Novel ketone compounds, radical scavengers, hyaluronidase inhibitors, PGE2 production inhibitors, tyrosinase inhibitors, elastase inhibitors, type I collagen production promoters, and hair papilla cell proliferation promoters

Abstract

To provide compounds that are obtained from extract of natural origin and have radical erasing action, hyaluronidase activity inhibitory action, PGE2 production inhibitory action, tyrosinase activity inhibitory action, elastase activity inhibitory action, collagen type I production promoting action and dermal papilla cell proliferation promoting action.SOLUTION: The present invention provides a ketone compound represented by the formula (1).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to novel ketone compounds, radical scavengers, hyaluronidase activity inhibitors, PGE2 production suppressors, tyrosinase activity inhibitors, elastase activity inhibitors, type I collagen production promoters, and hair papilla cell proliferation promoters. [Background technology]

[0002] In recent years, active oxygen and biological radicals have been attracting attention as factors that cause oxidation of biological components, and their adverse effects on the living body have become a problem. Active oxygen is generated during the energy metabolism process in living cells. For example, superoxide anion (·O2) is generated by the one-electron reduction of oxygen molecules. ― ), hydrogen peroxide (H2O2), singlet oxygen ( 1 These reactive oxygen species are essential for the bactericidal mechanism of phagocytes and play an important role in eliminating viruses and cancer cells.

[0003] However, excessive production of reactive oxygen species attacks biological molecules that make up membranes and tissues in the body, inducing various diseases. Superoxide, which is produced in the body and serves as the starting material for other reactive oxygen species, is normally eliminated sequentially by the catalytic action of superoxide dismutase (SOD) contained within cells. However, when superoxide production is excessive or SOD activity is impaired, superoxide elimination is insufficient, resulting in high superoxide concentrations. This is thought to be one of the causes of tissue damage such as rheumatoid arthritis and Behçet's disease, as well as myocardial infarction, stroke, cataracts, age spots, freckles, wrinkles, diabetes, arteriosclerosis, stiff shoulders, sensitivity to cold, and skin aging.

[0004] Among these, the skin is an organ that is susceptible to the production of superoxide because it is directly exposed to environmental factors such as ultraviolet rays, and therefore, when the concentration of superoxide increases, it can decompose, denature, or crosslink biological tissues such as collagen, and oxidize oils and fats to produce lipid peroxides that damage cells, resulting in problems such as the formation of wrinkles in the skin, aging such as decreased skin elasticity, inflammation, and skin pigmentation (see Non-Patent Document 1). Therefore, it is believed that inhibiting or suppressing the production of reactive oxygen species and biological radicals can prevent, treat, or improve various diseases involving reactive oxygen species, such as skin aging such as the formation of wrinkles and decreased elasticity, tissue disorders such as rheumatoid arthritis and Behçet's disease, myocardial infarction, stroke, cataracts, diabetes, arteriosclerosis, stiff shoulders, and sensitivity to cold.

[0005] Conventionally, extracts of Laeliocattleya drumbeat (Orchidaceae) (see Patent Document 1) and Laeliocattleya nigricans extract (see Patent Document 2) are known to have active oxygen scavenging activity, radical scavenging activity, or hydrogen peroxide scavenging activity.

[0006] Inflammatory diseases, such as contact dermatitis (rash), psoriasis, pemphigus vulgaris, and other skin diseases accompanied by rough skin, have diverse causes and onset mechanisms. Some of these causes are known to be due to increased hyaluronidase activity or increased production of prostaglandin E2 (PGE2).

[0007] Hyaluronidase is a hydrolase of hyaluronic acid. Hyaluronate, which maintains its affinity for body tissues, is decomposed by ultraviolet light, oxygen, etc. in aqueous systems, and its moisture-retaining effect decreases as its 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 their activation and acts as an inflammatory chemical mediator. Therefore, inhibiting hyaluronidase activity is expected to enhance moisture retention and prevent and improve inflammation.

[0008] Hitherto, substances known to have hyaluronidase activity inhibitory activity include, for example, hinokitiol and at least one extract selected from the group consisting of extracts of horsetail, hawthorn, and hamamelis (see Patent Document 3), and one or more plant extracts selected from white willow, loquat, lime, persimmon, and onion (see Patent Document 4).

[0009] Inflammation is a complex reaction that manifests with symptoms such as redness, edema, fever, pain, itching, and functional impairment. For example, when the skin is exposed to ultraviolet rays or comes into contact with an irritant, inflammatory cytokines are produced in the skin, causing skin inflammation. As a result, skin tissue is damaged, resulting in various symptoms such as rough skin, redness, edema, and pigmentation.

[0010] One of the inflammatory cytokines is prostaglandin E2 (PGE2). In the skin, PGE2 is produced, for example, by keratinocytes and causes skin inflammation. Therefore, inhibiting PGE2 production in keratinocytes is considered a method for preventing, treating, or ameliorating skin inflammation.

[0011] Hitherto, extracts of at least one raw material selected from the group consisting of kubi, hibiscus, japonica, and safflower (see Patent Document 5), pimento (Pimenta dioica), a plant belonging to the Myrtaceae family, and extracts thereof (see Patent Document 6) have been known to have an inhibitory effect on PGE2 production in keratinocytes.

[0012] In the skin, melanin protects the body from ultraviolet rays, but excessive production or uneven accumulation can cause skin darkening and blemishes. Melanin is generally produced by the enzyme tyrosinase, which is biosynthesized in pigment cells, converting tyrosine to dopa and then dopaquinone, followed by intermediates such as 5,6-dihydroxyindophenol. Therefore, inhibiting the melanin production process, i.e., inhibiting tyrosinase activity, is thought to be effective in preventing or improving skin darkening (cutaneous pigmentation).

