Prostaglandin d2 production inhibitors

Abstract

To provide a highly safe prostaglandin D2 production suppressing agent having excellent prostaglandin D2 production suppressing action for suppressing the production of prostaglandin D2 having hair growth inhibiting action and fibroblast growth factor-5 production promoting action.SOLUTION: The prostaglandin (PG) D2 production suppressing agent contains at least one selected from the group consisting of rutin, quercetin, isoquercetin and kaempferol.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an inhibitor of prostaglandin (PG) D2 (hereinafter sometimes referred to as "PGD2") production. [Background technology]

[0002] A healthy hair cycle consists of a growth phase (2-7 years), a regression phase (2-3 weeks), and a resting phase (2-3 months), with hair growth and hair loss occurring repeatedly. In patients with male pattern baldness (AGA), male hormones shorten the hair cycle, causing hair follicles to transition to the regression phase without sufficient growth, leading to miniaturization of hair follicles (see Non-Patent Document 1).

[0003] A comparative study of hair loss areas and non-hair loss areas of AGA patients revealed that prostaglandin (PG) D2 is produced in large amounts in hair loss tissue, and PGD2 has also been reported as a hair growth inhibitor involved in the onset of AGA (see Non-Patent Document 2). This report also reports that the expression of Ptgds, a PGD2 synthesizing enzyme, increases in the late hair growth phase, and that Fgf5, a factor involved in the regression phase, increases in dermal papilla cells at the same time as the transition to the regression phase.

[0004] Fibroblast growth factors (FGFs) are multifunctional secretory factors that not only have proliferation activity for fibroblasts but also play important roles as morphogenetic factors with cell proliferation and differentiation activity for various cells, tissue repair factors that act upon tissue injury, metabolic regulators that maintain homeostasis, etc. More than 20 types of FGFs are known to exist in the FGF family.

[0005] As mentioned above, hair grows and falls out repeatedly according to a cyclical hair cycle (hair growth period) consisting of an anagen phase, a catagen phase, and a telogen phase. If the anagen phase is short, the hair does not grow sufficiently and falls out while still thin and short. Furthermore, once the anagen phase is shortened, the next anagen phase also becomes shorter, and it is thought that the hair becomes even thinner and shorter. Among the above-mentioned FGFs, FGF-5 is known to be involved in the hair cycle, and it has been reported that it acts on the hair papilla to cause a transition from the anagen phase to the catagen phase in the hair cycle (see Non-Patent Document 3).

[0006] Therefore, if the function of FGF-5 can be suppressed, the transition from the growth phase to the regression phase in the hair cycle can be inhibited, thereby extending the growth phase of hair and preventing or improving thinning hair and hair loss, making it useful for hair growth and care.

[0007] Sebum components such as triglycerides, wax esters, squalene, and free fatty acids are present on the scalp and are considered necessary for protecting the scalp from bacteria and dryness. Squalene, in particular, has been reported to become squalene peroxide when oxidized by UV rays and other factors, and to act on epidermal cells to increase the expression of the inflammatory cytokine IL-1 and the prostanoid converting enzyme cyclooxygenase-2 (see Non-Patent Documents 4 and 5), as well as to decrease the expression of the KAP5.1 gene, which is involved in hair strength and body (see Non-Patent Document 4). Furthermore, it has been reported that the amount of squalene peroxide significantly increases in dandruff, a scalp problem (see Non-Patent Document 6), and that inflammation occurs in unhealthy scalps (see Non-Patent Document 2).

[0008] Therefore, there is a strong demand for a new material that is a highly safe, naturally-derived component that can suppress the production of PGD2, which has the effect of inhibiting hair growth and promoting FGF-5 production, and that can therefore be widely used as an ingredient in pharmaceuticals, quasi-drugs, external compositions such as cosmetics, foods and beverages, and research reagents, and there is currently a demand for its rapid development. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Maeda Noritoshi et al., The Frontline of Hair Regeneration, CMC Publishing, 2020 [Non-patent document 2] Sci Transl Med. 2012 March 21;4(126):126ra34 [Non-patent document 3] Cell,1994,vol.78,no.6,pp.1017-1025 [Non-patent document 4] J. Soc. Cosmet. Chem. Jpn. Vol.52, No.1 2018 [Non-patent document 5] Lipids (2010) 45:833-841 [Non-patent document 6] Arch Dermatol Res (2016) 308:153-163 Summary of the Invention [Problem to be solved by the invention]

