Pore ​​enlargement inhibitor

Abstract

To provide an agent for inhibiting the opening of facial pores, comprising extract from a hop oxidation reaction product, an agent for inhibiting the carbonylation of horny layer protein, and an NGAL production inhibitor.SOLUTION: The present invention provides an agent for inhibiting the opening of facial pores, comprising extract from a hop oxidation reaction product, an agent for inhibiting the carbonylation of horny layer protein, and an NGAL production inhibitor.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a pore-opening inhibitor containing an extract of a hop oxidation reaction product, a carbonylation inhibitor of keratin, and an NGAL production inhibitor.

Background Art

[0002] Hop (Humulus lupulus) is one of the main raw materials for beer and is related to the bitterness, aroma, preservation, etc. of beer. In addition, hops have been used as a folk medicine since ancient times and are known to have effects such as a sedative effect, a diuretic effect, a hair loss prevention effect, and an anti-allergic effect. Hops contain various components related to the above-mentioned effects, and research and utilization of the functional effects derived from hop-containing components in the fields of medicine, food, cosmetics, etc. have also been carried out. For example, Patent Document 1 (Japanese Patent Application Laid-Open No. 2003-183174) discloses a composition for sleep disorder containing hops, Patent Document 2 (Japanese Patent Application Laid-Open No. 2003-238383) discloses a cell growth promoter containing hops, and Patent Document 3 (Japanese Patent Application Laid-Open No. 2016-147917) proposes an autonomic nerve regulator containing hops.

[0003] Here, hops have a very strong bitterness and it is difficult to use them directly in oral pharmaceuticals and foods. As a means of reducing this bitterness, Patent Document 4 (Japanese Patent No. 5980687) and Non-Patent Document 1 report that by oxidizing (aging) hops, bitter acids such as iso-α-acid, α-acid, and β-acid, which are bitter components of hops, become matured hop bitter acids (MHBA), which are oxides, and that hop oxidation reaction products containing this matured hop bitter acid have a reduced bitterness and health functional effects such as an improved lipid metabolism function.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

[0005] [Non-Patent Document 1] Morimoto-Kobayashi, Y., Ohara, K., Ashigai, H. et al. Matured hop extract reduces body fat in healthy overweight humans: a randomized, double-blind, placebo-controlled parallel group study. Nutr J 15, 25 (2015). [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The objective is to provide a pore-opening inhibitor, a stratum corneum protein carbonylation inhibitor, and an NGAL production inhibitor containing an extract of hop oxidation reaction products. [Means for solving the problem]

[0007] The main means for solving the problems of the present invention are as follows: 1. A pore-enlarging inhibitor characterized by containing an extract of hop oxidation reaction products. 2. A stratum corneum protein carbonylation inhibitor characterized by containing an extract of the hop oxidation reaction product. 3. An NGAL production inhibitor characterized by containing an extract of the hop oxidation reaction product. 4. Extracts of hop oxidation reaction products and, A pore-enlarging inhibitor characterized by containing an aqueous polyol solution obtained by dissolving one or more polyols selected from the group consisting of pentylene glycol, 1,3-butylene glycol, dipropylene glycol, methyl gluceth-10, cyclohexylglycerin, and (eicosanedioic acid / tetradecanedioic acid) polyglyceryl-10 in water.

[0008] 5. Extract of hop oxidation reaction product and, A stratum corneum protein carbonylation inhibitor characterized by containing an aqueous polyol solution obtained by dissolving one or more polyols selected from the group consisting of pentylene glycol, 1,3-butylene glycol, dipropylene glycol, methyl gluceth-10, cyclohexylglycerin, and (eicosanedioic acid / tetradecanedioic acid) polyglyceryl-10 in water. 6. Extracts of hop oxidation reaction products and, An NGAL production inhibitor characterized by containing an aqueous polyol solution obtained by dissolving one or more polyols selected from the group consisting of pentylene glycol, 1,3-butylene glycol, dipropylene glycol, methyl gluceth-10, cyclohexylglycerin, and (eicosanedioic acid / tetradecanedioic acid) polyglyceryl-10 in water. Hereinafter, pore enlargement inhibitors, stratum corneum protein carbonylation inhibitors, and NGAL production inhibitors will be collectively referred to as the agents of the present invention. [Effects of the Invention]

