Whitening agent

Abstract

The objective is to identify compounds derived from natural products that exhibit superior whitening effects and to provide whitening agents that use these compounds as active ingredients. [Solution] The whitening agent of the present invention uses compound 1 and / or compound 2 represented by the following general formula (I) as active ingredients. TIFF2026063223000035.tif55153

Description

[Technical Field]

[0001] This invention relates to an anti-metabolic syndrome agent, a skin whitening agent, an anti-aging agent, a hair growth agent, an anti-inflammatory agent, and a liver function improving agent, all of which contain compounds derived from natural products as active ingredients. Furthermore, this invention also relates to oral compositions, skin cosmetics, and hair cosmetics containing such compounds. [Background technology]

[0002] In recent years, obesity has been increasing due to lifestyle factors such as overeating and lack of exercise, leading to an increase in body fat. This rise in obesity is not limited to humans; it is also seen in pets and livestock. Obesity is a major health problem, as it can cause metabolic syndrome, including hyperlipidemia and arteriosclerosis.

[0003] Here, cyclic AMP (cAMP) is known to be involved in lipolysis in the body. cAMP activates lipases present in the body, and activated lipases break down fats into fatty acids and glycerol. However, when cAMP phosphodiesterase is activated, the breakdown of cAMP is induced, and the activation of lipase is inhibited. Therefore, it is thought that inhibiting the activity of cAMP phosphodiesterase can increase the amount of cAMP in cells and promote lipolysis.

[0004] Furthermore, platelet aggregation, which triggers inflammatory responses, is related to the concentration of cyclic AMP (cAMP) in platelets. It is known that when cAMP is broken down by cAMP phosphodiesterase and its concentration decreases, platelets become more prone to aggregation. Therefore, it is thought that inhibiting the action of cAMP phosphodiesterase and preventing a decrease in cAMP concentration can prevent platelet aggregation, thereby preventing, treating, or improving allergic diseases, inflammatory diseases, etc. Tubeimoside I (see Patent Document 1) is known to have cAMP phosphodiesterase activity inhibitory effects.

[0005] Dipeptidyl peptidase IV (hereinafter also referred to as "DPP IV") is a serine protease that recognizes the second proline or alanine from the N-terminus and has enzymatic activity to cleave its C-terminal side. DPP IV is expressed on the cell surface of epithelial and endothelial cells in tissues such as the kidney, liver, intestines, and placenta, as well as T cells, and is thought to be involved in various physiological phenomena through its enzymatic activity.

[0006] One example of a substrate for DPP IV is a hormone called an incretin. Incretins are a general term for hormones secreted from the intestinal tract in response to nutrient stimulation that promote insulin secretion from pancreatic β-cells in a glucose-dependent manner, and GLP-1 and GIP are well known examples. These incretins not only promote glucose-dependent insulin secretion, but also have effects such as suppressing glucagon secretion from α-cells, lowering blood pressure, suppressing gastric emptying, and even suppressing appetite by acting on the hypothalamus (see Non-Patent Literature 1). However, incretins are degraded by DPP IV, and for example, the half-life of GLP-1 in vivo is known to be about 1.5 minutes. Therefore, if the enzymatic activity of DPP IV can be inhibited, the half-life of incretins in vivo can be extended, and it is expected that this will be useful in treating metabolic syndrome such as type 2 diabetes, obesity, hypertension, and insulin resistance through the aforementioned effects of incretins.

[0007] Furthermore, DPP IV is identical to CD26, one of the T cell activation markers, and is known to regulate the activity of many immunomodulatory peptides as substrates. Therefore, it is thought that controlling the activity of DPP IV may control immune responses such as autoimmune diseases like rheumatoid arthritis and transplant rejection. In addition, DPP IV is known to be involved in the metabolism of several neuropeptides and growth hormone; invasion, metastasis, and angiogenesis in cancer; and HIV infection of lymphocytes. Therefore, it is thought that inhibiting the activity of DPP IV may treat diseases such as pain, neurodegenerative diseases, and neuropsychiatric disorders (e.g., sciatica, Alzheimer's disease, depression, etc.); growth hormone deficiency and diseases for which growth hormone is used as treatment; cancer (e.g., T-cell lymphoma, acute lymphoblastic leukemia, thyroid cancer, basal cell carcinoma, breast cancer, etc.); and HIV infection (AIDS).

[0008] In the skin, melanin plays a role in protecting the body from ultraviolet rays, but excessive production or uneven accumulation can cause skin darkening and age spots. Generally, melanin is formed through the action of the enzyme tyrosinase, which is biosynthesized in pigment cells. Tyrosine is converted to dopa, dopa to dopaquinone, and then through intermediates such as 5,6-dihydroxyindophenol. Therefore, to prevent, treat, or improve skin darkening (cutaneous pigmentation), age spots, freckles, etc., it is thought that inhibiting the activity of tyrosinase, which is involved in melanin production, or suppressing melanin production may be effective.

[0009] Traditionally, the prevention, treatment, or improvement of skin pigmentation disorders, age spots, freckles, etc., has involved topical application of whitening agents containing chemically synthesized products such as hydroquinone as the active ingredient. However, chemically synthesized products such as hydroquinone may cause side effects such as skin irritation and allergies. Therefore, there is a need for the development of whitening agents using highly safe natural raw materials as active ingredients. Examples of substances with tyrosinase activity inhibitory effects include willow knotweed extract (see Patent Document 2). Examples of substances with melanin production inhibitory effects include extracts from plants of the genus Saussurea (see Patent Document 3).

[0010] The epidermis and dermis of the skin are composed of epidermal cells, fibroblasts, and extracellular matrix components such as collagen, elastin, and hyaluronic acid that support the skin structure outside of these cells. In young skin, fibroblast proliferation is active, and the interaction of skin tissues such as fibroblasts and extracellular matrix components maintains homeostasis, ensuring moisture retention, flexibility, and elasticity, and keeping the skin looking firm, radiant, and fresh.

[0011] However, exposure to certain external factors such as ultraviolet radiation, extreme dryness of the air, and excessive skin washing, as well as aging, can reduce the production of collagen, elastin, and hyaluronic acid, the main components of the extracellular matrix, and cause degradation and alteration. As a result, the skin's moisturizing function and elasticity decline, and abnormal exfoliation of the stratum corneum occurs, causing the skin to lose its firmness and luster, and exhibiting signs of aging such as rough skin and wrinkles. Thus, changes associated with skin aging, namely wrinkles, dullness, changes in texture, and decreased elasticity, are related to the decrease and alteration of matrix components such as collagen, elastin, and hyaluronic acid. Therefore, promoting the production of collagen, elastin, or hyaluronic acid is important for preventing, treating, or improving skin aging.

[0012] Here, among these extracellular matrix components, collagen is a fibrous protein that contributes to maintaining the structure and mechanical strength of skin tissue. If the production of collagen can be promoted, it is considered that it is possible to prevent, treat or improve skin aging symptoms such as the occurrence of wrinkles, sagging, roughness, loss of firmness, and decreased elasticity.

[0013] In addition, collagen is also abundantly present in bones, tendons, ligaments, corneas, blood vessels, etc., and it is known that a decrease in collagen production due to aging, etc. causes osteoporosis and the like. Furthermore, in the process of wound healing, it is known that the production amount of collagen increases and serves as a scaffold for fibroblasts and the like, thereby promoting wound healing. Therefore, promoting the production of collagen is also important from the viewpoints of preventing or treating osteoporosis and the like and promoting wound healing. As those having an action of promoting collagen production, for example, an extract from Phellodendron amurense Rupr. (see Patent Document 4) and the like are known.

[0014] On the other hand, among the extracellular matrix components described above, elastin is a fiber that gives elasticity to skin tissue. If the production of elastin can be promoted, it is considered that it is possible to prevent, treat or improve skin aging symptoms such as the occurrence of wrinkles, sagging, loss of firmness, and decreased elasticity. In addition, elastin is decomposed by an enzyme called elastase, and elastase is activated by ultraviolet irradiation, thereby accelerating the decomposition of elastin. Therefore, by inhibiting the activity of elastase, the decomposition of elastin is suppressed, and it is considered that skin aging symptoms such as loss of firmness and decreased elasticity can be prevented and improved.

[0015] Furthermore, elastin is widely expressed not only in skin tissue but also in tissues requiring elasticity in the body, such as the lungs and blood vessels. It is known that as we age, the amount of normal elastin in these tissues decreases, leading to a decline in elasticity in the lungs and blood vessels, and causing lung diseases such as emphysema, as well as vascular diseases such as hypertension and aneurysms. In addition, it is known that smoking and other factors increase the activity of elastase in the body, which may destroy the alveolar walls and lead to emphysema. Furthermore, it is thought that increased elastase activity may destroy pulmonary capillaries, potentially leading to acute respiratory syndromes (ARDS) such as pulmonary edema. Therefore, if elastin production can be promoted, it is thought that a decrease in elasticity in the lungs and blood vessels will be less likely to occur, and lung diseases such as emphysema, as well as vascular diseases such as hypertension and aneurysms, can be prevented and treated. Furthermore, if the activity of elastase in the body can be inhibited, it is thought that respiratory diseases such as emphysema and pulmonary edema can be prevented and treated. For example, extracts from plants belonging to the genus Hippophae in the family Elaeagnaceae are known to have elastin production-promoting effects (see Patent Document 5). Also, for example, star fruit extract is known to have elastase activity inhibitory effects (see Patent Document 6).

[0016] On the other hand, among the extracellular matrix components mentioned above, hyaluronic acid is a type of mucopolysaccharide that has the function of holding cells in place by filling the intercellular spaces, and also has numerous other functions such as retaining moisture in the intercellular spaces, providing lubrication and flexibility to tissues, and resisting external forces such as mechanical damage. It is believed that if the production of hyaluronic acid can be promoted, it may be possible to prevent, treat, or improve symptoms of skin aging such as rough skin, wrinkles, dullness, changes in texture, decreased elasticity, and decreased moisturizing function. Furthermore, it is believed that skin aging can be prevented, treated, or improved by promoting the expression of hyaluronic acid synthase 3 (HAS3), which is involved in promoting the synthesis of epidermal hyaluronic acid.

[0017] In addition to skin tissue, hyaluronic acid is also present in cartilage, synovial fluid, umbilical cord, vitreous humor, and other connective tissues. Of these, hyaluronic acid contained in synovial fluid covers the surface of articular cartilage and contributes to the smooth operation of joints through its lubricating function and its coating and protective function for cartilage. On the other hand, it is known that the concentration of hyaluronic acid in synovial fluid decreases in arthritis such as chronic rheumatoid arthritis. Therefore, it is thought that promoting the production of hyaluronic acid can prevent or treat arthritis such as chronic rheumatoid arthritis, osteoarthritis, suppurative arthritis, gouty arthritis, traumatic arthritis, or osteoarthritis. Furthermore, it is known that granulation tissue is formed during the healing process of wounds or burns, and that hyaluronic acid significantly increases in granulation tissue. Therefore, it is thought that promoting the production of hyaluronic acid can promote the healing of wounds or burns. Examples of substances that promote hyaluronic acid production include extracts from Quercus serrata (see Patent Document 4 mentioned above). Furthermore, licorice leaf extract (see Patent Document 7) and others are known to have an effect of promoting the expression of hyaluronic acid synthase 3 (HAS3).

[0018] On the other hand, the basement membrane exists at the boundary between the epidermis and dermis, which make up the skin. The basement membrane not only connects the epidermis and dermis but also plays an important role in maintaining skin function (see Non-Patent Literature 2). The main skeleton of the basement membrane is a mesh structure made of type IV collagen. Various glycoproteins, mainly composed of laminin-332, exist at the boundary between the basement membrane and the epidermis and connect the basement membrane and the epidermis. This laminin-332 is produced by epidermal keratinocytes present in the epidermis. In young skin, the interaction between the epidermis and dermis is maintained in a homeostatic state by the function of the basement membrane, ensuring moisture retention, flexibility, elasticity, etc., and the skin is maintained in a fresh state with firmness and luster.

[0019] However, when exposed to certain external factors such as ultraviolet radiation, extremely dry air, or excessive skin washing, or as aging progresses, laminin-332, a major component of the basement membrane, undergoes decomposition and alteration, destroying the basement membrane structure (see Non-Patent Literature 3). As a result, the skin loses its moisturizing function and elasticity, and the stratum corneum begins to peel abnormally, causing the skin to lose firmness and luster, and exhibiting aging symptoms such as roughness and wrinkles. Thus, changes associated with skin aging, namely wrinkles, dullness, changes in texture, and decreased elasticity, are related to a decrease in basement membrane components and structural changes in the basement membrane, and it is thought that promoting the production of laminin-332 can prevent and improve skin aging symptoms.

[0020] Laminins consist of various combinations of α, β, and γ chains, and currently 15 types (laminin 1 to laminin 15) are known. Of these, laminin-332 (α3β3γ2) is abundant in the basement membrane of epithelial tissues such as the skin, digestive tract, kidneys, and lungs. In genetic disorders caused by congenital abnormalities in the genes encoding each chain of laminin-332 (lethal congenital epidermolysis bullosa, Herlitz junctional epidermolysis bullosa), it is known to cause fatal symptoms of peeling of the epidermis throughout the body. Furthermore, laminin-332 is known to strongly adhere cells (high cell adhesion activity) and strongly promote cell movement (high cell motility activity) compared to other extracellular matrix molecules.

[0021] Thus, laminin-332 is known to promote cell migration in damaged skin and accelerate wound healing due to its high cell motility activity (see Patent Document 8). In other words, promoting the production of laminin-332 is important for accelerating the healing of skin damage that disrupts the structure of the basement membrane.

[0022] The epidermis functions to mitigate external stimuli and control the loss of internal components such as water. It consists of a four-layer structure, starting from the basal layer (the lowest layer), followed by the spinous layer, granular layer, and stratum corneum. The majority of cells in each layer are keratinocytes that differentiate from the basal layer. Keratinocytes that divide and proliferate in the basal layer differentiate as they pass through the spinous and granular layers to become corneocytes, which form the stratum corneum, composed of keratin protein fibers with strong cross-linking bonds. These corneocytes eventually slough off the stratum corneum as dead skin cells.

[0023] The stratum corneum is the outermost layer of the skin and acts as a physical barrier against external stimuli. To maintain this barrier function, the skin undergoes a cycle of keratinization, from the production of keratinocytes in the basal layer to their shedding as dead skin cells, which normally repeats every four weeks, thus carrying out epidermal metabolism. However, with age, the metabolic function of the stratum corneum also declines, leading to skin problems such as fine wrinkles, dullness, pigmentation, and rough skin. Therefore, it is believed that promoting the proliferation of keratinocytes and restoring the skin's metabolic function can improve skin aging such as fine wrinkles, dullness, and pigmentation. Conventionally, substances known to promote the proliferation of epidermal keratinocytes include oak extract (see Patent Document 9).

[0024] Furthermore, in order to promote cell proliferation, it is important to supply cells with the energy necessary for cell division. ATP is an example of an energy substance in living organisms, and it is thought that increasing the production of ATP promotes intracellular energy metabolism, leading to cell proliferation. However, as mentioned above, it has been reported that the amount of ATP, an energy substance, decreases in cells with reduced function or in senescent cells compared to normal cells (see Patent Document 10).

[0025] Therefore, it is believed that promoting ATP production in cells can activate those cells, stimulate cell division, and restore their proliferative capacity. In particular, promoting ATP production in skin cells is important for promoting skin turnover, restoring skin metabolic function, and preventing and improving skin aging such as wrinkles, dullness, and loss of texture. Conventionally, glycogen (see Patent Document 10) and extracts from natural products such as peaches (see Patent Document 11) have been known to have ATP production promoting effects.

[0026] Glutathione is a tripeptide composed of three amino acids: glutamic acid, cysteine, and glycine. It is a compound that contains the major cysteine ​​residue within cells. Within cells, glutathione plays a role in radical scavenging, regulating cellular function through oxidation-reduction, xenobiotic metabolism, and acting as an SH donor for various enzymes. It is also known as an antioxidant against reactive oxygen species. Its effects are thought to originate from the cysteine ​​residue. However, it has been reported that intracellular glutathione levels can become deficient or decreased due to excessive oxidative stress, the addition of foreign substances, and aging. This is thought to reduce the cell's ability to defend against oxidative stress and contribute to damage to cellular components such as DNA and proteins.