[0010] The present invention aims to solve the above-mentioned conventional problems and achieve the following object: That is, the present invention aims to provide a prostaglandin D2 production inhibitor that has an excellent prostaglandin D2 production inhibitory effect, suppressing the production of prostaglandin D2, which has the effect of inhibiting hair growth and promoting fibroblast growth factor-5 production, and is highly safe. [Means for solving the problem]

[0011] As a result of extensive research, the present inventors discovered that at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol, which are highly safe natural materials, has an excellent inhibitory effect on prostaglandin D2 production, and thus completed the present invention.

[0012] The present invention is based on the above findings of the present inventors, and the means for solving the above problems are as follows: <1> The prostaglandin (PG) D2 production inhibitor is characterized by containing at least one member selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol. [Effects of the Invention]

[0013] According to the present invention, the above-mentioned problems of the prior art can be solved, the above-mentioned objects can be achieved, and a prostaglandin D2 production inhibitor having excellent prostaglandin D2 production inhibitory activity, suppressing the production of prostaglandin D2, which has the effect of inhibiting hair growth and promoting the production of fibroblast growth factor-5, and which is highly safe can be provided. DETAILED DESCRIPTION OF THE INVENTION

[0014] (Prostaglandin (PG) D2 production inhibitor) The prostaglandin (PG) D2 production inhibitor according to this embodiment contains at least one active ingredient selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol, and further contains other ingredients as necessary.

[0015] As used herein, inhibiting the production of prostaglandin D2 means reducing the production of PGD2 compared to when the PGD2 production inhibitor is not administered.

[0016] It was not previously known that rutin, quercetin, isoquercetin, and kaempferol have excellent PGD2 production inhibitory effects and are useful as PGD2 production inhibitors, and this is a new discovery by the present inventors.

[0017] <At least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol> -Rutin- Rutin (CAS Registry Number: 153-18-4) is a compound represented by the following structural formula (1). [ka]

[0018] -Quercetin- Quercetin (CAS Registry Number: 117-39-5) is a compound represented by the following structural formula (2). [ka]

[0019] -Isoquercetin- Isoquercetin (CAS Registry Number: 482-35-9) is a compound represented by the following structural formula (3). [ka]

[0020] -Kaempferol- Kaempferol (CAS Registry Number: 520-18-3) is a compound represented by the following structural formula (4). [ka]

[0021] The rutin, quercetin, isoquercetin, and kaempferol are known compounds, and commercially available products may be used, or products produced by known methods may be used.

[0022] The rutin, quercetin, isoquercetin, and kaempferol are contained in plants such as licorice, and can also be prepared from plants by extraction methods generally used for plant extraction.

[0023] The rutin, quercetin, isoquercetin, and kaempferol may be used singly or in combination of two or more.

[0024] The total content of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol in the PGD2 production inhibitor is not particularly limited and can be appropriately selected depending on the purpose. Furthermore, when two or more of the rutin, quercetin, isoquercetin, and kaempferol are used in combination, the ratio of the amounts of each component to be blended is not particularly limited and can be appropriately selected depending on the purpose.

[0025] The PGD2 production inhibitor may consist of only at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol, or may be a formulation of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol.

[0026] At least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol can be formulated into any dosage form, such as powder, granules, tablets, or liquid, using a pharmaceutically acceptable carrier such as dextrin or cyclodextrin, or any other auxiliary agent, according to a conventional method. In this case, examples of the auxiliary agent that can be used include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, and the like. The PGD2 production inhibitor can be incorporated into other compositions (e.g., topical preparations such as topical preparations for hair and skin, or food and beverages), and can also be used as oral preparations such as tablets, powders, capsules, granules, extracts, and syrups; parenteral preparations such as injections, drips, and suppositories; ointments, eye drops, topical liquids, and patches.

[0027] The total content of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol in the formulated PGD2 production inhibitor is not particularly limited and can be selected appropriately depending on the purpose.

