration) dissolved in KGM medium was added to each petri dish and incubated at 37°C, 5% CO ₂ -95% air. It was cultured for 24 hours under the conditions of As a control, a KGM medium containing no sample was cultured in the same manner. After culturing, the medium was removed, total RNA was extracted with ISOGEN (manufactured by Nippon Gene, Cat. No. 311-02501), and the amount of each RNA was measured with a spectrophotometer so as to be 200 ng/μL. Total RNA was prepared.

Using this total RNA as a template, the mRNA expression levels of AQP3 and GAPDH as an internal standard were measured. Detection was performed by real-time 2-Step RT-PCR reaction using a real-time PCR device Smart Cycler (manufactured by Cepheid) and TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (manufactured by Takara Bio Inc., code No. RR063A). . The expression level of AQP3 mRNA was corrected with the value of GAPDH based on total RNA preparations prepared from cells cultured with "test sample added" and "sample not added". From the obtained values, the AQP3 mRNA expression promotion rate (%) was calculated according to the following formula.

AQP3 mRNA expression promotion rate (%) = A/B x 100
Each term in the formula represents the following.
A: Correction value with addition of test sample B: Correction value with no addition of sample Table 5 shows the results.

As shown in Table 5, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent effect of promoting AQP3 mRNA expression.

[Test Example 6] Tyrosinase activity inhibitory action test 4-Vinylguaiacol (Sample 1) was tested for tyrosinase activity inhibitory action as follows.

Test sample dissolved in 0.2 mL Mcllvaine buffer (pH 6.8), 0.06 mL 0.3 mg/mL tyrosine solution, 25% DMSO solution in a 48-well plate (Sample 1, see Table 6 below for sample concentrations) 0.18 mL was added and allowed to stand at 37° C. for 10 minutes. To this, 0.02 mL of 800 units/mL tyrosinase solution was added, followed by reaction at 37° C. for 15 minutes. After completion of the reaction, absorbance at a wavelength of 475 nm was measured.

In addition, as a blank, the same operation and absorbance measurement were performed in the case where no enzyme solution was added. Furthermore, as a control, the same measurement was performed when distilled water was added without adding the sample solution. From the obtained measurement results, the tyrosinase activity inhibition rate (%) was calculated according to the following formula.

Tyrosinase activity inhibition rate (%) = {1-(St-Sb)/(Ct-Cb)}×100
Each term in the formula represents the following.
St: Absorbance at wavelength 475 nm with test sample added/enzyme added Sb: Absorbance at wavelength 475 nm with test sample added/enzyme added Ct: Absorbance at wavelength 475 nm with sample not added/enzyme added Cb: Sample not added/enzyme Table 6 shows the absorbance results at a wavelength of 475 nm without addition.

As shown in Table 6, 4-vinylguaiacol (Sample 1) was found to have excellent tyrosinase activity inhibitory activity.

[Test Example 7] Melanin production inhibitory action test on B16 melanoma cells For 4-vinylguaiacol (sample 1), melanin production inhibitory action on B16 melanoma cells was tested as follows.

After culturing B16 melanoma cells using 10% FBS-containing Dulbecco's MEM medium, the cells were collected by trypsinization. After diluting the collected cells with Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline to a cell density of 24.0×10 ⁴ cells/mL, 300 μL per well was seeded in a 48-well plate. cultured for hours.

After completion of the culture, 300 μL of a test sample (Sample 1, see Table 7 below for sample concentrations) dissolved in Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline was added to each well and cultured for 4 days. As a control, Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline to which no sample was added was used and cultured in the same manner. After the culture was completed, the medium was removed, 200 μL of a 2 mol/L NaOH solution was added, the cells were disrupted with an ultrasonicator, and the absorbance at a wavelength of 475 nm was measured. The amount of melanin was calculated from the measured absorbance values based on a calibration curve prepared using synthetic melanin (manufactured by SIGMA).

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

Melanin production suppression 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 wavelength 540 nm without addition of sample D: Absorbance at wavelength 540 nm with addition of test sample Table 7 shows the results.

As shown in Table 7, 4-vinylguaiacol (Sample 1) was found to have an excellent melanin production inhibitory effect.

