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WO2006106992A1 - Melanin production inhibitory agent - Google Patents

Melanin production inhibitory agent Download PDF

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Publication number
WO2006106992A1
WO2006106992A1 PCT/JP2006/306976 JP2006306976W WO2006106992A1 WO 2006106992 A1 WO2006106992 A1 WO 2006106992A1 JP 2006306976 W JP2006306976 W JP 2006306976W WO 2006106992 A1 WO2006106992 A1 WO 2006106992A1
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WO
WIPO (PCT)
Prior art keywords
acid
hibiscus
melanin production
whitening
active ingredient
Prior art date
Application number
PCT/JP2006/306976
Other languages
French (fr)
Japanese (ja)
Inventor
Kyoko Ishiguro
Naoe Oku
Naoko Harada
Katsuyuki Fujimura
Original Assignee
Kobayashi Pharmaceutical Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kobayashi Pharmaceutical Co., Ltd. filed Critical Kobayashi Pharmaceutical Co., Ltd.
Publication of WO2006106992A1 publication Critical patent/WO2006106992A1/en

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Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/02Preparations for care of the skin for chemically bleaching or whitening the skin
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/49Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
    • A61K8/4973Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom

Definitions

  • the present invention relates to a melanin production inhibitor containing hibiscus acids as active ingredients, and a whitening agent, a whitening cosmetic, a whitening food and an oral composition containing the melanin production inhibitor.
  • active oxygen scavengers such as ascorbic acid and tyrosinase activity inhibitors such as kojic acid and arbutin have been proposed as active ingredients for whitening agents! Speak (see Patent Document 1 etc.).
  • the substances proposed as the active ingredients of conventional whitening agents act only on part of the melanin production process, and melanin is also produced by other processes. The power was not satisfactory.
  • the conventional whitening ingredients have not been able to satisfy the viewpoint of sustaining the effect.
  • Patent Document 2 includes 3 hydroxy-3,4 dicarboxy 1,4-butanolide or derivatives thereof. Cosmetics characterized in that they contain a body are described. Patent Document 3 discloses a glycosidase inhibitor and a therapeutic agent for diabetes containing hibiscus acids as active ingredients. However, Hibiscus acids are effective in inhibiting melanin production and whitening! None before had it been known.
  • Patent Document 1 JP-A-1-85907
  • Patent Document 2 Japanese Patent Application Laid-Open No. 200-154134
  • Patent Document 3 Japanese Patent Laid-Open No. 2000-239164
  • the present invention has an excellent melanin production inhibitory action, has a persistent melanin production inhibitory action, and can be used safely, a melanin production inhibitor, a whitening agent, a whitening cosmetic,
  • the main purpose is to provide a food for whitening and a composition for oral cavity.
  • the present invention relates to the following melanin production inhibitor, whitening agent, cosmetic, food and oral composition.
  • a melanin production inhibitor comprising as an active ingredient one or more compounds selected from the group consisting of hibiscus acids represented by the formula (1) and a salt power thereof.
  • Item 2 The melanin production inhibitor according to Item 1, wherein at least one of R 1 and R 2 of the hibiscus acids represented by the formula (1) is a substituted or unsubstituted alkyl group.
  • IT and R 2 each represent a hydrogen atom, a substituted or unsubstituted alkyl group or a benzyl group, and at least one of R 1 and R 2 is a substituted or unsubstituted alkyl group.
  • a melanin production inhibitor comprising as an active ingredient at least one compound selected from the group consisting of bis and biscus acids.
  • a melanin production inhibitor comprising as an active ingredient one or two or more compounds selected from the group power of hibiscus monoalkyl ester and bisbisester dialkyl esterka.
  • melanin production inhibition comprising one or two or more compounds selected from the group power consisting of bismuth acid monomethyl ester, bis, biscus acid monoethyl ester, hibiscus acid dimethyl ester and hibiscus acid dimethyl ester as active ingredients Agent.
  • Item 3 A whitening agent comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient.
  • Item 4 A whitening cosmetic comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient.
  • Item 5 A whitening roro comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient.
  • Item 6 An oral composition comprising the melanin production inhibitor according to any one of Items 12 and 12 as an active ingredient.
  • the melanin production inhibitor of the present invention has the following formula (1)
  • R 1 and R 2 each represent a hydrogen atom, a substituted or unsubstituted alkyl group, or a benzyl group.
  • One or more compounds are active ingredients.
  • the type of the alkyl group is not particularly limited as long as the effect of the present invention is exhibited, and may be linear or branched.
  • the number of carbon atoms of the alkyl group can be set as appropriate within the range in which the effects of the present invention are exerted.
  • alkyl group examples include a methyl group, an ethyl group, a propyl group, a butyl group, an isopropyl group, an isobutyl group, and a hexyl group.
  • the alkyl group or benzyl group may have a substituent, and the type of the substituent may be appropriately set within the range where the effect of the present invention is exerted. And the like.
  • hibiscus acids used in the present invention include, for example, hibiscus acid R 1 and R 2 in the formula are hydrogen atoms, one of R 1 and R 2 in the formula is substituted And Hibiscus acid diester wherein R 1 and R 2 in the formula are both substituted.
  • Hibiscus acid monoesters include, for example, hibiscus monoalkyl esters such as hibiscus monomethyl ester and bisbis monoethyl ester.
  • Hibiscus acid diesters include, for example, bismuth acid dialkyl esters such as bismuth acid dimethyl ester, and biscus acid decyl ester.
  • the salts of hibiscus acids used in the present invention include the above-mentioned salts of hibiscus acids, for example, when R 1 and Z or R 2 are hydrogen atoms, they are monovalent or divalent metal bases. This includes alkali metal salts and alkaline earth metal salts that have been replaced.
  • alkali metal salt examples include sodium salt and potassium salt.
  • alkaline earth metal salts examples include calcium salts and magnesium salts.
  • the salt also includes those forming a complex.
  • an example of a complex is R
  • Examples include alkaline earth metal complexes formed by replacing 1 and Z or R 2 with calcium or magnesium.
  • the means for producing the hibiscus acids and salts thereof used in the present invention are not particularly limited, and may be obtained by force separation such as a plant body or an extract thereof which may be obtained by chemical synthesis. But you can.
  • the ester or salt may be obtained by esterifying a hibiscus acid obtained by separating from a plant or its extract according to a conventional method or reacting with various bases.
  • a method for separating hibiscus acids from a plant body or an extract thereof is not particularly limited, and a known method can be appropriately used.
  • a plant that contains hibiscus acid may be extracted with a suitable extraction solvent and then separated by means such as chromatography.
  • Examples of plants containing hibiscus acids include plants belonging to the genus Fuyu.
  • Examples of plants belonging to the genus Fuyou include the annual annual Roselle (Hibiscus sabdarif fa) native to the tropics and the bushy geese (Hibiscus) of the tropical shrub.
  • Roselle Roselle, Bussouge and Mukuge, and Roselle and Bussouge are particularly preferable.
  • the extraction site of the plant is not particularly limited, and for example, the whole plant of the plant or a part of the plant such as a flower, a bud, a leaf, a stem, a branch, a branch tip, a root, a rhizome, and a seed. Can be mentioned. Among these, Preferably, a flower and a leaf can be mentioned.
  • the extracted raw material may be a pulverized product obtained by using plant bodies as they are, or a product obtained by drying, chopping, crushing, pressing, boiling or fermenting them.
  • the type of the solvent used for extraction can be set as appropriate, and examples thereof include water, various organic solvents, and a mixture of water and an organic solvent.
  • the organic solvent include methanol, ethanol, n -propanol, isopropanol, butanol, isobutanol, hexanol, propylene glycol, 1,3-butylene glycol, acetone, ethyl acetate, black mouth form, pentane, hexane, Including heptane, hydrochloric acid, etc. alone or in combination of two or more.
  • preferable extraction solvents include lower alcohols such as methanol, ethanol, and isopropanol, or a mixture of water and lower alcohols.
  • water can be removed by performing a drying treatment such as reduced-pressure drying or freeze-drying to use as a dried product.
  • Specific conditions for extraction can also be appropriately set according to a conventional method without particular limitation.
  • a dried roselle flower is crushed, and about 10 parts by weight of 1.5% hydrochloric acid-ethanol aqueous solution is added to 1 part by weight of the extraction target, stirred at room temperature, and then solids are removed by filtration.
  • a method of performing purification by HPLC can be mentioned.
  • commercially available bis and ibiscus acids commercially available ones can also be used.
  • the ester can be obtained from commercially available hibiscus acid by subjecting it to esterification according to a known method.
  • the compound used as the active ingredient of the melanin production inhibitor of the present invention may be a single compound selected from the above-mentioned hibiscus acids or a group power of salt power thereof, or a mixture of two or more compound powers. May be.
  • the mixture consisting of two or more kinds of compounds contains biscus acid or biscus acid which may be obtained by mixing various hibiscus acids or salts thereof obtained by synthesis or isolation from plants. It may be a mixture or a fraction obtained from a plant or an extract thereof.
  • the melanin production inhibitor of the present invention contains, as an active ingredient, one or more compounds selected from the group consisting of the above-mentioned hibiscus acids and their salt strength, and preferably a hibiscus monoalkyl ester and hibiscus It contains one or more compounds selected from the group consisting of acid dialkyl esters as active ingredients.
  • it includes one or more compounds selected as an active ingredient, which are selected from bismuth acid monomethyl ester, bis, biscus acid monodimethyl ester, hibiscus acid dimethyl ester, and hibiscus acid dimethyl ester. .
  • the melanin production inhibitor of the present invention contains, as an active ingredient, one or two or more compounds selected from the group consisting of the above-mentioned hibiscus acids and salts thereof, and has a high and sustained tyrosinase activity inhibiting action and a high tyrosinase. It has a production inhibitory action and exhibits an effective and continuous melanin production inhibitory action.
  • the whitening agent of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
  • the whitening agent of the present invention may consist of the melanin production inhibitor itself, or it may contain other ingredients such as pharmaceutically or sanitary acceptable carriers or additives as an active ingredient. It could be something! /
  • the types and blending amounts of the strong carriers or additives can be appropriately selected and adjusted according to the dosage form of the whitening agent or the application target, as long as the effects of the present invention are not impaired.