[0027] Diseases in which a decrease or deficiency of intracellular glutathione is known to be associated with the pathological condition include a group of diseases induced by oxidative stress, such as the formation of age spots on the skin, as well as liver damage (caused by excessive alcohol consumption or ingestion of foreign substances such as heavy metals and chemicals). In other words, it is thought that promoting glutathione production can enhance the cell's ability to defend against oxidative stress and prevent or treat the above-mentioned group of diseases caused by a decrease or deficiency of intracellular glutathione. Liquiritigenin (see Patent Document 12) is known to have a glutathione production-promoting effect.

[0028] Furthermore, among the basal layer, spinous layer, granular layer, and stratum corneum that make up the epidermis, in particular, in the granular layer, the cell membrane thickens to form a thickened cell membrane, and through the action of transglutaminase-1, glutamyl-lysine crosslinks are formed between protein molecules, creating strong keratin protein fibers. In addition, ceramides and other molecules covalently bond to some of these fibers, adopting a hydrophobic structure, which provides the foundation for the lamellar structure of intercellular lipids and forms the basis for the stratum corneum barrier function.

[0029] However, as we age, the production of transglutaminase-1 in the epidermis decreases, leading to a decline in the stratum corneum barrier function and the skin's moisturizing function. This can result in skin aging symptoms such as rough skin and dry skin, as well as the development of dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.). Therefore, it is believed that promoting the production of transglutaminase-1 in the epidermis can prevent, treat, or improve skin aging symptoms and dry skin diseases. Extracts from Hunan sweet tea (see Patent Document 13) are known to have transglutaminase-1 expression-promoting effects.

[0030] Ceramides are produced during the keratinization process of epidermal cells, based on serine and palmitoyl-CoA, through the action of enzymes, including serine palmitoyltransferase (SPT), which is known as the rate-limiting enzyme for ceramide synthesis. Ceramides are specifically present as the main component of intercellular lipids that cover the outermost layer of the skin, and play an important role in maintaining the skin's natural function as a barrier between the body and the outside world.

[0031] The structure of the stratum corneum can be likened to bricks and mortar, with intercellular lipids binding together keratinocytes stacked in about 15 layers to form a strong barrier membrane. Keratinocytes retain moisture by containing natural moisturizing factors, mainly composed of amino acids, within the cell. Meanwhile, the intercellular lipids of the keratinocytes are mainly composed of ceramides (about 50%), along with amphiphilic lipids such as cholesterol and fatty acids, and are characterized by a lamellar structure in which hydrophobic and hydrophilic parts alternate.

[0032] A decline in the skin's barrier function due to various internal and external factors increases transepidermal water loss, leading to dryness, desquamation, itching, and other symptoms, resulting in what is known as dry skin. Furthermore, a decline in the skin's barrier function increases skin inflammation, creating a vicious cycle where the skin's defense against various external stimuli is reduced. Recent studies have reported a decrease or change in the composition of keratinocyte ceramide components (so-called intercellular lipids) due to aging or in patients with atopic dermatitis, which is known as a barrier disorder (see Non-Patent Literature 4), and it has become widely known that ceramide is important for maintaining and improving the skin's barrier function. Methods for improving the skin's barrier function include supplementing ceramide from external sources (see Non-Patent Literature 5) and increasing the skin's ability to produce ceramide within the skin (see Non-Patent Literature 6).

[0033] In skin cells, aquaporins, known as water channels, are expressed on the cell membrane and play a role in taking in water and other low-molecular-weight substances from the intercellular space into the cell. In humans, 13 types of aquaporins (AQP0 to AQP12) are known to exist. In epidermal cells, AQP3 is mainly present and is thought to play a role in taking in not only water but also low-molecular-weight compounds such as glycerol and urea, which are involved in water retention.

[0034] However, AQP3 levels decrease with age, and this is suggested to be one of the causes of the decline in water retention function. Therefore, it is thought that promoting AQP3 expression can control age-related water retention function and barrier function (see Non-Patent Document 7). Examples of substances that promote AQP3 expression include extracts from star fruit leaves (see Patent Document 14).

[0035] Filaggrin is a component of the skin and is thought to be involved in the skin's barrier function, preventing the invasion of allergens, toxins, and infectious organisms. A decrease in filaggrin function due to mutations in the filaggrin gene is associated with an increased risk of developing atopic diseases, including atopic dermatitis (eczema, skin inflammation, itching, etc.), allergies, and asthma, and in more severe cases, it is known to lead to skin diseases such as ichthyosis vulgaris (see Non-Patent Literature 8).

[0036] On the other hand, the amino acids that are the main components of natural moisturizing factors (NMF) are produced when filaggrin, derived from keratohyalin granules, is broken down in the stratum corneum. This filaggrin is expressed as profilaggrin in epidermal keratinocytes located in the granular layer just below the stratum corneum. Subsequently, it is immediately phosphorylated and accumulated in keratohyalin granules, where it is broken down into filaggrin through dephosphorylation and hydrolysis, and then migrates to the stratum corneum, where it is known to enhance the aggregation efficiency of keratin filaments and participate in the internal structure of corneocytes (see Non-Patent Literature 9). In recent years, it has become known that this filaggrin is extremely important and indispensable for maintaining moisture in the skin, and that the ability to synthesize filaggrin decreases under conditions such as dryness, leading to a decrease in the amount of amino acids in the stratum corneum (see Non-Patent Literature 10).

[0037] Therefore, it is believed that promoting filaggrin (profilaggrin) expression in epidermal keratinocytes can prevent, treat, or improve atopic diseases, including atopic dermatitis (eczema, skin inflammation, itching, etc.), allergies, and asthma. Furthermore, it is expected that promoting filaggrin expression and thereby increasing the amount of amino acids in the stratum corneum can fundamentally improve the moisture environment of the stratum corneum. Artemisia princeps extract (see Patent Document 15) is known to have filaggrin expression-promoting effects.

[0038] Traditionally, the skin's barrier function was thought to be solely the responsibility of the stratum corneum. However, in recent years, it has been discovered that genetically deficient proteins in tight junctions (hereinafter sometimes referred to as "TJs") present in the epidermal granular layer cause the skin's barrier function to collapse, suggesting that TJs also play an important role in the skin's barrier function (see Non-Patent Literature 11). TJs are intercellular adhesion structures that not only tightly connect adjacent cells but also seal the gaps between cells, thereby controlling the permeability of substances. TJs are composed of cell membrane proteins such as claudin and occludin, and supporting proteins such as ZO-1 and ZO-2. These proteins are thought to constitute the backbone of the TJ strand and control the barrier function of TJs (see Non-Patent Literature 12). If the expression of claudin or occludin decreases for any reason, structural destruction of TJs occurs, and they cease to function as a barrier for the permeability of substances. This is thought to contribute to skin symptoms such as dry skin, rough skin, atopic dermatitis, and various infections.

[0039] Therefore, it is believed that promoting the production of claudins and occludins in the epidermis can stimulate the formation of tight junctions (TJs) in epidermal keratinocytes, thereby enhancing the skin's barrier function and moisture retention function, and preventing or improving skin conditions such as dry skin, rough skin, atopic dermatitis, and various infections. Asparagus linearis extract (Patent Document 16) is known to have claudin production-promoting and occludin production-promoting effects.

[0040] Carbohydrates are extremely important as an energy source for living organisms, including humans. However, carbohydrates are also known to undergo glycation reactions with proteins. Glycation is a series of reactions that begin with a non-enzymatic reaction between the carbonyl group of a carbohydrate and the amino group of a protein, etc., and proceed through Schiff bases and Amadori compounds to finally form advanced glycation end products (AGEs). As a result of glycation, proteins are modified by sugars in a non-enzymatic manner, which leads to denaturation of the protein and cross-linking between proteins, ultimately reducing the function of the protein.

[0041] Glycation reactions directly damage extracellular matrix proteins such as collagen by modifying and altering their structure, and also have other effects, such as triggering cellular responses when glycated proteins are recognized by receptors that use them as ligands. The effects of glycation reactions are particularly serious for diabetic patients with high blood glucose levels. Diabetic complications such as diabetic neuropathy, diabetic retinopathy, and diabetic nephropathy are known to be partly caused by protein glycation. Furthermore, glycation reactions in blood vessel walls are known to lead to the progression of arteriosclerosis through damage to endothelial cells and the accumulation of denatured proteins. In addition, extracellular matrix components, including collagen, account for more than half of the dry weight of tissues such as bone and skin. Therefore, for example, when collagen is glycated and abnormally cross-linked, it can lead to osteoporosis and osteoarthritis in bone and cartilage tissue, and a decrease in elasticity and dullness due to yellowing in the skin. Furthermore, abnormally cross-linked collagen becomes less susceptible to degradation by collagenases, which can induce the expression of collagenases and lead to problems such as the degradation of normal collagen.

[0042] Therefore, if glycation reactions can be suppressed in some way, for example, by inhibiting the formation of AGEs or promoting the breakdown of AGEs, it is expected to be useful in preventing or treating the aforementioned diseases, namely diabetic complications, arteriosclerosis, osteoporosis, and osteoarthritis. Furthermore, it is expected to be effective in preventing or improving decreased skin elasticity and dullness. As substances that have AGEs formation inhibitory effects and AGEs breakdown promoting effects, Osmanthus fragrans extract (see Patent Document 17) is known.

[0043] Many steroid hormones exert their effects by binding to receptors in their molecular form, secreted from the organs that produce them. However, in the case of male hormones collectively called androgens, for example, testosterone enters the cells of target organs, is reduced to 5α-dihydrotestosterone (5α-DHT) by testosterone 5α-reductase, and then binds to receptors to exert its androgenic effects.

[0044] Androgens are important hormones, but when they act excessively, they can induce a variety of undesirable symptoms, such as male pattern baldness, hirsutism, seborrhea, acne, benign prostatic hyperplasia, prostate tumors, and precocious puberty in boys. Therefore, methods have been known to suppress the effects of excessive androgens in order to improve these various symptoms, specifically by inhibiting the action of testosterone 5α-reductase, which reduces testosterone to active 5α-DHT, thereby suppressing the production of active 5α-DHT. To date, extracts from Japanese perilla (see Patent Document 18) have been known to have testosterone 5α-reductase inhibitory activity.

[0045] Hair grows and falls out repeatedly according to a cyclical hair cycle consisting of a growth phase, a regression phase, and a resting phase. Of this hair cycle, the stage from the resting phase to the growth phase, in which new hair follicles are formed, is considered to be the most important for hair growth, and dermal papilla cells are thought to play a crucial role in the proliferation and differentiation of hair follicle epithelial cells during this stage. Dermal papilla cells are located in the root portion of the hair root, enclosed by the basement membrane, inside the hair follicle epithelial cells which consist of outer root sheath cells and matrix cells near the hair root, and play an important role in the proliferation and differentiation of hair follicle epithelial cells and in hair formation, such as by acting on hair follicle epithelial cells to promote their proliferation (see Non-Patent Literature 13).

[0046] Thus, dermal papilla cells play an important role in the proliferation and differentiation of hair follicle epithelial cells and in hair formation, and it is thought that alopecia can be prevented or improved by promoting the proliferation of dermal papilla cells. To date, substances known to promote the proliferation of dermal papilla cells include, for example, wild thyme extract (see Patent Document 19).

[0047] The causes and mechanisms of inflammatory diseases, such as contact dermatitis, psoriasis, pemphigus vulgaris, atopic dermatitis, and other various skin inflammatory diseases accompanied by skin irritation, as well as rheumatoid arthritis, osteoarthritis, and asthma, are diverse. The causes are mainly known to be due to excessive production of nitric oxide (NO), histamine release, increased hyaluronidase activity, and prostaglandin E2 (PGE2) production.

[0048] Nitric oxide (NO) is a nitrogen oxide that contributes to air pollution and acid rain, but in recent years, it has been discovered that nitric oxide is a physiologically active substance that exhibits diverse functions in the body, including vascular endothelial relaxation factor (EDRF), neurotransmitter, and inhibitory factor for microorganisms and tumor cells in biological defense. In biological defense, nitric oxide produced by macrophages in particular protects against bacterial and viral infections.

[0049] However, when large amounts of nitric oxide are biosynthesized, it is not harmless to the body, causing the destruction of self-tissues and contributing to conditions such as inflammation, rheumatism, and diabetes. It is also known that large amounts of biosynthesized nitric oxide lead to relaxation of vascular smooth muscle and excessive increase in permeability, causing a significant drop in blood pressure and leading to endotoxin shock.

[0050] Therefore, in inflammatory diseases, it is important to suppress the excessive production of nitric oxide. Examples of substances that have an inhibitory effect on nitric oxide production include Angelica acutiloba, Aralia cordata root bark, Citrus aurantium, Plantago asiatica, Dioscorea japonica, Madder root, Scutellaria baicalensis, Sophora japonica flower, Sichuan pepper (see Non-Patent Document 14), extracts from plants belonging to the genus Hydrocotyle (see Patent Document 20), and malturosylarginine (see Patent Document 21).

[0051] Histamine release is the phenomenon in which histamine from mast cells is released outside the cell, and the released histamine triggers an inflammatory response. Therefore, attempts have been made to prevent or treat allergic and inflammatory diseases by using substances that inhibit or suppress histamine release. However, it is difficult to directly evaluate histamine release, and histamine release can be evaluated using the release of hexosaminidase, which has been confirmed to be released simultaneously with histamine, as an indicator. Therefore, it is thought that suppressing the release of hexosaminidase can simultaneously suppress histamine release, thereby being effective in preventing, treating, or improving inflammatory diseases.

[0052] Furthermore, histamine acts as a local neurotransmitter, mediating intercellular communication. In the digestive system, it is known to increase gastric acid secretion, and in the central nervous system, it functions as a neurotransmitter, contributing to the maintenance of wakefulness. However, excessive histamine release can cause ulcers due to excess gastric acid in the digestive system and contribute to sleep disorders in the central nervous system. As mentioned above, inhibiting the release of hexosaminidase can simultaneously inhibit the release of histamine, and it is thought that this can prevent, treat, or improve gastric ulcers, sleep disorders, etc., caused by excess gastric acid. Examples of substances that have a hexosaminidase release inhibitory effect include extracts from wisteria tea (see Patent Document 22).

[0053] Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid. Hyaluronic acid salts, which maintain affinity to body tissues, are broken down by ultraviolet light, oxygen, etc., in a hydrated system, and their water-retaining effect decreases as their molecular weight decreases. Hyaluronic acid also exists as intercellular tissue in the body and is involved in vascular permeability. Furthermore, hyaluronidase is present in mast cells, but is released by degranulation caused by its activation and acts as an inflammatory chemical mediator. Therefore, inhibiting the activity of hyaluronidase is expected to enhance moisturizing and prevent / reduce inflammation. Examples of substances that have hyaluronidase activity inhibitory effects include extracts from plants of the genus Osbecchia (see Patent Document 23).

[0054] Inflammation is a complex reaction that manifests as symptoms such as redness, swelling, fever, pain, itching, and functional impairment. For example, in the skin, exposure to ultraviolet light or contact with irritating substances can trigger the production of inflammatory cytokines and other substances within the skin, leading to skin inflammation. As a result, skin tissue is damaged, leading to various symptoms such as rough skin, redness, swelling, and hyperpigmentation.

[0055] One example of an inflammatory cytokine is prostaglandin E2 (PGE2). In the skin, PGE2 is produced, for example, in keratinocytes, and is a cause of skin inflammation. It has been shown that cyclooxygenase-2 (COX-2), an inducible type of cyclooxygenase, is mainly involved in the production of prostaglandins during inflammation. Therefore, as a method of treating or preventing skin inflammation, it is conceivable to suppress the production of PGE2 in keratinocytes or inhibit the activity of COX-2. Pentaerythritol and others are known to have an inhibitory effect on PGE2 production in keratinocytes (see Patent Document 24).

[0056] The liver is a vital organ essential for maintaining life, playing a central role in metabolism. Its main functions include the metabolism of sugars, proteins, lipids, and hormones, detoxification of harmful substances, bile production, and blood storage. Although liver damage is often difficult to detect, impaired liver function due to alcohol consumption, overnutrition, drug abuse, hepatitis viruses, etc., can still cause fatigue, malaise, loss of appetite, and even jaundice. As liver dysfunction progresses, it can lead to lifestyle-related diseases such as hepatitis and cirrhosis. Therefore, improving liver function and protecting it from damage is extremely important for maintaining a healthy life.

[0057] Glutathione is known as a biological component that protects the liver. Glutathione is a tripeptide composed of three amino acids: glutamic acid, cysteine, and glycine, and is a compound that has the major cysteine ​​residue in cells. Glutathione is mainly produced in the liver and supplied throughout the body. Within cells, it plays a role in radical scavenging, regulating cellular function through oxidation-reduction, and acting as an SH donor for various enzymes. In particular, in the liver, it is known to be involved in detoxification mechanisms and protect liver function. Liver glutathione is consumed in large quantities during the metabolism of ethanol due to alcohol consumption, or during the metabolism and detoxification of drugs. A decrease in liver glutathione levels can cause liver dysfunction such as acute or chronic alcoholic hepatitis and drug-induced hepatitis. Furthermore, a decrease in systemic glutathione levels is known to cause a variety of symptoms, including cataracts, Parkinson's disease, and the formation of age spots on the skin.