[Test Example 8] Testosterone 5α-reductase inhibitory activity test 4-Vinylguaiacol (Sample 1) was tested for testosterone 5α-reductase inhibitory activity as follows.

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

Furthermore, 80 μL of a test sample (sample 1, see Table 8 below for sample concentration) solution prepared with 50% ethanol and 75 μL of S-9 (manufactured by Oriental Yeast Co., Ltd., rat liver homogenate) were added and mixed, Incubated for 30 minutes at 37°C. After that, 1 mL of methylene chloride was added to stop the reaction. This was centrifuged (1600×g, 10 minutes), the methylene chloride layer was separated, and the separated methylene chloride layer was subjected to gas chromatographic analysis under the following conditions to obtain - Dihydrotestosterone (5α-DHT) and testosterone concentrations were determined. As a control, the same amount (80 μL) of the sample solvent was used instead of the test sample solution, and the sample was treated in the same manner and subjected to gas chromatography analysis.

<Gas chromatography conditions>
Apparatus 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

The concentrations of 3α-androstanediol, 5α-DHT and testosterone were quantified by the following methods.
Standards of 3α-androstanediol, 5α-DHT and testosterone were dissolved in methylene chloride, and the solution was subjected to gas chromatography analysis. The corresponding relationship with the concentration of is obtained in advance. Then, the concentrations per peak area of 3α-androstanediol, 5α-DHT, and testosterone after the reaction between testosterone and S-9 are calculated using the correspondence relationship obtained in advance, using the following formula (1) sought 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 D: peak area of standard

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

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: concentration of testosterone (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) x 100 (3)
Each term in the formula represents the following.
H: Conversion rate with addition of test sample I: Conversion rate with no addition of sample Table 8 shows the results.

As shown in Table 8, 4-vinylguaiacol (Sample 1) was confirmed to have excellent testosterone 5α-reductase inhibitory activity.

[Test Example 9] Androgen Receptor Binding Inhibitory Activity Test 4-Vinylguaiacol (Sample 1) was tested for androgen receptor binding inhibitory activity as follows.

Androgen-dependent mouse breast cancer cells (SC-3 cells) cloned from mouse spontaneous breast cancer (Shionogi cancer; SC115) were grown in 10 ^-8 mol/L testosterone-containing MEM medium (MEM/2) containing 2% DCC-FBS. After culturing using , the cells were harvested by trypsinization. After diluting the recovered cells with MEM/2 medium to a cell density of 1.0×10 ⁵ cells/μL, 100 μL of each well was seeded in a 96-well plate and cultured overnight.

Thereafter, the medium was removed, and 100 μL of the test sample (Sample 1, see Table 9 below for sample concentrations) dissolved in 0.5% BSA-containing HamF12+MEM medium (HMB) containing 1.0×10 ⁻⁹ mol/L DHT. was added and cultured for an additional 48 hours. After completion of the culture, the medium was removed, 100 μL of 0.4 mg/mL MTT prepared in MEM/2 medium containing 2% DCC-FBS was added, and after further culturing for 2 hours, blue formazan produced in the cells was treated with 2-propanol. Extracted in 200 μL. The absorbance of this extract was measured at 570 nm where the absorption maximum of blue formazan exists. At the same time, absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was taken as the amount of blue formazan produced.

In parallel with the above, in order to examine the effect of the sample alone on SC-3 cells, the same culture and measurement were performed in the case where only the test sample was added to the HMB medium without adding DHT. In addition, as controls, the same measurements were carried out in the case of culturing in HMB medium without addition of sample and DHT, and in the case of culturing in HMB medium with only DHT added without addition of sample. From the measurement results, the androgen receptor binding inhibition rate (%) was calculated according to the following formula.

Androgen receptor binding inhibition rate (%) = {1-(C-D)/(A-B)} x 100
Each term in the formula represents the following.
A: Blue formazan production amount with no sample addition/DHT addition B: Blue formazan production amount with no sample addition/DHT addition C: Blue formazan production amount with test sample addition/DHT addition D: Test sample addition/DHT Table 9 shows the amount of blue formazan produced without addition.