  • the form of the whitening agent is not particularly limited, and the dosage form and form of the product to be applied, Depending on the application, etc., it can be arbitrarily prepared into liquids, emulsions, creams, powders, granules, pills, ointments and the like.
  • the whitening agent of the present invention may be an external preparation or an internal preparation.
  • the whitening agent of the present invention inorganic pigments, ultraviolet absorbers, other whitening components, tyrosinase activity inhibitors or melanin dye reducing agents, surfactants, cell activation agents.
  • Additives such as agents, anti-inflammatory agents, antibacterial agents, moisturizers, fragrances, and coloring agents may be added as appropriate.
  • the types of external preparations to which the whitening agent of the present invention can be applied are not particularly limited.
  • external medicines and external quasi drugs skin external preparations, etc.
  • basic cosmetics skin lotion, emulsion, Tarim, ointment, lotion, oil, pack, etc.
  • makeup cosmetics foundation, lipstick, scarlet, mascara, etc.
  • other skin cosmetics face wash, skin cleanser, massage agent, cleansing agent, etc.
  • Oral compositions bath preparations, and the like.
  • the amount of the external preparation varies depending on the compounding target, dosage form, expected effect, etc., and cannot be uniformly defined.
  • Hibiscus acids and salts thereof are 0.00001 to 10% by weight, preferably 0.00005 to 5% by weight, more preferably about 0.0001 to 3% by weight, based on the whole agent.
  • a binder In formulating the whitening agent of the present invention as an internal preparation, a binder, a disintegrant, a lubricant, a wetting agent, a buffering agent, an excipient, an extender, and a preservative are used as necessary. In addition, additives such as fragrances can be combined. In addition, when formulating the whitening agent of the present invention as an internal preparation, other pharmaceutically active ingredients can be further blended.
  • the type of internal preparation to which the whitening agent of the present invention can be applied is not particularly limited, and examples thereof include oral preparations such as powders, tablets, granules, capsules, and syrups.
  • the blending ratio of the internal preparation varies depending on the administration target, dosage form, administration method, etc., and cannot be uniformly defined.
  • the amount of hibiscus acids and salts thereof is 0.00001 to 100% by weight, preferably 0.0001 to 50% by weight, more preferably about 0.001 to 30% by weight, based on the whole internal preparation.
  • the whitening agent of the present invention comprises, as an active ingredient, a melanin production inhibitor containing as an active ingredient one or more compounds selected from the above-mentioned hibiscus acids and the group power of their salt strength.
  • a melanin production inhibitor containing as an active ingredient one or more compounds selected from the above-mentioned hibiscus acids and the group power of their salt strength.
  • it has a continuous and excellent melanin production inhibitory action including a high and sustained tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and exhibits a continuous and excellent whitening action.
  • the whitening cosmetic composition of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
  • the type of cosmetic is not particularly limited, and can be applied to various cosmetics.
  • various types of cosmetics such as rinsing and other hair cosmetics, sarcophagus, beauty nails, eau de cologne, and bath salts.
  • the form of the cosmetic is not particularly limited, either, solution, emulsion, ointment, sol, gel, powder.
  • aqueous component a powder component
  • a moisturizer a surfactant
  • a preservative an increase agent
  • Adhesives ultraviolet absorbers, fragrances and the like can be blended.
  • the method for producing the cosmetic of the present invention is not particularly limited, and can be suitably produced according to a known method.
  • the method of applying the cosmetic composition of the present invention is not particularly limited, but may be applied to, for example, an easily or affected part such as a stain on the face, neck, arm or hand, or freckles, or an easily tanned or tanned part. Apply an appropriate amount once or several times a day.
  • the content ratio of the melanin production inhibitor in the cosmetic of the present invention can be appropriately set within the range in which the effect of the present invention can be exerted.
  • a hibiscus acid and its salt with respect to the entire cosmetic composition 0.00001 -10% by weight, preferably 0.00005-5% by weight, more preferably 0.0001-3% by weight.
  • the whitening cosmetic composition of the present invention includes a melanin production inhibitor containing, as an active ingredient, the above-mentioned hibiscus acid and one or more compounds selected from the group power of its salt strength as an active ingredient, and is high and durable.
  • Tyrosinase activity inhibitory activity and high tyrosiner It has a continuous and excellent melanin production-inhibiting action, including a zeogenesis-inhibiting action, and has a continuous and excellent whitening effect.
  • the whitening food of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
  • the whitening food of the present invention can be used as, for example, health foods, food additives, functional foods, nutritional functional foods, health functional foods, and the like.
  • the method for producing the whitening food is not particularly limited, and can be suitably produced according to a known method. For example, it can be obtained by mixing the whitening agent with various foods or food materials.
  • the type of food is not particularly limited, and can be appropriately set from various known foods and drinks.
  • it can be used as cookies, cereals, candy, tablets, jelly, marine products, seasonings, bread, soft drinks, carbonated drinks, mineral water, fruit juice drinks, teas, nutrition drinks, drinks, and the like.
  • the food of the present invention can be blended with other components used in ordinary foods and drinks within a range not impairing the effects of the present invention.
  • other ingredients include various vitamins, minerals, animal and plant ingredients, excipients, sweeteners, flavoring agents, coloring agents, preservatives, disintegrating agents, lubricants, wetting agents, bulking agents, and flavoring agents. It is done.
  • the content of the melanin production inhibitor in the food of the present invention can be set as appropriate within the range in which the effects of the present invention can be exerted, but 0.00001 to 10 as hibiscus acids and salts thereof with respect to the entire food composition. % By weight, preferably 0.00005 to 5% by weight, more preferably about 0.0001 to 3% by weight.
  • the amount of intake when eating the whitening food of the present invention can be appropriately set according to the age, weight, symptoms, etc. of the subject of application, but is generally 0.00001 per day for adults as hibiscus acids and salts thereof. About 50 mg, preferably about 0.0001-20 mg, more preferably about 0.001-10 mg.
  • the whitening food of the present invention effectively comprises a melanin production inhibitor containing as an active ingredient one or more compounds selected from the above-mentioned hibiscus acids and the group power of their salt strength. It has a continuous and excellent melanin production inhibitory action including a high and sustained tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and has a continuous and excellent whitening action.
  • composition for oral cavity of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
  • the type of oral composition is not particularly limited, and can be applied to various oral compositions.
  • toothpaste such as toothpaste, mouthwash, mouth rinse, oral paste, troche, gum massage cream and the like.
  • the form of the oral composition is not particularly limited, and can be used in various forms such as a solution, emulsion, ointment, sol, gel, paste, tablet, powder, spray, sheet, and film.
  • composition for oral cavity of the present invention various components generally blended in the composition for oral cavity can be blended within the range where the effects of the present invention are exhibited.
  • Various components include, for example, an adhesive, a refreshing agent, a binder, a sweetener, a flavoring agent, a disintegrant, a lubricant, a colorant, a sustained release regulator, a surfactant, a solubilizer, a wetting agent, and a pH adjuster.
  • An agent etc. can be mentioned.
  • the method for producing the composition for oral cavity of the present invention is not particularly limited, and can be suitably produced according to a known method.
  • the method for applying the oral composition of the present invention is not particularly limited. For example, an appropriate amount may be applied once or several times a day to the affected area or the entire oral cavity.
  • the content ratio of the melanin production inhibitor in the oral composition of the present invention can be appropriately set within the range in which the effect of the present invention can be exerted, but the hibiscus acids and salts thereof with respect to the entire oral composition. As 0.00001 to 10% by weight, preferably 0.00005 to 5% by weight, and more preferably about 0.0001 to 3% by weight.
  • the composition for oral cavity of the present invention includes a melanin production inhibitor containing, as an active ingredient, the above-mentioned hibiscus acids and one or more compounds selected from the group power of its salt strength as an active ingredient, and is high and durable. It has a long-lasting and excellent melanin production-inhibiting action, including a typical tyrosinase activity-inhibiting action and a high tyrosinase-inhibiting action, and is deposited on the gum The pigment
  • a melanin production inhibitor comprising as an active ingredient one or more compounds selected from the group strength of hibiscus acid having a specific structure and its salt strength.
  • the compound as an active ingredient of the melanin production inhibitor of the present invention has an excellent melanin production inhibitory action including a high tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and has an excellent whitening effect. It is what you play. Furthermore, the whitening effect is achieved for a long time. However, the compound is also highly safe.
  • the melanin production inhibitor of the present invention comprising such a compound as an active ingredient has an excellent whitening effect and has a long-lasting whitening effect, and is effective in skin cells, oral mucosa and the like. It effectively suppresses the melanin produced and effectively suppresses the deposition of pigments such as spots and freckles. Furthermore, the melanin production inhibitor of the present invention can be used with peace of mind.
  • the melanin production inhibitor can be used as an active ingredient in whitening agents, whitening cosmetics, whitening foods and oral compositions, and has an excellent whitening action according to the present invention.
  • a whitening agent, a whitening cosmetic, a whitening food, and a composition for oral cavity that have a long-lasting whitening effect and can be used safely are also provided.
  • the present invention having such characteristics can be suitably used in the cosmetic industry, the pharmaceutical industry, the food industry, and the like, which have recently been particularly interested in the whitening effect.
  • the obtained fraction is also referred to as “no biscus acid monoethyl ester fraction”.
  • Hibiscus acid obtained in Production Example 1 was dissolved in a mixed solvent of 1.8 ml of methyl-t-butyl ether (MTBE) and 1.8 ml of methanol. This solution was stirred while cooling in an ice-water bath, and 3.5 ml of trimethylsilyl diazomethane 2.0M jetyl ether solution was added little by little. The resulting precipitate was filtered, washed with MTBE, and dried under reduced pressure to obtain 300 mg of bis (bisbisamic acid) dimethyl ester.
  • MTBE methyl-t-butyl ether
  • a biscus extract (extract) was produced by the following method.
  • Mouse-derived B16 melanoma cells (D16, manufactured by Dainippon Pharmaceutical Co., Ltd., B16F0) were cultured for 24 hours in an 8 ml culture medium in a 25 cm 2 flask. Were added at 12.5, 25, 50 or 100 ml, respectively, and cultured for 72 hours.
  • the cells were pulverized to extract proteins, and subjected to electrophoresis.