[0009] The pore-enlarging inhibitor of the present invention can reduce the volume of pores and make them less noticeable. The carbonylation inhibitor for stratum corneum proteins of the present invention can suppress carbonylation of stratum corneum proteins. By suppressing carbonylation of stratum corneum proteins, it is expected that the invention can prevent a decrease in the skin's barrier function and dryness. The NGAL production inhibitor of the present invention can suppress the production of NGAL (Neutrophil gelatinase-associated lipocalin), which is known to increase in inflammatory diseases and the like. By suppressing the production of NGAL, it is expected to suppress skin inflammation. The polyol aqueous solution of the hop oxidation reaction product is excellent in stability and hardly forms precipitates. Therefore, the agent of the present invention containing this polyol aqueous solution can be stably stored for a long period of time.

Brief Description of the Drawings

[0010] [Figure 1] Measurement results of pore volume before and after combined use [Figure 2] Measurement results of carbonylation of stratum corneum protein before and after combined use [Figure 3] Measurement results of the change rate of NGAL expression level before and after combined use

Modes for Carrying Out the Invention

[0011] The present invention has been made by finding that an extract of a hop oxidation reaction product has an effect of suppressing pore enlargement, an effect of suppressing carbonylation of stratum corneum protein, and an effect of suppressing NGAL production. The present invention relates to a pore enlargement inhibitor, a stratum corneum protein carbonylation inhibitor, and an NGAL production inhibitor containing an extract of a hop oxidation reaction product.

[0012] ·Hop oxidation reaction product The hop oxidation reaction product refers to a product obtained by subjecting hop or its processed product (such as hop pellets) to an oxidation treatment. The hop oxidation reaction product of the present invention can be obtained by bringing hop into contact with oxygen in the air and oxidizing it. The oxidation treatment method in the present invention is not particularly limited, but it is preferable to heat hop in the air.

[0013] In the oxidation treatment method of the present invention, the heating temperature is not particularly limited, but the preferable upper limit is 100 °C, and the more preferable upper limit is 80 °C. When the heating temperature is 100 °C or lower, it is advantageous for oxidation to proceed preferentially over isomerization. Also, the preferable lower limit of the heating temperature is 40 °C, but oxidation may be carried out at room temperature and low temperatures. The more preferable lower limit of the heating temperature is 60 °C, and the particularly preferable lower limit of the heating temperature is 65 °C. When the heating temperature is 60 °C or higher, it is advantageous for the oxidation reaction to proceed efficiently. Also, the reaction time is not particularly limited and can be appropriately determined according to the variety of hops and the reaction temperature. The oxidation treatment can be carried out, for example, under the conditions of 60 to 80 °C for 8 to 120 hours. If the temperature is 60 °C, 48 to 120 hours is preferable, and if it is 80 °C, 8 to 24 hours is preferable. Furthermore, the form of the hops is not particularly limited as long as it can contact oxygen in the air, but preferably in powder form to shorten the reaction time. Also, it may be stored in a high-humidity environment.

[0014] In the present invention, hops (Humulus lupulus L.) may be in any form as long as they contain lupulin, and those before harvesting and drying, those harvested and dried, compressed, pulverized, or processed into pellets may be used. Preferably, they are in the form of hop pellets. Commercially available products may be used for the hop pellets. Examples include those obtained by compressing hop strobiles into pellets (Type 90 pellets), pellets in which the lupulin portion is selectively concentrated (Type 45 pellets), or isomerized hop pellets (e.g., Isomerized Pellets (hopsteiner)).