As shown in Table 9, 4-vinylguaiacol (Sample 1) was confirmed to have excellent androgen receptor binding inhibitory activity.

[Test Example 10] Dermal papilla cell growth promoting action test 4-Vinylguaiacol (Sample 1) was tested for dermal papilla cell growth promoting action as follows.

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

Thereafter, the medium was removed, and 200 μL of the test sample (Sample 1, see Table 10 below for sample concentration) dissolved in serum-free DMEM medium was added to each well and cultured for 4 more days. As a control, cells were cultured in the same manner using a serum-free DMEM medium to which no sample was added. After completion of the culture, the dermal papilla cell proliferation-promoting action was measured by MTT assay. Specifically, the medium was removed, 200 μL of 0.4 mg/mL MTT prepared in serum-free DMEM medium was added, and the cells were further cultured for 2 hours, after which blue formazan produced in cells was extracted with 100 μL of 2-propanol. The absorbance of this extract was measured at 570 nm where the absorption maximum of blue formazan exists. At the same time, absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was taken as the amount of blue formazan produced. From the measurement results, the dermal papilla cell proliferation promotion rate (%) was calculated based on the following formula.

Dermal papilla cell proliferation promotion rate (%) = A/B x 100
Each term in the formula represents the following.
A: Amount of blue formazan produced with test sample added B: Amount of blue formazan produced with no sample added Table 10 shows the results.

As shown in Table 10, 4-vinylguaiacol (Sample 1) was found to have an excellent dermal papilla cell proliferation-promoting effect.

[Test Example 11] Tumor necrosis factor (TNF-α) production inhibitory action test Mouse macrophage cells (RAW264.7) were cultured in 10% FBS-containing Dulbecco's MEM medium, and the cells were collected with a cell scraper. After diluting the collected cells with the above medium to a cell density of 1.0×10 ⁶ cells/mL, 100 μL of each well was seeded in a 96-well microplate and cultured for 4 hours.

After the culture was completed, the medium was removed, and 100 μL of a test sample (Sample 1, see Table 11 below for sample concentrations) dissolved in Dulbecco's MEM medium containing 10% FBS with a final concentration of 0.5% DMSO was added to each well. 100 μL of lipopolysaccharide (LPS) (DIFCO, E.coli 0111; B4) dissolved in 10% FBS-containing Dulbecco's MEM at a final concentration of 1 μg/mL was added and cultured for 24 hours. As a control, Dulbecco's MEM medium containing 10% FBS containing a final concentration of 0.5% DMSO to which no sample was added was used in the same manner. After completion of the culture, the amount of TNF-α in the culture supernatant of each well was measured using the sandwich ELISA method. From the obtained results, the TNF-α production inhibition rate (%) was calculated by the following formula.

TNF-α production suppression rate (%) = {(B-A) / B} × 100
Each term in the formula represents the following.
A: amount of TNF-α with addition of test sample B: amount of TNF-α with no addition of sample Table 11 shows the results.

As shown in Table 11, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent TNF-α production inhibitory effect.

[Test Example 12] Hyaluronidase activity inhibitory action test A test sample (sample 1, see Table 12 below for sample concentration) dissolved in 0.1 mol / L acetate buffer (pH 3.5) was added to 0.2 mL of a hyaluronidase solution (SIGMA Company, Type IV-S, from bovine testes, 400 NF units/mL) 0.1 mL was added and allowed to stand at 37°C for 20 minutes. Furthermore, 0.2 mL of 2.5 mmol/L calcium chloride was added as an activator, and the mixture was 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 after cooling, 0.2 mL of boric acid solution was added to each reaction solution and boiled for 3 minutes. After cooling with ice, 6 mL of p-DABA reagent was added and reacted at 37° C. for 20 minutes. After that, absorbance at a wavelength of 585 nm was measured.

In addition, as a blank, the same operation and absorbance measurement were performed in the case where no enzyme solution was added. Furthermore, 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 according to 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 with test sample added/enzyme added Sb: Absorbance at wavelength 585 nm with test sample added/enzyme added Ct: Absorbance at wavelength 585 nm with sample not added/enzyme added Cb: Sample not added/enzyme Table 12 shows the absorbance results at a wavelength of 585 nm without addition.