[0013] Conventionally, substances known to have tyrosinase activity inhibitory activity include, for example, an extract of Cassia garrettiana (see Patent Document 7), an ethanol-soluble component extracted from Cucumis melo ver. conomon (Cucumis melo ver. conomon) as the extraction material, or the juice of the fruit of Cucumis melo or a hydrolysate thereof (see Patent Document 8).

[0014] The dermis and epidermis of the skin are composed of epidermal cells, fibroblasts, and the dermal extracellular matrix, such as elastin and type I collagen, which are located outside these cells and support the skin structure. In young skin, these skin tissues maintain homeostasis, ensuring moisturizing function, flexibility, elasticity, etc., and the skin is maintained in a firm, glossy, and moist appearance.

[0015] However, external factors such as ultraviolet radiation, extremely dry air, and excessive skin cleansing, as well as aging, reduce the production of elastin, type I collagen, and other major components of the dermal extracellular matrix, leading to their denaturation and degradation. As a result, the stratum corneum begins to peel abnormally, the skin loses firmness and luster, and symptoms of aging such as rough skin and wrinkles appear. These changes associated with skin aging, such as wrinkle formation, loss of firmness, and decreased elasticity, are related to the reduction and denaturation of dermal extracellular matrix components such as elastin and type I collagen.

[0016] It is known that elastase, an enzyme that breaks down elastin, is activated by ultraviolet light, accelerating the breakdown of elastin and causing the skin to lose its firmness and elasticity. Therefore, it is thought that inhibiting the activity of elastase will suppress the breakdown of elastin, thereby preventing and improving skin aging symptoms such as loss of firmness and loss of elasticity.

[0017] It is also known that smoking and other factors increase elastase activity in the body. Because the lung matrix is ​​composed of collagen and elastin, increased elastase activity due to smoking and other factors can destroy the alveolar walls and lead to emphysema and other conditions. Furthermore, increased elastase activity can destroy pulmonary capillaries, potentially leading to acute respiratory distress syndrome (ARDS) such as pulmonary edema. Therefore, inhibiting elastase activity in the body could potentially prevent and improve respiratory diseases such as emphysema and pulmonary edema.

[0018] Conventionally, substances known to have an elastase activity inhibitory effect include, for example, an extract of Actinidia polygama extracted from the fruit of Actinidiaceae (Actinidia polygama) (see Patent Document 9), and an enzymatic hydrolyzate, extract, extraction residue, or sprout of ripe seeds of okra (Abelmoschus esculentus) (see Patent Document 10).

[0019] Among collagens, type I collagen is the most abundant collagen found in the body, and is also found in large amounts in the dermis of the skin, where it is known to play a role in ensuring the skin's moisturizing function, flexibility, elasticity, etc. However, as the production of type I collagen decreases with age, the skin's moisturizing function and elasticity decrease, resulting in skin aging symptoms such as wrinkle formation and loss of firmness. Therefore, it is thought that promoting the production of type I collagen can prevent or improve skin aging symptoms such as wrinkle formation, loss of firmness, and loss of elasticity.

[0020] Conventionally, known substances that have the effect of promoting type I collagen production include, for example, plants of Ryukyu bamboo, Ficus batatas, and Tsuruna japonica or extracts thereof (see Patent Document 11), and extracts of Scenedesmus phytoplankton (microalgae) in a water-containing solvent (see Patent Document 12).

[0021] Hair grows and falls out repeatedly according to a cyclical hair cycle (hair cycle) consisting of an anagen phase, a regressive phase, and a telogen phase. The stage of this hair cycle, in which a new hair follicle is formed from the telogen phase to the anagen phase, is considered to be the most important for hair growth, and dermal papilla cells are thought to play an important role in the proliferation and differentiation of hair follicle epithelial cells during this stage. Dermal papilla cells are located inside hair follicle epithelial cells, which are composed of outer root sheath cells and matrix cells near the hair root, in the basement membrane-enclosed shaft portion of the hair root. They act on hair follicle epithelial cells to promote their proliferation, and thus play an important role in the proliferation and differentiation of hair follicle epithelial cells and hair formation (see Non-Patent Document 2).

[0022] As such, dermal papilla cells play the most important role in the proliferation and differentiation of hair follicle epithelial cells and in hair formation, and a method has been proposed in the past in which a target substance is brought into contact with cultured dermal papilla cells and the presence or absence and / or strength of the proliferation activity of the cells is determined to assay the hair growth effect of the target substance (see Patent Document 13).

[0023] Hitherto, extracts of three or more seaweeds selected from the families Laminaria, Pyrrhocoridae, Lessoniaceae, Fucalaceae, Sargassum, Mirrana, Gigarciniaceae, Thalleriaceae, Agalyaceae, Gracilaria, and Ulva (see Patent Document 14), extracts of Sargassum fasciatus (see Patent Document 15), and the like have been known to have the effect of promoting the proliferation of hair papilla cells. [Prior art documents] [Patent documents]

[0024] [Patent Document 1] Japanese Patent Application Laid-Open No. 2006-282536 [Patent Document 2] Japanese Patent Application Laid-Open No. 2008-088072 [Patent Document 3] Patent Publication No. 2021-187849 [Patent Document 4] Japanese Patent Application Laid-Open No. 2006-104098 [Patent Document 5] Japanese Patent Application Publication No. 2018-065763 [Patent Document 6] Japanese Patent Application Laid-Open No. 2014-118349 [Patent Document 7] Japanese Patent Application Laid-Open No. 2013-184952 [Patent Document 8] Japanese Patent Application Laid-Open No. 2011-256147 [Patent Document 9] Japanese Patent Application Laid-Open No. 2011-195542 [Patent Document 10] Japanese Patent Publication No. 2020-050601 [Patent Document 11] Japanese Patent Application Laid-Open No. 2011-195505 [Patent Document 12] Japanese Patent Application Laid-Open No. 2007-186471 [Patent Document 13] Japanese Patent Application Publication No. 10-229978 [Patent Document 14] Japanese Patent Application Laid-Open No. 2007-131571 [Patent Document 15] Japanese Patent Application Laid-Open No. 2008-247784 [Non-patent literature]