[0015] Hop varieties are not particularly limited, but examples include Galaxy, Saaz, Cascade, Hallertau Tradition, Bullion, Brewers Gold, Chinook, Cluster, East Kent Golding, Fuggles, Hallertau, Mount Hood, and Northern Brewer. Brewer, Perle, Styrian, Target, Tettnanger, Willamette, Hersbrucker, Bravo, Columbus, Herkules, Magnum, Millennium, Nugget, Summit, Tomahawk, Warrior, Zeus, Hallertau Perle Examples include Perle, and from the standpoint of a stable supply of raw materials, it is preferable to use relatively inexpensive bitter varieties. Specific examples include Bravo, Columbus, Herkules, Magnum, Millennium, Nugget, Summit, Tomahawk, Warrior, and Zeus.

[0016] Hops contain resin-derived bitter components such as alpha acids (humulones), beta acids (luprons), and iso-alpha acids (isohumulones). In this invention, "alpha acids (humulones)" is used to mean humulone, adhumulone, cohumulone, posthumulone, and prehumulone. Also, in this invention, "beta acids (luprons)" is used to mean luprone, adolprone, colprone, postlupron, and prelupron. Furthermore, in this invention, "iso-alpha acid (isohumulones)" is used to mean including isohumulone, isoadhumulone, isocohumulone, isoposthumulone, isoprehumulone, Rho-isohumulone, Rho-isoadhumulone, Rho-isocohumulone, Rho-isoposthumulone, Rho-isoprehumulone, tetrahydroisohumulone, tetrahydroisoadhumulone, tetrahydroisocohumulone, tetrahydroisoprehumulone, tetrahydroisoposthumulone, hexahydroisohumulone, hexahydroisoadhumulone, hexahydroisocohumulone, hexahydroisoposthumulone, and hexahydroisoprehumulone. Note that iso-alpha acid has cis and trans stereoisomers, but unless otherwise specified, the term is used to mean including both.

[0017] By subjecting hops or their processed products (such as hop pellets) to an oxidation treatment, a "hop oxidation reaction product" is obtained in which the content of alpha acids, beta acids, and iso-alpha acids is reduced, and the content of other components increases. The hop oxidation reaction product of the present invention preferably contains a high level of matured hop bitter acids (MHBA), which are oxides of bitter components such as α-acids, β-acids, and iso-α-acids. Compared to α-acids, β-acids, iso-α-acids, MHBA has a very low bitterness and possesses health-promoting effects (Patent Document 4, Biosci. Biotech. Biochem. 2015;79:1684-1694), making it advantageous for imparting health functions to the hop oxidation reaction product. Suitable examples of components constituting such MHBA include the compounds described in the following formulas (1a-c) to (8a-c). Therefore, the hop oxidation reaction product of the present invention preferably contains at least one compound selected from the following formulas (1a-c) to (8a-c), and preferably contains two or more or all of the compounds. The quantitative determination of MHBA can be carried out according to the method described in Test Example 1 of Japanese Patent Application Publication No. 2018-199639.

[0018] [ka]

[0019] The hop oxidation reaction product preferably contains "tricyclooxyisohumulones." "Tricyclooxyisohumulones" refers to tricyclooxyisocohumulone A (TCOIcoH A: formula (5a), IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(2-methylpropanoyl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione) and tricyclooxyisohumulone A (TCOIH A: Formula (5b), IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(3-methylbutyryl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione), tricyclooxyisoadhumulone A(tricyclooxyisoadhumulone A)(TCOIadH A: Formula (5c), IUPAC name: (3aS,5aS,7S,8aS)-3,3a-dihydroxy-7-(1-hydroxy-1-methylethyl)-6,6-dimethyl-2-(2-methylbutanoyl)-5a,6,7,8-tetrahydro-3aH,5H-cyclopenta[c]pentalene-1,4-dione).