As shown in Table 12, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent hyaluronidase activity inhibitory action. It was also confirmed that the degree of hyaluronidase activity inhibitory action can be adjusted by the concentration of 4-vinylguaiacol.

[Test Example 13] Hexosaminidase release inhibitory activity test 4-Vinylguaiacol (Sample 1) was tested for hexosaminidase release inhibitory activity as follows.

After culturing rat basophilic leukemia cells (RBL-2H3) using 15% FBS-containing S-MEM medium, the cells were collected by trypsinization. The recovered 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. After that, 100 μL of each well was seeded in a 96-well plate and cultured overnight.

After culturing, the medium was removed and the cells were washed twice with 100 μL of Silaganian buffer. Next, 10 μL of the test sample (Sample 1, see Table 13 below for the sample concentration) dissolved in the same buffer 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 performed using 40 μL of Silaganian buffer solution to which no sample was added. Subsequently, 10 μL of 100 ng/mL DNP-BSA solution was added, and left at 37° C. for 15 minutes to release hexosaminidase.

Release was then stopped by placing the 96-well plate on ice. Collect 10 μL of cell supernatant from each well into a new 96-well plate, add 10 μL of 1 mmol/L p-NAG (p-nitrophenyl-N-acetyl-β-D-glucosaminide) solution to each well, and place at 37°C. was reacted 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, absorbance at wavelengths of 415 nm and 650 nm was measured, and the value obtained by subtracting the absorbance at 650 nm from the absorbance at 415 nm was used as a correction value. As a blank, a mixture of 10 μL of cell supernatant and 250 μL of 0.1 mol/L Na ₂ CO ₃ /NaHCO ₃ was measured for absorbance at wavelengths of 415 nm and 650 nm, and corrected values were calculated. From the obtained measurement results, the hexosaminidase release inhibition rate (%) was calculated according to the following formula.

Hexosaminidase release inhibition rate (%) = {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 and no addition of p-NAG Table 13 shows the results.

As shown in Table 13, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent hexosaminidase release inhibitory action.

[Test Example 14] COX-2 activity inhibitory action test in mouse macrophages (PGE ₂ production inhibitory action test)
After culturing mouse macrophage cells (RAW264.7) using 10% FBS-containing Dulbecco's MEM medium, the cells were collected with a cell scraper. The recovered cells were diluted with 10% FBS-containing Dulbecco's MEM medium to a concentration of 2.0×10 ⁵ cells/mL, then seeded in 96-well plates at 100 μL per well and cultured for 18 hours.

After completion of the culture, the medium was changed to a medium containing 500 μmol/L aspirin and cultured for 4 hours in order to acetylate and inactivate existing COX-1 and a small amount of COX-2. After that, the cells were washed three times with PBS (-) buffer, and the test sample dissolved in Dulbecco's MEM medium containing 10% FBS containing a final concentration of 0.5% DMSO (Sample 1, see Table 14 below for sample concentrations) to each well, 100 μL of lipopolysaccharide (LPS) (DIFCO, E. coli 0111; B4) dissolved in Dulbecco's MEM containing 10% FBS at a final concentration of 1 μg/mL was added and cultured for 16 hours. bottom. As a control, Dulbecco's MEM medium containing 10% FBS containing a final concentration of 0.5% DMSO to which no sample was added was used and cultured in the same manner. After completion of the culture, the amount of prostaglandin E ₂ in the culture supernatant of each well was quantified using PGE ₂ EIA Kit (manufactured by Cayman Chemical). From the obtained results, the COX-2 activity inhibition rate (%, PGE ₂ production inhibition rate) was calculated according to the following formula.

Macrophage COX-2 activity inhibition rate (%) = {1-(A-C)/(B-C)} x 100
Each term in the formula represents the following.
A: Prostaglandin _E2 amount with test sample added/LPS stimulation B: Prostaglandin _E2 amount with no sample added/LPS stimulation C: Prostaglandin _E2 amount with no sample added/LPS stimulation Results is shown in Table 14.