[0058] Therefore, if glutathione production in the liver (hepatocytes) can be promoted, it is expected that liver function can be improved, and various disorders caused by glutathione deficiency can be prevented, treated, or improved. Liquiritin (see Patent Document 11 mentioned above) is known to have a glutathione production-promoting effect in hepatocytes.

[0059] Adenosine triphosphate (ATP) is known as an ingredient that improves liver function. ATP is produced through the metabolism of glucose and fat and is used as an energy source. In the liver, ATP is used as an energy source in many chemical reactions that occur in the liver, such as the metabolism and detoxification of ethanol and drugs. It is thought that a decrease in ATP production leads to a decrease in the efficiency of these metabolic and detoxification processes, as well as a lack of energy necessary for activities such as work and exercise, resulting in fatigue. Therefore, if ATP production in the liver can be promoted, it is thought that it may be possible to prevent or improve symptoms caused by ethanol intake (e.g., hangovers), prevent, treat or improve liver damage caused by drugs, and promote recovery from fatigue. Long pepper extract (see Patent Document 25) is known to have an ATP production promoting effect in liver cells. [Prior art documents] [Patent Documents]

[0060] [Patent Document 1] Japanese Patent Publication No. 2006-056855 [Patent Document 2] Japanese Patent Publication No. 2004-083488 [Patent Document 3] Japanese Patent Publication No. 2002-201122 [Patent Document 4] Japanese Patent Publication No. 2003-146837 [Patent Document 5] Japanese Patent Publication No. 2005-022993 [Patent Document 6] Japanese Patent Publication No. 2003-300893 [Patent Document 7] Japanese Patent Publication No. 2010-090035 [Patent Document 8] Japanese Patent Publication No. 2006-063033 [Patent Document 9] Japanese Patent Publication No. 2006-056854 [Patent Document 10] Japanese Patent Publication No. 2003-321373 [Patent Document 11] Japanese Patent Publication No. 2009-256272 [Patent Document 12] Japanese Patent Publication No. 2009-269889 [Patent Document 13] Japanese Patent Publication No. 2007-099698 [Patent Document 14] Japanese Patent Publication No. 2009-191039 [Patent Document 15] Japanese Patent Publication No. 2012-219047 [Patent Document 16] Japanese Patent Publication No. 2009-256244 [Patent Document 17] Japanese Patent Publication No. 2013-023487 [Patent Document 18] Japanese Patent Publication No. 2010-184915 [Patent Document 19] Japanese Patent Publication No. 2006-219407 [Patent Document 20] Japanese Patent Publication No. 2006-28036 [Patent Document 21] Japanese Patent Publication No. 2010-90076 [Patent Document 22] Japanese Patent Publication No. 2003-012532 [Patent Document 23] Japanese Patent Publication No. 2003-055242 [Patent Document 24] Japanese Patent Publication No. 2015-214533 [Patent Document 25] Japanese Patent Publication No. 2011-184381 [Non-patent literature]

[0061] [Non-Patent Document 1] "Japanese Journal of Pharmacology," 2005, Vol. 125, No. 6, pp. 379-384. [Non-Patent Document 2] J. Cell Biol., 1992, Vol. 119, No. 3, p. 695-703 [Non-Patent Document 3] J. Invest. Dermatol., 1979, Vol. 73, No. 1, p. 59-66 [Non-Patent Document 4] J. Dermatol.,1993,Vol.20,No.1,p.1-6 [Non-Patent Document 5] "Fragrance Journal," 2004, Vol. 32, No. 11, pp. 23-32 [Non-Patent Document 6] Br. J. Dermatol.,2000,Vol.143,Issue 3,p.524-531 [Non-Patent Document 7] "Fragrance Journal," 2006, Vol. 34, No. 10, pp. 19-23 [Non-Patent Document 8] "Nat Genet.",2006,Vol.38,No.4,p.441-446 [Non-Patent Document 9] "Fragrance Journal Special Issue," 2000, Vol. 17, pp. 14-19 [Non-Patent Document 10] "Arch. Dermatol. Res.", 1996, Vol. 288, p. 442-446 [Non-Patent Document 11] J. Cell Biol.,2002,vol.156,pp.1099-1111 [Non-Patent Document 12] Journal of the Japanese Society of Cosmetic Science, 2007, vol.31, pp.296-301. [Non-Patent Document 13] Trends Genet.,1992,Vol.8,Issue 2,p.55-61 [Non-Patent Document 14] "Journal of Japanese and Chinese Medicine," 1998, Vol. 15, pp. 302-303. [Overview of the Initiative] [Problems that the invention aims to solve]

[0062] The present invention aims to identify compounds derived from natural products that exhibit excellent effects in anti-metabolic syndrome, skin whitening, anti-aging, hair growth, anti-inflammatory, or liver function improvement, and to provide anti-metabolic syndrome agents, skin whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, and liver function improving agents that contain these compounds as active ingredients. Furthermore, the present invention aims to provide oral compositions, skin cosmetics, and hair cosmetics that are suitable for anti-metabolic syndrome applications, skin whitening applications, anti-aging applications, hair growth applications, anti-inflammatory applications, or liver function improvement applications, by incorporating naturally derived compounds that have excellent effects in anti-metabolic syndrome, skin whitening, anti-aging, hair growth, anti-inflammatory, or liver function improvement applications. [Means for solving the problem]

[0063] To solve the above problems, the present invention's anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, and liver function improving agent is characterized by containing one or more compounds selected from the group consisting of compounds 1 to 3 represented by the following general formula (I) as active ingredients. Furthermore, the oral composition, skin cosmetic, and hair cosmetic of the present invention are characterized by containing one or more compounds selected from the group consisting of compounds 1 to 3 represented by the following general formula (I).

[0064] [ka] [Effects of the Invention]

[0065] According to the present invention, by using one or more compounds selected from the group consisting of compounds 1 to 3 represented by the above general formula (I) as an active ingredient, it is possible to provide an anti-metabolic syndrome agent, a skin whitening agent, an anti-aging agent, a hair growth agent, an anti-inflammatory agent, and a liver function improving agent having excellent effects. Furthermore, by incorporating one or more compounds selected from the group consisting of compounds 1 to 3 represented by the above general formula (I), oral compositions, skin cosmetics, and hair cosmetics suitable for anti-metabolic syndrome applications, skin whitening applications, anti-aging applications, hair growth applications, anti-inflammatory applications, or liver function improvement applications can be provided. [Modes for carrying out the invention]

[0066] Embodiments of the present invention will be described below.

[0067] [Phenylpropionic acids] The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, and liver function improving agent according to this embodiment are characterized by containing one or more compounds selected from the group consisting of compounds 1 to 3 represented by the following general formula (I) as active ingredients. Furthermore, the oral composition, skin cosmetic, and hair cosmetic according to this embodiment are characterized by containing one or more compounds selected from the group consisting of compounds 1 to 3 represented by the following general formula (I).

[0068] [ka]

[0069] The compounds represented by the above general formula (I) are all phenylpropionic acid derivatives. Compound 1 is 3,4-dihydroxyhydrocinnamic acid. Compound 2 is 3-(4-hydroxyphenyl)propionic acid, and Compound 3 is 3-phenylpropionic acid. Hereinafter, in this specification, the compound represented by formula (I) above may be referred to as "phenylpropionic acid."

[0070] The above-mentioned phenylpropionic acids can be produced, for example, by isolating and purifying them from plant extracts containing phenylpropionic acids. In this case, such plant extracts containing phenylpropionic acids can be obtained by methods commonly used for plant extraction. Examples of plants containing phenylpropionic acids include rice, barley, wheat, soybeans, adzuki beans, and corn.

[0071] Furthermore, the above-mentioned phenylpropionic acids can also be produced by fermenting, for example, 3,4-dihydroxycinnamic acid, 4-hydroxycinnamic acid, or cinnamic acid (hereinafter, these three compounds may be collectively referred to as "cinnamic acids"), or compositions containing these (for example, crushed plant material or extracts, etc.), with microorganisms possessing phenolic acid reductase to convert the cinnamic acids into phenylpropionic acids, and then extracting, isolating, and purifying the resulting fermented product. Examples of compositions containing cinnamic acids include crushed and extracted plants such as coffee, wheat, corn, tomato, mate, mugwort, and burdock. In addition, since cinnamic acids are components of lignin in woody and herbaceous plants, lignin or compositions containing it may be used as fermentation raw materials. On the other hand, examples of microorganisms possessing phenolic acid reductase include lactic acid bacteria such as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus crispatus, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus delbrueckii, Lactobacillus buchneri, Lactobacillus kefiranofaciens, Lactobacillus gallinarum, and Enterococcus faecalis.

[0072] The method for extracting, isolating, and purifying the phenylpropionic acids from the above-mentioned plants or fermented products is not particularly limited and can be carried out according to conventional methods. For example, the extraction process can be carried out by drying the above-mentioned plants or fermented products as extraction raw materials, then crushing them as is or using a crusher, and then subjecting them to extraction with an extraction solvent. Drying can be carried out in the sun or using a commonly used drying oven. Alternatively, the raw materials may be pretreated by degreasing with a non-polar solvent such as hexane before being used as extraction raw materials. Pretreatment such as degreasing allows for more efficient extraction with a polar solvent.

[0073] As the extraction solvent, it is preferable to use a polar solvent, such as water or a hydrophilic organic solvent. It is preferable to use these individually or in combination of two or more at room temperature or below the boiling point of the solvent.

[0074] Water that can be used as an extraction solvent includes pure water, tap water, well water, mineral water, mineral water, hot spring water, spring water, fresh water, etc., as well as water that has undergone various treatments. Treatments applied to water include, for example, purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, etc. Therefore, in this embodiment, water that can be used as an extraction solvent also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate-buffered physiological saline, etc.

[0075] Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin; and lower aliphatic ketones, such as acetone and methyl ethyl ketone.

[0076] When using a mixture of two or more polar solvents as an extraction solvent, the mixing ratio is arbitrary and can be adjusted as appropriate. For example, when using a mixture of water and a hydrophilic organic solvent as an extraction solvent, it can be mixed in any ratio, i.e., between 0:100 and less than 100:0 (volume ratio, hereafter expressed similarly), and can be adjusted as appropriate. For example, when using a mixture of water and a lower aliphatic alcohol as an extraction solvent, the mixing ratio (volume ratio) of water to the lower aliphatic alcohol can be 9:1 or higher, or even 7:3 or higher, or 1:9 or lower, or even 2:8 or lower. Furthermore, when using a mixture of water and a polyhydric alcohol, the mixing ratio of water to the polyhydric alcohol can be 8:2 or higher, or 1:9 or lower, and when using a mixture of water and a lower aliphatic ketone, the mixing ratio of water to the lower aliphatic ketone can be 9:1 or higher, or 2:8 or lower.

[0077] The extraction process is not particularly limited as long as it allows the soluble components contained in the raw material to be eluted into the extraction solvent, and can be carried out according to conventional methods. For example, the raw material can be immersed in an extraction solvent in an amount (by mass ratio) of 5 to 15 times the amount of the raw material, and the soluble components can be extracted at room temperature or under reflux heating. After extraction, the extract can be obtained by filtering to remove the extraction residue. By distilling off the solvent from the obtained extract, a paste-like concentrate can be obtained, and by further drying this concentrate, a dried product can be obtained.

[0078] The method for isolating and purifying the phenylpropionic acids from the extract, concentrate, or dried extract obtained as described above is not particularly limited and can be carried out by conventional methods. For example, the extract can be 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 recover the fraction containing phenylpropionic acids. In this case, the developing solvent can be appropriately selected according to the stationary phase used, but for example, when separating the extract by normal-phase chromatography using silica gel as the stationary phase, a chloroform:methanol ratio of 95:5 can be used as the developing solvent. Furthermore, the fraction containing phenylpropionic acids obtained by column chromatography may be purified using any organic compound purification method such as reverse-phase silica gel chromatography using ODS, recrystallization, liquid-liquid countercurrent extraction, or column chromatography using ion exchange resin.

[0079] [Anti-metabolic syndrome agents, skin whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, liver function enhancers] The phenylpropionic acids obtained as described above have excellent effects in anti-metabolic syndrome, skin whitening, anti-aging, hair growth, anti-inflammatory, and liver function improvement. Therefore, they can be used as active ingredients in anti-metabolic syndrome agents, skin whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, and liver function improving agents. Furthermore, the phenylpropionic acids can be used to manufacture anti-metabolic syndrome agents, skin whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents. The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, and liver function improving agent of this embodiment can be used in a wide range of applications, including pharmaceuticals, quasi-drugs, and cosmetics.

[0080] Here, the anti-metabolic syndrome effect of the above-mentioned phenylpropionic acids is preferably exerted based on cyclic AMP (cAMP) phosphodiesterase activity inhibition and / or dipeptidyl peptidase IV (DPP IV) activity inhibition. However, the anti-metabolic syndrome effect of the above-mentioned phenylpropionic acids is not limited to the anti-metabolic syndrome effect exerted based on the above-mentioned action. Furthermore, the phenylpropionic acids described above can be used for cAMP phosphodiesterase activity inhibition or DPP IV activity inhibition, respectively, by utilizing their cAMP phosphodiesterase activity inhibitory effect or DPP IV activity inhibitory effect. In other words, the anti-metabolic syndrome agent of this embodiment can also be used as a cAMP phosphodiesterase activity inhibitor or a DPP IV activity inhibitor containing the phenylpropionic acids described above as active ingredients.

[0081] The skin-whitening effect of the above-mentioned phenylpropionic acids is preferably exerted based on tyrosinase activity inhibition and / or melanin production suppression. However, the skin-whitening effect of the above-mentioned phenylpropionic acids is not limited to the skin-whitening effect exerted based on the above-mentioned effect. Furthermore, the phenylpropionic acids described above can be used for tyrosinase activity inhibition or melanin production inhibition purposes, respectively, by utilizing their tyrosinase activity inhibitory effect or melanin production inhibitory effect. In other words, the whitening agent of this embodiment can also be used as a tyrosinase activity inhibitor or melanin production inhibitor containing the phenylpropionic acids described above as an active ingredient.

[0082] The anti-aging effects of the above-mentioned phenylpropionic acids are preferably exerted based on one or more actions selected from the group consisting of type I collagen production promoting effect, elastin production promoting effect, hyaluronic acid production promoting effect, elastase activity inhibitory effect, HAS3 mRNA expression promoting effect, laminin-332 production promoting effect, epidermal keratinocyte proliferation promoting effect, ATP production promoting effect, glutathione production promoting effect, transglutaminase-1 (TGM1) mRNA expression promoting effect, serine palmitoyltransferase (SPT) mRNA expression promoting effect, aquaporin 3 (AQP3) mRNA expression promoting effect, filaggrin mRNA expression promoting effect, claudin-1 mRNA expression promoting effect, claudin-4 mRNA expression promoting effect, occludin mRNA expression promoting effect, inhibition of advanced glycation end products (AGEs) formation effect, and promotion of advanced glycation end product (AGEs) degradation effect. However, the anti-aging effects of the above-mentioned phenylpropionic acids are not limited to those exerted based on the above-mentioned effects. Furthermore, the above-mentioned phenylpropionic acids are used to promote type I collagen production, elastin production, hyaluronic acid production, elastase activity inhibition, HAS3 mRNA expression, laminin-332 production, epidermal keratinocyte proliferation, ATP production, glutathione production, TGM1 mRNA expression, SPT mRNA expression, AQP3 mRNA expression, filaggrin mRNA expression, claudin-1 mRNA expression, claudin-4 mRNA expression, occludin mRNA expression, AGEs formation inhibition, or AGEs degradation, respectively, for the following purposes: promoting type I collagen production, promoting elastin production, promoting hyaluronic acid production, inhibiting elastase activity, promoting HAS3 mRNA expression, promoting laminin-332 production, promoting epidermal keratinocyte proliferation, promoting ATP production, promoting glutathione production, promoting TGM1 mRNA expression, and promoting SPT It can be used for mRNA expression promotion, AQP3 mRNA expression promotion, filaggrin mRNA expression promotion, claudin-1 mRNA expression promotion, claudin-4 mRNA expression promotion, occludin mRNA expression promotion, AGEs formation suppression, or AGEs degradation promotion. In other words, the anti-aging agent of this embodiment can also be used as a type I collagen production promoter, elastin production promoter, hyaluronic acid production promoter, elastase activity inhibitor, HAS3 mRNA expression promoter, laminin-332 production promoter, epidermal keratinocyte proliferation promoter, ATP production promoter, glutathione production promoter, TGM1 mRNA expression promoter, SPT mRNA expression promoter, AQP3 mRNA expression promoter, filaggrin mRNA expression promoter, claudin-1 mRNA expression promoter, claudin-4 mRNA expression promoter, occludin mRNA expression promoter, AGEs formation inhibitor, or AGEs degradation promoter, with the above-mentioned phenylpropionic acids as the active ingredient.