[0020] Regarding the MHBA content in the hop oxidation reaction product of the present invention, a suitable lower limit can be 0.1% by mass, 0.5% by mass, or 1% by mass, relative to the total dry mass of the hop oxidation reaction product, and a suitable upper limit can be 15% by mass, 20% by mass, 30% by mass, 50% by mass, 70% by mass, 90% by mass, 96% by mass, or 98% by mass. Furthermore, the range of MHBA content in the hop oxidation reaction product of the present invention is, for example, 0.1 to 98% by mass, more preferably 0.1 to 90% by mass, even more preferably 0.1 to 70% by mass, even more preferably 0.1 to 30% by mass, and even more preferably 1 to 20% by mass.

[0021] Furthermore, in the hop oxidation reaction product of the present invention, the content of α-acids, β-acids, and iso-α-acids is reduced by oxidizing the hops. The range of bitter component content in the hop oxidation reaction product of the present invention, when converted to the total content of α-acids, β-acids, and iso-α-acids relative to the total dry mass of hops, can be preferably a lower limit of 0.01% by mass, 0.02% by mass, 0.05% by mass, or 0.1% by mass, and a preferred upper limit of 0.5% by mass, 1% by mass, 5% by mass, or 10% by mass. The range of bitter component content in the hop oxidation reaction product of the present invention, as converted values, is, for example, 0.01 to 5% by mass, preferably 0.05 to 5% by mass, and more preferably 0.1 to 1% by mass. The content of such bitter components can be determined, for example, by converting it to the total content of α-acids, β-acids, and iso-α-acids according to the method described in Test Example 1 of Japanese Patent Application Publication No. 2018-199639.

[0022] Furthermore, the MHBA content in the hop oxidation reaction product is preferably equal to or greater than the bitter components (total amount of α-acids, β-acids, and iso-α-acids), more preferably 4 times or more, even more preferably 50 times or more, even more preferably 90 times or more, and even more preferably 100 times or more.

[0023] The hop oxidation reaction product is used as an extract obtained by extracting with a solvent containing water or various organic solvents, or by supercritical extraction using supercritical carbon dioxide. The solvent extract is a liquid in which useful components derived from the hop oxidation reaction product are dissolved, and is preferable because it has excellent handling properties, permeability, and odor reduction. The solvent extraction temperature is not particularly limited, but from the viewpoint of odor removal, it is preferably 60°C or lower, and considering the extraction efficiency, 50-60°C is more preferable. Examples of organic solvents include lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, and butanol; lower alkyl esters such as ethyl acetate; glycols such as ethylene glycol, butylene glycol, propylene glycol, and glycerin; other polar solvents such as acetone and acetic acid; hydrocarbons such as benzene and hexane; and non-polar solvents such as ethers such as ethyl ether and petroleum ether. Since the agent of the present invention is applied to the skin, it is preferably an aqueous medium, more preferably containing water, and even more preferably just water. That is, in the present invention, the extract of the hop oxidation reaction product is preferably an aqueous extract, and more preferably a water extract.

[0024] • Water extract of hop oxidation reaction product The aqueous extract of the hop oxidation reaction product of the present invention is an extract obtained by subjecting the above-mentioned hop oxidation reaction product to extraction with water. The extraction method is not particularly limited, and for example, the method described in Non-Patent Document 1 (Nutr J. 2015;15:25.) can be used. The unpleasant odor of the hop oxidation reaction product can be reduced by aqueous extraction. The extraction temperature is not particularly limited, but from the viewpoint of odor removal, it is preferably 60°C or lower, and considering the extraction efficiency, 50-60°C is more preferable.

[0025] In the present invention, the aqueous extract of the hop oxidation reaction product can be used as a dried solid, a concentrated liquid, or an aqueous extract as is, but using it as an aqueous extract as is is preferable from the viewpoint of ease of handling. In addition to drying or concentration as described above, heat sterilization treatment can also be performed. Specifically, boiling, vacuum degassing, vacuum concentration, spray drying, freeze-drying, high-temperature short-time heat treatment, high-temperature short-time sterilization, or a combination thereof can be used. The aqueous extract of the hop oxidation reaction product is a clear solution at high temperatures immediately after extraction, but at low temperatures, the dissolved components precipitate out, so it is usually a suspension. In the present invention, the aqueous extract of the hop oxidation reaction product may be used as a suspension, or it may be treated with filtration, activated carbon or adsorbents such as polyvinylpolypyrrolidone (PVPP), filter aids, fining agents, or descaling agents as needed. However, since an aqueous polyol solution obtained by adding a polyol to the aqueous extract of the hop oxidation reaction product can dissolve much of the insoluble components in the aqueous extract of the hop oxidation reaction product, it is preferable to use it as a suspension from the viewpoint of utilizing the active ingredients contained in these insoluble components.