  • the protein is transferred to a polyvinylidene difluoride membrane (PVDF membrane), reacted with a goat anti-tyrosinase antibody, further reacted with an alkaline phosphatase-labeled anti-goat antibody, and finally a chromogenic substrate is obtained.
  • PVDF membrane polyvinylidene difluoride membrane
  • a goat anti-tyrosinase antibody further reacted with an alkaline phosphatase-labeled anti-goat antibody
  • a chromogenic substrate is obtained.
  • tyrosinase was detected.
  • the same test was carried out using a bismuth acid-free additive.
  • FIG. 1 shows the obtained results.
  • FIG. 1 shows a PVDF membrane image showing the expression level of tyrosinase in B16 melanoma cells cultured in the presence or absence of neubiscus acid.
  • lane I shows no addition of hibiscus acid solution
  • lane II adds 100 ⁇ 1 of 5% hibiscus acid solution
  • lane III adds 50 ⁇ 1 of 5% hibiscus acid solution
  • lane IV shows 5% hibiscus acid solution 25
  • Lane 1 shows the results for a sample with 1 ⁇ l of 5% bismuth acid added.
  • Mouse-derived B16 melanoma cells (D16, manufactured by Dainippon Pharmaceutical Co., Ltd., B16F0) were seeded in a 96-well microphone mouth plate at 5 ⁇ 10 4 cells / well, and cultured in sputum to establish the cells. Next, remove the culture solution from the culture plate with an aspirator, add 1Z Triton X-100-containing PBS (phosphate buffered saline) to lZwell, destroy the cell membrane structure, and remove the crude tyrosinase solution. It was.
  • Control 1 B16 melanoma cell seeding 'Hibiscus acid solution free supplemented solution (PBS added instead)
  • Control 2 B 16 melanoma cell-free seeding 'Hibiscus acid solution free additive (added PBS instead)
  • FIG. Figure 2 illustrates the relationship between the added concentration of bismuth, ibiscus acid and hibiscus extract and the inhibition rate (%) of tyrosinase activity! / Speak.
  • Example 2 the final concentration of bismuth acid monoethyl ester was set to 0.25 by using the bismuth acid monoethyl ester fraction obtained in Production Example 2 instead of bis and biscus acid. Evaluation was performed in the same manner as in Example 2 except that the concentration was adjusted to ⁇ 5 mg / ml.
  • Example 2 As in Example 2, for comparison, instead of the hibiscus monoethyl ester fraction, the hibiscus (Hibiscus sabdarilfa) obtained in Production Example 4 was used.
  • FIG. 3 shows the obtained results.
  • FIG. 3 shows the relationship between the added concentration of the hibiscus monoethyl ester fraction and the nobiscus extract and the inhibition rate (%) of tyrosinase activity.
  • Example 2 instead of bis and biscus acids, the bismuth acid dimethyl ester fraction obtained in Production Example 3 was used, and the final concentration of bismuth acid dimethyl ester was adjusted to 0.25 to 5 mg / ml. Evaluation was performed in the same manner as in Example 2 except that.
  • FIG. Fig. 4 illustrates the relationship between the concentration of bismuth acid dimethyl ester and hibiscus extract and the inhibition rate (%) of tyrosinase activity.
  • the tyrosinase activity inhibition rate (%) was calculated from the measured absorbance value based on the following equation (2).
  • Blank B16 melanoma cell non-seeding: Hibiscus acid solution-free caro
  • Control 2 B 16 melanoma cell-free seeding 'Hibiscus acid solution-free potassium (added PBS instead)
  • Tyrosinase activity inhibition rate 1-(Absorbance of control, Absorbance of control 2 ) ⁇ J 00
  • arbutin-added sample a test was conducted in the same manner except that arbutin was used in place of hibiscus acid, and the tyrosinase activity inhibition rate (%) of arbutin was calculated (hereinafter referred to as an arbutin-added sample).
  • Fig. 5 shows the obtained results.
  • ⁇ and ⁇ indicate the tyrosinase activity inhibition rate (%) of the sample added with hibiscus acid
  • ⁇ and ⁇ indicate the tyrosinase activity inhibition rate (%) of the sample added with arbutin.
  • the dotted line is the measurement result after 20 hours, and the solid line is 108 hours. Later measurement results
  • hibiscus acid has an inhibitory effect on the production of tyrosinase involved in the production of melanin and an inhibitory effect on tyrosinase activity. Even the effect was evaluated.
  • the cells were seeded to 10 5 cellsZwell and cultured overnight to fix the cells. Next, a solution obtained by diluting the nobiscus acid obtained in Production Example 1 with PBS so that the final concentration of the hibiscus acid in the well is 0.2 to 2 mgZml (in the following formula 3, the target sample is also included). ) was added, and after 4 days of culture, the cells were detached using a 0.25% trypsin solution, and the cells were collected by centrifugation. After visually confirming the color difference of the collected cells, the membrane structure of the cells was destroyed with 5N NaOH to elute melanin, and the absorbance at 405 nm was measured.
  • the amount of protein in the sample was measured using a quantification kit, corrected with the amount of unit protein, and the melanin production rate (%) was based on Equation 3 below. Calculated.
  • FIG. Figure 6 shows the melanin production rate (%) at each additive concentration of bis and biscus acids.
  • Example 6 tests and evaluations were performed in the same manner as in Example 6 except that the bisbiscus monoethyl ester fraction obtained in Production Example 2 was used in place of bis and biscus acids.
  • FIG. 7 shows the melanin production rate (%) at each addition concentration of the hibiscus monoethyl ester fraction.
  • Example 6 tests and evaluations were carried out in the same manner as in Example 6 except that the bis-biscus acid dimethyl ester obtained in Production Example 3 was used instead of bis-biscus acid.
  • FIG. 8 shows the obtained results.
  • FIG. 8 shows the melanin production rate (%) at each addition concentration of hibiscus dimethyl ester.
  • the extract obtained from plant power may contain the isomers of bis and biscus acids.
  • the optical isomers of hibiscus acid represented by the following formula (2) galsinic acid represented by the following formula (3) is considered as a naturally occurring one.
  • the cells were seeded to 10 5 cellsZwell and cultured in anther culture to establish the cells. Next, a solution diluted with PBS so that the final concentration of noci, biscus acid or garcinia acid in the well is 0.5 mg / ml (the “target sample” in Equation 4 below). After culturing for 4 days, the cells were detached using a 0.25% tribsin solution, and the cells were collected by centrifugation. After visually confirming the color difference of the collected cells, the membrane structure of the cells was destroyed with 5N NaOH to elute melanin, and the absorbance at 405 nm was measured.
  • non-added sample As a control, the measurement was carried out in the same manner with respect to a system not added with hibiscus acid or garcinia acid (hereinafter referred to as "non-added sample” in formula 4 below).
  • the amount of protein in the sample was measured using a quantification kit, corrected with the amount of unit protein, and the melanin production rate (%) was calculated based on the following equation. .
  • FIG. 9 shows the obtained results.
  • FIG. 9 shows the melanin production rate (%) and the number of cells of garcinia acid and neurobiscus acid at a concentration of 0.5%. BK indicates a blank (no additive sample).
  • the raw materials were blended at the blending ratios shown in the following table, and a lotion was produced according to a conventional method.
  • the raw materials were blended at the blending ratios shown in the table below, and the emulsion was produced according to a conventional method (raw materials) (wt%)
  • Nonfat dry milk 1. 0
  • the ingredients were blended at the blending ratios shown in the table below, and a drink was produced according to a conventional method.
  • the raw materials were blended at the blending ratios shown in the table below, and toothpastes were produced according to conventional methods.
  • the raw materials were blended at the blending ratios shown in the table below, and a mouthwash was produced according to a conventional method.
  • FIG. 1 is a PVDF image showing the expression level of tyrosinase in B16 melanoma cells cultured in Example 1 in the presence or absence of hibiscus acid.
  • Lane I is without hibiscus acid; Lane II is with 5% hibiscus acid solution 100 ⁇ 1; Lane III is with 5% hibiscus acid solution 50 ⁇ 1; Lane IV is 5% hibiscus acid solution 25 ⁇ m
  • Lane 1 is a diagram showing a sample of 12.5 ⁇ l-added 5% biscus acid solution.
  • FIG. 2 shows the concentration of hibiscus acid or hibiscus extract and tyrosin in Example 2. It is a figure which shows the relationship of the enzyme activity inhibition rate (%).
  • FIG. 3 is a graph showing the relationship between the added concentration of hibiscus monoethyl ester fraction or hibiscus extract and the tyrosinase activity inhibition rate (%) in Example 3.
  • FIG. 4 shows the relationship between the concentration of hibiscus dimethyl ester or hibiscus extract and the inhibition rate (%) of tyrosinase activity in Example 4.
  • FIG. 5 is a graph showing the tyrosinase activity inhibition rate (%) in Example 5 after 20 hours and 108 hours of incubation in the hibiscus acid added group and arbutin added group.
  • FIG. 6 is a graph showing the relationship between the concentration of hibiscus acid added and the melanin production rate (%) in Example 6.
  • FIG. 7 is a graph showing the relationship between the added concentration of the hibiscus monoethyl ester fraction and the melanin production rate (%) in Example 7.
  • FIG. 8 shows the relationship between the concentration of bismuth acid dimethyl ester added and the melanin production rate (%) in Example 8.
  • FIG. 9 is a graph showing the number of cells and melanin production rate (%) when the concentration of hibiscus acid and garcinic acid in the well is 0.5 mgZml in a test example.

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Abstract

[PROBLEMS] To provide a skin-whitening agent, a skin-whitening cosmetic, a skin-whitening food and a composition for oral use having sustained, excellent melanin production inhibitory effect or skin whitening effect. [MEANS FOR SOLVING PROBLEMS] A melanin production inhibitory agent comprising one or more compounds selected from the group consisting of hibiscus acid and hibiscus acid derivatives represented by the general formula (1) and salts thereof as the active ingredient(s); and a skin-whitening agent, a skin-whitening cosmetic, a skin-whitening food or a composition for oral use comprising the melanin production inhibitory agent as the active ingredient. (1) wherein R1 and R2 independently represent a hydrogen atom, a substituted or unsubstituted alkyl group or a benzyl group.