[0028] <Other ingredients> The other components are not particularly limited as long as they do not impair the effects of the present invention and can be appropriately selected depending on the form of use of the PGD2 production inhibitor, and examples include the components that can be used when formulating the above-mentioned formulations. Furthermore, if necessary, a component having a PGD2 production inhibitory effect other than at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol may be blended. The other components may be used alone or in combination of two or more.

[0029] The content of the other components in the PGD2 production inhibitor is not particularly limited and can be appropriately selected depending on the purpose.

[0030] The uses of the PGD2 production inhibitor are not particularly limited and can be selected appropriately depending on the purpose, and can be used for a wide range of purposes, such as pharmaceuticals, quasi-drugs, cosmetics, food and beverages, and research reagents.

[0031] The method of use of the PGD2 production inhibitor is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include oral, parenteral, and external use.

[0032] The dosage form of the PGD2 production inhibitor is not particularly limited, and any known dosage form can be appropriately selected depending on the purpose. The method for producing the PGD2 production inhibitor in any dosage form is not particularly limited, and any known method can be appropriately selected.

[0033] There are no particular restrictions on the administration method, dosage, administration site, administration period, and administration interval (hereinafter sometimes referred to as "usage method, dosage, administration site, administration period, and administration interval") of the PGD2 production inhibitor, and these can be selected appropriately depending on the purpose.

[0034] The PGD2 production inhibitor can inhibit PGD2 production through the PGD2 production inhibitory activity of rutin, quercetin, isoquercetin, and kaempferol. This inhibits the hair growth inhibitory activity of PGD2, thereby preventing and ameliorating thinning hair and hair loss. It can also inhibit the function of FGF-5, inhibiting the transition from the anagen phase to the catagen phase in the hair cycle, thereby extending the anagen phase and preventing and ameliorating thinning hair and hair loss. However, the PGD2 production inhibitor of this embodiment can also be used for all other applications where exerting a PGD2 production inhibitory activity is meaningful.

[0035] The PGD2 production inhibitor has an excellent PGD2 production inhibitory effect and is highly safe, and therefore can be incorporated into external preparations such as external preparations for hair and skin, or into foods and beverages. In this case, at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol may be incorporated as is, or a formulation of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol may be incorporated.

[0036] The topical preparations are not limited to any particular category and include a wide range of cosmetics, quasi-drugs, pharmaceuticals, etc. that are used transdermally.

[0037] There are no particular limitations on the topical preparations that can be formulated with the PGD2 production inhibitor, and they can be selected appropriately depending on the purpose. Examples include topical hair and skin preparations such as hair tonic, hair lotion, hair cream, hair liquid, pomade, shampoo, rinse, and treatment, as well as ointments, creams, emulsions, lotions, skin lotions, beauty serums, gels, beauty oils, packs, jellies, lip balms, lipsticks, foundations, bath additives, soaps, body soaps, and astringents.

[0038] When the PGD2 production inhibitor is incorporated into an external preparation, the amount to be incorporated is not particularly limited and can be adjusted as appropriate depending on the type of external preparation, etc. However, the total amount of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol is preferably 0.0001% by mass to 20% by mass, and more preferably 0.0001% by mass to 10% by mass.

[0039] The topical preparation may contain, in addition to the main ingredient, auxiliary agent, or other ingredient typically used in the manufacture of topical preparations, such as astringents, bactericides / 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, fragrances, etc., so long as the effects of the present invention are not impaired. Such combinations may result in more general-purpose products, and the synergistic effect with the other active ingredients used in combination may bring about effects superior to those normally expected.

[0040] The method for producing the topical agent is not particularly limited, and any known method can be appropriately selected.

[0041] The administration method, dosage, administration site, administration period, administration interval, etc. of the topical preparation are not particularly limited and can be appropriately selected depending on the purpose.

[0042] The topical preparation can be suitably used for inhibiting PGD2 production, and for hair growth based on the PGD2 production inhibitory effect.