[0083] The hair growth effect of the above-mentioned phenylpropionic acids is preferably exerted based on their testosterone 5α-reductase activity inhibitory effect and / or hair papilla cell proliferation promoting effect. However, the hair growth effect of the above-mentioned phenylpropionic acids is not limited to the hair growth effect exerted based on the above-mentioned effect. Furthermore, the above-mentioned phenylpropionic acids can be used for testosterone 5α-reductase inhibitory activity or dermal papilla cell proliferation promoting activity, respectively, by utilizing their testosterone 5α-reductase inhibitory activity or dermal papilla cell proliferation promoting activity. In other words, the hair growth agent of this embodiment can also be used as a testosterone 5α-reductase inhibitor or dermal papilla cell proliferation promoter containing the above-mentioned phenylpropionic acids as an active ingredient.

[0084] The anti-inflammatory effect of the above-mentioned phenylpropionic acids is preferably exerted based on one or more actions selected from the group consisting of nitric oxide (NO) production inhibition, hyaluronidase activity inhibition, hexosaminidase release inhibition, and prostaglandin E2 (PGE2) activity inhibition. However, the anti-inflammatory effect of the above-mentioned phenylpropionic acids is not limited to the anti-inflammatory effect exerted based on the above actions. Furthermore, the above-mentioned phenylpropionic acids can be used for NO production inhibition, hyaluronidase activity inhibition, hexosaminidase release inhibition, or PGE2 production promotion, respectively, by utilizing their NO production inhibitory effect, hyaluronidase activity inhibition effect, hexosaminidase release inhibition effect, or PGE2 production promotion effect. In other words, the anti-inflammatory agent of this embodiment can also be used as an NO production inhibitor, hyaluronidase activity inhibitor, hexosaminidase release inhibitor, or PGE2 production promoter, with the above-mentioned phenylpropionic acids as the active ingredient.

[0085] The liver function-enhancing effect of the above-mentioned phenylpropionic acids is preferably exerted based on the promotion of hepatocyte glutathione production and / or hepatocyte adenosine triphosphate (ATP) production. However, the liver function-enhancing effect of the above-mentioned phenylpropionic acids is not limited to the liver function-enhancing effect exerted based on the above-mentioned effect. Furthermore, the phenylpropionic acids described above can be used to promote hepatocyte glutathione production or hepatocyte ATP production, respectively, by utilizing their hepatocyte glutathione production-promoting or hepatocyte ATP production-promoting effects. In other words, the liver function improving agent of this embodiment can also be used as a hepatocyte glutathione production-promoting agent or a hepatocyte ATP production-promoting agent containing the phenylpropionic acids described above as an active ingredient.

[0086] Furthermore, in this embodiment, a composition containing phenylpropionic acids may be used as the active ingredient of the anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent, instead of the isolated phenylpropionic acids. Here, the "composition containing phenylpropionic acids" in this embodiment includes extracts obtained using plants containing phenylpropionic acids as extraction raw materials, fermented products containing phenylpropionic acids, and extracts obtained using the fermented products as extraction raw materials. In addition, the "extract" includes extracts obtained by extraction treatment, diluted or concentrated extracts, or dried products obtained by drying the extract.

[0087] When using a composition containing the above-mentioned phenylpropionic acids as an active ingredient in an anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent according to this embodiment, it is preferable that the composition contains 0.1% by mass or more of phenylpropionic acids, more preferably 5% by mass or more, and particularly preferably 50% by mass or more. By using a highly purified form of phenylpropionic acids as an active ingredient, an anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent with even greater efficacy can be obtained.

[0088] The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent of this embodiment may consist solely of the above-mentioned phenylpropionic acids or compositions containing phenylpropionic acids, or it may be a formulation of a composition containing phenylpropionic acids or phenylpropionic acids.

[0089] The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent of this embodiment can be formulated into any dosage form such as powder, granules, tablets, or liquid by conventional methods using pharmaceutically acceptable carriers such as dextrin and cyclodextrin, or other optional auxiliary agents. In this case, auxiliary agents such as excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, and deodorizing agents can be used. The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, and liver function improving agent can be used in combination with other compositions (for example, oral compositions and skin cosmetics described later), or they can be used as topical solutions, patches, etc.

[0090] When formulating an anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent according to this embodiment, the content of the above-mentioned phenylpropionic acids or the composition containing phenylpropionic acids is not particularly limited and can be set as appropriate depending on the purpose.

[0091] Furthermore, the anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent of this embodiment may, if necessary, be used as an active ingredient by combining it with other natural extracts, etc., that have anti-metabolic syndrome effects, whitening effects, anti-aging effects, hair growth effects, anti-inflammatory effects, or liver function improving effects, together with the phenylpropionic acids or compositions containing phenylpropionic acids.

[0092] The anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent of this embodiment can be administered to patients by oral administration, transdermal administration, intraperitoneal administration, intravenous administration, subcutaneous administration, etc., but the method best suited for prevention or treatment should be appropriately selected depending on the type of disease. Furthermore, the dosage of the anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent of this embodiment can be appropriately increased or decreased depending on the type and severity of the disease, individual differences in the patient, method of administration, duration of administration, etc.

[0093] The anti-metabolic syndrome agent of this embodiment can promote the breakdown of fat through its anti-metabolic syndrome effect, preferably through its cAMP phosphodiesterase activity inhibitory effect, and as a result, it can prevent and improve obesity and various lifestyle-related diseases associated with it, such as arteriosclerosis, diabetes, and metabolic syndrome. Furthermore, the anti-metabolic syndrome agent of this embodiment can prevent and treat type 2 diabetes, obesity, hypertension, insulin resistance, etc., through its anti-metabolic syndrome effect, preferably through its DPP IV activity inhibitory effect. However, the anti-metabolic syndrome agent of this embodiment can be used in any application other than those mentioned above where it is meaningful to exhibit an anti-metabolic syndrome effect, preferably through its cAMP phosphodiesterase activity inhibitory effect or DPP IV activity inhibitory effect.

[0094] For example, the anti-metabolic syndrome agent of this embodiment or the aforementioned cAMP phosphodiesterase activity inhibitor can suppress platelet aggregation by inhibiting the degradation of cAMP, thereby preventing, treating, or improving allergic diseases and various inflammatory diseases. Furthermore, the anti-metabolic syndrome agent of this embodiment or the aforementioned DPP IV activity inhibitor can prevent and treat autoimmune diseases such as rheumatoid arthritis and transplant rejection reactions through its DPP IV activity inhibitory effect, as well as prevent and treat diseases such as pain, neurodegenerative diseases, and neuropsychiatric disorders (e.g., sciatica, Alzheimer's disease, depression, etc.); growth hormone deficiency and diseases for which growth hormone is used in treatment; cancer (e.g., T-cell lymphoma, acute lymphoblastic leukemia, thyroid cancer, basal cell carcinoma, breast cancer, etc.); and HIV infection (AIDS).

[0095] The whitening agent of this embodiment can prevent and improve skin darkening, age spots, freckles, and other pigmentation through its whitening effect, preferably its tyrosinase activity inhibitory effect and / or its melanin production inhibitory effect. However, the whitening agent of this embodiment can be used in any application other than these where it is meaningful to exhibit a whitening effect, preferably its tyrosinase activity inhibitory effect or melanin production inhibitory effect.

[0096] The anti-aging agent of this embodiment can prevent, treat, or improve skin aging symptoms such as wrinkle formation, decreased elasticity, and decreased moisturizing function through its anti-aging effects, preferably one or more effects selected from the group consisting of type I collagen production promoting effect, elastin production promoting effect, hyaluronic acid production promoting effect, elastase activity inhibitory effect, HAS3 mRNA expression promoting effect, laminin-332 production promoting effect, epidermal keratinocyte proliferation promoting effect, ATP production promoting effect, glutathione production promoting effect, TGM1 mRNA expression promoting effect, SPT mRNA expression promoting effect, AQP3 mRNA expression promoting effect, filaggrin mRNA expression promoting effect, claudin-1 mRNA expression promoting effect, claudin-4 mRNA expression promoting effect, occludin mRNA expression promoting effect, AGEs formation inhibitory effect, and AGEs degradation promoting effect. However, the anti-aging agent of this embodiment can be used in any application where it is meaningful to exhibit anti-aging effects, preferably type I collagen production promoting effects, elastin production promoting effects, hyaluronic acid production promoting effects, elastase activity inhibitory effects, HAS3 mRNA expression promoting effects, laminin-332 production promoting effects, epidermal keratinocyte proliferation promoting effects, ATP production promoting effects, glutathione production promoting effects, TGM1 mRNA expression promoting effects, SPT mRNA expression promoting effects, AQP3 mRNA expression promoting effects, filaggrin mRNA expression promoting effects, claudin-1 mRNA expression promoting effects, claudin-4 mRNA expression promoting effects, occludin mRNA expression promoting effects, AGEs formation inhibitory effects, or AGEs degradation promoting effects, in addition to the applications described above.

[0097] For example, the anti-aging agent of this embodiment or the aforementioned type I collagen production promoter can be used, through its type I collagen production promoting effect, for the prevention, treatment, or improvement of diseases caused by decreased collagen production, such as osteoporosis; to promote the regeneration of damaged tendons and ligaments; to promote the healing of wounds or burns; and so on. Furthermore, the anti-aging agent of this embodiment or the aforementioned elastin production promoter or elastase activity inhibitor can be used, through its elastin production promoting effect or elastase activity inhibitory effect, for the prevention, treatment, or improvement of lung diseases such as emphysema; vascular diseases such as hypertension and aneurysms; and so on. In addition, the anti-aging agent of this embodiment or the aforementioned hyaluronic acid production promoter can be used, through its hyaluronic acid production promoting effect, for the prevention, treatment, or improvement of arthritis such as rheumatoid arthritis, osteoarthritis, suppurative arthritis, gouty arthritis, traumatic arthritis, and osteoarthritis; to promote the healing of wounds or burns; and so on.

[0098] In addition to the uses described above, the anti-aging agent of this embodiment or the laminin-332 production promoter described above can induce the reconstruction of the basement membrane structure through its laminin-332 production promoting effect, thereby treating and improving wounds in the skin. Furthermore, the anti-aging agent of this embodiment or the laminin-332 production promoter described above can be used as a preventive or therapeutic agent for diseases (such as epidermolysis bullosa) caused by laminin-332 deficiency (deficiency).

[0099] In addition to the uses described above, the anti-aging agent of this embodiment or the epidermal keratinocyte proliferation promoter described above can be used in applications such as preventing, treating, or improving fine wrinkles, dullness, and pigmentation; regenerative medicine; by promoting the proliferation of epidermal keratinocytes, it can restore skin metabolism. Furthermore, the anti-aging agent of this embodiment or the ATP production promoter described above can promote cell turnover through its ATP production-promoting effect, preventing and improving skin aging symptoms such as wrinkles, loss of texture, and decreased elasticity. It can also prevent and improve symptoms such as dullness and pigmentation by restoring skin metabolic function, such as shedding keratinocytes with abnormal melanin accumulation from the stratum corneum. Moreover, the anti-aging agent of this embodiment or the ATP production promoter or glutathione production promoter described above can also be used in applications based on their respective hepatocyte ATP production-promoting effects or hepatocyte glutathione production-promoting effects, which will be described later.

[0100] In addition to the uses described above, the anti-aging agent of this embodiment, or the aforementioned TGM1 mRNA expression promoter, SPT mRNA expression promoter, or filaggrin mRNA expression promoter, can strengthen the skin's barrier function through its TGM1 mRNA expression promoting effect, SPT mRNA expression promoting effect, or filaggrin mRNA expression promoting effect, thereby preventing, treating, or improving skin roughness, dry skin, and other dry skin diseases (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.). Furthermore, the anti-aging agent of this embodiment, or the aforementioned filaggrin mRNA expression promoter or AQP3 mRNA expression promoter, can improve age-related moisture retention and barrier functions through its filaggrin mRNA expression promoting effect or AQP3 mRNA expression promoting effect.

[0101] In addition to the uses described above, the anti-aging agent of this embodiment, or the aforementioned claudin-1 mRNA expression promoter, claudin-4 mRNA expression promoter, or occludin mRNA expression promoter, can promote the formation of tight junctions in epidermal keratinocytes through their claudin-1 mRNA expression promoting effect, claudin-4 mRNA expression promoting effect, or occludin mRNA expression promoting effect. This enhances the skin's barrier function and moisture retention function, and can prevent or improve skin conditions such as dry skin, rough skin, atopic dermatitis, and various infections. Furthermore, the anti-aging agent of this embodiment, or the aforementioned claudin-1 mRNA expression promoter, claudin-4 mRNA expression promoter, or occludin mRNA expression promoter, can improve the barrier function in the gastrointestinal tract and can prevent or improve inflammatory bowel disease, food allergies, and various infections originating from the gastrointestinal tract.

[0102] In addition to the uses described above, the anti-aging agent of this embodiment, or the aforementioned AGEs formation inhibitor or AGEs decomposition accelerator, can prevent or treat diabetic complications such as diabetic neuropathy, diabetic retinopathy, and diabetic nephropathy; arteriosclerosis caused by protein glycation reactions; and osteoporosis and osteoarthritis caused by protein glycation reactions, etc., through their AGEs formation inhibitory or AGEs decomposition promoting effects. Furthermore, the anti-aging agent of this embodiment or the aforementioned AGEs formation inhibitor can prevent or suppress hair damage caused by protein glycation reactions through its AGEs formation inhibitory effect, thereby preventing or suppressing hair stiffness, restoring hair elasticity and suppleness, and giving hair firmness and body.

[0103] The hair growth agent of this embodiment can prevent, treat, or improve hair loss such as male pattern baldness, alopecia areata, and trichotillomania through its hair growth action, preferably testosterone 5α-reductase activity inhibitory action and / or hair papilla cell proliferation promoting action, and is particularly suitable for the prevention, treatment, or improvement of male pattern baldness. However, the hair growth agent of the present invention can be used in any application other than these where it is meaningful to exhibit a hair growth action, preferably testosterone 5α-reductase activity inhibitory action or hair papilla cell proliferation promoting action.

[0104] For example, the hair growth agent of this embodiment or the aforementioned testosterone 5α-reductase activity inhibitor can prevent, treat, or improve diseases related to male hormones, such as hirsutism, seborrhea, acne, benign prostatic hyperplasia, prostate tumors, and precocious puberty in boys, through its testosterone 5α-reductase activity inhibitory effect. Furthermore, the hair growth agent of this embodiment or the aforementioned dermal papilla cell proliferation promoter can also be used in the field of regenerative medicine, such as hair regeneration using dermal papilla cells, through its dermal papilla cell proliferation promoting effect.

[0105] The anti-inflammatory agent of this embodiment can prevent, treat, or improve various skin inflammatory diseases associated with skin roughness, such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, atopic dermatitis, and other skin irritations, through its anti-inflammatory action, preferably one or more actions selected from the group consisting of NO production inhibition, hyaluronidase activity inhibition, hexosaminidase release inhibition, and PGE2 production promotion. However, the anti-inflammatory agent of this embodiment can be used in any application other than these where it is meaningful to exhibit an anti-inflammatory action, preferably NO production inhibition, hyaluronidase activity inhibition, hexosaminidase release inhibition, or PGE2 production promotion.

[0106] For example, the anti-inflammatory agent of this embodiment, or the aforementioned NO production inhibitor, hyaluronidase activity inhibitor, hexosaminidase release inhibitor, or PGE2 production promoter, can prevent, treat, or improve rheumatoid arthritis, osteoarthritis, asthma, etc., through their NO production inhibitory effect, hyaluronidase activity inhibitory effect, hexosaminidase release inhibitory effect, or PGE2 production promoting effect. Furthermore, the anti-inflammatory agent of this embodiment, or the aforementioned hexosaminidase release inhibitor, can prevent, treat, or improve gastric ulcers, sleep disorders, etc., caused by excessive stomach acid, through their hexosaminidase release inhibitory effect.