As shown in Table 14, 4-vinylguaiacol (Sample 1) was confirmed to have excellent COX-2 activity inhibitory action in macrophages.

[Test Example 15] COX-2 activity inhibitory action test in keratinocytes (PGE ₂ production inhibitory action test)
After culturing normal human neonatal epidermal keratinocytes (NHEK) using a normal human epidermal keratinocyte growth medium (KGM), the cells were collected by trypsinization. The collected cells were diluted with KGM medium to a cell density of 12.5×10 ⁴ cells/mL, and then seeded at 200 μL per well on a collagen-coated 48-well plate (2.5×10 ⁴ cells/well). ), cultured overnight. After confirming that the cells had settled, the cells were cultured in 200 μL of KGM medium without hydrocortisone for 24 hours.

After culturing, 200 μL of 500 μmol/L aspirin-containing KGM medium (without hydrocortisone) was added to acetylate and inactivate existing COX-1 and small amount of COX-2, and cultured for 4 hours. After culturing, the cells were washed three times with PBS(-) buffer, added with 100 μL of PBS(-) buffer, irradiated with UV-B (50 mJ/cm ² ), and then dissolved in KGM medium (without hydrocortisone). 400 μL of the prepared test sample (Sample 1, see Table 15 below for sample concentration) was added to each well and cultured for 24 hours under conditions of 37° C. and 5% CO ₂ . As a control, a sample-free KGM medium (without hydrocortisone) was cultured in the same manner. After completion of the culture, the amount of prostaglandin E ₂ in the culture supernatant of each well was quantified using PGE ₂ EIA Kit (manufactured by Cayman Chemical). From the obtained results, the COX-2 activity inhibition rate (%, PGE ₂ production inhibition rate) was calculated according to the following formula.

Keratinocyte COX-2 activity inhibition rate (%) = {1-(A-C)/(B-C)} x 100
Each term in the formula represents the following.
_{Result _ _} is shown in Table 15.

As shown in Table 15, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent COX-2 inhibitory effect on keratinocytes.

[Test Example 16] Cyclic AMP phosphodiesterase activity inhibitory action test 4-Vinylguaiacol (Sample 1) was tested for cyclic AMP phosphodiesterase activity inhibitory action as follows.

0.1 mL of 2.5 mg/mL bovine serum albumin solution and 0.1 mg/mL cyclic AMP phosphodiesterase solution in 0.2 mL of 50 mmol/L Tris-HCl buffer (pH 7.5) containing 5 mmol/L magnesium chloride. 0.1 mL and 0.05 mL of test sample solution (Sample 1, see Table 16 below for sample concentration) were added, and the mixture was allowed to stand at 37° C. for 5 minutes. After that, 0.05 mL of 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, centrifuged (2260×g, 10 minutes, 4° C.), and the reaction substrate in the supernatant, cyclic AMP, was removed as follows. Analysis was performed under high performance liquid chromatography conditions. As a control, the same operation was performed by adding only the solvent without the sample.

<High performance liquid chromatography conditions>
Product name: Chromatocorder 12 (manufactured by SYSTEM INSTRUMENTS)
Stationary phase: Wakosil C ₁₈ -ODS 5 μm (manufactured by Wako Pure Chemical Industries, Ltd.)
Column length: 250mm
Mobile phase: 1 mmol/L TBAP _in 25 mmol/L _KH2PO4 : _CH3CN =90:10
Mobile phase flow rate: 1.0 mL/min
Detection: 260nm

Next, the peak area of the cyclic AMP standard (A), the peak area of the supernatant of the reaction solution of the cyclic AMP standard and cyclic AMP phosphodiesterase when no sample was added (B1), and the peak area when the test sample was added The peak area (B2) of the supernatant of the reaction solution of the click AMP standard and cyclic AMP phosphodiesterase was determined. From the obtained results, the decomposition rate (C) of the cyclic AMP standard product without addition of the sample and the decomposition rate (D) of the cyclic AMP standard product with the addition of the test sample were calculated by the following formulas.