Description

メラニン生成抑制剤  Melanin production inhibitor
技術分野  Technical field
[0001] 本発明は、ハイビスカス酸類を有効成分とするメラニン生成抑制剤、及び該メラニン 生成抑制剤を含む美白剤、美白用化粧料、美白用食品及び口腔用組成物に関する 背景技術  TECHNICAL FIELD [0001] The present invention relates to a melanin production inhibitor containing hibiscus acids as active ingredients, and a whitening agent, a whitening cosmetic, a whitening food and an oral composition containing the melanin production inhibitor.
[0002] 従来、シミやそばかす等の色素の沈着は、皮膚細胞内でメラニンが生成し、蓄積す ること〖こよって生じることが知られている。また、皮膚だけでなく口腔内粘膜内でもメラ ニンが生成すると 、われて!/、る。  Conventionally, it is known that the deposition of pigments such as stains and freckles is caused by the formation and accumulation of melanin in skin cells. In addition, when melanin is generated not only in the skin but also in the mucous membrane of the oral cavity, it will be broken!
[0003] メラニンの生成過程については未だ十分に解明されているとはいえないが、その主 要な生成過程の一つとして以下に示す過程が明ら力となっている。即ち、(1)皮膚が 紫外線を浴びることによって皮膚の表皮細胞に活性酸素が発生する、(2)活性酸素 によって細胞が損傷を受け、種々の情報伝達物質が生成 *放出される、(3)種々の 情報伝達物質によって、チロシナーゼの生成促進および活性化が引き起こされ、メラ ニンが生成する。  [0003] The production process of melanin has not yet been fully elucidated, but the following process has become clear as one of its main production processes. That is, (1) active oxygen is generated in the epidermis cells of the skin when the skin is exposed to ultraviolet rays, (2) cells are damaged by active oxygen, and various information-transmitting substances are generated and released, (3) Various signaling substances cause the promotion and activation of tyrosinase and produce melanin.
[0004] 従って、メラニンの生成を抑制して肌を白く保つには、皮膚上の活性酸素を消去す ること、チロシナーゼの生成を抑制すること、或いはチロシナーゼ活性を阻害すること 等が有効な手段として考えられて 、る。  [0004] Therefore, in order to suppress the production of melanin and keep the skin white, it is effective to eliminate active oxygen on the skin, inhibit the production of tyrosinase, or inhibit tyrosinase activity. It is considered as
[0005] これまでにァスコルビン酸等の活性酸素消去剤ゃコウジ酸やアルブチン等のチロ シナーゼ活性阻害剤が美白剤の有効成分として提案されて!ヽる (特許文献 1等参照 )。し力しながら、従来の美白剤の有効成分として提案されている物質は、メラニンの 生成過程の一部分に作用するに過ぎず、メラニンは他の過程によっても生成するた め、そのメラニン生成抑制効果は満足できるものではな力つた。さらに、従来の美白 剤成分では、効果の持続性の観点力もも満足できるものではな力つた。  [0005] To date, active oxygen scavengers such as ascorbic acid and tyrosinase activity inhibitors such as kojic acid and arbutin have been proposed as active ingredients for whitening agents! Speak (see Patent Document 1 etc.). However, the substances proposed as the active ingredients of conventional whitening agents act only on part of the melanin production process, and melanin is also produced by other processes. The power was not satisfactory. Furthermore, the conventional whitening ingredients have not been able to satisfy the viewpoint of sustaining the effect.
[0006] 一方、ハイビスカス酸類を含有させたィ匕粧料や医薬品が知られて 、る。例えば、特 許文献 2には、 3 ヒドロキシ—3, 4 ジカルボキシ 1, 4ーブタノリド又はその誘導 体を含有することを特徴とする化粧料が記載されている。また、特許文献 3には、ハイ ビスカス酸類を有効成分として含有するグリコシダーゼ阻害剤及び糖尿病治療剤が 記載されている。しかし、ハイビスカス酸類のメラニン生成抑制作用及び美白作用に つ!ヽては今まで知られて 、なかった。 [0006] On the other hand, cosmetics and pharmaceuticals containing hibiscus acids are known. For example, Patent Document 2 includes 3 hydroxy-3,4 dicarboxy 1,4-butanolide or derivatives thereof. Cosmetics characterized in that they contain a body are described. Patent Document 3 discloses a glycosidase inhibitor and a therapeutic agent for diabetes containing hibiscus acids as active ingredients. However, Hibiscus acids are effective in inhibiting melanin production and whitening! Never before had it been known.
特許文献 1:特開平 1— 85907号公報  Patent Document 1: JP-A-1-85907
特許文献 2:特開 200 — 154134号公報  Patent Document 2: Japanese Patent Application Laid-Open No. 200-154134
特許文献 3 :特開 2000— 239164号公報  Patent Document 3: Japanese Patent Laid-Open No. 2000-239164
発明の開示  Disclosure of the invention
発明が解決しょうとする課題  Problems to be solved by the invention
[0007] 本発明は、優れたメラニン生成抑制作用を有し、し力もメラニン生成抑制作用の持 続性があり、且つ、安全に使用し得るメラニン生成抑制剤、美白剤、美白用化粧料、 美白用食品及び口腔用組成物を提供することを主な目的とする。 [0007] The present invention has an excellent melanin production inhibitory action, has a persistent melanin production inhibitory action, and can be used safely, a melanin production inhibitor, a whitening agent, a whitening cosmetic, The main purpose is to provide a food for whitening and a composition for oral cavity.
課題を解決するための手段  Means for solving the problem
[0008] 本発明者らは、上記課題を解決することを目的として鋭意検討を重ねた結果、特定 の構造を有する化合物が優れたメラニン生成抑制作用を有することを見出し、更に 鋭意検討を重ねて本発明を完成するに至った。 [0008] As a result of intensive studies aimed at solving the above-mentioned problems, the present inventors have found that a compound having a specific structure has an excellent melanin production inhibitory effect, and further intensive studies. The present invention has been completed.
[0009] 即ち、本発明は、以下のメラニン生成抑制剤、美白剤、化粧料、食品及び口腔用 組成物に関する。 [0009] That is, the present invention relates to the following melanin production inhibitor, whitening agent, cosmetic, food and oral composition.
[0010] 項 1 :下式(1) [0010] Item 1: Formula (1) below
[0011] [化 1] [0011] [Chemical 1]
Figure imgf000004_0001
Figure imgf000004_0001
[0012] (但し、式中の R1及び R2は、それぞれ水素原子、置換又は非置換のアルキル基又は ベンジル基を示す。 )で表されるハイビスカス酸類及びその塩力 なる群力 選ばれ る 1又は 2種以上の化合物を有効成分とするメラニン生成抑制剤。 (Where R 1 and R 2 are each a hydrogen atom, a substituted or unsubstituted alkyl group, or A benzyl group is shown. A melanin production inhibitor comprising as an active ingredient one or more compounds selected from the group consisting of hibiscus acids represented by the formula (1) and a salt power thereof.
[0013] 項 2 :式(1)で表されるハイビスカス酸類の R1及び R2の少なくとも 1つが置換又は非 置換のアルキル基である項 1に記載のメラニン生成抑制剤。 [0013] Item 2: The melanin production inhibitor according to Item 1, wherein at least one of R 1 and R 2 of the hibiscus acids represented by the formula (1) is a substituted or unsubstituted alkyl group.
[0014] 好ましくは、下式(1)  [0014] Preferably, the following formula (1)
[0015] [化 2] [0015] [Chemical 2]
Figure imgf000005_0001
Figure imgf000005_0001
[0016] (但し、式中の IT及び R2は、それぞれ水素原子、或いは置換又は非置換のアルキル 基又はベンジル基を示し、且つ、 R1及び R2の少なくとも 1つが置換又は非置換のァ ルキル基である。 )で表されるノ、ィビスカス酸類力もなる群力も選ばれる 1又は 2種以 上の化合物を有効成分とするメラニン生成抑制剤。 (In the formula, IT and R 2 each represent a hydrogen atom, a substituted or unsubstituted alkyl group or a benzyl group, and at least one of R 1 and R 2 is a substituted or unsubstituted alkyl group. A melanin production inhibitor comprising as an active ingredient at least one compound selected from the group consisting of bis and biscus acids.
[0017] 好ましくは、ハイビスカス酸モノアルキルエステル及びハイビスカス酸ジアルキルェ ステルカ なる群力 選ばれる 1又は 2種以上の化合物を有効成分とするメラニン生 成抑制剤。  [0017] Preferably, a melanin production inhibitor comprising as an active ingredient one or two or more compounds selected from the group power of hibiscus monoalkyl ester and bisbisester dialkyl esterka.
[0018] 更に好ましくは、ハイビスカス酸モノメチルエステル、ノ、ィビスカス酸モノェチルエス テル、ハイビスカス酸ジメチルエステル及びハイビスカス酸ジェチルエステルからなる 群力 選ばれる 1又は 2種以上の化合物を有効成分とするメラニン生成抑制剤。  [0018] More preferably, melanin production inhibition comprising one or two or more compounds selected from the group power consisting of bismuth acid monomethyl ester, bis, biscus acid monoethyl ester, hibiscus acid dimethyl ester and hibiscus acid dimethyl ester as active ingredients Agent.
[0019] 項 3 :項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む美白 剤。  [0019] Item 3: A whitening agent comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient.
[0020] 項 4:項 1〜2の 、ずれかに記載のメラニン生成抑制剤を有効成分として含む美白 用化粧料。  [0020] Item 4: A whitening cosmetic comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient.
[0021] 項 5 :項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む美白 用 ロロ。 [0022] 項 6 :項 1 2のいずれかに記載のメラニン生成抑制剤を有効成分として含む口腔 用組成物。 Item 5: A whitening roro comprising the melanin production inhibitor according to any one of Items 1 to 2 as an active ingredient. [0022] Item 6: An oral composition comprising the melanin production inhibitor according to any one of Items 12 and 12 as an active ingredient.
[0023] 以下、本発明について、更に詳細に説明する。  [0023] Hereinafter, the present invention will be described in more detail.