[0043] Food and beverage products refer to products that are unlikely to be harmful to human health and that are taken orally or by administration through the digestive tract in normal social life, and are not limited to administrative classifications such as food, medicine, or quasi-drug. Therefore, in this embodiment, "food and beverage products" broadly includes general foods taken orally, health foods (functional food and beverage products), health functional foods (foods for specified health uses, foods with nutrient functions), quasi-drugs, medicines, etc. Preferably, the food and beverage products are those that can display the favorable effects of the active ingredient of this embodiment on the food and beverage product or its packaging, and are particularly preferably health functional foods (foods for specified health uses, foods with functional claims, foods with nutrient functions), quasi-drugs, and medicines.

[0044] When the PGD2 production inhibitor is incorporated into food and beverages, the amount to be incorporated is not particularly limited and can be adjusted appropriately depending on the type of food and beverage. However, the total amount of at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol is preferably 0.0001% by mass to 20% by mass, and more preferably 0.0001% by mass to 10% by mass.

[0045] The method for producing the food or drink is not particularly limited, and any known method can be appropriately selected.

[0046] The method, amount, period, and interval of ingestion of the food and drink are not particularly limited and can be appropriately selected depending on the purpose.

[0047] The food and drink products can be suitably used for inhibiting PGD2 production, and for hair growth based on the PGD2 production inhibitory effect.

[0048] Although the PGD2 production inhibitor of this embodiment is suitable for use in humans, it can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as it has the effect of inhibiting PGD2 production.

[0049] Furthermore, the PGD2 production inhibitor according to this embodiment can also be suitably used as a reagent for research into the mechanism of PGD2 production inhibitory action.

[0050] As described above, the PGD2 production inhibitor of this embodiment has an excellent PGD2 production inhibitory effect. Therefore, the present invention also relates to a method for inhibiting PGD2 production, which comprises administering the PGD2 production inhibitor to an individual. [Example]

[0051] Test examples and formulation examples of the present invention will be explained below, but the present invention is not limited to these test examples and formulation examples.

[0052] (Test Example 1: Prostaglandin (PG) D2 production promoting effect test) The PGD2 production promoting effect of squalene peroxide was tested by the following method.

[0053] 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 collected at a concentration of 7.5 × 10 4 After diluting with KGM to a concentration of 1.5 × 10 cells / mL, 200 μL of the solution was seeded per well of a collagen-coated 48-well plate (1.5 × 10 cells / mL). 4 After confirming that the cells had settled, the cells were cultured in 200 μL of basal medium (KBM) for 24 hours. After incubation, the supernatant was removed, and a squalene peroxide solution (prepared by irradiating an ethanol solution containing 100 mmol / L squalene with UVA light (see Table 1 for the amount of UVA light) was dissolved in KBM. 400 μL of this solution was added to each well, and the cells were incubated at 37°C under 5% CO for 24 hours. As a control, KBM without squalene peroxide was incubated in the same manner. After the culture was completed, the amount of PGD2 in the culture supernatant of each well was quantified using a PGD2 ELISA Kit (Cayman Chemical). From the obtained results, the PGD2 production promotion rate was calculated using the following formula. The results are shown in Table 1 below. In the following formula, the PGD2 production promotion rate when squalene peroxide is not added is 100%. PGD2 production promotion rate (%)=A / B×100 In the above formula, A and B respectively represent the following. A: PGD2 production amount when squalene peroxide was added B: PGD2 production without squalene peroxide

[0054] [Table 1]

[0055] As shown in Table 1, it was confirmed that squalene peroxide has the effect of promoting PGD2 production.

[0056] (Test Example 2: PGD2 production inhibitory effect test) The following compounds were used as test samples and tested for their inhibitory effect on PGD2 production by the following test method.

[0057] <Test sample> (1) Rutin (Fujifilm Wako Pure Chemical Industries, Ltd.) (2) Quercetin (Fujifilm Wako Pure Chemical Industries, Ltd.) (3) Isoquercetin (Fujifilm Wako Pure Chemical Industries, Ltd.) (4) Kaempferol (Fujifilm Wako Pure Chemical Industries, Ltd.)