[0025] [Non-Patent Document 1] "Fragrance Journal" Special Issue No. 14, p. 156, 1995 [Non-patent document 2] Trends Genet, 1992, Vol. 8, pp. 56-61 Summary of the Invention [Problem to be solved by the invention]

[0026] The present invention aims to provide compounds that are obtained from highly safe extracts of natural origin and have radical scavenging activity, hyaluronidase activity inhibitory activity, PGE2 production inhibitory activity, tyrosinase activity inhibitory activity, elastase activity inhibitory activity, type I collagen production promoting activity, and dermal papilla cell proliferation promoting activity, as well as new PGE2 production inhibitors, tyrosinase activity inhibitors, elastase activity inhibitors, type I collagen production promoters, and dermal papilla cell proliferation promoters that contain said compounds. [Means for solving the problem]

[0027] The present inventors have succeeded in isolating and identifying novel ketone compounds from licorice extract that have radical scavenging activity, hyaluronidase activity inhibitory activity, PGE2 production inhibitory activity, tyrosinase activity inhibitory activity, elastase activity inhibitory activity, type I collagen production promotion activity, and hair papilla cell proliferation promotion activity, thereby completing the present invention.

[0028] Specifically, the present invention provides a ketone compound represented by the following structural formula (1), a radical scavenger, a hyaluronidase activity inhibitor, a PGE2 production suppressor, a tyrosinase activity inhibitor, an elastase activity inhibitor, a type I collagen production promoter, and a hair papilla cell proliferation promoter, each of which contains the ketone compound as an active ingredient.

[0029] [ka] [Effects of the Invention]

[0030] According to the present invention, it is possible to provide compounds that have radical scavenging activity, hyaluronidase activity inhibitory activity, PGE2 production inhibitory activity, tyrosinase activity inhibitory activity, elastase activity inhibitory activity, type I collagen production promoting activity, and hair papilla cell proliferation promoting activity, from highly safe extracts of natural origin, as well as radical scavengers, hyaluronidase activity inhibitors, PGE2 production inhibitors, tyrosinase activity inhibitors, elastase activity inhibitors, type I collagen production promoters, and hair papilla cell proliferation promoters that contain these compounds. DETAILED DESCRIPTION OF THE INVENTION

[0031] An embodiment of the present invention will be described. The ketone compound according to this embodiment is represented by the following structural formula (1).

[0032] [ka]

[0033] It has been confirmed in the Examples below that the ketone compound has the structure of 1-(2,4-dihydroxyphenyl)-2-(5-hydroxy-2,2-dimethyl-2H-chromen-6-yl)ethanone represented by the above structural formula (1), and is a novel ketone compound that can be isolated from licorice. Note that although the ketone compound can be isolated from licorice, it may also be present in plants other than licorice. Therefore, the source of isolation of the ketone compound is not limited to licorice, and any plant containing the ketone compound may be used.

[0034] The above-mentioned ketone compounds can be obtained, for example, by subjecting an oil-soluble licorice extract to purification procedures such as liquid-liquid partition extraction, various chromatographies, membrane separation, etc., and then treating the fraction containing the above-mentioned ketone compounds with preparative HPLC and recycling HPLC, etc.

[0035] Licorice is a perennial herb belonging to the genus Glycyrrhiza in the family Fabaceae. Licorice includes Glychyrrhiza glabra, Glychyrrhiza inflata, Glychyrrhiza uralensis, Glychyrrhiza aspera, Glychyrrhiza eurycarpa, Glychyrrhiza pallidiflora, Glychyrrhiza yunnanensis, Glychyrrhiza lepidota, Glychyrrhiza echinata, and Glychyrrhiza acanthocarpa. There are various types of licorice, including Glycyrrhiza acanthocarpa, and any of these may be used as the extraction raw material, but it is particularly preferable to use Glychyrrhiza glabra as the extraction raw material. Licorice parts that can be used as the extraction raw material include, for example, leaves, branches, bark, trunks, stems, fruits, seeds, flowers, roots, or mixtures thereof, with the roots being preferred.

[0036] Licorice that can be used as the extraction raw material can be obtained by drying, then either using it as is or crushing it using a crusher, and then extracting it with an extraction solvent. Drying can be carried out in the sun or using a commonly used dryer. Alternatively, dried roots or dried rhizomes, which are used as the extraction raw material for industrially produced glycyrrhizin, or the water extraction residue after extraction with water to obtain glycyrrhizin, etc., can also be used as the raw material.

[0037] The licorice extraction method is not particularly limited and can be selected appropriately depending on the purpose. For example, licorice, which is the extraction raw material, is placed in a treatment tank filled with an extraction solvent, and the soluble components are eluted while stirring as necessary, and then the extract is obtained by filtering to remove solids. The licorice may also be used as an extraction raw material after pretreatment such as degreasing with a nonpolar solvent such as hexane. Pretreatment such as degreasing allows for efficient extraction of the plant with a polar solvent.

[0038] As the solvent used for extraction, it is preferable to use an organic solvent at room temperature or a temperature below the boiling point of the solvent.

[0039] Examples of organic solvents that can be used as extraction solvents include benzene, toluene, xylene, ethyl ether, methyl ethyl ketone, methyl isobutyl ketone, dichloromethane, dichloroethane, chloroform, ethyl acetate, propyl acetate, butyl acetate, acetone, methanol, ethanol, propanol, aqueous methanol, aqueous ethanol, aqueous propanol, etc. Furthermore, carbon dioxide can also be used as a supercritical fluid.