[0024] メラニン生成抑制剤 [0024] Melanin production inhibitor
本発明のメラニン生成抑制剤は、下式(1)  The melanin production inhibitor of the present invention has the following formula (1)
[0025] [化 3] [0025] [Chemical 3]
Figure imgf000006_0001
Figure imgf000006_0001
[0026] (但し、式中の R1及び R2は、それぞれ水素原子、置換又は非置換のアルキル基又は ベンジル基を示す。 )で表されるハイビスカス酸類及びその塩力 なる群力 選ばれ る 1又は 2種以上の化合物を有効成分とする。 [0026] (In the formula, R 1 and R 2 each represent a hydrogen atom, a substituted or unsubstituted alkyl group, or a benzyl group.) One or more compounds are active ingredients.
[0027] アルキル基の種類は、本発明の効果が奏される範囲であれば、特に限定されず、 直鎖状でもよぐ分岐鎖状でもよい。 [0027] The type of the alkyl group is not particularly limited as long as the effect of the present invention is exhibited, and may be linear or branched.
[0028] アルキル基の炭素数は、本発明の効果が奏される範囲で適宜設定し得るが、好ま しくは炭素数 1 6程度、特に 1 3程度である。 [0028] The number of carbon atoms of the alkyl group can be set as appropriate within the range in which the effects of the present invention are exerted.
[0029] アルキル基としては、具体的には、メチル基、ェチル基、プロピル基、ブチル基、ィ ソプロピル基、イソブチル基、へキシル基等が挙げられる。 [0029] Specific examples of the alkyl group include a methyl group, an ethyl group, a propyl group, a butyl group, an isopropyl group, an isobutyl group, and a hexyl group.
[0030] アルキル基又はベンジル基は置換基を有して 、てもよ 置換基の種類は、本発 明の効果が奏される範囲で適宜設定し得るが、例えば、炭素数 1 4のアルキル基 等が挙げられる。 [0030] The alkyl group or benzyl group may have a substituent, and the type of the substituent may be appropriately set within the range where the effect of the present invention is exerted. And the like.
[0031] 本発明で使用されるハイビスカス酸類の具体例としては、例えば、式中の R1及び R2 が共に水素原子であるハイビスカス酸、式中の R1及び R2の一方が置換されて 、るハ ィビスカス酸モノエステル、式中の R1及び R2が共に置換されて 、るハイビスカス酸ジ エステルが挙げられる。 [0032] ハイビスカス酸モノエステルには、例えば、ハイビスカス酸モノメチルエステル、ハイ ビスカス酸モノェチルエステル等のハイビスカス酸モノアルキルエステルなどが含ま れる。 [0031] Specific examples of hibiscus acids used in the present invention include, for example, hibiscus acid R 1 and R 2 in the formula are hydrogen atoms, one of R 1 and R 2 in the formula is substituted And Hibiscus acid diester wherein R 1 and R 2 in the formula are both substituted. [0032] Hibiscus acid monoesters include, for example, hibiscus monoalkyl esters such as hibiscus monomethyl ester and bisbis monoethyl ester.
[0033] ハイビスカス酸ジエステルには、例えば、ハイビスカス酸ジメチルエステル、ノ、イビス カス酸ジェチルエステル等のハイビスカス酸ジアルキルエステルなどが含まれる。  [0033] Hibiscus acid diesters include, for example, bismuth acid dialkyl esters such as bismuth acid dimethyl ester, and biscus acid decyl ester.
[0034] 本発明で使用されるハイビスカス酸類の塩には、上記ハイビスカス酸類の塩、例え ば、 R1及び Z又は R2が水素原子である場合にそれらが 1価や 2価の金属塩基で置 換されているアルカリ金属塩やアルカリ土類金属塩が含まれる。 [0034] The salts of hibiscus acids used in the present invention include the above-mentioned salts of hibiscus acids, for example, when R 1 and Z or R 2 are hydrogen atoms, they are monovalent or divalent metal bases. This includes alkali metal salts and alkaline earth metal salts that have been replaced.
[0035] アルカリ金属塩としては、例えば、ナトリウム塩やカリウム塩が挙げられる。また、ァ ルカリ土類金属塩としては、例えば、カルシウム塩やマグネシウム塩などが挙げられ る。また、塩には、錯体を形成しているものも含まれる。例えば、錯体の例としては、 R [0035] Examples of the alkali metal salt include sodium salt and potassium salt. Examples of alkaline earth metal salts include calcium salts and magnesium salts. The salt also includes those forming a complex. For example, an example of a complex is R
1及び Z又は R2がカルシウムやマグネシウムで置換されて形成されたアルカリ土類金 属錯体が挙げられる。 Examples include alkaline earth metal complexes formed by replacing 1 and Z or R 2 with calcium or magnesium.
[0036] 本発明で使用されるハイビスカス酸類及びその塩の生産手段は、特に限定されず 、化学的に合成して得られるものでもよぐ植物体やその抽出物など力 分離して得 られるものでもよい。また、エステル体や塩は、植物体やその抽出物から分離して得 られるハイビスカス酸類を、常法に従ってエステルイ匕したり、各種塩基と反応させたり することにより得られるちのでもよい。  [0036] The means for producing the hibiscus acids and salts thereof used in the present invention are not particularly limited, and may be obtained by force separation such as a plant body or an extract thereof which may be obtained by chemical synthesis. But you can. In addition, the ester or salt may be obtained by esterifying a hibiscus acid obtained by separating from a plant or its extract according to a conventional method or reacting with various bases.
[0037] 植物体やその抽出物からハイビスカス酸類を分離する方法は、特に限定されず、 公知の方法を適宜用いることができる。例えばハイビスカス酸を含有して ヽる植物体 を、適当な抽出溶媒で、抽出した後、クロマトグラフィー等の手段を用いて分離すれ ばよい。  [0037] A method for separating hibiscus acids from a plant body or an extract thereof is not particularly limited, and a known method can be appropriately used. For example, a plant that contains hibiscus acid may be extracted with a suitable extraction solvent and then separated by means such as chromatography.
[0038] ハイビスカス酸類を含有している植物には、例えば、フヨゥ属に属する植物が挙げ られる。  [0038] Examples of plants containing hibiscus acids include plants belonging to the genus Fuyu.
[0039] フヨゥ属に属する植物としては、例えば熱帯原産の一年草ロゼル(Hibiscus sabdarif faし)、熱帯性低木のブッソゥゲ(Hibiscus  [0039] Examples of plants belonging to the genus Fuyou include the annual annual Roselle (Hibiscus sabdarif fa) native to the tropics and the bushy geese (Hibiscus) of the tropical shrub.
rosa- sinensis L.)、中国原産のムクゲ(Hibiscus syriacus L.)、アフリカ原産のケナフ( Hibiscus cannabinus L.)、北米原産のモミジァオイ (Hibiscus coccineus L.)、 日本原産のハマボウ(Hibiscus hamabo)、中国原産のフヨウ(H¾iscus rosa- sinensis L.), Chinese mugwort (Hibiscus syriacus L.), African native kenaf (Hibiscus cannabinus) L.), North American-grown Momiji (Hibiscus coccineus L.), Japanese-grown hawk (Hibiscus hamabo), Chinese-grown Fuyou (H¾iscus
mutabilis L.)などを挙げることができる。このうち、ロゼル、ブッソゥゲ、ムクゲを用いる ことが好ましぐ特に、ロゼル、ブッソゥゲが、好ましい。  mutabilis L.). Of these, it is preferable to use Roselle, Bussouge and Mukuge, and Roselle and Bussouge are particularly preferable.
[0040] 植物体の抽出部位については特に制限されず、例えば該植物の全草、或いは花、 芽、葉、茎、枝、枝先、根、根茎、及び種子等の該植物の一部を挙げることができる。 これらの中で、好ましくは、花、葉を挙げることができる。  [0040] The extraction site of the plant is not particularly limited, and for example, the whole plant of the plant or a part of the plant such as a flower, a bud, a leaf, a stem, a branch, a branch tip, a root, a rhizome, and a seed. Can be mentioned. Among these, Preferably, a flower and a leaf can be mentioned.
[0041] また抽出原料は、植物体をそのまま用いてもよぐその粉砕物、或いは、それらを乾 燥、細切、破砕、圧搾、煮沸或いは発酵処理したものを用いてもよい。  [0041] The extracted raw material may be a pulverized product obtained by using plant bodies as they are, or a product obtained by drying, chopping, crushing, pressing, boiling or fermenting them.
[0042] 抽出に使用される溶媒の種類は、適宜設定し得るが、例えば、水又は各種有機溶 媒、或いは水と有機溶媒の混合物が挙げられる。有機溶媒には、例えば、メタノール 、エタノール、 n—プロパノール、イソプロパノール、ブタノール、イソブタノール、へキ サノール、プロピレングリコール、 1, 3—ブチレングリコール、アセトン、酢酸ェチル、 クロ口ホルム、ペンタン、へキサン、ヘプタン、塩酸等の単独或いは 2種以上の組み合 わせが含まれる。このうち、好ましい抽出溶媒としては、メタノール、エタノール、イソ プロパノール等の低級アルコール、あるいは水と低級アルコールの混合物を挙げるこ とがでさる。 [0042] The type of the solvent used for extraction can be set as appropriate, and examples thereof include water, various organic solvents, and a mixture of water and an organic solvent. Examples of the organic solvent include methanol, ethanol, n -propanol, isopropanol, butanol, isobutanol, hexanol, propylene glycol, 1,3-butylene glycol, acetone, ethyl acetate, black mouth form, pentane, hexane, Including heptane, hydrochloric acid, etc. alone or in combination of two or more. Among these, preferable extraction solvents include lower alcohols such as methanol, ethanol, and isopropanol, or a mixture of water and lower alcohols.
[0043] 抽出後は、溶解、再抽出、分液、傾斜、濾過、濃縮、蒸留、昇華、結晶化、薄層クロ マトグラフィー、カラムクロマトグラフィー、ガスクロマトグラフィー、高速液体クロマトグ ラフィーなどの各種クロマトグラフィーを用いた分離等の各種方法を単独で又は組み 合わせて、適宜精製を行うことができる。  [0043] After extraction, various chromatographies such as dissolution, re-extraction, separation, gradient, filtration, concentration, distillation, sublimation, crystallization, thin-layer chromatography, column chromatography, gas chromatography, and high-performance liquid chromatography. Various methods such as separation using a graph can be purified appropriately, alone or in combination.