[0026] In the aqueous extract of the hop oxidation reaction product, the content of the aqueous extract of the hop oxidation reaction product is 1 to 30% by mass, and the MHBA content is preferably 0.1 to 10% by mass, more preferably 0.5 to 8% by mass, and particularly preferably 1 to 5% by mass, from the viewpoint of functional effect. Furthermore, the Brix value of the aqueous extract of the hop oxidation reaction product is preferably 5 to 30°Bx, and more preferably 10 to 20°Bx, from the viewpoint of ease of handling.

[0027] • Polyol The polyol can be one or more selected from the group consisting of pentylene glycol, 1,3-butylene glycol, dipropylene glycol, methyl gluceth-10, cyclohexylglycerin, and (eicosanedioic acid / tetradecanedioic acid) polyglyceryl-10, and two or more can be mixed and used within a range that does not hinder the effects of the present invention. Among these, pentylene glycol is preferred because of its solubility of solids, ease of handling, and minimal change in odor. Furthermore, aqueous solvents other than the polyols mentioned above can also be included within a range that does not hinder the effects of the present invention.

[0028] In the aqueous polyol solution of the present invention, when pentylene glycol is used as the polyol, the amount of pentylene glycol is preferably 8% by mass or more and 22% by mass or less, and more preferably 8% by mass or more and 20% by mass or less. When the amount of pentylene glycol is within this range, the solubility is excellent, the amount of insoluble matter can be reduced, and in some cases, complete dissolution is possible.

[0029] In the aqueous polyol solution of the present invention, the ratio of the aqueous extract of the hop oxidation reaction product to the polyol is not particularly limited as long as it does not hinder the effects of the present invention. However, when the amount of aqueous extract of the hop oxidation reaction product is 30% by mass or less, the ratio of the amount of polyol to the amount of aqueous extract of the hop oxidation reaction product is preferably 0.1 to 45, more preferably 0.2 to 30, and even more preferably 0.3 to 20.

[0030] ·water In the aqueous polyol solution of the present invention, the ratio of polyol to water such as purified water is not particularly limited as long as it does not hinder the effects of the present invention. However, the weight ratio of polyol to water (polyol:water) is preferably in the range of 5:95 to 40:60, more preferably in the range of 10:90 to 30:70, and even more preferably in the range of 10:90 to 25:75.

[0031] • Polyol aqueous solution The polyol aqueous solution of the hop oxidation reaction product of the present invention uses both polyol and water as solvents. The insoluble portion of the aqueous extract (or suspension) of the hop oxidation reaction product does not completely dissolve in polyol alone or in water alone. By adding polyol and water to the aqueous extract (or suspension) of the hop oxidation reaction product and heating as necessary, much of the insoluble portion in water at room temperature can be dissolved and reduced, and in some cases, it can be completely dissolved. The temperature at which heating is performed is not particularly limited as long as it is below the boiling point of the polyol aqueous solution used, but it is preferably below 100°C. Adding water after adding polyol is preferable from the viewpoint of improving solubility. Furthermore, heating is preferable from the viewpoint of improving solubility when performed after adding polyol and water.