[0040] The method for obtaining a licorice extract from the water-extracted residue as the extraction raw material using the above-mentioned organic solvent is not particularly limited, but examples include a method in which 2 to 10 times the amount of organic solvent is added to the water-extracted residue as the extraction raw material, and extraction is performed at room temperature while stirring, and a method in which extraction is performed by heating under reflux.

[0041] The purification can be carried out by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, etc. Since the licorice extract has a unique smell and taste, it can be purified for the purpose of decolorization, deodorization, etc., within a range that does not result in a decrease in its physiological activity.

[0042] The extract thus obtained may be subjected to solid-liquid separation such as centrifugation or filtration to remove insoluble matter, and then may be subjected to treatments such as dilution, concentration, drying, purification, etc. in accordance with conventional methods to obtain a diluted or concentrated extract, a dried extract, or a crude or purified product thereof.

[0043] The licorice extract obtained as described above is adsorbed onto an adsorbent and then eluted with water, a water-soluble solvent, or a mixed solvent thereof. The hydrophilic organic solvent that may be contained in the licorice extract is optionally removed by distillation before adsorption onto the adsorbent. The licorice extract from which the hydrophilic organic solvent has been removed is dissolved or suspended in, for example, water, a water-soluble solvent, or a mixed solvent thereof, and then adsorbed onto an adsorbent. After adsorption of the licorice extract onto the adsorbent, it is eluted with water, a water-soluble solvent, or a mixed solvent thereof.

[0044] Examples of hydrophilic organic solvents that can be used for dissolution, suspension, or elution include lower aliphatic alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin. Lower aliphatic alcohols having 1 to 5 carbon atoms are preferred as the dissolution, suspension, or elution solution, and methanol or ethanol are more preferred. When using a mixed solvent of water and a water-soluble solvent, the mixing ratio can be adjusted appropriately. For example, it is preferred to mix 10 parts by volume of water with 1 to 90 parts by volume of the lower aliphatic alcohol.

[0045] The adsorbent is not particularly limited as long as it can adsorb the ketone compounds represented by the above structural formula (1), and known adsorbents such as ion exchange resins, synthetic adsorption resins, activated carbon, chelating resins, silica gel, alumina gel-based adsorbents, and porous glass can be used alone or in combination.However, a porous synthetic adsorbent such as Diaion HP-20 (manufactured by Mitsubishi Chemical Corporation), which is a porous synthetic adsorption resin, is used, and the licorice extract is applied to column chromatography using the porous synthetic adsorbent as the filler, and eluted with an eluent.

[0046] The ketone compound represented by the structural formula (1) can be isolated from the eluate obtained as described above. The ketone compound represented by the structural formula (1) can be isolated from the eluate by, for example, combining various chromatographic treatments such as reverse-phase column chromatography, normal-phase column chromatography, gel permeation chromatography (GPC), and high-performance liquid chromatography (HPLC). The order of the various chromatographic treatments is not particularly limited.

[0047] The developing solvent or mobile phase in various types of chromatography can be water, a water-soluble solvent, a low-polarity solvent, a mixed solvent of these, or a non-polar solvent. The water-soluble solvent can be the hydrophilic solvents described above or acetonitrile, but it is preferable to use a lower alcohol having 1 to 5 carbon atoms or a lower ketone having 1 to 5 carbon atoms, and it is more preferable to use methanol, ethanol, or acetone.

[0048] Examples of packings that can be used in various types of chromatography include octadecylated silica gel (ODS), phenylated silica gel, silica gel, hydroxypropylated dextran, polyvinyl alcohol (PVA) polymers, etc. In reversed-phase column chromatography, it is preferable to use ODS or phenylated silica gel, in normal-phase column chromatography, it is preferable to use silica gel, and in gel permeation chromatography (GPC), it is preferable to use hydroxypropylated dextran or PVA polymers.

[0049] The purified ketone compound can be isolated by fractionating the fraction containing the ketone compound represented by the above structural formula (1) using various types of chromatography.

[0050] The ketone compound represented by the structural formula (1) obtained as described above has a radical scavenging effect, a hyaluronidase activity inhibitory effect, a PGE2 production inhibitory effect, a tyrosinase activity inhibitory effect, an elastase activity inhibitory effect, a type I collagen production promoter effect, and a hair papilla cell proliferation promoter effect, and therefore, by utilizing these effects, it can be used as the active ingredient of a radical scavenger, a hyaluronidase activity inhibitor, a PGE2 production inhibitor, a tyrosinase activity inhibitor, an elastase activity inhibitor, a type I collagen production promoter, and a hair papilla cell proliferation promoter.

[0051] The radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and hair papilla cell proliferation promoter according to the present embodiment can be formulated into any dosage form, such as powder, granules, or liquid, using a pharmaceutically acceptable carrier such as dextrin or cyclodextrin, or any other auxiliary agent, according to standard methods. In this case, examples of auxiliary agents that can be used include excipients, stabilizers, and odor masking agents. Examples of the formulated radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and hair papilla cell proliferation promoter include ointments and topical solutions.

[0052] The radical scavenger of this embodiment can prevent and improve skin aging symptoms such as wrinkle formation and loss of elasticity, as well as pigmentation such as darkening of the skin, age spots, and freckles, through the radical scavenging activity of the ketone compound represented by the structural formula (1) as its active ingredient. Furthermore, the radical scavenger of this embodiment can prevent, treat, or improve various diseases such as tissue disorders such as rheumatoid arthritis and Behçet's disease, various arteriosclerosis diseases (ischemic heart disease, myocardial infarction, cerebral ischemia, etc.), neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, Huntington's disease, etc.), cancer, lung diseases caused by smoking, cataracts, diabetes, stiff shoulders, and sensitivity to cold, through the radical scavenging activity of the ketone compound represented by the structural formula (1) as its active ingredient. However, the radical scavenger of this embodiment can also be used for any other purpose where exerting radical scavenging activity is meaningful. The radical means a molecule or atom having one or more unpaired electrons, and includes superoxide, hydroxyl radical, DPPH (diphenyl-p-picrylhydrazyl), and the like.