[0044] また必要に応じて、さらに、減圧乾燥や凍結乾燥等の乾燥処理を施すことにより水 分を取り除き、乾燥品として用いることもできる。  [0044] Further, if necessary, water can be removed by performing a drying treatment such as reduced-pressure drying or freeze-drying to use as a dried product.
[0045] 抽出の際の具体的条件も、特に制限されるものではなぐ常法に従って適宜設定 することができる。例えば、乾燥したロゼルの花を粉砕し、これに抽出対象物 1重量部 に対して 10重量部程度の 1.5%塩酸—エタノール水溶液をカ卩え、室温で撹拌した 後、濾過により固形分を取り除き、更に HPLCにて精製を行う方法を挙げることができ る。 [0046] また、ノ、イビスカス酸類は、簡便には、商業的に入手したものを用いることもできる。 また、エステル体は、商業的に入手したハイビスカス酸を公知の方法に従って、エス テノレイ匕すること〖こより、得ることちできる。 [0045] Specific conditions for extraction can also be appropriately set according to a conventional method without particular limitation. For example, a dried roselle flower is crushed, and about 10 parts by weight of 1.5% hydrochloric acid-ethanol aqueous solution is added to 1 part by weight of the extraction target, stirred at room temperature, and then solids are removed by filtration. Furthermore, a method of performing purification by HPLC can be mentioned. [0046] In addition, as commercially available bis and ibiscus acids, commercially available ones can also be used. Further, the ester can be obtained from commercially available hibiscus acid by subjecting it to esterification according to a known method.
[0047] 本発明のメラニン生成抑制剤の有効成分として用いる化合物は、上記ハイビスカス 酸類又はその塩力 なる群力 選ばれる 1種単独の化合物であってもよぐ 2種以上 の化合物力もなる混合物であってもよ 、。  [0047] The compound used as the active ingredient of the melanin production inhibitor of the present invention may be a single compound selected from the above-mentioned hibiscus acids or a group power of salt power thereof, or a mixture of two or more compound powers. May be.
[0048] 2種以上の化合物力 なる混合物は、合成又は植物体から単離して得られる各種 ハイビスカス酸類又はその塩を混合して得られるものであってもよぐノ、ィビスカス酸 類を含有する植物体又はその抽出物から得られる混合物や分画物であってもよい。  [0048] The mixture consisting of two or more kinds of compounds contains biscus acid or biscus acid which may be obtained by mixing various hibiscus acids or salts thereof obtained by synthesis or isolation from plants. It may be a mixture or a fraction obtained from a plant or an extract thereof.
[0049] 本発明のメラニン生成抑制剤は、上記ハイビスカス酸類及びその塩力もなる群から 選ばれる 1又は 2以上の化合物を有効成分として含むものであり、好ましくは、ハイビ スカス酸モノアルキルエステル及びハイビスカス酸ジアルキルエステルからなる群か ら選ばれる 1又は 2以上の化合物を有効成分として含むものである。  [0049] The melanin production inhibitor of the present invention contains, as an active ingredient, one or more compounds selected from the group consisting of the above-mentioned hibiscus acids and their salt strength, and preferably a hibiscus monoalkyl ester and hibiscus It contains one or more compounds selected from the group consisting of acid dialkyl esters as active ingredients.
[0050] また、更に好ましくは、ハイビスカス酸モノメチルエステル、ノ、ィビスカス酸モノジメチ ルエステル、ハイビスカス酸ジメチルエステル及びハイビスカス酸ジェチルエステル 力 なる群力 選ばれる 1又は 2種以上の化合物を有効成分として含むものである。  [0050] Further, more preferably, it includes one or more compounds selected as an active ingredient, which are selected from bismuth acid monomethyl ester, bis, biscus acid monodimethyl ester, hibiscus acid dimethyl ester, and hibiscus acid dimethyl ester. .
[0051] 本発明のメラニン生成抑制剤は、上記ハイビスカス酸類及びその塩力もなる群から 選ばれる 1又は 2以上の化合物を有効成分として含み、高く且つ持続的なチロシナ ーゼ活性阻害作用及び高いチロシナーゼ生成抑制作用を有し、効果的で且つ持続 的なメラニン生成抑制作用を奏するものである。  [0051] The melanin production inhibitor of the present invention contains, as an active ingredient, one or two or more compounds selected from the group consisting of the above-mentioned hibiscus acids and salts thereof, and has a high and sustained tyrosinase activity inhibiting action and a high tyrosinase. It has a production inhibitory action and exhibits an effective and continuous melanin production inhibitory action.
[0052]  [0052]
本発明の美白剤は、上記本発明のメラニン生成抑制剤を有効成分として含む。  The whitening agent of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
[0053] 本発明の美白剤は、メラニン生成抑制剤そのものからなるものであってもよいし、そ れを有効成分として薬学上又は衛生上許容される担体又は添加物等の他の成分が 配合されて 、るものであってもよ!/、。 [0053] The whitening agent of the present invention may consist of the melanin production inhibitor itself, or it may contain other ingredients such as pharmaceutically or sanitary acceptable carriers or additives as an active ingredient. It could be something! /
[0054] 力かる担体又は添加物の種類及び配合量は、本発明の効果を損なわないことを限 度として、美白剤の剤型又は適用対象に応じて、適宜選択調整することができる。 [0054] The types and blending amounts of the strong carriers or additives can be appropriately selected and adjusted according to the dosage form of the whitening agent or the application target, as long as the effects of the present invention are not impaired.
[0055] また、美白剤の形態についても特に制限されず、適用される製品等の剤型、形態、 用途等に応じて、液状、乳液状、クリーム状、粉末状、顆粒、丸剤、軟膏等に任意に 調製することができる。 [0055] Further, the form of the whitening agent is not particularly limited, and the dosage form and form of the product to be applied, Depending on the application, etc., it can be arbitrarily prepared into liquids, emulsions, creams, powders, granules, pills, ointments and the like.
[0056] また、本発明の美白剤は、外用剤とすることも、また内用剤とすることもできる。  [0056] Further, the whitening agent of the present invention may be an external preparation or an internal preparation.
[0057] 本発明の美白剤を外用剤として処方するにあたっては、必要に応じて、無機顔料、 紫外線吸収剤、他の美白成分、チロシナーゼ活性阻害剤又はメラニン色素還元剤、 界面活性剤、細胞賦活剤、抗炎症剤、抗菌剤、保湿剤、香料、着色剤等の添加剤を 適宜添カ卩してもよい。 [0057] In formulating the whitening agent of the present invention as an external preparation, if necessary, inorganic pigments, ultraviolet absorbers, other whitening components, tyrosinase activity inhibitors or melanin dye reducing agents, surfactants, cell activation agents. Additives such as agents, anti-inflammatory agents, antibacterial agents, moisturizers, fragrances, and coloring agents may be added as appropriate.
[0058] 本発明の美白剤を適用できる外用剤の種類は、特に制限されないが、例えば、外 用医薬品類や外用医薬部外品類 (皮膚外用剤等)、基礎化粧料 (化粧水、乳液、タリ ーム、軟膏、ローション、オイル、パック等)、メークアップ化粧料(ファンデーション、口 紅、頰紅、マスカラ等)、その他皮膚用化粧料 (洗顔料、皮膚洗浄料、マッサージ剤、 クレンジング剤等)、口腔用組成物、あるいは浴用剤等が挙げられる。  [0058] The types of external preparations to which the whitening agent of the present invention can be applied are not particularly limited. For example, external medicines and external quasi drugs (skin external preparations, etc.), basic cosmetics (skin lotion, emulsion, Tarim, ointment, lotion, oil, pack, etc.), makeup cosmetics (foundation, lipstick, scarlet, mascara, etc.), other skin cosmetics (face wash, skin cleanser, massage agent, cleansing agent, etc.) ), Oral compositions, bath preparations, and the like.
[0059] 本発明の美白剤を外用剤として処方する場合、その外用剤の配合量は、配合対象 、剤型、期待される効果等によって異なり、一律に規定することはできないが、通常、 外用剤全体に対し、ハイビスカス酸類及びその塩として、 0.00001〜10重量%、好ま しくは 0.00005〜5重量%、更に好ましくは 0.0001〜3重量%程度である。  [0059] When the whitening agent of the present invention is formulated as an external preparation, the amount of the external preparation varies depending on the compounding target, dosage form, expected effect, etc., and cannot be uniformly defined. Hibiscus acids and salts thereof are 0.00001 to 10% by weight, preferably 0.00005 to 5% by weight, more preferably about 0.0001 to 3% by weight, based on the whole agent.
[0060] 本発明の美白剤を内用剤として処方するにあたっては、必要に応じて、結合剤、崩 壊剤、滑沢剤、湿潤化剤、緩衝剤、賦形剤、増量剤、保存剤、香料等の添加剤を配 合することができる。また、本発明の美白剤を内用剤として処方するにあたって、更に 他の薬学的活性成分を配合することもできる。  [0060] In formulating the whitening agent of the present invention as an internal preparation, a binder, a disintegrant, a lubricant, a wetting agent, a buffering agent, an excipient, an extender, and a preservative are used as necessary. In addition, additives such as fragrances can be combined. In addition, when formulating the whitening agent of the present invention as an internal preparation, other pharmaceutically active ingredients can be further blended.
[0061] 本発明の美白剤を適用できる内用剤の種類は、特に制限されないが、例えば、散 剤、錠剤、顆粒剤、カプセル剤、あるいはシロップ剤等の経口用剤が挙げられる。  [0061] The type of internal preparation to which the whitening agent of the present invention can be applied is not particularly limited, and examples thereof include oral preparations such as powders, tablets, granules, capsules, and syrups.
[0062] 本発明の美白剤を内用剤として処方する場合、その内用剤の配合割合は、投与対 象、剤型、投与方法等によっても異なり、一律に規定することはできないが、例えば、 内用剤全体に対し、ハイビスカス酸類及びその塩として、 0.00001〜100重量%、好ま しくは 0.0001〜50重量%、更に好ましくは 0.001〜30重量%程度である。  [0062] When the whitening agent of the present invention is formulated as an internal preparation, the blending ratio of the internal preparation varies depending on the administration target, dosage form, administration method, etc., and cannot be uniformly defined. The amount of hibiscus acids and salts thereof is 0.00001 to 100% by weight, preferably 0.0001 to 50% by weight, more preferably about 0.001 to 30% by weight, based on the whole internal preparation.