[0032] In the aqueous polyol solution of the present invention, the concentration of the aqueous extract of the hop oxidation reaction product is preferably 3% by mass or less. If the concentration of the aqueous extract of the hop oxidation reaction product exceeds 3% by mass, insoluble matter tends to remain, and storage stability decreases, making it easier for precipitates to form over time. The concentration of the aqueous extract of the hop oxidation reaction product is more preferably 2% by mass or less, and even more preferably 1.5% by mass or less. Furthermore, the lower limit of the aqueous extract concentration of the hop oxidation reaction product is preferably 0.01% by mass or more, more preferably 0.02% by mass or more, and even more preferably 0.05% by mass or more.

[0033] In the aqueous polyol solution of the present invention, the MHBA content is preferably 0.001% by mass or more, more preferably 0.005% by mass or more, even more preferably 0.01% by mass or more, and also preferably 3.0% by mass or less, more preferably 1.0% by mass or less, and even more preferably 0.5% by mass or less, from the viewpoint of functionality.

[0034] The polyol aqueous solution of the present invention may or may not contain insoluble matter. Insoluble matter in the polyol aqueous solution can be removed by filtration or centrifugation. By removing insoluble matter, which acts as a nucleus for aggregation, the polyol aqueous solution of the present invention can be made less prone to precipitation over time, thereby improving its storage stability. When the polyol aqueous solution of the present invention contains insoluble matter, the proportion of insoluble matter is preferably 0.3% by mass or less, more preferably 0.2% by mass or less, and even more preferably 0.15% by mass or less, relative to the polyol aqueous solution.

[0035] • Pore enlargement inhibitor, stratum corneum protein carbonylation inhibitor, NGAL production inhibitor The agent of the present invention may contain an extract of the hop oxidation reaction product, and its dosage form is not limited. For example, it can be applied in the form of liquid preparations such as aqueous solutions, emulsions, and suspensions; semi-solid preparations such as gels and creams; and solid preparations such as powders, granules, capsules, and solids. The agent of the present invention may contain polyhydric alcohols, oils and fats such as vegetable oils, higher fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, thickeners, metal ion chelating agents, polymers such as water-soluble polymers, UV absorbers, UV blockers, moisturizers such as hyaluronic acid and amino acids, fragrances, pH adjusters, drying agents, etc., within limits that do not impair the effects of the present invention. It may also contain other pharmacoactive and physiologically active ingredients such as vitamins, skin activators, blood circulation promoters, resident bacteria control agents, reactive oxygen species scavengers, anti-inflammatory agents, whitening agents, and bactericides. [Examples]

[0036] The present invention will be described in detail below based on examples, but the present invention is not limited thereto. • Aqueous extract of hop oxidation reaction product Aged hop extract (manufactured by Kirin Holdings Co., Ltd., solids content 11% by mass, MHBA 2.65% by mass)

[0037] • Polyol aqueous solution of hop oxidation reaction product 1. Aged hop extract 10.0% 2. Pentylene glycol 20.0% 3.Purified water remainder Furthermore, the aged hop extract was heated and dissolved to remove insoluble components by adding pentylene glycol and purified water.

[0038] Using an aqueous polyol solution of the hop oxidation reaction product, emulsions of Example 1 and Comparative Example 1 containing the components shown below were prepared by standard methods to the final concentrations shown below, and used in the following human continuous use tests. "Example 1: Emulsion" 1.Purified water remainder 2. Glycerin 4.0% 3. Dipropylene glycol 2.0% 4,1,3-Butylene glycol 4.0% 5. Diglycerin 2.0% 6. Pentylene glycol 3.0% 7. Betaine 4.0% 8. Squalane 3.0% 9. Pentaerythrityl tetraethylhexanoate 3.0% 10. Jojoba seed oil 2.0% 11. Dimethicone 2.0% 12. (Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate) Copolymer 0.25% 13. Sorbitan isostearate 0.3% 14. Polysorbate 60 0.65% 15. Hydrogenated lecithin 0.4% 16. Behenyl alcohol 0.5% 17. Xanthan gum 0.05% 18. Carboxyvinyl polymer 0.1% 19. Potassium hydroxide 0.015% 20. Aged hop extract 0.5% "Comparative Example 1: Emulsion" Examples 1 to 20. Emulsion from which aged hop extract has been removed.