Degradation rate of standard product without addition of sample (C,%) = (1-B1/A) x 100
Degradation rate (D,%) of standard product by addition of test sample = (1-B2/A) x 100

After that, based on each decomposition rate (C, D) calculated by the above formula, the cyclic AMP phosphodiesterase activity inhibition rate (%) was calculated by the following formula.
cAMP phosphodiesterase activity inhibition rate (%) = (1-D / C) × 100
The results are shown in Table 16.

As shown in Table 16, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent cyclic AMP phosphodiesterase activity inhibitory action.

[Test Example 17] Glutathione depletion inhibitory action test using normal human hepatocytes Normal human hepatocytes were cultured using Dulbecco's MEM medium containing 10% FBS, and then the cells were collected by trypsinization. The recovered cells were diluted with 10% FBS-containing Dulbecco's MEM medium to a cell density of 10×10 ⁴ cells/mL, seeded in 48-well plates at 200 μL per well, and cultured overnight.

After culturing, 200 μL of the test sample dissolved in 1% FBS-containing Dulbecco's MEM (Sample 1, see Table 17 below for sample concentrations) and 200 μL of galactosamine hydrochloride at a final concentration of 5 mM were added to each well, and incubated for another 24 hours. cultured. In addition, after similarly seeding the cells, the cells not treated with galactosamine hydrochloride and the cells treated with galactosamine hydrochloride after seeding and without adding the test sample were measured in the same manner, and were assigned to a non-treated group and a treated group, respectively. After the culture was completed, the medium was removed from each well, washed with 400 μL of PBS(-) buffer, and the cells were lysed using 150 μL of M-PER (PIERCE).

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

Glutathione decrease suppression rate (%) = {(Nt-C) - (Nt-Sa)} / (Nt-C) × 100
Each term in the formula represents the following.
Nt: Glutathione amount per total protein amount in galactosamine hydrochloride non-treated (untreated group) C: Glutathione amount per total protein amount in sample-free/galactosamine hydrochloride-treated (treated group) Sa: Test sample added/galactosamine Glutathione amount per total protein amount in hydrochloride treatment Table 17 shows the results.

As shown in Table 17, 4-vinylguaiacol (Sample 1) was confirmed to have an excellent glutathione reduction inhibitory effect.

[Test Example 18] DPPIV activity inhibitory activity test 4-Vinylguaiacol (Sample 1) was tested for dipeptidyl peptidase IV (DPPIV) inhibitory activity as follows.

In a 96-well plate, 25 μL of the test sample (Sample 1, see Table 18 below for final concentration) prepared in 25 mM Tris-HCl buffer (pH 8.0) and 0.4 μg/ 25 μL of mL DPPIV (rhCD26, manufactured by R&D Systems) solution was mixed and pre-incubated at 37° C. for 5 minutes. After that, 50 μL of 0.5 mM Gly-Pro-p-NA·Tos (manufactured by Peptide Institute) prepared with the above buffer solution was added and allowed to react 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-(C-D)/(A-B)} x 100
Each term in the formula represents the following.
A: Absorbance at a wavelength of 415 nm with no sample added/enzyme added B: Absorbance at a wavelength of 415 nm with no sample added/enzyme added C: Absorbance at a wavelength of 415 nm with test sample added/enzyme added D: Test sample added/enzyme Table 18 shows the absorbance results at a wavelength of 415 nm without addition.

As shown in Table 18, 4-vinylguaiacol (Sample 1) exhibited excellent DPPIV inhibitory activity.

[Formulation Example 1]
A milky lotion having the following composition was prepared by a conventional method.
0.01 g of 4-vinyl guaiacol
Jojoba oil 4.00g
1,3-butylene glycol 3.00 g
Arbutin 3.00g
Polyoxyethylene cetyl ether (20E.O.) 2.50g
2.00 g olive oil
2.00 g of squalane
Cetanol 2.00g
Glyceryl monostearate 2.00g
Polyoxyethylene sorbitan oleate (20E.O.) 2.00g
Methyl paraoxybenzoate 0.15g
Stearyl glycyrrhizinate 0.10 g
Huangji extract 0.10g
Dipotassium glycyrrhizinate 0.10g
Ginkgo biloba extract 0.10g
Conchiolin 0.10g
Phellodendron bark extract 0.10g
Chamomile extract 0.10g
Perfume 0.05g
Remainder of purified water (total amount is 100 g)