[0063] 本発明の美白剤は、上記ハイビスカス酸類及びその塩力 なる群力 選ばれる 1又 は 2種以上の化合物を有効成分として含有するメラニン生成抑制剤を有効成分とし て含み、高く且つ持続的なチロシナーゼ活性阻害作用及び高いチロシナーゼ生成 抑制作用を含む、持続的で優れたメラニン生成抑制作用を有し、持続的で優れた美 白作用を奏するものである。 [0063] The whitening agent of the present invention comprises, as an active ingredient, a melanin production inhibitor containing as an active ingredient one or more compounds selected from the above-mentioned hibiscus acids and the group power of their salt strength. In addition, it has a continuous and excellent melanin production inhibitory action including a high and sustained tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and exhibits a continuous and excellent whitening action.
[0064] &m^M  [0064] & m ^ M
本発明の美白用化粧料は、上記本発明のメラニン生成抑制剤を有効成分として含 む。  The whitening cosmetic composition of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
[0065] 化粧料の種類は、特に限定されず、種々の化粧料に適用できる。たとえば、タリー ム、乳液、化粧水、ノ ック剤、洗顔料、ノ《ツチなどの各種基礎ィ匕粧料、ファンデーンョ ン、ほほ紅、白粉などの各種メーキャップ料、洗髪料、養毛剤、シャンプー、リンスな どの各種頭髪用化粧料、石鹼、美爪料、オーデコロン、入浴剤等の各種化粧料に適 用できる。  [0065] The type of cosmetic is not particularly limited, and can be applied to various cosmetics. For example, creams, emulsions, lotions, knocking agents, facial cleansers, various basic cosmetics such as powders, foundations, cheeks, white powders, makeup products, hair washing products, hair nourishing agents, shampoos, etc. It can be applied to various types of cosmetics such as rinsing and other hair cosmetics, sarcophagus, beauty nails, eau de cologne, and bath salts.
[0066] 化粧料の形態も特に限定されず、溶液、ェマルジヨン、軟膏、ゾル、ゲル、パウダー [0066] The form of the cosmetic is not particularly limited, either, solution, emulsion, ointment, sol, gel, powder.
、スプレー、シート、フィルムなどの種々の形態で用いることができる。 , Sprays, sheets, films, and the like.
[0067] 本発明の化粧料には、本発明の効果を奏する範囲内で、一般に化粧料に配合さ れる各種成分、例えば、水性成分、粉末成分、保湿剤、界面活性剤、防腐剤、増粘 剤、紫外線吸収剤、香料等を配合することができる。 [0067] In the cosmetic of the present invention, various components that are generally blended in cosmetics, for example, an aqueous component, a powder component, a moisturizer, a surfactant, a preservative, an increase agent, within the scope of the effects of the present invention. Adhesives, ultraviolet absorbers, fragrances and the like can be blended.
[0068] 本発明の化粧料の製造方法は特に限定されず、公知の方法に従って、適宜製造 することができる。 [0068] The method for producing the cosmetic of the present invention is not particularly limited, and can be suitably produced according to a known method.
[0069] 本発明の化粧料の適用方法も特に限定されないが、例えば、顔、首、腕及び手等 のシミ、ソバカス等のでき易い部位もしくは患部、 日焼けし易い部位もしくは日焼けし た部位等に、 1日 1回乃至数回、適当量塗布すれば良い。  [0069] The method of applying the cosmetic composition of the present invention is not particularly limited, but may be applied to, for example, an easily or affected part such as a stain on the face, neck, arm or hand, or freckles, or an easily tanned or tanned part. Apply an appropriate amount once or several times a day.
[0070] 本発明の化粧料におけるメラニン生成抑制剤の含有割合は、本発明の効果が奏し 得る範囲で適宜設定し得るが、化粧料組成物全体に対し、ハイビスカス酸類及びそ の塩として、 0.00001〜10重量%、好ましくは 0.00005〜5重量%、更に好ましくは 0.00 01〜3重量%程度である。  [0070] The content ratio of the melanin production inhibitor in the cosmetic of the present invention can be appropriately set within the range in which the effect of the present invention can be exerted. However, as a hibiscus acid and its salt with respect to the entire cosmetic composition, 0.00001 -10% by weight, preferably 0.00005-5% by weight, more preferably 0.0001-3% by weight.
[0071] 本発明の美白用化粧料は、上記ハイビスカス酸類及びその塩力 なる群力 選ば れる 1又は 2種以上の化合物を有効成分として含有するメラニン生成抑制剤を有効 成分として含み、高く且つ持続的なチロシナーゼ活性阻害作用及び高いチロシナー ゼ生成抑制作用を含む、持続的で優れたメラニン生成抑制作用を有し、持続的で優 れた美白作用を奏するものである。 [0071] The whitening cosmetic composition of the present invention includes a melanin production inhibitor containing, as an active ingredient, the above-mentioned hibiscus acid and one or more compounds selected from the group power of its salt strength as an active ingredient, and is high and durable. Tyrosinase activity inhibitory activity and high tyrosiner It has a continuous and excellent melanin production-inhibiting action, including a zeogenesis-inhibiting action, and has a continuous and excellent whitening effect.
[0072] 塞白用食品  [0072] Food for whitening
本発明の美白用食品は、有効成分として上記本発明のメラニン生成抑制剤を含む  The whitening food of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
[0073] 本発明の美白用食品は、例えば、健康食品、食品添加剤、機能性食品、栄養機能 食品、保健機能食品などとして用いることができる。 [0073] The whitening food of the present invention can be used as, for example, health foods, food additives, functional foods, nutritional functional foods, health functional foods, and the like.
[0074] 美白用食品の製造方法は特に限定されず、公知の方法に従って、適宜製造するこ とができる。例えば、上記美白剤を各種食品又は食品素材と混合することにより、得 ることがでさる。 [0074] The method for producing the whitening food is not particularly limited, and can be suitably produced according to a known method. For example, it can be obtained by mixing the whitening agent with various foods or food materials.
[0075] 食品の種類は、特に限定されず、公知の各種飲食品から適宜設定することができ る。例えばクッキー、シリアル、キャンディー、タブレット、ゼリー、水産練り製品、調味 料、パン、清涼飲料、炭酸飲料、ミネラルウォーター、果汁飲料、茶類、栄養飲料、ド リンク剤等として用いることができる。  [0075] The type of food is not particularly limited, and can be appropriately set from various known foods and drinks. For example, it can be used as cookies, cereals, candy, tablets, jelly, marine products, seasonings, bread, soft drinks, carbonated drinks, mineral water, fruit juice drinks, teas, nutrition drinks, drinks, and the like.
[0076] また、本発明の食品には、本発明の効果を損なわない範囲で、通常の飲食品に用 いられる他の成分を配合することができる。他の成分としては、例えば、各種ビタミン 、ミネラル、動植物成分、賦形剤、甘味料、香味剤、着色剤、防腐剤、崩壊剤、滑沢 剤、湿潤化剤、増量剤、香料などが挙げられる。  [0076] In addition, the food of the present invention can be blended with other components used in ordinary foods and drinks within a range not impairing the effects of the present invention. Examples of other ingredients include various vitamins, minerals, animal and plant ingredients, excipients, sweeteners, flavoring agents, coloring agents, preservatives, disintegrating agents, lubricants, wetting agents, bulking agents, and flavoring agents. It is done.
[0077] 本発明の食品におけるメラニン生成抑制剤の含有割合は、本発明の効果が奏し得 る範囲で適宜設定し得るが、食品組成物全体に対し、ハイビスカス酸類及びその塩 として、 0.00001〜10重量%、好ましくは 0.00005〜5重量%、更に好ましくは 0.0001〜 3重量%程度である。 [0077] The content of the melanin production inhibitor in the food of the present invention can be set as appropriate within the range in which the effects of the present invention can be exerted, but 0.00001 to 10 as hibiscus acids and salts thereof with respect to the entire food composition. % By weight, preferably 0.00005 to 5% by weight, more preferably about 0.0001 to 3% by weight.
[0078] 本発明の美白用食品を食する際の摂取量は、適用対象の年齢、体重、症状などに 応じて適宜設定し得るが、通常、成人 1日当たり、ハイビスカス酸類及びその塩として 、 0.00001〜50mg程度、好ましくは 0.0001〜20mg程度、さらに好ましくは 0.001〜10m g程度である。  [0078] The amount of intake when eating the whitening food of the present invention can be appropriately set according to the age, weight, symptoms, etc. of the subject of application, but is generally 0.00001 per day for adults as hibiscus acids and salts thereof. About 50 mg, preferably about 0.0001-20 mg, more preferably about 0.001-10 mg.
[0079] 本発明の美白用食品は、上記ハイビスカス酸類及びその塩力 なる群力 選ばれ る 1又は 2種以上の化合物を有効成分として含有するメラニン生成抑制剤を有効成 分として含み、高く且つ持続的なチロシナーゼ活性阻害作用及び高いチロシナーゼ 生成抑制作用を含む、持続的で優れたメラニン生成抑制作用を有し、持続的で優れ た美白作用を奏するものである。 [0079] The whitening food of the present invention effectively comprises a melanin production inhibitor containing as an active ingredient one or more compounds selected from the above-mentioned hibiscus acids and the group power of their salt strength. It has a continuous and excellent melanin production inhibitory action including a high and sustained tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and has a continuous and excellent whitening action.
[0080] 口腔用組成物  [0080] Oral composition
本発明の口腔用組成物は、上記本発明のメラニン生成抑制剤を有効成分として含 む。  The composition for oral cavity of the present invention contains the melanin production inhibitor of the present invention as an active ingredient.
[0081] 口腔用組成物の種類は、特に限定されず、種々の口腔用組成物に適用できる。  [0081] The type of oral composition is not particularly limited, and can be applied to various oral compositions.
[0082] 例えば、練り歯磨き等の歯磨き剤、洗口液、マウスリンス、口腔用パスタ、トローチ、 歯茎のマッサージクリーム等の各種口腔用組成物に適用できる。 [0082] For example, it can be applied to various oral compositions such as toothpaste such as toothpaste, mouthwash, mouth rinse, oral paste, troche, gum massage cream and the like.