[0039] • Human continuous use test Seventeen subjects were instructed to apply either a lotion containing an extract of hop oxidation reaction products (Example 1) or a lotion without an extract of hop oxidation reaction products (Comparative Example 1) to the left and right halves of their faces daily for four weeks, after washing their faces in the morning and evening and applying their usual toner. The application of the lotion to the left and right halves of the face was determined randomly.

[0040] The following items were evaluated before and after use. The evaluation results are shown in Figures 1-3. • Pore volume Using ANTERA 3D (Miravex), images were taken of the left and right cheeks of the subjects, and the pore volume in a certain area was measured. A smaller value after 4 weeks of continuous use compared to the value before continuous use indicates that pore enlargement was suppressed.

[0041] • Carbonylation of stratum corneum proteins Stratum corneum cells were collected from the face using the tape stripping method. The stratum corneum sampling tape was cut into eight equal parts, and one piece was placed in a 24-well plate. The stratum corneum sampling tape was immersed in a solution of Fluorescein-5-thiosemicarbazide (Adipogen Life Sciences), a fluorescent labeling agent for carbonyl groups, diluted with MES Buffer (WAKO) to a concentration of 20 μM, and allowed to stand at 25°C for 1 hour in the dark to allow the reaction to occur. Washed three times with 2 mL of PBS(-)(WAKO) and thoroughly drained. Stratum corneum sampling tape was attached to a glass slide (Matsunami Glass Industry Co., Ltd.), and fluorescence images were captured using an in-cell analyzer (Cytiva). The luminance value per unit area of ​​the stratum corneum portion in the images was quantified using an in-cell developer (Cytiva). Because fluorescein-5-thiosemicarbazide has a hydrazino group that binds to the carbonyl group, the carbonylation level can be evaluated by fluorescence intensity.

[0042] • NGAL expression level Stratum corneum cells were collected from the face using the tape stripping method. Stratum corneum collection tapes containing glass beads and 500 μl of T-PER buffer (Thermo SCIETIFIC) were placed in a tube, and the tube was shaken using the bead crushing method to extract stratum corneum proteins.

[0043] 100 μl / well of immobilized antibody diluted in PBS(-)(WAKO) was dispensed into a 96-well plate (COSTAR), and the antibody was incubated overnight at 25°C to allow it to be fixed. After washing three times with 300 μl of Wash buffer (PBS containing 0.05% Tween20), 200 μl / well of Reagent Diluent (R&D systems) was added, and the samples were blocked at 25°C for 1 hour. After washing again, 100 μl / well of stratum corneum extract sample was added and reacted at 25°C for approximately 2 hours. Purified protein antigens of each biomarker were also added in a concentration gradient and reacted to create a calibration curve.

[0044] Next, after washing, 100 μl / well of a solution prepared by diluting the detection antibody with Reagent Diluent to a concentration of 25 ng / ml was added, and the mixture was reacted at 25°C for 2 hours. After the reaction was complete, the wells were washed, and then Streptavidin-HRP (R&D Systems) was diluted 200-fold with Reagent Diluent. 100 μl of this solution was added to each well, and the reaction was carried out at 25°C for 20 minutes. For the immobilized antibody, purified protein antigen, and detection antibody, we used DY1757 from R&D Corporation.

[0045] After the reaction was complete, the wells were washed, and the mixture was colored with 100 μl / well of TMB reagent (Promega) (25°C, 7 minutes 30 seconds). The reaction was then stopped with 1N sulfuric acid, and the absorbance at 450 nm was measured using a SPECTRA MAX 190 (Molecular Device). NGAL expression levels were calculated from the aforementioned calibration curve. The expression levels were corrected for the total protein content in the stratum corneum extract and expressed as an amount per unit protein. The total protein content was measured using the BCA Protein Assay kit (Thermo Fisher Scientific).