[0107] The liver function improving agent of this embodiment can improve liver function through its liver function improving effect, preferably through its effect of promoting hepatocyte glutathione production and / or hepatocyte ATP production. Uses included in liver function improving include: prevention, treatment, or improvement of symptoms caused by ethanol intake (e.g., hangovers); promotion of metabolism and breakdown of sugars and fats; prevention, treatment, or improvement of fatty liver, hepatitis such as alcoholic hepatitis and drug-induced hepatitis, cirrhosis, etc. However, the liver function improving agent of this embodiment can be used in any application other than those described above where it is meaningful to exhibit a liver function improving effect, preferably a hepatocyte glutathione production promoting effect or a hepatocyte ATP production promoting effect.

[0108] For example, the liver function improving agent of this embodiment or the hepatocyte glutathione production promoting agent described above can be used for the prevention, treatment, or improvement of diseases associated with decreased glutathione concentration in the body, such as cataracts and Parkinson's disease, through its hepatocyte glutathione production promoting effect; and for the prevention, treatment, or improvement of pigmentation such as skin darkening, age spots, and freckles. Furthermore, the liver function improving agent of this embodiment or the hepatocyte ATP production promoting agent described above can be used for purposes such as the recovery from fatigue and malaise, through its hepatocyte ATP production promoting effect.

[0109] Furthermore, the anti-metabolic syndrome agent, skin whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, and liver function improving agent of this embodiment have excellent effects in anti-metabolic syndrome, skin whitening, anti-aging, hair growth, anti-inflammatory, and liver function improving effects, respectively, and can therefore be suitably used as reagents for research on their mechanisms of action.

[0110] [Oral composition] The above-mentioned phenylpropionic acids have excellent effects in anti-metabolic syndrome, skin whitening, anti-aging, hair growth, anti-inflammatory, and liver function improving effects, making them suitable for incorporation into oral compositions. In this case, the above-mentioned phenylpropionic acids or compositions containing phenylpropionic acids may be incorporated as is, or formulations derived from phenylpropionic acids, such as anti-metabolic syndrome agents, skin whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents, may be incorporated.

[0111] By incorporating the above-mentioned phenylpropionic acids or compositions containing phenylpropionic acids, or by formulating anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents from phenylpropionic acids or compositions containing phenylpropionic acids into an oral composition, oral compositions suitable for anti-metabolic syndrome, whitening, anti-aging, hair growth, anti-inflammatory, or liver function improving applications can be obtained. Among these, the anti-metabolic syndrome effect and the liver function improving effect are preferred because their effects are easily exerted when imparted to an oral composition.

[0112] Here, "oral composition" refers to a substance that poses little risk to human health and is ingested orally or via the gastrointestinal tract in normal social life, and is not limited to administrative classifications such as food, pharmaceuticals, or quasi-drugs. Therefore, "oral composition" in this embodiment broadly includes general foods, animal feed, health foods, functional foods (foods for specified health uses, foods with nutritional function claims, functional foods and beverages), quasi-drugs, pharmaceuticals, etc., that are ingested orally. The oral composition according to this embodiment is preferably an oral composition that can display the desirable effects of the phenylpropionic acids on the oral composition or its packaging, and is particularly preferably a functional food (foods for specified health uses, foods with nutritional function claims, functional foods), a quasi-drug, or a pharmaceutical.

[0113] When incorporating the above-mentioned phenylpropionic acids or compositions containing phenylpropionic acids, or anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents formulated from phenylpropionic acids or compositions containing phenylpropionic acids, into an oral composition, the amount of active ingredients can be appropriately changed considering the intended use, symptoms, gender, etc. However, considering the general intake of the oral composition to which the phenylpropionic acids are added, it is preferable to ensure that the daily intake of phenylpropionic acids for adults is approximately 1 to 1000 mg. Furthermore, when the oral composition to be added is in the form of granules, tablets, or capsules, the amount of phenylpropionic acid or a composition containing phenylpropionic acid, or an anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, or liver function improving agent formulated from a composition containing phenylpropionic acid or phenylpropionic acid, is usually 0.1 to 100% by mass, preferably 5 to 100% by mass, relative to the oral composition to be added.

[0114] The oral composition of this embodiment may be an oral composition into which the above-mentioned phenylpropionic acids are incorporated without interfering with their activity, or it may be a nutritional supplement mainly composed of the above-mentioned phenylpropionic acids.

[0115] When preparing the oral composition of this embodiment, any auxiliary agents such as sugars like dextrin and starch; proteins like gelatin, soy protein, and corn protein; amino acids like alanine, glutamine, and isoleucine; polysaccharides like cellulose and gum arabic; and oils and fats like soybean oil and medium-chain triglycerides can be added to create an oral composition of any shape.

[0116] Oral compositions that may contain the above-mentioned phenylpropionic acids are not particularly limited, but specific examples include: beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit drinks, and lactic acid drinks (including concentrated stocks and powders for adjusting these beverages); frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as soba, udon, vermicelli, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; confectionery such as candy, chewing gum, candy, gum, chocolate, tablets, snacks, biscuits, jelly, jam, cream, and baked goods; and Examples of phenylpropionic acids include processed seafood and livestock products such as fish cakes, ham, and sausages; dairy products such as processed milk and fermented milk; oils and fats and processed oils such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressings; seasonings such as sauces and dips; soups, stews, salads, prepared foods, and pickles; and various other forms of health and nutritional supplements; as well as tablets, capsules, and drinks. When incorporating the above-mentioned phenylpropionic acids into these oral compositions, commonly used auxiliary ingredients and additives can be used in combination.

[0117] [Skin cosmetics, hair cosmetics] The above-mentioned phenylpropionic acids have excellent effects in anti-metabolic syndrome, whitening, anti-aging, hair growth, anti-inflammatory, and liver function improving effects, making them suitable for incorporation into skin cosmetics or hair cosmetics. In this case, the above-mentioned phenylpropionic acids may be incorporated as is, or anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents formulated from the above-mentioned phenylpropionic acids may be incorporated.

[0118] By incorporating the above-mentioned phenylpropionic acids or the above-mentioned anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents, it is possible to impart anti-metabolic syndrome effects, whitening effects, anti-aging effects, hair growth effects, anti-inflammatory effects, or liver function improving effects to skin cosmetics or hair cosmetics, and the resulting skin cosmetics or hair cosmetics can be used for anti-metabolic syndrome applications, whitening applications, anti-aging applications, hair growth applications, anti-inflammatory applications, or liver function improving applications.

[0119] Among these, the whitening, anti-aging, and anti-inflammatory effects are easily exhibited when incorporated into skin cosmetics, meaning that skin cosmetics particularly suitable for whitening, anti-aging, or anti-inflammatory purposes can be created. Furthermore, the hair growth, anti-aging, and anti-inflammatory effects are easily exhibited when incorporated into hair cosmetics, meaning that hair cosmetics particularly suitable for hair growth, anti-aging, or anti-inflammatory purposes can be created.

[0120] The types of skin cosmetics or hair cosmetics that may contain the above-mentioned phenylpropionic acids, or the above-mentioned anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents are not particularly limited. Examples of skin cosmetics include ointments, creams, emulsions, lotions, gels, beauty oils, masks, and foundations, while examples of hair cosmetics include hair tonics, hair creams, hair liquids, shampoos, pomades, and conditioners.

[0121] When the above-mentioned phenylpropionic acids, or the above-mentioned anti-metabolic syndrome agents, whitening agents, anti-aging agents, hair growth agents, anti-inflammatory agents, or liver function improving agents are incorporated into skin cosmetics or hair cosmetics, the amount incorporated can be appropriately adjusted depending on the type of skin cosmetic or hair cosmetic, but the preferred blending ratio is about 0.0001 to 10% by mass, and the particularly preferred blending ratio is about 0.001 to 1% by mass when converted to a standard extract.

[0122] The skin cosmetic or hair cosmetic of this embodiment may be used in combination with main ingredients, auxiliary ingredients, or other components commonly used in the manufacture of skin cosmetic or hair cosmetic, such as astringents, bactericidal / antibacterial agents, whitening agents, UV absorbers, moisturizers, cell activators, anti-inflammatory / anti-allergic agents, antioxidants / reactive oxygen species scavengers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, etc., as long as they do not interfere with the anti-metabolic syndrome effect, whitening effect, anti-aging effect, hair growth effect, anti-inflammatory effect, or liver function improving effect of the phenylpropionic acids mentioned above. By using such combinations, a more general product can be produced, and the synergistic effect between the combined active ingredients may result in superior effects beyond what is normally expected.

[0123] The skin cosmetic of this embodiment exhibits the following effects of phenylpropionic acids: whitening effect, tyrosinase activity inhibitory effect, melanin production inhibitory effect; anti-aging effect, type I collagen production promoting effect, elastin production promoting effect, hyaluronic acid production promoting effect, elastase activity inhibitory effect, HAS3 mRNA expression promoting effect, laminin-332 production promoting effect, epidermal keratinocyte proliferation promoting effect, ATP production promoting effect, glutathione production promoting effect, TGM1 mRNA expression promoting effect, SPT mRNA expression promoting effect, AQP3 mRNA expression promoting effect, filaggrin mRNA expression promoting effect, claudin-1 mRNA expression promoting effect, claudin-4 mRNA expression promoting effect, and occluding mRNA expression promoting effect, AGEs formation inhibitory effect, AGEs degradation promoting effect; hair growth effect, testosterone 5α-reductase activity inhibitory effect, hair papilla cell proliferation promoting effect; anti-inflammatory effect, NO production inhibitory effect, hyaluronidase activity inhibitory effect, hexosaminidase release inhibitory effect, PGE2 production promoting effect; anti-metabolic syndrome effect, cAMP phosphodiesterase activity inhibitory effect, DPP Through one or more actions selected from the group consisting of IV activity inhibitory action, liver function improving action, hepatocyte glutathione production promoting action, and hepatocyte ATP production promoting action, it can prevent, treat or improve pigmentation such as skin darkening, age spots, and freckles; prevent, treat or improve skin aging symptoms such as wrinkle formation, decreased elasticity, and decreased moisturizing function; promote the healing of wounds or burns; prevent, treat or improve dry skin diseases such as rough skin and dry skin (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.); prevent, treat or improve diseases involving male hormones such as seborrhea and acne; prevent, treat or improve contact dermatitis (rash), psoriasis, pemphigus vulgaris, and various other inflammatory skin diseases associated with rough skin; and prevent, treat or improve obesity and lifestyle-related diseases such as arteriosclerosis, diabetes, and metabolic syndrome.

[0124] Furthermore, the hair cosmetic of this embodiment exhibits the hair growth effect, testosterone 5α-reductase activity inhibitory effect, hair papilla cell proliferation promoting effect; anti-inflammatory effect, NO production inhibitory effect, hyaluronidase activity inhibitory effect, hexosaminidase release inhibitory effect, PGE2 production promoting effect; anti-aging effect, type I collagen production promoting effect, elastin production promoting effect, hyaluronic acid production promoting effect, elastase activity inhibitory effect, HAS3 mRNA expression promoting effect, laminin-332 production promoting effect, epidermal keratinocyte proliferation promoting effect, ATP production promoting effect, glutathione production promoting effect, TGM1 mRNA expression promoting effect, SPT mRNA expression promoting effect, AQP3 mRNA expression promoting effect, filaggrin mRNA expression promoting effect, claudin-1 mRNA expression promoting effect, claudin-4 mRNA expression promoting effect, and occludin Through one or more actions selected from the group consisting of mRNA expression promotion, AGEs formation inhibition, AGEs degradation promotion; anti-metabolic syndrome effect, cAMP phosphodiesterase activity inhibition, DPP IV activity inhibition; skin whitening effect, tyrosinase activity inhibition, melanin production inhibition; liver function improvement effect, hepatocyte glutathione production promotion effect, hepatocyte ATP production promotion effect, etc., it can prevent, treat or improve alopecia such as male pattern baldness, alopecia areata, and trichotillomania; prevent, treat or improve various inflammatory skin diseases associated with contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other skin irritations; and treat dry skin diseases such as rough skin and dry skin (e.g., atopic dermatitis, psoriasis, ichthyosis, etc.).

[0125] Furthermore, while the anti-metabolic syndrome agent, whitening agent, anti-aging agent, hair growth agent, anti-inflammatory agent, liver function improving agent, oral composition, skin cosmetic, and hair cosmetic of this embodiment are suitably applied to humans, they can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as their respective effects are achieved. [Examples]

[0126] The present invention will be specifically described below with reference to test examples, but the present invention is not limited in any way to the following examples. In these test examples, the following commercially available compounds were used as test samples.

[0127] [Table 1]

[0128] [Test Example 1] Cyclic AMP phosphodiesterase activity inhibition test The inhibitory effect of compound 1 (sample 1) on cyclic AMP phosphodiesterase activity was tested as follows.

[0129] To 0.2 mL of 50 mmol / L Tris-HCl buffer (pH 7.5) containing 5 mmol / L magnesium chloride, 0.1 mL of 2.5 mg / mL bovine serum albumin solution, 0.1 mL of 0.1 mg / mL cyclic AMP phosphodiesterase solution, and 0.05 mL of the test sample solution (Sample 1, see Table 2 below for final concentration) were added and allowed to stand at 37°C for 5 minutes. Then, 0.05 mL of 0.5 mg / mL cyclic AMP solution was added and the mixture was reacted at 37°C for 60 minutes. After the reaction was complete, the reaction was stopped by boiling in a boiling water bath for 3 minutes, and the mixture was centrifuged (2260 × g, 10 minutes, 4°C). The cyclic AMP, the reaction substrate in the supernatant, was analyzed under the high-performance liquid chromatography conditions described below. As a control, the same procedure was performed with only the solvent without the sample added.

[0130] <High-performance liquid chromatography conditions> Product Name: Chromatographer 12 (Manufactured by SYSTEM INSTRUMENTS) Stationary phase: Wakosil C 18 -ODS 5μm (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) Column length: 250 mm Mobile phase: 1mmol / L TBAP in 25mmol / L KH2PO4:CH3CN=90:10 Mobile phase flow rate: 1.0mL / min Detection: 260nm

[0131] Next, the peak area of ​​the cyclic AMP standard (A), the peak area of ​​the supernatant of the reaction solution between the cyclic AMP standard and cyclic AMP phosphodiesterase without the sample (B1), and the peak area of ​​the supernatant of the reaction solution between the cyclic AMP standard and cyclic AMP phosphodiesterase with the test sample added (B2) were determined. From the obtained results, the degradation rate of the cyclic AMP standard without the sample (C) and the degradation rate of the cyclic AMP standard with the test sample added (D) were calculated using the following formula.

[0132] Decomposition rate of standard without sample (C,%) = (1 - B1 / A) × 100 Decomposition rate of the standard after adding the test sample (D,%) = (1 - B2 / A) × 100

[0133] Subsequently, based on the respective degradation rates (C, D) calculated using the above formula, the cyclic AMP phosphodiesterase activity inhibition rate (%) was calculated using the following formula. cAMP phosphodiesterase activity inhibition rate (%) = (1 - D / C) × 100 The results are shown in Table 2.

[0134] [Table 2]

[0135] As shown in Table 2, compound 1 (sample 1) was confirmed to have excellent inhibitory activity on cyclic AMP phosphodiesterase activity.

[0136] [Test Example 2] DPP IV Activity Inhibition Test The inhibitory effect of dipeptidyl peptidase IV (DPP IV) activity on compounds 1-3 (samples 1-3) was tested as follows.

[0137] In a 96-well plate, 25 μL of the test samples (samples 1-3, see Table 3 below for final concentrations) prepared in 25 mM Tris-HCl buffer (pH 8.0) was mixed with 25 μL of a 0.4 μg / m LDP-IV (R&D Systems, rhCD26) solution prepared in the same buffer, and the mixture was pre-incubated at 37°C for 5 minutes. Subsequently, 50 μL of 0.5 mM Gly-Pro-p-NA·Tos (Peptide Research Institute) prepared in the same buffer was added, and the mixture was reacted at 37°C for 90 minutes. After the reaction was complete, the absorbance at a wavelength of 415 nm was measured. From the obtained results, the DPP-IV activity inhibition rate (%) was calculated using the following formula.

[0138] DPP IV activity inhibition rate (%) = {1 - (CD) / (AB)} × 100 Each term in the formula represents the following: A: Absorbance at a wavelength of 415 nm with no sample added and with enzyme added. B: Absorbance at a wavelength of 415 nm without sample or enzyme addition. C: Absorbance at a wavelength of 415 nm after adding the test sample and enzyme. D: Absorbance at a wavelength of 415 nm with test sample added but without enzyme added. The results are shown in Table 3.