[0053] The hyaluronidase activity inhibitor of this embodiment can enhance moisturizing function and prevent, treat, and improve inflammatory diseases (contact dermatitis, psoriasis, pemphigus vulgaris, and various other skin diseases accompanied by rough skin, etc.) through the hyaluronidase activity inhibitory effect of the ketone compound represented by the above structural formula (1), which is the active ingredient. However, in addition to these uses, the hyaluronidase activity inhibitor of this embodiment can also be used for all uses in which exerting a hyaluronidase activity inhibitory effect is significant.

[0054] The PGE2 production inhibitor of this embodiment can prevent, treat, or ameliorate inflammatory diseases (contact dermatitis, psoriasis, pemphigus vulgaris, and various other skin diseases accompanied by rough skin) through the PGE2 production inhibitory effect of the ketone compound represented by the structural formula (1) as an active ingredient. Furthermore, the PGE2 production inhibitor of this embodiment can prevent, treat, or ameliorate skin inflammation, such as redness, erythema, and pigmentation caused by ultraviolet exposure, through the PGE2 production inhibitory effect of the ketone compound represented by the structural formula (1) as an active ingredient. However, the PGE2 production inhibitor of this embodiment can also be used for all other applications where exerting a PGE2 production inhibitory effect is meaningful.

[0055] The tyrosinase activity inhibitor of this embodiment can prevent, treat, or improve skin darkening (cutaneous pigmentation) through the tyrosinase activity inhibitory effect of the ketone compound represented by the structural formula (1) as an active ingredient. However, in addition to these uses, the tyrosinase activity inhibitor of this embodiment can also be used for all uses in which exerting a tyrosinase activity inhibitory effect is significant.

[0056] The elastase activity inhibitor of this embodiment can prevent, treat, or ameliorate skin aging symptoms such as loss of firmness and reduced elasticity through the elastase activity inhibitory effect of the ketone compound represented by the structural formula (1) as an active ingredient. Furthermore, the elastase activity inhibitor of this embodiment can prevent, treat, or ameliorate respiratory diseases such as emphysema and pulmonary edema through the elastase activity inhibitory effect of the ketone compound represented by the structural formula (1) as an active ingredient. However, the elastase activity inhibitor of this embodiment can also be used for all other applications in which exerting an elastase activity inhibitory effect is significant.

[0057] The type I collagen production promoter according to this embodiment can prevent, treat, or improve skin aging symptoms such as wrinkle formation, loss of firmness, and decreased elasticity through the type I collagen production-promoting activity of the ketone compound represented by the structural formula (1) as an active ingredient. However, in addition to these uses, the type I collagen production promoter according to this embodiment can also be used for all uses in which exerting the type I collagen production-promoting activity is meaningful.

[0058] The dermal papilla cell proliferation promoter according to this embodiment can activate dermal papilla cells and promote the proliferation and differentiation of hair follicle epithelial cells and hair formation through the dermal papilla cell proliferation-promoting activity of the ketone compound represented by structural formula (1) as the active ingredient. This can promote hair growth and / or hair care, and can prevent and improve various types of alopecia (e.g., androgenetic alopecia, alopecia areata, etc.). However, in addition to these uses, the dermal papilla cell proliferation promoter according to this embodiment can be used for all uses in which it is meaningful to exert the dermal papilla cell proliferation-promoting activity.

[0059] Methods of administering the radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and dermal papilla cell proliferation promoter according to the present embodiments to a patient include subcutaneous administration, intramuscular administration, intravenous administration, oral administration, transdermal administration, etc., and a method suitable for the prevention, treatment, etc. of the disease may be appropriately selected depending on the type of disease. Furthermore, the dosage of the radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and dermal papilla cell proliferation promoter according to the present embodiments may be increased or decreased as appropriate depending on the type and severity of the disease, individual patient differences, administration method, administration period, etc.

[0060] Furthermore, the radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and hair papilla cell proliferation promoter according to the present embodiment have excellent radical scavenging activity, hyaluronidase activity inhibitory activity, PGE2 production inhibitory activity, tyrosinase activity inhibitory activity, elastase activity inhibitory activity, type I collagen production promoting activity, and hair papilla cell proliferation promoting activity, and are therefore suitable for incorporation into cosmetics such as scalp cosmetics and hair cosmetics.

[0061] Cosmetics that can be formulated with a radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and dermal papilla cell proliferation promoter, each containing a ketone compound represented by structural formula (1) as an active ingredient, include, for example, hair tonic, hair lotion, hair cream, hair liquid, shampoo, rinse, pomade, etc. When the radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and dermal papilla cell proliferation promoter are formulated in a cosmetic, the amount blended can be adjusted appropriately depending on the type of cosmetic, but a preferred blending ratio is about 0.0001 to 10% by mass, calculated as a standard extract, and a particularly preferred blending ratio is about 0.001 to 1% by mass, calculated as a standard extract. Cosmetics can be used in combination with main ingredients, auxiliaries, and other ingredients typically used in the manufacture of cosmetics, such as astringents, germicides / antibacterial agents, whitening agents, ultraviolet absorbers, moisturizers, cell activators, anti-inflammatory / antiallergic agents, antioxidants / active oxygen scavengers, oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, and fragrances, as long as they do not interfere with the radical scavenging activity, hyaluronidase activity inhibitory activity, PGE2 production inhibitory activity, tyrosinase activity inhibitory activity, elastase activity inhibitory activity, type I collagen production promotion activity, and hair papilla cell proliferation promotion activity of the ketone compound represented by structural formula (1) above. Such combinations can result in more versatile products, and the synergistic effect with the other active ingredients used in combination can sometimes bring about effects superior to those normally expected.