[0058] <Test Method> The inhibitory effect of squalene peroxide on PGD2 production was tested as follows. Normal human neonatal epidermal keratinocytes or normal human adult epidermal keratinocytes were cultured in normal human epidermal keratinocyte growth medium and then harvested by trypsinization. The harvested cells were collected at a concentration of 7.5 × 10 4 After diluting with KGM to a concentration of 1.5 × 10 cells / mL, 200 μL of the solution was seeded per well of a collagen-coated 48-well plate (1.5 × 10 cells / mL).4 After confirming that the cells had settled, the cells were cultured in 200 μL of basal medium (KBM) for 24 hours. After the incubation, the supernatant was removed and the cells were placed in an ethanol solution containing 100 mmol / L squalene and exposed to UVA irradiation (60 J / cm 2 The malondialdehyde content in the squalene peroxide solution prepared by the above procedure was kept constant for each test run, and the test sample was dissolved in KBM to the required concentration (see Tables 2 and 3 below for the final test sample concentrations). 200 to 400 μL was added to each well and cultured at 37°C under 5% CO2 for 24 hours. As a control, KBM without the test sample or squalene peroxide was used and cultured in the same manner. After incubation, the amount of PGD2 in the culture supernatant was quantified using a PGD2 ELISA Kit (Cayman Chemical Co.). At the same time, the number of cells in each well was counted, and the amount of extracellular PGD2 produced per cell was calculated by dividing the count by the number of cells. From the obtained results, the PGD2 production inhibition rate was calculated using the following formula. The results are shown in Tables 2 and 3. In the formula below, the PGD2 production inhibition rate without the addition of squalene peroxide and without the addition of the test sample is 100%, and the PGD2 production inhibition rate with the addition of squalene peroxide and without the addition of the test sample is 0%. PGD2 production suppression rate (%)={1-(CE) / (DE)}×100 In the above formula, C to E respectively represent the following. C: PGD2 production per cell after addition of test sample and squalene peroxide D: PGD2 production per cell without test sample and with squalene peroxide added E: PGD2 production per cell without the addition of test sample or squalene peroxide

[0059] The results of the test using normal human neonatal epidermal keratinocytes are shown in Table 2, and the results of the test using normal human adult epidermal keratinocytes are shown in Table 3.

[0060] [Table 2]

[0061] [Table 3]

[0062] The results in Tables 2 and 3 confirmed that the test sample had an excellent inhibitory effect on PGD2 production.

[0063] (Combination example 1) A treatment having the following composition was prepared by a conventional method. Rutin 1.0% by mass 1,3-propylene glycol 0.5% by mass Pentaerythritol tetra-2-ethylhexanate 1.2% by mass 95% ethanol 60.0% by mass · Appropriate amount of fragrance · DME / LPG(95 / 5) appropriate amount Remaining purified water

[0064] (Combination example 2) A rinse having the following composition was prepared by a conventional method. 0.5g Quercetin 1.5g stearyltrimethylammonium chloride Polyoxyethylene cetyl ether 1.0g Cetyl alcohol 2.0g Octyldodecanol 1.0g Cationic cellulose 0.5g Propylene glycol 5.0g Soapberry extract 0.2g Phellodendron bark extract 0.5g Rosemary extract 0.5g · Fragrance 3.0g Purified water, balance (total amount 100g)

[0065] (Combination example 3) A hair tonic having the following composition was prepared by a conventional method. Isoquercetin 0.4g Tocopherol acetate (appropriate amount) Cephalatin 0.002g Isopropylmethylphenol 0.1g Sodium hyaluronate 0.15g 15.0g glycerin 15.0g ethanol · Appropriate amount of fragrance Chelating agent (sodium edetate) appropriate amount Preservative (hinokitiol) appropriate amount Solubilizer (polyoxyethylene cetyl ether) appropriate amount Purified water, balance (total amount 100g)

[0066] (Combination example 4) A shampoo having the following composition was prepared by a conventional method. Kaempferol 1.5g Plum fruit extract 0.2g Sodium coconut oil fatty acid methyl taurate 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 · Appropriate amount of fragrance Purified water, balance (total amount 100g)

Claims

【Request Item 1】 Prostaglandin (PG) D, characterized by containing at least one selected from the group consisting of rutin, quercetin, isoquercetin, and kaempferol ２ Production inhibitor.

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