[Formulation Example 2]
A cream having the following composition was produced by a conventional method.
0.05 g of 4-vinyl guaiacol
Kujin extract 0.1g
Scutellaria root extract 0.1g
Liquid paraffin 5.0g
Bleached beeswax 4.0g
Squalane 10.0g
Cetanol 3.0g
Lanolin 2.0g
1.0 g of stearic acid
Polyoxyethylene sorbitan oleate (20E.O.) 1.5g
Glyceryl monostearate 3.0g
Oil-soluble licorice extract 0.1g
1,3-butylene glycol 6.0 g
Methyl paraoxybenzoate 1.5g
Perfume 0.1g
Remainder of purified water (total amount is 100 g)

[Formulation Example 3]
A beauty essence having the following composition was prepared by a conventional method.
0.01 g of 4-vinyl guaiacol
Chamomile extract 0.1g
carrot extract 0.1g
xanthan gum 0.3g
0.1 g of hydroxyethyl cellulose
Carboxyvinyl polymer 0.1 g
1,3-butylene glycol 4.0 g
Dipotassium glycyrrhizinate 0.1g
2.0 g of glycerin
Potassium hydroxide 0.25g
Perfume 0.01g
Preservative (methyl paraoxybenzoate) 0.15g
2.0 g of ethanol
Remainder of purified water (total amount is 100 g)

[Formulation Example 4]
A hair tonic having the following composition was produced by a conventional method.
0.4 g of 4-vinyl guaiacol
Tocopherol acetate Appropriate amount Cephalatine 0.002g
Isopropylmethylphenol 0.1g
Sodium hyaluronate 0.15g
15.0 g of glycerin
Ethanol 15.0g
Perfume Appropriate amount Chelating agent (edetate sodium) Appropriate amount Preservative (hinokitiol) Appropriate amount Solubilizer (polyoxyethylene cetyl ether) Appropriate amount Purified water Balance

[Formulation Example 5]
A shampoo having the following composition was produced by a conventional method.
0.5 g of 4-vinyl guaiacol
Marjoram extract 1.0g
Plum fruit part extract 0.2g
Coconut fatty acid methyl taurate sodium 10.0 g
Coconut fatty acid amidopropyl betaine 10.0g
Sodium polyoxyethylene alkyl ether sulfate 20.0 g
Coconut fatty acid diethanolamide 4.0 g
Propylene glycol 2.0 g
Fragrance Appropriate amount Purified water Remainder (Total amount is 100g)

[Formulation Example 6]
A tablet having the following composition was produced by a conventional method.
4-vinyl guaiacol 5.0 mg
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]
An oral liquid preparation having the following composition was prepared by a conventional method.
<Composition in 1 ampoule (100 mL per bottle)>
4-vinyl guaiacol 0.3% by mass
Sorbit 12.0% by mass
Sodium benzoate 0.1% by mass
Perfume 1.0% by mass
Calcium sulfate 0.5% by mass
Remainder of purified water (100% by mass)

The anti-aging agent, whitening agent, hair restorer, anti-androgenic agent, anti-inflammatory agent, anti-obesity agent, cAMP phosphodiesterase activity inhibitor, glutathione lowering inhibitor and DPPIV activity inhibitor of the present invention are useful for preventing skin aging symptoms, Promoting healing of wounds or burns; Prevention, treatment or improvement of dry skin diseases; Prevention or improvement of pigmentation such as skin darkening, age spots and freckles; Alopecia, especially male pattern baldness. prevention, treatment or improvement of various skin inflammatory diseases; prevention, treatment or improvement of obesity; improvement or improvement of liver function; prevention or treatment of type 2 diabetes, obesity, hypertension, insulin resistance, etc. or improvement;

Claims (1)

A food or drink containing 4-vinylguaiacol,
Food and drink for use in one or more anti-skin aging applications selected from the group consisting of promotion of epidermal keratinocyte proliferation, promotion of serine palmitoyltransferase mRNA expression and promotion of aquaporin 3 mRNA expression.