[0062] The radical scavenger, hyaluronidase activity inhibitor, PGE2 production inhibitor, tyrosinase activity inhibitor, elastase activity inhibitor, type I collagen production promoter, and hair papilla cell proliferation promoter according to the present embodiment are preferably applied to humans, but can also be applied to animals other than humans as long as their respective action effects are exerted. [Example]

[0063] The present invention will be explained in more detail below by showing production examples, test examples, etc., but the present invention is not limited to the following production examples, test examples, etc.

[0064] [Production Example 1] Production of licorice extract 1.0 kg of crushed licorice root was added to 2.5 L of chloroform, and reflux extraction was carried out at 60°C for 30 minutes. The extract was then filtered while hot to obtain an extract. 2.5 L of chloroform was then added to the extraction residue, and the same procedure was carried out to obtain an extract. The resulting extracts were combined, concentrated under reduced pressure, and then dried under reduced pressure to obtain a licorice extract (14 g).

[0065] [Production Example 2] Production of novel ketone compounds 14 g of the licorice extract obtained in Production Example 1 was applied to a porous synthetic adsorption resin (product name: Diaion HP-20, manufactured by Mitsubishi Chemical Corporation), and distilled water (0.5 L), 80% methanol (1.5 L, v / v), methanol (1.5 L), and acetone (1.5 L) were passed through the resin in this order, yielding a water eluate (233.7 mg), an 80% methanol eluate (1.6 g), a methanol eluate (6.0 g), and an acetone eluate (5.1 g). Of this, the methanol eluate (6.0 g) was fractionated by column chromatography (mobile phase: n-hexane:ethyl acetate = 2:1 → 1:1 → methanol) using silica gel (product name: Silica gel 60, manufactured by Merck Millipore) as the packing material to obtain Fraction 1 (306 mg), Fraction 2 (1.0 g), Fraction 3 (1.6 g), Fraction 4 (1.0 g), Fraction 5 (1.0 g), and Fraction 6 (1.2 g).

[0066] Fraction 2 (1.0 g) was purified by column chromatography (mobile phase: methanol:water=60:40) using ODS (product name: CHROMATREX ODS-DM1020T, manufactured by Fuji Silysia Chemical Ltd.) as the packing material, followed by recycling preparative HPLC (stationary phase: JAIGEL-GS310 × 2 (manufactured by Japan Analytical Industry Co., Ltd.), mobile phase: methanol) to obtain a purified product (5 mg) (Sample 1).

[0067] Column chromatography was performed on Fr. 3 (1.6 g) using ODS (product name: CHROMATREX ODS-DM1020T, manufactured by Fuji Silysia Chemical Ltd.) as the packing material (mobile phase: methanol:water = 60:40 → methanol), yielding Fr. 3-1 (167 mg), Fr. 3-2 (316 mg), Fr. 3-3 (227 mg), Fr. 3-4 (76 mg), Fr. 3-5 (63 mg), Fr. 3-6 (217 mg), and Fr. 3-7 (514 mg).

[0068] Fr.3-1 (167 mg) was purified by preparative HPLC (stationary phase: YMC-Actus Triart C18 (YMC), mobile phase: acetonitrile:water = 50:50), followed by recycling preparative HPLC (stationary phase: JAIGEL-GS310 x 2 (Japan Analytical Industry Co., Ltd.), mobile phase: methanol) to obtain a purified product (3 mg) (sample 2).

[0069] Mass spectrometry of the obtained test samples (sample 1 and sample 2); 1 H-NMR analysis and 13 The structure was determined by C-NMR analysis.

[0070] <Mass spectrum> HR negative-ion ESI-MS:calculated for C 19 H 17 O5: m / z 325.1076 [MH] - ,found:325.1059

[0071] < 1 H-NMR (400 MHz, CDCl3) δ H > 1.38(3H,s,11′-H,12′-H),4.04(2H,s,2-H),5.57(1H,d,J=10.4,3′-H),6.33(1H,d,J=8.0,8′-H),6.35(1H,d,J=2 .0,3″-H),6.43(1H,d,J-9.2,5″-H),6.72(1H,d,J=9.6,4′-H),6.86(1H,d,J=8.4,7′-H),7.85(1H,d,J=7.2,6″-H)

[0072] < 13 C-NMR (100 MHz, CDCl3) δ C > 27.8(C-11′,C-12′),40.1(C-2),76.0(C-2′),103.9(C-3″),108.5(C-5″),109.0(C-8′),111.5(C-10′),112.7(C-6′,C-1″), 117.0(C-4′),129.3(C-3′),130.1(C-7′),133.5(C-6″),151.7(C-5′),153.5(C-9′),164.0(C-4″),166.6(C-2″),204.3(C-1)

[0073] From the above results, it was confirmed that the compound isolated from the licorice extract was 1-(2,4-dihydroxyphenyl)-2-(5-hydroxy-2,2-dimethyl-2H-chromen-6-yl)ethanone represented by the following structural formula (1).

[0074] [ka]

[0075] [Test Example 1] DPPH radical scavenging activity test 3 mL of the test sample solution obtained in the above manufacturing example was added to 3 mL of 150 μmol / L DPPH (diphenyl-p-picrylhydrazyl) ethanol solution, and the container was then sealed, shaken, and left to stand for 30 minutes. After standing, the absorbance at a wavelength of 520 nm was measured. As a blank, 3 mL of the test sample solution was added to 3 mL of ethanol, and the absorbance at a wavelength of 520 nm was immediately measured. As a control, a DPPH ethanol solution without the test sample was also measured in the same way. The DPPH radical scavenging rate (%) was calculated from the obtained measurement results using the following formula. The results are shown in Table 1.

[0076] DPPH radical scavenging rate (%) = {A-(BC)} / A x 100 In the formula, "A" represents the absorbance of the control, "B" represents the absorbance of the solution to which the test sample was added, and "C" represents the absorbance of the blank.