[0139] [Table 3]

[0140] As shown in Table 3, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) all exhibited excellent DPP IV inhibitory activity.

[0141] [Test Example 3] Tyrosinase Activity Inhibition Test The tyrosinase activity inhibitory effect of compound 3 (sample 3) was tested as follows.

[0142] In a 48-well plate, 0.2 mL of McIlvaine buffer (pH 6.8), 0.06 mL of 0.3 mg / mL tyrosine solution, and 0.18 mL of the test sample (Sample 3, see Table 4 below for final concentration) dissolved in 25% DMSO solution were added and allowed to stand at 37°C for 10 minutes. Then, 0.02 mL of 1000 units / mL tyrosinase solution was added and the mixture was reacted at 37°C for another 15 minutes. After the reaction was complete, the absorbance at a wavelength of 475 nm was measured.

[0143] Furthermore, as a blank, the same procedure and absorbance measurements were performed without adding the enzyme solution. In addition, as a control, the same measurements were performed when a 25% DMSO solution was added without adding the sample solution. From the obtained measurement results, the tyrosinase activity inhibition rate (%) was calculated using the following formula.

[0144] Tyrosinase activity inhibition rate (%) = {1 - (AB) / (CD)} × 100 Each term in the formula represents the following: A: Absorbance at a wavelength of 475 nm after adding the test sample and enzyme. B: Absorbance at a wavelength of 475 nm with test sample added but without enzyme added. C: Absorbance at a wavelength of 475 nm with no sample added and with enzyme added. D: Absorbance at a wavelength of 475 nm without sample or enzyme addition. The results are shown in Table 4.

[0145] [Table 4]

[0146] As shown in Table 4, compound 3 (sample 3) was found to have excellent tyrosinase activity inhibitory activity.

[0147] [Test Example 4] Test of melanin production inhibitory effect on B16 melanoma cells The inhibitory effect of compounds 1-3 (samples 1-3) on melanin production in B16 melanoma cells was tested as follows.

[0148] B16 melanoma cells were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then harvested by trypsin treatment. The harvested cells were 24.0 × 10⁴ 4 After diluting the cells with 10% FBS and 1 mmol / L theophylline-containing DMEM to achieve a cell density of cells / mL, 300 μL was seeded per well in a 48-well plate and cultured for 6 hours.

[0149] After the culture period, 300 μL of the test samples (Samples 1-3, see Table 5 below for final concentrations) dissolved in 10% FBS and 1 mmol / L theophylline-containing DMEM were added to each well and cultured for 4 days. As a control, cells were cultured similarly using 10% FBS and 1 mmol / L theophylline-containing DMEM without any sample additions. After the culture period, the culture medium was removed, 200 μL of 2 mol / L NaOH solution was added, and the cells were destroyed using an ultrasonic disruptor. The absorbance at a wavelength of 475 nm was measured. From the measured absorbance values, the amount of melanin was calculated based on a calibration curve created using synthetic melanin (SIGMA).

[0150] Furthermore, to measure cell viability, the cells were cultured in the same manner as described above, the culture medium was removed, and the cells were washed with 400 μL of PBS(-) buffer. 200 μL of neutral red dissolved in 10% FBS-containing DMEM at a final concentration of 0.05 mg / mL was added to each well, and the cells were cultured for 2.5 hours. After culturing, the neutral red solution was removed, and 200 μL of ethanol-acetic acid solution (ethanol:acetic acid:water = 50:1:49) was added to each well to extract the pigment. After extraction, the absorbance at a wavelength of 540 nm was measured. From the obtained results, the melanin production inhibition rate (%) corrected for cell viability was calculated using the following formula.

[0151] Melanin production inhibition rate (%) = {1 - (B / D) / (A / C)} × 100 Each term in the formula represents the following: A: Melanin content without sample addition B: Melanin content in test samples C: Absorbance at a wavelength of 540 nm without sample addition D: Absorbance at a wavelength of 540 nm with the test sample added. The results are shown in Table 5.

[0152] [Table 5]

[0153] As shown in Table 5, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were all found to have excellent melanin production inhibitory effects.

[0154] [Test Example 5] Test on the effect of promoting type I collagen production The type I collagen production promoting effect of compound 1 (sample 1) was tested as follows.

[0155] Normal human dermal fibroblasts (NB1RGB) were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were 1.6 × 10⁶ 5 After diluting with 0.25% FBS-containing DMEM to achieve a cell density of cells / mL, 100 μL was seeded per well in a 96-well microplate and cultured overnight.

[0156] After the culture period, 100 μL of the test sample (Sample 1, see Table 6 below for final concentration), dissolved in 0.25% FBS-containing DMEM, was added to each well and cultured for 3 days. As a control, culture was performed similarly using 0.25% FBS-containing DMEM without the sample. After culturing, the amount of type I collagen in the culture medium of each well was measured by ELISA. From the measurement results, the type I collagen production promotion rate (%) was calculated using the following formula.

[0157] Type I collagen production promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of type I collagen when added to the test sample B: Amount of type I collagen without sample addition The results are shown in Table 6.

[0158] [Table 6]

[0159] As shown in Table 6, compound 1 (sample 1) was confirmed to have excellent type I collagen production promoting activity.

[0160] [Test Example 6] Elastin Production Promoting Effect Test The elastin production-promoting effect of compound 2 (sample 2) and compound 3 (sample 3) was tested as follows.

[0161] Normal human dermal fibroblasts (NB1RGB) were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were 2.2 × 10⁶ 5 After diluting the culture medium to achieve a cell density of cells / mL, 100 μL was seeded per well in a 96-well microplate and cultured overnight.

[0162] After the culture period, the culture medium was removed, and 150 μL of the test samples (Samples 2 and 3; see Table 7 below for final concentrations) dissolved in 0.25% FBS-containing DMEM was added to each well and cultured for 5 days. As a control, culture was performed similarly using 0.25% FBS-containing DMEM without any sample added. After the culture period, the supernatant was collected, and the amount of elastin released into the culture supernatant was measured by ELISA. From the measurement results, the elastin production promotion rate (%) was calculated using the following formula.

[0163] Elastin production promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of elastin when test samples are added B: Amount of elastin without sample addition The results are shown in Table 7.

[0164] [Table 7]

[0165] As shown in Table 7, both compound 2 (sample 2) and compound 3 (sample 3) exhibited excellent elastin production-promoting effects.

[0166] [Test Example 7] Elastase activity inhibition test The elastase activity inhibitory effect was tested for compounds 1-3 (samples 1-3) as follows.

[0167] In a 96-well microplate, 50 μL of the test samples (samples 1-3, see Table 8 below for final concentrations) prepared in 0.2 mol / L Tris-HCl buffer (pH 8.0) were mixed with 50 μL of 20 μg / mL elastase type III (SIGMA-Aldrich) solution. Then, 100 μL of 0.4514 mg / mL N-succinyl-Ala-Ala-Ala-p-nitroanilide (SIGMA-Aldrich), prepared in the above buffer, was added, and the mixture was reacted at 25°C for 15 minutes. After the reaction, the absorbance at a wavelength of 415 nm was measured. A blank test without enzyme addition was also performed in the same manner for correction. From the obtained results, the elastase activity inhibition rate (%) was calculated using the following formula.

[0168] Elastase activity inhibition rate (%) = {1 - (CD) / (AB)} × 100 Each term in the formula represents the following: A: Absorbance at a wavelength of 415 nm with no sample added and with enzyme added. B: Absorbance at a wavelength of 415 nm without sample or enzyme addition. C: Absorbance at a wavelength of 415 nm after adding the test sample and enzyme. D: Absorbance at a wavelength of 415 nm with test sample added but without enzyme added. The results are shown in Table 8.

[0169] [Table 8]

[0170] As shown in Table 8, compounds 1-3 (samples 1-3) were found to have excellent elastase activity inhibitory effects.

[0171] [Test Example 8] Hyaluronic Acid Production Promotion Test The hyaluronic acid production-promoting effect was tested for compound 1 (sample 1) and compound 2 (sample 2) as follows.

[0172] Normal human dermal fibroblasts (NB1RGB) were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were 1.6 × 10⁶ 5 After diluting with 0.25% FBS-containing DMEM to achieve a cell density of cells / mL, 100 μL was seeded per well in a 96-well plate and cultured overnight.

[0173] After the culture period, 100 μL of the test samples (Samples 1 and 2; see Table 9 below for final concentrations) dissolved in 0.25% FBS-containing DMEM was added to each well and cultured for 3 days. As a control, culture was performed similarly using 0.25% FBS-containing DMEM without any added samples. After culturing, the amount of hyaluronic acid in the culture medium of each well was measured using a sandwich method with hyaluronic acid-binding protein (HABP). From the obtained results, the hyaluronic acid production promotion rate (%) was calculated using the following formula.

[0174] Hyaluronic acid production promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of hyaluronic acid added to the test sample B: Amount of hyaluronic acid without sample addition The results are shown in Table 9.

[0175] [Table 9]

[0176] As shown in Table 9, compound 1 (sample 1) and compound 2 (sample 2) were confirmed to have excellent hyaluronic acid production promoting effects.

[0177] [Test Example 9] Hyaluronic acid synthase 3 (HAS3) mRNA expression promotion test The HAS3 mRNA expression-promoting effect of compound 1 (sample 1) was tested as follows.

[0178] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0179] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test sample (Sample 1, see Table 10 below for final concentration) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without the sample was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each well was measured using a spectrophotometer, and the total RNA was prepared to a concentration of 150 ng / μL.

[0180] Using this total RNA as a template, mRNA expression levels for HAS3 and the internal standard GAPDH were measured. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green(R) The two-step real-time RT-PCR reaction was performed using Fast qPCR Mix (manufactured by Takara Bio Inc.). Primers manufactured by Takara Bio Inc. were used. The expression level of HAS3 mRNA was corrected with the value of GAPDH based on the total RNA samples prepared from the cells cultured with "test sample added" and "no sample added", respectively. From the obtained values, the promotion rate (%) of HAS3 mRNA expression was calculated according to the following formula.

[0181] Promotion rate (%) of HAS3 mRNA expression = A / B × 100 Each term in the formula represents the following respectively. A: Corrected value with test sample added B: Corrected value with no sample added The results are shown in Table 10.

[0182]

Table 10

[0183] As shown in Table 10, Compound 1 (Sample 1) had an excellent effect of promoting HAS3 mRNA expression.

[0184] 〔Test Example 10〕Test for the effect of promoting laminin-332 production For Compound 1 (Sample 1), the effect of promoting laminin-332 production was tested as follows.

[0185] Normal human neonatal epidermal keratinocytes (NHEK) were pre-cultured in a 75 cm 2 flask using a growth medium for normal human epidermal keratinocytes (KGM), and the cells were collected by trypsin treatment. The collected cells were diluted with a medium (KGM-BPE) obtained by removing BPE from KGM to a cell density of 1.0 × 10 5 cell / mL, and then seeded at 500 μL per well in a 24-well plate and cultured for one day.

[0186] After the culture period, the culture medium was removed, and 500 μL of the test sample (Sample 1, see Table 11 below for the final concentration) dissolved in KGM-BPE was added to each well and cultured for 48 hours. As a control, the same culture was performed using KGM-BPE without the sample. After the culture period, 100 μL of the culture medium supernatant was transferred to an ELISA plate and adsorbed onto the plate at 37°C for 2 hours. The amount of adsorbed laminin-332 was then measured by ELISA. From the obtained measurement results, the laminin-332 production promotion rate (%) was calculated using the following formula.

[0187] Laminin-332 production promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of laminin-332 when added to the test sample B: Amount of laminin-332 without sample addition The results are shown in Table 11.

[0188] [Table 11]

[0189] As shown in Table 11, compound 1 (sample 1) was confirmed to have excellent laminin-332 production promoting activity.

[0190] [Test Example 11] Test for promoting the proliferation of epidermal keratinocytes The effect of compound 1 (sample 1) on promoting epidermal keratinocyte proliferation was tested as follows.

[0191] Normal human neonatal epidermal keratinocytes (NHEKs) were cultured in normal human epidermal keratinocyte growth medium (KGM), and then the cells were harvested by trypsin treatment. The harvested cells were 3.0 × 10⁶ 4After diluting the sample with KGM to a cell density of cells / mL, 100 μL was seeded per well in a collagen-coated 96-well plate and cultured overnight. After culturing, 100 μL of the test sample (Sample 1, see Table 12 below for final concentration), dissolved in KGM, was added to each well and cultured for 3 days. As a control, the same culture was performed using KGM without the sample.

[0192] The proliferative effect on epidermal keratinocyte proliferation was measured using the MTT assay method. After 3 days of culture, the culture 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 2 hours of culture, the blue formazan produced in the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. Simultaneously, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. From the obtained results, the epidermal keratinocyte proliferation promotion rate (%) was calculated using the following formula.

[0193] Epidermal keratinocyte proliferation promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of blue formazan produced when the test sample is added. B: Amount of blue formazan produced without sample addition The results are shown in Table 12.

[0194] [Table 12]

[0195] As shown in Table 12, compound 1 (sample 1) was found to have excellent epidermal keratinocyte proliferation-promoting activity.

[0196] [Test Example 12] ATP production promotion test (epidermal keratinocytes) The ATP production-promoting effect of compound 1 (sample 1) and compound 2 (sample 2) was tested as follows.

[0197] Normal human neonatal epidermal keratinocytes (NHEK) were cultured using normal human epidermal keratinocyte growth medium (KGM), and then the cells were recovered by trypsin treatment. The recovered cells were diluted with KGM to a cell density of 2.0×10 5 cells / mL, and then seeded at 100 μL per well in a collagen-coated 96-well plate and cultured overnight. After the culture, the medium was removed, and 100 μL of KGM supplemented with the test samples (Samples 1 and 2, see Table 13 below for the final concentrations) was added to each well and cultured for 2 hours. As a control, the same culture was performed using KGM without the sample.

[0198] The effect of promoting ATP production was measured by the firefly luciferase luminescence method to measure the amount of ATP in the cells. That is, after culturing for 2 hours, 100 μL of an ATP measurement reagent (manufactured by Toyo B-net Co., Ltd., trade name “ATP Measurement Reagent for Cells”) was added to each well, and a chemiluminescence reaction by luciferase was performed. After the reaction, the chemiluminescence amount proportional to the amount of intracellular ATP was measured using a chemiluminescence measurement device (manufactured by Thermo Fisher Scientific, product name: Varioskan LUX multimode microplate reader). From the obtained results, the ATP production promotion rate (%) was calculated by the following formula.

[0199] ATP production promotion rate (%) = A / B × 100 Each term in the formula represents the following respectively. A: Chemiluminescence amount with the addition of the test sample B: Chemiluminescence amount without the addition of the sample The results are shown in Table 13.

[0200]

Table 13

[0201] As shown in Table 13, Compound 1 (Sample 1) and Compound 2 (Sample 2) were found to have an excellent effect of promoting ATP production.

[0202] [Test Example 13] Glutathione production promoting effect test (skin fibroblasts) The glutathione production-promoting effect in skin fibroblasts was tested for compounds 1-3 (samples 1-3) as follows.

[0203] Normal human dermal fibroblasts (NB1RGB) were cultured in α-modified Eagle's Minimum Essential Medium (α-MEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were measured in 2.0 × 10⁶ units. 5 After diluting with 10% FBS-containing α-MEM to achieve a cell density of cells / mL, 200 μL was seeded per well in a 48-well plate and cultured for 48 hours.

[0204] After culturing, the culture medium was removed, and 200 μL of the test samples (Samples 1-3, see Table 14 below for final concentrations) dissolved in 1% FBS-containing Dulbecco's modified Eagle medium (DMEM) was added to each well and cultured for 24 hours. As a control, cells were cultured similarly in 1% FBS-containing DMEM without the test samples. After culturing, the culture medium was removed from each well, washed with 400 μL of PBS(-) buffer, and then the cells were lysed using 150 μL of M-PER (PIERCE).

[0205] Of this, 100 μL was used to quantify total glutathione. Specifically, 100 μL of lysed cell extract, 50 μL of 0.1 mol / L phosphate buffer, 25 μL of 2 mmol / L NADPH, and 25 μL of 3.2 unit / mL glutathione reductase were added to a 96-well plate. After heating at 37°C for 10 minutes, 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 determine ΔOD / min. The total glutathione concentration was calculated based on a calibration curve created using oxidized glutathione (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). After correcting the obtained values ​​to glutathione amount per total protein amount, the glutathione production promotion rate (%) was calculated using the following formula.