[0046] To evaluate the test results, a t-test was used to examine the statistical significance of each evaluation item. The significance levels were set as P<0.01:** and P<0.05:*. As a result, significant differences were observed in all evaluation items, and it was revealed that continuous use of the emulsion containing the extract of hop oxidation reaction products suppressed pore enlargement (Figure 1), suppressed carbonylation of stratum corneum proteins (Figure 2), and suppressed NGAL expression levels (Figure 3).

[0047] Examples of prescriptions are shown below. Prescription example 1: Lotion 1.Purified water remainder 2. Glycerin 5.0% 3. Diglycerin 7.0% 4,1,3-Butylene glycol 5.0% 5. Methyl Gluceth-10 1.0% 6. Pentylene glycol 2.2% 7. Aged hop extract 0.05%

[0048] Prescription Example 2: Beauty Gel 1.Purified water remainder 2. Dipropylene glycol 6.0% 3. PEG-75 0.5% 4. Raffinose 2.0% 5. Pentylene glycol 1.1% 6. Xanthan gum 0.1% 7. Squalane 3.0% 8. SIMULGEL NS* 2.5% 9. Aged hop extract 0.1% 10. Ethanol 2.0% *SIMULGEL NS (manufactured by SEPPIC; a mixture of (hydroxyethyl acrylate / sodium acryloyldimethyl taurate) copolymer, squalane, polysorbate 60, water, and sorbitan isostearate)

[0049] Prescription Example 3: Sheet Mask Serum 1.Purified water remainder 2,1,3-Butylene glycol 5.0% 3. Glycerin 3.0% 4. Betaine 1.5% 5. Pentylene glycol 1.1% 6. Phenoxyethanol 0.1% 7. Carboxyvinyl polymer 0.1% 8. Potassium hydroxide 0.02% 9. Sodium Hyaluronate 0.001% 10. Aged hop extract 0.3%

[0050] Prescription example 4: Cream 1.Purified water remainder 2. Squalane 7.0% 3. Olive oil 5.0% 4. Behenyl alcohol 1.5% 5. Beeswax 1.0% 6. Lauroyl glutamate di(phytosteryl / octyldodecyl) 1.0% 7. Polyglyceryl-10 stearate 2.0% 8. Glycerin 6.0% 9. Dipropylene glycol 6.0% 10,1,3-Butylene glycol 3.0% 11. Pentylene glycol 2.0% 12. Xanthan gum 0.05% 13. Carboxyvinyl polymer 0.2% 14. Potassium hydroxide 0.03% 15. Aged hop extract 0.01%

Claims

1. Contains an extract of the hop oxidation reaction product, The aforementioned extract contains 1 to 20% by mass of matured hop bitter acids (MHBA), and is characterized in that it contains 50 times or more MHBA relative to the total amount of bitter components (α-acids, β-acids, and iso-α-acids).

2. Contains an extract of the hop oxidation reaction product, The aforementioned extract contains 1 to 20% by mass of matured hop bitter acids (MHBA), and is characterized by containing MHBA at a ratio of 50 times or more relative to the bitter components (total amount of α-acids, β-acids, and iso-α-acids), thereby inhibiting carbonylation of stratum corneum proteins.

3. Contains an extract of the hop oxidation reaction product, An NGAL production inhibitor characterized in that the extract contains 1 to 20% by mass of matured hop bitter acids (MHBA), and contains 50 times or more MHBA relative to the total amount of bitter components (α-acids, β-acids, and iso-α-acids).

4. Extract of hop oxidation reaction product and, The product contains an aqueous polyol solution obtained by dissolving one or more polyols selected from the group consisting of pentylene glycol, 1,3-butylene glycol, dipropylene glycol, methyl gluceth-10, cyclohexylglycerin, and (eicosanedioic acid / tetradecanedioic acid) polyglyceryl-10 in water. The aforementioned extract contains 1 to 20% by mass of matured hop bitter acids (MHBA), and is characterized in that it contains 50 times or more MHBA relative to the total amount of bitter components (α-acids, β-acids, and iso-α-acids).

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