[0077] [Table 1]

[0078] As shown in Table 1, the ketone compound represented by the above structural formula (1) exhibited a high DPPH radical scavenging rate. This result confirmed that the ketone compound represented by the above structural formula (1) has excellent radical scavenging activity.

[0079] [Test Example 2] Hyaluronidase activity inhibitory effect test The test sample obtained in the above Preparation Example was dissolved in 0.2 mL of 0.1 mol / L acetate buffer (pH 3.5), and 0.1 mL of hyaluronidase solution (Type IV-S (from bovine testis), 400 NF units / mL, manufactured by SIGMA) was added and allowed to react for 20 minutes at 37°C. 0.2 mL of 2.5 mmol / L calcium chloride solution was then added as an activator, and the reaction was allowed to continue for 20 minutes at 37°C. 0.5 mL of 0.8 mg / mL sodium hyaluronate solution (from rooster comb) was then added, and the reaction was allowed to continue for 40 minutes at 37°C. 0.2 mL of 0.4 mol / L sodium hydroxide solution was then added to stop the reaction. After cooling, 0.2 mL of boric acid solution was added to the reaction solution, and the mixture was boiled for 3 minutes. After cooling on ice, 6 mL of p-DABA reagent was added, and the reaction was allowed to continue for 20 minutes at 37°C. The absorbance at a wavelength of 585 nm was then measured.

[0080] Similar measurements were also performed using a control solution containing distilled water instead of the test sample solution. Furthermore, similar measurements were performed using test sample solutions and control solutions without the addition of hyaluronidase solution to create blanks for the test sample solution and the control solution. The hyaluronidase activity inhibition rate (%) was calculated from the obtained measurement results using the following formula. The results are shown in Table 2.

[0081] Hyaluronidase activity inhibition rate (%) = {1-(DE) / (FG)} x 100 In the formula, "D" represents the absorbance of the test sample solution, "E" represents the absorbance of the test sample solution blank, "F" represents the absorbance of the control solution, and "G" represents the absorbance of the control solution blank.

[0082] [Table 2]

[0083] As shown in Table 2, the ketone compound represented by the above structural formula (1) exhibited a high hyaluronidase activity inhibitory rate. These results confirmed that the ketone compound represented by the above structural formula (1) has excellent hyaluronidase activity inhibitory activity.

[0084] [Test Example 3] PGE2 production inhibitory effect test Normal human neonatal epidermal keratinocytes (NHEK) were cultured in normal human epidermal keratinocyte growth medium (KGM) and then harvested by trypsinization. The harvested cells were diluted to 1.0 × 10 5 After diluting the cells to a density of 2.0 × 10 cells / mL, 200 μL of the cells were seeded per well on a collagen-coated 48-well plate (2.0 × 10 cells / mL). 4 After confirming that the cells had settled, the medium was replaced with 200 μL of basal medium (KBM) and the cells were cultured for an additional 24 hours.

[0085] After the incubation, the medium was removed, and 200 μL of 500 μmol / L aspirin-containing KBM was added to inactivate the existing COX-1 and the small amount of COX-2 expressed by acetylation. After 4 hours, the cells were washed three times with PBS(-) buffer, added with 100 μL of PBS(-) buffer, and exposed to UVB irradiation (60 mJ / cm). 2) was performed, and then 400 μL of the test sample obtained in the above Preparation Example, dissolved in KBM to the required concentration, was added to each well and cultured at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample was measured in the same manner.

[0086] After the incubation, the amount of prostaglandin E2 (PGE2) in the culture supernatant of each well was quantified using a PGE2 EIA Kit (Cayman Chemical). The PGE2 production inhibition rate (%) was calculated from the quantification results using the following formula. The results are shown in Table 3.

[0087] Hyaluronidase activity inhibition rate (%) = {1-(HJ) / (IJ)} x 100 In the formula, "H" represents the amount of PGE2 in cells with the test sample added when irradiated with UV rays, "I" represents the amount of PGE2 in cells without the test sample added when irradiated with UV rays, and "J" represents the amount of PGE2 in cells without the test sample added when not irradiated with UV rays.

[0088] [Table 3]

[0089] As shown in Table 3, the ketone compound represented by the above structural formula (1) exhibited a high PGE2 production inhibitory rate. From these results, it was confirmed that the ketone compound represented by the above structural formula (1) has an excellent PGE2 production inhibitory effect.

[0090] [Test Example 4] Tyrosinase activity inhibitory effect test To 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 a 25% DMSO solution containing the test sample obtained in the above Preparation Example were added and allowed to stand at 37°C for 10 minutes. 0.02 mL of an 800 unit / mL tyrosinase solution was added, followed by a 15-minute reaction at 37°C. After completion of the reaction, the absorbance at 475 nm was measured.

[0091] Similar measurements were also performed on a control solution containing distilled water instead of the test sample solution. Furthermore, similar measurements were performed on test sample solutions and control solutions without the addition of hyaluronidase solution to create blanks for the test sample solution and the control solution. The tyrosinase activity inhibition rate (%) was calculated from the obtained measurement results using the following formula. The results are shown in Table 4.

[0092] Tyrosinase activity inhibition rate (%) = {1-(KL) / (MN)} x 100 In the formula, "K" represents the absorbance of the test sample solution, "L" represents the absorbance of the test sample solution blank, "M" represents the absorbance of the control solution, and "N" represents the absorbance of the control solution blank.

[0093] [Table 4]

[0094] As shown in Table 4, the ketone compound represented by the above structural formula (1) exhibited a high tyrosinase activity inhibitory rate. From these results, it was confirmed that the ketone compound represented by the above structural formula (1) has an excellent tyrosinase activity inhibitory effect.