[0206] Glutathione production promotion rate (%) = B / A × 100 Each term in the formula represents the following: A: Glutathione content per unit of total protein in the sample without additives B: Amount of glutathione per unit of total protein in the test sample The results are shown in Table 14.

[0207] [Table 14]

[0208] As shown in Table 14, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were all found to have excellent glutathione production-promoting effects in fibroblasts.

[0209] [Test Example 14] Transglutaminase-1 (TGM1) mRNA expression promotion test The TGM1 mRNA expression-promoting effect was tested for compounds 1-3 (samples 1-3) as follows.

[0210] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0211] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test samples (Samples 1-3, see Table 15 below for final concentrations) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without any added samples was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each sample was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0212] Using this total RNA as a template, mRNA expression levels for TGM1 and the internal standard GAPDH were measured. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers with the sequences 5'-AGGTGGAGCTTAGCCCTGTG-3' and 5'-GCAAGTGAAGACTGACTCCCTCTC-3' were used. The expression level of TGM1 mRNA was corrected using the GAPDH value, based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the TGM1 mRNA expression enhancement rate (%) was calculated using the following formula.

[0213] TGM1 mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 15.

[0214] [Table 15]

[0215] As shown in Table 15, all of Compound 1 (Sample 1), Compound 2 (Sample 2) and Compound 3 (Sample 3) had excellent promoting effects on TGM1 mRNA expression.

[0216] [Test Example 15] Test for Promoting Effect on Serine Palmitoyltransferase (SPT) mRNA Expression Regarding Compound 1 (Sample 1), the promoting effect on SPT mRNA expression was tested as follows.

[0217] Normal human neonatal epidermal keratinocytes (NHEK) were pre-cultured using a growth medium for normal human epidermal keratinocytes (KGM), and the cells were recovered by trypsin treatment. The recovered cells were diluted with KGM to a cell density of 15×10 4 cells / mL, and then 2 mL each was seeded into 6-well plates (30×10 4 cells / well) and cultured overnight under the conditions of 37 °C and 5% CO2. After culturing, the medium was replaced with normal human epidermal keratinocyte basal medium (KBM, which is the above KGM without the addition of growth additives (hEGF, BPE, insulin, antibacterial agent, hydrocortisone)), and further cultured for 24 hours.

[0218] After culturing for 24 hours, the medium was removed, and the test sample (Sample 1, the final concentration is shown in Table 16 below) dissolved in KBM was added 2 mL each to each well and cultured for 24 hours under the conditions of 37 °C and 5% CO2. As a control, the same culture was performed using KBM without the addition of the sample. After culturing, the medium was removed, and total RNA was extracted with ISOGEN II (manufactured by Nippon Gene), the amount of each RNA was measured with a spectrophotometer, and the total RNA was prepared to be 150 ng / μL.

[0219] Using this total RNA as a template, the expression levels of mRNA for SPT and GAPDH, which is an internal standard, were measured. Detection was performed using a real-time PCR device Thermal Cycler Dice Real Time System III (manufactured by Takara Bio Inc.) with PrimeScript TMRT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. SPT mRNA expression levels were corrected using GAPDH values ​​based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the SPT mRNA expression enhancement rate (%) was calculated using the following formula.

[0220] SPT mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 16.

[0221] [Table 16]

[0222] As shown in Table 16, compound 1 (sample 1) exhibited excellent SPT mRNA expression-promoting activity.

[0223] [Test Example 16] Aquaporin 3 (AQP3) mRNA Expression Enhancement Test The AQP3 mRNA expression-promoting effect of compound 1 (sample 1) was tested as follows.

[0224] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0225] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test sample (Sample 1, see Table 17 below for final concentration) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without the sample was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each well was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0226] Using this total RNA as a template, mRNA expression levels for AQP3 and the internal standard GAPDH were measured. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. The expression level of AQP3 mRNA was corrected using the GAPDH value, based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the AQP3 mRNA expression enhancement rate (%) was calculated using the following formula.

[0227] AQP3 mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 17.

[0228] [Table 17]

[0229] As shown in Table 17, compound 1 (sample 1) exhibited excellent AQP3 mRNA expression-promoting activity.

[0230] [Test Example 17] Filaggrin (FLG) mRNA Expression Enhancement Test The FLG mRNA expression-promoting effect was tested for compounds 1-3 (samples 1-3) as follows.

[0231] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0232] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test samples (Samples 1-3, see Table 18 below for final concentrations) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without any added samples was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each sample was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0233] Using this total RNA as a template, mRNA expression levels were measured for FLG and the internal standard GAPDH. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. FLG mRNA expression levels were corrected using GAPDH values ​​based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the FLG mRNA expression enhancement rate (%) was calculated using the following formula.

[0234] FLG mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 18.

[0235] [Table 18]

[0236] As shown in Table 18, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) all exhibited excellent FLG mRNA expression-promoting activity.

[0237] [Test Example 18] Claudin-1 (CLDN1) mRNA expression enhancement test The CLDN1 mRNA expression-promoting effect was tested for compound 1 (sample 1) and compound 3 (sample 3) as follows.

[0238] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0239] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test samples (Samples 1 and 3, see Table 19 below for final concentrations) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without any added samples was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each sample was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0240] Using this total RNA as a template, mRNA expression levels for CLDN1 and the internal standard GAPDH were measured. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. CLDN1 mRNA expression levels were corrected using GAPDH values ​​based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the CLDN1 mRNA expression enhancement rate (%) was calculated using the following formula.

[0241] CLDN1 mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 19.

[0242] [Table 19]

[0243] As shown in Table 19, both compound 1 (sample 1) and compound 3 (sample 3) exhibited excellent CLDN1 mRNA expression-promoting activity.

[0244] [Test Example 19] Claudin-4 (CLDN4) mRNA Expression Enhancement Test The CLDN4 mRNA expression-promoting effect of compound 1 (sample 1) was tested as follows.

[0245] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0246] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test sample (Sample 1, see Table 20 below for final concentration) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without the sample was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each well was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0247] Using this total RNA as a template, mRNA expression levels for CLDN4 and the internal standard GAPDH were measured. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R) The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. CLDN4 mRNA expression levels were corrected using GAPDH values ​​based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the CLDN4 mRNA expression enhancement rate (%) was calculated using the following formula.

[0248] CLDN4 mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 20.

[0249] [Table 20]

[0250] As shown in Table 20, compound 1 (sample 1) exhibited excellent CLDN4 mRNA expression-promoting activity.

[0251] [Test Example 20] Ocludin (OCLN) mRNA Expression Enhancement Test The OCLN mRNA expression-promoting effect was tested for compound 1 (sample 1) and compound 3 (sample 3) as follows.

[0252] Normal human neonatal epidermal keratinocytes (NHEKs) were pre-cultured in normal human epidermal keratinocyte growth medium (KGM), and the cells were harvested by trypsin treatment. The harvested cells were 15 × 10⁴ 4 After diluting with KGM to achieve a cell density of cells / mL, seed 2 mL into each 6-well plate (30 × 10 4 Cells were cultured overnight in wells at 37°C and 5% CO2. After culturing, the culture medium was replaced with normal human epidermal keratinocyte basal medium (KBM, the same as KGM above but without growth additives (hEGF, BPE, insulin, antibacterial agents, hydrocortisone)), and the cells were cultured for a further 24 hours.

[0253] After 24 hours of incubation, the culture medium was removed, and 2 mL of the test samples (Samples 1 and 3, see Table 21 below for final concentrations) dissolved in KBM was added to each well. The samples were then incubated at 37°C and 5% CO2 for 24 hours. As a control, KBM without any added samples was cultured in the same manner. After incubation, the culture medium was removed, and total RNA was extracted using ISOGEN II (Nippon Gene Co., Ltd.). The amount of RNA in each sample was measured using a spectrophotometer, and the total RNA was adjusted to 150 ng / μL.

[0254] Using this total RNA as a template, mRNA expression levels were measured for OCLN and the internal standard GAPDH. Detection was performed using a real-time PCR instrument, Thermal Cycler Dice Real Time System III (Takara Bio Inc.), and PrimeScript. TM RT Master Mix (Perfect Real Time) (Takara Bio Co., Ltd.) TB Green (R)The reaction was performed using a two-step real-time RT-PCR reaction with Fast qPCR Mix (Takara Bio Inc.). Primers from Takara Bio Inc. were used. OCLN mRNA expression levels were corrected using GAPDH values ​​based on total RNA preparations prepared from cells cultured with and without the test sample. From the obtained values, the OCLN mRNA expression enhancement rate (%) was calculated using the following formula.

[0255] OCLN mRNA expression enhancement rate (%) = A / B × 100 Each term in the formula represents the following: A: Correction value when adding test sample B: Corrected value without sample addition The results are shown in Table 21.

[0256] [Table 21]

[0257] As shown in Table 21, both compound 1 (sample 1) and compound 3 (sample 3) exhibited excellent OCLN mRNA expression-promoting activity.

[0258] [Test Example 21] Test for inhibitory effect on the formation of advanced glycation end products (AGEs) The inhibitory effect on AGEs formation was tested for compound 1 (sample 1) and compound 2 (sample 2) as follows.

[0259] A 100 μL mixture of 0.2MD(-)-ribose prepared in PBS(-) buffer and test samples (Samples 1 and 2; see Table 22 below for final concentrations) was added to a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.), and the mixture was left to stand at 37°C for 20 days to allow AGEs to form. A negative control was prepared using only PBS(-) buffer, and a positive control was prepared using a 0.2MD(-)-ribose solution prepared in PBS(-) buffer, both of which were left to stand in the same manner. After 20 days, the amount of AGEs was measured by ELISA using an anti-AGEs antibody (Transgenic Co., Ltd.), and the inhibitory effect on AGEs formation was evaluated. From the obtained results, the AGEs formation inhibition rate (%) was calculated using the following formula.

[0260] AGEs formation inhibition rate (%)={(BC) / (BA)}×100 Each term in the formula represents the following: A: Absorbance at a wavelength of 405 nm in the negative control B: Absorbance at a wavelength of 405 nm in the positive control C: Absorbance at a wavelength of 405 nm with the test sample added. The results are shown in Table 22.

[0261] [Table 22]

[0262] As shown in Table 22, compound 1 (sample 1) and compound 2 (sample 2) exhibited excellent inhibitory effects on AGEs formation.

[0263] [Test Example 22] Test on the effect of promoting the degradation of advanced glycation end products (AGEs) The AGEs degradation-promoting effect of compound 1 (sample 1) was tested as follows.

[0264] 100 μL of 0.2 MD (-)-ribose solution prepared in PBS(-) buffer was added to a 96-well type I collagen-coated plate (Asahi Glass Co., Ltd.), and the plates were left to stand at 37°C for 2 weeks to allow AGEs to form. A negative control was prepared by adding only PBS(-) buffer and leaving it to stand in the same manner. After 2 weeks, 100 μL of the test sample (Sample 1, see Table 23 below for the final concentration) prepared in PBS(-) buffer was added to each plate, and the plates were left to stand at 37°C for another 20 days. A positive control was prepared by using only PBS(-) buffer instead of the test sample, and a negative control was prepared by continuing to use only PBS(-) buffer, both of which were left to stand in the same manner. After 20 days, the amount of AGEs was measured by ELISA using an anti-AGEs antibody (Transgenic Co., Ltd.), and the AGEs degradation promoting effect was evaluated. From the obtained results, the AGEs degradation promotion rate (%) was calculated using the following formula.

[0265] AGEs degradation promotion rate (%)={(BC) / (BA)}×100 Each term in the formula represents the following: A: Absorbance at a wavelength of 405 nm in the negative control B: Absorbance at a wavelength of 405 nm in the positive control C: Absorbance at a wavelength of 405 nm with the test sample added. The results are shown in Table 23.

[0266] [Table 23]

[0267] As shown in Table 23, compound 1 (sample 1) exhibited excellent AGEs degradation promoting activity.

[0268] [Test Example 23] Test for Inhibition of Testosterone 5α-Reductase The testosterone 5α-reductase inhibitory effect was tested for compounds 1-3 (samples 1-3) as follows.

[0269] In a capped V-bottom test tube, 20 μL of a 4.2 mg / mL testosterone solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) prepared with propylene glycol was mixed with 825 μL of 5 mmol / L Tris-HCl (pH 7.13) buffer containing 1 mg / mL NADPH.

[0270] Furthermore, 80 μL of the test sample solution (samples 1-3, see Table 24 below for final concentrations) prepared in 80% ethanol was added to 75 μL of S-9 (Oriental Yeast Co., Ltd., rat liver homogenate) and mixed, and incubated at 37°C for 60 minutes. Then, 1 mL of methylene chloride was added to stop the reaction. This was centrifuged (1600 × g, 10 minutes), the methylene chloride layer was separated, and the separated methylene chloride layer was subjected to gas chromatography analysis under the following conditions to quantify the concentrations of 3α-androstanediol, 5α-dihydrotestosterone (5α-DHT), and testosterone. As a control, the same amount (80 μL) of sample solvent was used instead of the test sample solution and treated in the same manner, and subjected to gas chromatography analysis.

[0271] <Gas chromatography conditions> Equipment used: Shimadzu GC-2010 (manufactured by Shimadzu Corporation) Column: DB-1701 (Inner diameter: 0.53 mm, Length: 30 m, Film thickness: 1.0 μm) (Manufactured by J&W Scientific) Column temperature: 240℃ Inlet temperature: 300℃ Detector: FID Sample injection volume: 1 μL Split ratio: 1:2 Carrier gas: Nitrogen gas Carrier gas flow rate: 12 mL / min

[0272] The concentrations of 3α-androstanediol, 5α-DHT, and testosterone were quantified using the following method. Standard samples of 3α-androstanediol, 5α-DHT, and testosterone were dissolved in ethanol, and the solutions were subjected to gas chromatography analysis. The correspondence between the peak area and the concentration of these compounds was determined in advance from the concentrations (μg / mL) and peak areas of the compounds. Then, the concentrations per peak area of ​​3α-androstanediol, 5α-DHT, and testosterone after the reaction of testosterone with S-9 were determined based on the previously determined correspondence using the following formula (1).

[0273] 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 the standard sample D: Peak area of ​​standard product

[0274] Using the compound concentrations calculated based on equation (1), the conversion rate (the ratio of the concentrations of 3α-androstanediol and 5α-DHT produced by the reduction of testosterone by testosterone 5α-reductase to the initial concentration of testosterone) was calculated based on equation (2) below.

[0275] Conversion rate = (E + F) / (E + F + G) ... (2) Each term in the formula represents the following: E: Concentration of 3α-androstanediol (μg / mL) F: Concentration of 5α-DHT (μg / mL) G: Testosterone concentration (μg / mL)

[0276] Using the conversion rate calculated based on equation (2), the testosterone 5α-reductase inhibition rate (%) was calculated based on equation (3) below. Testosterone 5α-reductase inhibition rate (%) = (1 - H / I) × 100 ... (3) Each term in the formula represents the following: H: Conversion rate when test sample is added I: Conversion rate without sample addition The results are shown in Table 24.

[0277] [Table 24]

[0278] As shown in Table 24, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were confirmed to have excellent testosterone 5α-reductase inhibitory activity.

[0279] [Test Example 24] Test on the effect of promoting hair papilla cell proliferation The hair papilla cell proliferation promoting effect of compound 1 (sample 1) was tested as follows.

[0280] Normal human hair papilla cells (HFDPC, derived from male scalp) were cultured in hair papilla cell growth medium (PCGM, Toyobo Co., Ltd.) containing 1% FCS and growth additives, and then the cells were harvested by trypsin treatment. The harvested cells were then subjected to 1.0 × 10⁶ filtration in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS. 4 After diluting to a cell density of cells / mL, 200 μL was seeded per well in a collagen-coated 96-well plate and cultured for 3 days.

[0281] Subsequently, the culture medium was removed, and 200 μL of the test sample (Sample 1, see Table 25 below for final concentration) dissolved in serum-free DMEM was added to each well, and the cells were cultured for a further 4 days. As a control, cells were cultured similarly using serum-free DMEM without the sample. After the culture period, the hair papilla cell proliferation promoting effect was measured by the MTT assay. Specifically, the culture medium was removed, 100 μL of 0.4 mg / mL MTT prepared in serum-free DMEM was added to each well, and the cells were cultured for a further 2 hours. Then, the blue formazan produced in the cells was extracted with 100 μL of 2-propanol. The absorbance of this extract at 570 nm, where the absorption maximum of blue formazan is located, was measured. Simultaneously, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. From the measurement results, the hair papilla cell proliferation promotion rate (%) was calculated based on the following formula.

[0282] Hair papilla cell proliferation promotion rate (%) = A / B × 100 Each term in the formula represents the following: A: Amount of blue formazan produced when the test sample is added. B: Amount of blue formazan produced without sample addition The results are shown in Table 25.

[0283] [Table 25]

[0284] As shown in Table 25, compound 1 (sample 1) was found to have excellent hair papilla cell proliferation promoting activity.

[0285] [Test Example 25] Nitric Oxide (NO) Production Inhibitory Effect Test The inhibitory effect of compound 1 (sample 1) on nitric oxide (NO) production was tested as follows.

[0286] Mouse macrophage cells (RAW264.7) were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested using a cell scraper. The harvested cells were 3.0 × 10⁶. 6After diluting the cells in phenol red-free DMEM containing 10% FBS to a cell density of cells / mL, 100 μL was seeded per well in a 96-well plate and cultured for 4 hours.

[0287] After incubation, the culture medium was removed, and 100 μL of the test sample (Sample 1, see Table 26 below for final concentration) dissolved in phenol red-free DMEM containing 10% FBS and 0.5% DMSO was added to each well. Then, 100 μL of lipopolysaccharide (LPS, final concentration 1 μg / mL, E. coli 0111:B4, DIFCO) dissolved in phenol red-free DMEM containing 10% FBS was added, and the culture was incubated for 48 hours. As a control, instead of the test sample solution, phenol red-free DMEM containing phenol red-free and 0.5% DMSO with no sample added was used, and the same LPS treatment was performed.

[0288] Nitric oxide (NO) production is related to nitrite ions (NO2) - The amount was measured as an indicator. After the culture was completed, the same amount of Gris reagent (5% phosphoric acid solution containing 1% sulfanilamide and 0.1% N-1-naphthyl ethylendiamine dihydrochloride) as the culture supernatant was added to the culture medium in each well, and the mixture was reacted at room temperature for 10 minutes. After the reaction, the absorbance at a wavelength of 540 nm was measured. The nitric oxide (NO) production inhibition rate (%) was calculated using the following formula based on the amount of nitric oxide (NO) produced when no sample was added (control).

[0289] NO production suppression rate (%)={(BA) / B}×100 Each term in the formula represents the following: A: NO amount when test sample is added B: NO amount when test sample is added The results are shown in Table 26.

[0290] [Table 26]

[0291] As shown in Table 26, compound 1 (sample 1) was confirmed to have excellent nitric oxide production inhibitory activity.

[0292] [Test Example 26] Hyaluronidase activity inhibition test The hyaluronidase activity inhibitory effect was tested for compounds 1-3 (samples 1-3) as follows.

[0293] To 0.2 mL of the test samples (samples 1-3, see Table 27 below for final concentrations) dissolved in 0.1 mol / L acetate buffer (pH 3.5), 0.1 mL of hyaluronidase solution (SIGMA, Type IV-S, from bovine testes, 400 NF units / mL) was added and the mixture was allowed to stand at 37°C for 20 minutes. Furthermore, 0.2 mL of 2.5 mmol / L calcium chloride was added as an activator and the mixture was allowed to stand at 37°C for 20 minutes. 0.5 mL of 0.8 mg / mL sodium hyaluronate solution (from rooster comb) was then added and the mixture was reacted at 37°C for 40 minutes. Afterward, 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 on ice, 6 mL of p-DABA reagent was added and the mixture was reacted at 37°C for 20 minutes. The absorbance at a wavelength of 585 nm was then measured.

[0294] Furthermore, as a blank, the same procedure and absorbance measurements were performed in a case where no enzyme solution was added. In addition, as a control, the same measurements were performed in a case where distilled water was added instead of the sample solution. From the obtained results, the hyaluronidase activity inhibition rate (%) was calculated using the following formula. Hyaluronidase activity inhibition rate (%) = {1 - (AB) / (CD)} × 100 Each term in the formula represents the following: A: Absorbance at a wavelength of 585 nm after adding the test sample and enzyme. B: Absorbance at a wavelength of 585 nm with test sample added but no enzyme added. C: Absorbance at a wavelength of 585 nm with no sample added and with enzyme added. D: Absorbance at a wavelength of 585 nm without sample or enzyme addition. The results are shown in Table 27.

[0295] [Table 27]

[0296] As shown in Table 27, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were all confirmed to have excellent hyaluronidase activity inhibitory effects.

[0297] [Test Example 27] Hexosaminidase release inhibition test The hexosaminidase release inhibitory effect of compound 2 (sample 2) was tested as follows.

[0298] Rat basophilic leukemia cells (RBL-2H3) were cultured in a modified Eagle's Minimum Essential Medium (S-MEM) spinner containing 15% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were 4.0 × 10⁶ 5 The cells were diluted with 15% FBS-containing S-MEM to achieve a cell density of cells / mL, and DNP-specific IgE was added to a final concentration of 0.5 μg / mL. The cells were then seeded in 96-well plates at a rate of 100 μL per well and cultured overnight.

[0299] After incubation, the culture medium was removed and the wells were washed twice with 100 μL of Silaganan buffer. Next, 30 μL of the same buffer and 10 μL of the test sample (Sample 2, see Table 28 below for final concentration) dissolved in the same buffer were added to each well, and the mixture was allowed to stand at 37°C for 10 minutes. As a control, the same procedure was performed using 40 μL of Silaganan buffer without any sample added. Subsequently, 10 μL of 400 ng / mL DNP-BSA solution was added, and the mixture was allowed to stand at 37°C for 15 minutes to release hexosaminidase.

[0300] Subsequently, the release was stopped by placing the 96-well plate on ice. 10 μL of the cell supernatant from each well was collected in a new 96-well plate, and 10 μL of 1 mmol / L p-nitrophenyl-N-acetyl-β-D-glucosaminid (p-NAG) solution was added to each well. The mixture was reacted at 37°C for 1 hour.

[0301] After the reaction was complete, 250 μL of 0.1 mol / L Na2CO3 / NaHCO3 was added to each well, and the absorbance at wavelengths of 415 nm and 650 nm was measured. The corrected value was obtained by subtracting the absorbance at 650 nm from the absorbance at 415 nm. From the obtained measurement results, the hexosaminidase release inhibition rate (%) was calculated using the following formula.

[0302] Hexosaminidase release inhibition rate (%) = {1 - (B / A)} × 100 Each term in the formula represents the following: A: Absorbance at wavelengths of 415-650 nm without sample addition B: Absorbance at wavelengths of 415-650 nm with the test sample added. The results are shown in Table 28.

[0303] [Table 28]

[0304] As shown in Table 28, compound 2 (sample 2) was confirmed to have excellent hexosaminidase release inhibitory activity.

[0305] [Test Example 28] Test of the inhibitory effect on prostaglandin E2 (PGE2) production in mouse macrophages The PGE2 production inhibitory effect of compound 1 (sample 1) was tested as follows.

[0306] Mouse macrophage cells (RAW264.7) were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested using a cell scraper. The harvested cells were 2.0 × 10⁶. 5After diluting to a concentration of cells / mL using 10% FBS-containing DMEM, 100 μL was seeded per well in a 96-well plate and incubated for 18 hours.

[0307] After culturing, the culture medium was replaced with 500 μmol / L aspirin-containing medium and cultured for 4 hours to acetylate and inactivate the already present COX-1 and the small amount of COX-2 being expressed. Subsequently, the cells were washed three times with PBS(-) buffer, and 100 μL of the test sample (Sample 1, see Table 29 below for final concentration) dissolved in 10% FBS-containing DMEM with a final concentration of 0.5% DMSO was added to each well. Then, 100 μL of lipopolysaccharide (LPS) (DIFCO, E. coli 0111; B4) dissolved in 10% FBS-containing DMEM at a final concentration of 1 μg / mL was added, and the cells were cultured for 16 hours. As a control, cells were cultured similarly using 10% FBS-containing DMEM with a final concentration of 0.5% DMSO without any sample added. After culturing, the amount of prostaglandin E2 in the culture supernatant of each well was quantified using the PGE2EIA Kit (Cayman Chemical). Based on the results obtained, the PGE2 production inhibition rate (%) was calculated using the following formula.

[0308] PGE2 production suppression rate (%)={1-(AC) / (BC)}×100 Each term in the formula represents the following: A: PGE2 quantity upon addition of test sample and LPS stimulation B: PGE2 amount in sample-free, LPS-stimulated state C: PGE2 amount without sample addition and LPS irritation. The results are shown in Table 29.

[0309] [Table 29]

[0310] As shown in Table 29, compound 1 (sample 1) was confirmed to have excellent PGE2 production inhibitory activity in macrophages.

[0311] [Test Example 29] Glutathione production promotion test (hepatocytes) The glutathione production-promoting effect in hepatocytes was tested for compounds 1-3 (samples 1-3) as follows.

[0312] Normal human hepatocytes were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were divided into 10 × 10⁶ cells. 4 After diluting the cells in 10% FBS-containing DMEM to achieve a cell density of cells / mL, 200 μL was seeded per well in a 48-well plate and cultured overnight.

[0313] After culturing, the culture medium was removed, and 200 μL of the test samples (Samples 1-3, see Table 30 below for final concentrations) dissolved in 1% FBS-containing DMEM was added to each well, and the cells were cultured for a further 24 hours. As a control, cells were cultured similarly using 1% FBS-containing DMEM without any added samples. After culturing, the culture medium was removed from each well, washed with 400 μL of PBS(-), and then the cells were lysed with 150 μL of M-PER (PIERCE).

[0314] Of this, 100 μL was used to quantify total glutathione. Specifically, 100 μL of cell extract dissolved in a 96-well plate, 50 μL of 0.1 mol / L phosphate buffer, 25 μL of 2 mmol / L NADPH, and 25 μL of 3.2 units / mL glutathione reductase were added and incubated at 37°C for 10 minutes. Then, 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 determine ΔOD / min. The total glutathione concentration was calculated based on a calibration curve created using oxidized glutathione (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.). After correcting the obtained values ​​to glutathione amount per total protein amount, the glutathione production promotion rate (%) was calculated based on the following formula.

[0315] Glutathione production promotion rate (%) = B / A × 100 Each term in the formula represents the following: A: Glutathione content per unit of total protein in the sample without additives B: Amount of glutathione per unit of total protein in the test sample The results are shown in Table 30.

[0316] [Table 30]

[0317] As shown in Table 30, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were all found to have excellent glutathione production-promoting effects in hepatocytes.

[0318] [Test Example 30] Test on the effect of promoting ATP production in hepatocytes The ATP production-promoting effect in hepatocytes was tested for compounds 1-3 (samples 1-3) as follows.

[0319] Normal human hepatocytes were cultured in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS, and then the cells were harvested by trypsin treatment. The harvested cells were 2.0 × 10⁶ 5 After diluting with 10% FBS-containing DMEM to achieve a cell density of cells / mL, 100 μL was seeded per well in a 96-well plate and cultured overnight.

[0320] After the culture period, the culture medium was removed, and 100 μL of the test samples (Samples 1-3, see Table 31 below for final concentrations) dissolved in 10% FBS-containing DMEM was added to each well and cultured for 2 hours. As a control, cultures were performed similarly using 10% FBS-containing DMEM without any added samples.

[0321] The ATP production-promoting effect was evaluated by measuring the amount of ATP in cells using the firefly luciferase luminescence method. Specifically, after 2 hours of incubation, 100 μL of ATP measurement reagent (Toyo B-Net Co., Ltd., product name: "ATP Measurement Reagent for Cells") was added to each well, and a chemiluminescence reaction by luciferase was performed. After the reaction, the amount of chemiluminescence proportional to the amount of ATP in cells was measured using a chemiluminescence analyzer (Thermo Fisher Scientific, product name: Varioskan LUX multimode microplate reader). From the obtained results, the ATP production promotion rate (%) was calculated using the following formula.

[0322] ATP production promotion rate (%)=A / B×100 Each term in the formula represents the following: A: Amount of chemiluminescence upon addition of test sample B: Chemiluminescence without sample addition The results are shown in Table 31.

[0323] [Table 31]

[0324] As shown in Table 31, compound 1 (sample 1), compound 2 (sample 2), and compound 3 (sample 3) were all found to have excellent ATP production-promoting effects in hepatocytes.

[0325] [Formulation example 1] Tablets having the following composition were manufactured by conventional methods. Compound 1 5.0mg Dolomite (containing 20% ​​calcium and 10% magnesium) 83.4 mg Casein phosphopeptide 16.7 mg Vitamin C 33.4mg Maltitol 136.8mg Collagen 12.7mg Sucrose fatty acid ester 12.0 mg

[0326] [Formulation example 2] An oral liquid formulation having the following composition was manufactured by a conventional method. <Composition per ampoule (100 mL)> Compound 2 0.3% by mass Sorbitol 12.0% by mass Sodium benzoate 0.1% by mass Fragrance 1.0% by mass Calcium sulfate 0.5% by mass Purified water remainder (100% by mass)

[0327] [Formulation example 3] Capsules having the following composition were manufactured by conventional methods. No. 1 hard gelatin capsules were used as the capsules. <Composition per capsule (1 tablet, 200mg)> Compound 3 10.0mg Corn starch 70.0 mg Lactose 100.0mg Calcium lactate 10.0 mg Hydroxypropylcellulose (HPC-L) 10.0 mg

[0328] [Formulation example 4] An emulsion was prepared by conventional methods according to the following composition. Compound 1 0.01g Jojoba oil 4.00g 1,3-Butylene glycol 3.00g Arbutin 3.00g Polyoxyethylene cetyl ether (20 E.O.) 2.50g Olive oil 2.00g Squalane 2.00g Cetanol 2.00g Glyceryl monostearate 2.00g Polyoxyethylene sorbitan oleate (20E.O.) 2.00g Methyl parahydroxybenzoate 0.15g Stearyl glycyrrhetinate 0.10g Scutellaria baicalensis extract 0.10g Dipotassium glycyrrhizinate 0.10g Ginkgo biloba extract 0.10g Conchiolin 0.10g Phellodendron bark extract 0.10g Chamomile extract 0.10g Fragrance 0.05g Purified water, remaining amount (assuming a total volume of 100g)

[0329] [Formulation example 5] A cream with the following composition was manufactured by conventional methods. Compound 2 0.05g Sophora flavescens extract 0.1g Scutellaria baicalensis extract 0.1g Liquid paraffin 5.0g 4.0g of bleached beeswax Squalane 10.0g Cetanol 3.0g Lanolin 2.0g Stearic acid 1.0g Polyoxyethylene sorbitan oleate (20E.O.) 1.5g Glyceryl monostearate 3.0g Oil-soluble licorice extract 0.1g 1,3-Butylene glycol 6.0g Methyl parahydroxybenzoate 1.5g Fragrance 0.1g Purified water, remaining amount (assuming a total volume of 100g)

[0330] [Formulation example 6] A beauty serum with the following composition was manufactured by conventional methods. Compound 3 0.01g Chamomile extract 0.1g Carrot extract 0.1g Xanthan gum 0.3g Hydroxyethylcellulose 0.1g Carboxyvinyl polymer 0.1g 1,3-Butylene glycol 4.0g Dipotassium glycyrrhizinate 0.1g Glycerin 2.0g Potassium hydroxide 0.25g Fragrance 0.01g Preservative (methyl parahydroxybenzoate) 0.15g Ethanol 2.0g Purified water, remaining amount (assuming a total volume of 100g)

[0331] [Formulation example 7] A hair tonic with the following composition was manufactured by conventional methods. Compound 1 0.2g Compound 2 0.1g Compound 3 0.1g Tocopherol acetate (appropriate amount) Cephalatin 0.002g Isopropylmethylphenol 0.1g Sodium hyaluronate 0.15g Glycerin 15.0g Ethanol 15.0g Fragrance (appropriate amount) Chelating agent (sodium edetate) appropriate amount Preservative (hinokitiol) appropriate amount Solubilizer (polyoxyethylene cetyl ether) appropriate amount Purified water, remaining amount (assuming a total volume of 100g)

[0332] [Formulation example 8] A shampoo with the following composition was manufactured by conventional methods. Compound 1 0.2g Compound 2 0.2g Compound 3 0.2g Marjoram extract 1.0g Plum fruit extract 0.2g Sodium methyl taurate (coconut oil fatty acid) 10.0g Coconut oil fatty acid amidopropyl betaine 10.0g Sodium polyoxyethylene alkyl ether sulfate 20.0g Coconut oil fatty acid diethanolamide 4.0g Propylene glycol 2.0g Fragrance (appropriate amount) Purified water, remaining amount (assuming a total volume of 100g)

Claims

[Claim 1] A skin whitening agent characterized by containing compound 1 and / or compound 2 represented by the following general formula (I) as an active ingredient. 【Chemistry 1】

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