[0095] [Test Example 5] Elastase activity inhibitory effect test 50 μL of test sample solution prepared in 0.1 mol / L HEPES buffer (pH 7.5) and 25 μL of 6 μg / mL elastase (Human Leukocyte) solution were mixed in a 96-well plate. 25 μL of N-METHOXYSUCCINYL-ALA-ALA-PRO-VAL-p-NITRO-ANILIDE solution prepared in the above buffer to a concentration of 2 mmol / L was then added and reacted at 25°C for 15 minutes. After the reaction was completed, the absorbance at 415 nm was measured.

[0096] As a control solution, an elastase solution without the addition of the test sample solution was subjected to the same measurement. Furthermore, as a blank for the test sample solution and the control solution, measurements were also performed in the same manner for the test sample solution and the control solution without adding any elastase solution. The elastase activity inhibition rate (%) was calculated from the obtained measurement results using the following formula. The results are shown in Table 5.

[0097] Elastase activity inhibition rate (%) = {1-(OP) / (QR)} x 100 In the formula, "O" represents the absorbance of the test sample solution, "P" represents the absorbance of the test sample solution blank, "Q" represents the absorbance of the control solution, and "R" represents the absorbance of the control solution blank.

[0098] [Table 5]

[0099] As shown in Table 5, the ketone compound represented by the above structural formula (1) exhibited a high elastase activity inhibitory rate. From these results, it was confirmed that the ketone compound represented by the above structural formula (1) has an excellent elastase activity inhibitory effect.

[0100] [Test Example 6] Type I collagen production promotion test Normal human skin fibroblasts (NB1RGB) were cultured in Dulbecco's MEM medium containing 10% FBS, and then the cells were harvested by trypsinization. The harvested cells were diluted to 1.6 × 10 in Dulbecco's MEM medium containing 0.25% FBS. 5 After dilution to a cell density of 100 cells / mL, 100 μL of the solution was seeded into each well of a 96-well microplate and cultured overnight.

[0101] After incubation, the medium was removed, and 100 μL of the test sample dissolved in 0.25% FBS-containing Dulbecco's MEM medium was added to each well and incubated for 3 days. After incubation, the amount of type I collagen in the medium in each well was measured by ELISA. As a control, Dulbecco's MEM medium containing FBS without the test sample was also measured in the same manner.

[0102] The type I collagen production promotion rate (%) was calculated from the measurement results using the following formula. The results are shown in Table 6.

[0103] Type I collagen production promotion rate (%) = S / T x 100 In the formula, "S" represents the amount of type I collagen when the test sample was added, and "T" represents the amount of type I collagen when the test sample was not added.

[0104] [Table 6]

[0105] As shown in Table 6, the ketone compound represented by the structural formula (1) exhibited a high rate of promoting type I collagen production. This result confirmed that the ketone compound represented by the structural formula (1) has an excellent effect of promoting type I collagen production.

[0106] [Test Example 7] Test of dermal papilla cell proliferation promoting effect Normal human hair dermal papilla cells were cultured in a dermal papilla cell growth medium containing 1% FCS and growth additives, and then the cells were harvested by trypsinization. The harvested cells were then diluted to 1.0 × 10 in DMEM (Dulbecco's modified minimal essential medium) containing 10% FBS. 4 After dilution to a cell density of 100 cells / mL, 200 μL of the solution was seeded per well on a collagen-coated 96-well plate and cultured for 3 days.

[0107] After incubation, the medium was removed, and 200 μL of the test sample dissolved in serum-free DMEM was added to each well, followed by incubation for an additional 4 days. As a control, serum-free DMEM without the sample was incubated in the same manner.

[0108] The proliferation of dermal papilla cells was measured using an MTT assay. Specifically, after the completion of the culture, the medium was removed, and 100 μL of MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium Bromide) dissolved in serum-free DMEM to a final concentration of 0.4 mg / mL was added to each well. After 2 hours of culture, the blue formazan produced within the cells was extracted with 100 μL of 2-propanol. After extraction, the absorbance at a wavelength of 570 nm was measured. At the same time, the absorbance at a wavelength of 650 nm was measured as turbidity, and the difference between the two was used as the amount of blue formazan produced. The proliferation promotion rate of dermal papilla cells was then calculated according to the following formula.

[0109] Dermal papilla cell proliferation promotion rate (%) = (U / V) x 100 In the formula, "U" represents the amount of blue formazan produced when a test sample is added, and "V" represents the amount of blue formazan produced when no test sample is added.

[0110] [Table 7]

[0111] As shown in Table 7, the ketone compound represented by the above structural formula (1) exhibited a high rate of promoting proliferation of dermal papilla cells. These results confirmed that the ketone compound represented by the above structural formula (1) has an excellent effect of promoting proliferation of dermal papilla cells. [Industrial Applicability]

[0112] The novel ketone compounds, radical scavengers, hyaluronidase activity inhibitors, PGE2 production suppressors, tyrosinase activity inhibitors, elastase activity inhibitors, type I collagen production promoters, and hair papilla cell proliferation promoters of the present invention can be suitably used as components of cosmetics and the like, and further as research reagents.

Claims

1. A ketone compound represented by the following structural formula (1): 【Chemistry 1】

2. A radical scavenger containing the ketone compound according to claim 1 as an active ingredient.

3. A hyaluronidase activity inhibitor containing the ketone compound of claim 1 as an active ingredient.

4. A PGE containing the ketone compound according to claim 1 as an active ingredient. ２ Production inhibitor.

5. A tyrosinase activity inhibitor containing the ketone compound according to claim 1 as an active ingredient.

6. An elastase activity inhibitor comprising the ketone compound according to claim 1 as an active ingredient.

7. A type I collagen production promoter comprising the ketone compound according to claim 1 as an active ingredient.

8. A hair papilla cell proliferation promoter comprising the ketone compound according to claim 1 as an active ingredient.

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