[0083] 口腔用組成物の形態も特に限定されず、溶液、ェマルジヨン、軟膏、ゾル、ゲル、 ペースト、錠剤、パウダー、スプレー、シート、フィルムなどの種々の形態で用いること ができる。 [0083] The form of the oral composition is not particularly limited, and can be used in various forms such as a solution, emulsion, ointment, sol, gel, paste, tablet, powder, spray, sheet, and film.
[0084] 本発明の口腔用組成物には、本発明の効果を奏する範囲内で、一般に口腔用組 成物に配合される各種成分を配合することができる。各種成分としては、例えば、粘 着剤、清涼剤、結合剤、甘味料、着香料、崩壊剤、滑沢剤、着色料、徐放調整剤、 界面活性剤、溶解剤、湿潤剤、 pH調整剤等を挙げることができる。  [0084] In the composition for oral cavity of the present invention, various components generally blended in the composition for oral cavity can be blended within the range where the effects of the present invention are exhibited. Various components include, for example, an adhesive, a refreshing agent, a binder, a sweetener, a flavoring agent, a disintegrant, a lubricant, a colorant, a sustained release regulator, a surfactant, a solubilizer, a wetting agent, and a pH adjuster. An agent etc. can be mentioned.
[0085] 本発明の口腔用組成物の製造方法は特に限定されず、公知の方法に従って、適 宜製造することができる。  [0085] The method for producing the composition for oral cavity of the present invention is not particularly limited, and can be suitably produced according to a known method.
[0086] 本発明の口腔用組成物の適用方法も特に限定されず、例えば、患部もしくは口腔 内全体等に、 1日 1回乃至数回、適当量適用すれば良い。  [0086] The method for applying the oral composition of the present invention is not particularly limited. For example, an appropriate amount may be applied once or several times a day to the affected area or the entire oral cavity.
[0087] 本発明の口腔用組成物におけるメラニン生成抑制剤の含有割合は、本発明の効 果が奏し得る範囲で適宜設定し得るが、口腔用組成物全体に対し、ハイビスカス酸 類及びその塩として、 0.00001〜10重量%、好ましくは 0.00005〜5重量%、更に好ま しくは 0.0001〜3重量%程度である。  [0087] The content ratio of the melanin production inhibitor in the oral composition of the present invention can be appropriately set within the range in which the effect of the present invention can be exerted, but the hibiscus acids and salts thereof with respect to the entire oral composition. As 0.00001 to 10% by weight, preferably 0.00005 to 5% by weight, and more preferably about 0.0001 to 3% by weight.
[0088] 本発明の口腔用組成物は、上記ハイビスカス酸類及びその塩力 なる群力 選ば れる 1又は 2種以上の化合物を有効成分として含有するメラニン生成抑制剤を有効 成分として含み、高く且つ持続的なチロシナーゼ活性阻害作用及び高いチロシナー ゼ生成抑制作用を含む、持続的で優れたメラニン生成抑制作用を有し、歯茎に沈着 した色素を除去し、優れた美白作用を奏するものである。 [0088] The composition for oral cavity of the present invention includes a melanin production inhibitor containing, as an active ingredient, the above-mentioned hibiscus acids and one or more compounds selected from the group power of its salt strength as an active ingredient, and is high and durable. It has a long-lasting and excellent melanin production-inhibiting action, including a typical tyrosinase activity-inhibiting action and a high tyrosinase-inhibiting action, and is deposited on the gum The pigment | dye which carried out is removed and there exists an outstanding whitening effect | action.
発明の効果  The invention's effect
[0089] 本発明によれば、特定の構造を有するハイビスカス酸及びその塩力 なる群力 選 ばれる 1種又は 2種以上の化合物を有効成分とするメラニン生成抑制剤が提供され る。  [0089] According to the present invention, there is provided a melanin production inhibitor comprising as an active ingredient one or more compounds selected from the group strength of hibiscus acid having a specific structure and its salt strength.
[0090] 本発明のメラニン生成抑制剤の有効成分となる化合物は、高いチロシナーゼ活性 阻害作用及び高いチロシナーゼ生成抑制作用を含む、優れたメラニン生成抑制作 用を有し、且つ、優れた美白効果を奏するものである。更に該美白効果は長時間に わたって奏される。し力ゝも該化合物は安全性も高い。  [0090] The compound as an active ingredient of the melanin production inhibitor of the present invention has an excellent melanin production inhibitory action including a high tyrosinase activity inhibitory action and a high tyrosinase production inhibitory action, and has an excellent whitening effect. It is what you play. Furthermore, the whitening effect is achieved for a long time. However, the compound is also highly safe.
[0091] このような化合物を有効成分とする本発明のメラニン生成抑制剤は、優れた美白作 用を有し、しかも美白作用の持続性を有するものであり、皮膚細胞や口腔内粘膜等 で発生するメラニンを有効に抑制し、シミやそばかす等の色素の沈着を効果的に抑 えることができる。更に本発明のメラニン生成抑制剤は安心して使用し得るものである  [0091] The melanin production inhibitor of the present invention comprising such a compound as an active ingredient has an excellent whitening effect and has a long-lasting whitening effect, and is effective in skin cells, oral mucosa and the like. It effectively suppresses the melanin produced and effectively suppresses the deposition of pigments such as spots and freckles. Furthermore, the melanin production inhibitor of the present invention can be used with peace of mind.
[0092] そして、該メラニン生成抑制剤は、美白剤、美白用化粧料、美白用食品及び口腔 用組成物の有効成分として使用することができ、本発明によって、優れた美白作用を 有し、しかも美白作用の持続性を有し、かつ安全に使用し得る美白剤、美白用化粧 料、美白用食品及び口腔用組成物も提供される。 [0092] The melanin production inhibitor can be used as an active ingredient in whitening agents, whitening cosmetics, whitening foods and oral compositions, and has an excellent whitening action according to the present invention. In addition, a whitening agent, a whitening cosmetic, a whitening food, and a composition for oral cavity that have a long-lasting whitening effect and can be used safely are also provided.
[0093] このような特徴を有する本発明は、近年、特に美白効果に関する関心が高まってい る化粧品産業や医薬品産業、食品産業等において、好適に利用し得るものである。 発明を実施するための最良の形態  [0093] The present invention having such characteristics can be suitably used in the cosmetic industry, the pharmaceutical industry, the food industry, and the like, which have recently been particularly interested in the whitening effect. BEST MODE FOR CARRYING OUT THE INVENTION
[0094] 以下、実施例、製造例及び試験例により本発明を詳細に説明するが、本発明はこ れらに限定されるものではない。  [0094] Hereinafter, the present invention will be described in detail with reference to Examples, Production Examples, and Test Examples, but the present invention is not limited thereto.
[0095] 製诰例 1 (ハイビスカス酸)  [0095] Example 1 (Hibiscus acid)
乾燥した Hibiscus sabdariffa L.の花 800gをミルで粉砕し、これに 1.5%塩酸-メタノー ル水溶液 8Lを加え、室温で撹拌しながら 3時間抽出処理した。次いで、濾過により固 形分を取り除き、濾液をエバポレーター (45°C)にて濃縮した。この濾液を ΗΡ-20 ( φ = 70mm、 h=500mm)カラムクロマトグラフィーにて精製、濃縮し、粗生成物 118gを得 た。 800 g of dried Hibiscus sabdariffa L. flowers were pulverized with a mill, and 8 L of 1.5% hydrochloric acid-methanol aqueous solution was added thereto, followed by extraction treatment with stirring at room temperature for 3 hours. Subsequently, the solid content was removed by filtration, and the filtrate was concentrated with an evaporator (45 ° C.). This filtrate was purified and concentrated by ΗΡ-20 (φ = 70 mm, h = 500 mm) column chromatography to obtain 118 g of a crude product. It was.
[0096] 得られた粗生成物を ODS ( φ = 110mm, h= 1200mm)カラムクロマトグラフィーにて 数回精製濃縮を行い、その後結晶化を経て、ノ、イビスカス酸 34.8gを得た。  [0096] The obtained crude product was purified and concentrated several times by ODS (φ = 110 mm, h = 1200 mm) column chromatography, and then crystallized to obtain 34.8 g of biscus acid.
[0097] 確認のため得られた物質を分析したところ、マススペクトルによる分子量は 190であ つた。またプロトン NMRは次のとおりであった。  [0097] The substance obtained for confirmation was analyzed. As a result, the molecular weight according to the mass spectrum was 190. Proton NMR was as follows.
[0098] プロトン NMR ^ベクトル(400MHz) (アセトン一 d6) δ (ppm) : 5.35 (1H, S) , 3.26 (1H , d, J = 17.2Hz) , 2.78 (1H, d, J=17.2Hz) 0 [0098] Proton NMR ^ vector (400MHz) (acetone d6) δ (ppm): 5.35 (1H, S), 3.26 (1H, d, J = 17.2Hz), 2.78 (1H, d, J = 17.2Hz) 0
これらの値は、 Bollら(Acta Chemica Scandinavica,1969年, Vol.23, 286- 293頁)によ つて報告されたノ、ィビスカス酸の値と一致した。  These values were consistent with the values for bis and biscus acids reported by Boll et al. (Acta Chemica Scandinavica, 1969, Vol. 23, pp. 286-293).
[0099] 製诰例 2 (ノヽィビスカス酸モノェチルエステル)  [0099] Example 2 (Nobiscus monoethyl ester)
乾燥した Hibiscus sabdariffa L.の花 100gをミルで粉砕し、これに 30%エタノール水 溶液 500mLを加え、室温で撹拌しながら 3日間抽出処理した。次いで、酢酸ェチル抽 出を行い、得られた水層を更にブタノールで抽出し、その後、シリカゲルクロマトダラ フィー (溶出液:酢酸ェチル—メタノール溶液)にて、精製を行い、分画物を得た。  100 g of dried Hibiscus sabdariffa L. flowers were pulverized with a mill, added with 500 mL of 30% aqueous ethanol, and extracted for 3 days with stirring at room temperature. Next, extraction with ethyl acetate was performed, and the obtained aqueous layer was further extracted with butanol, and then purified with silica gel chromatography (eluent: ethyl acetate-methanol solution) to obtain a fraction. .
[0100] 得られた分画物について分析したところ、マススペクトルによる分子量は 218であつ た。また、プロトン NMRは、いくつかのシグナルの重なりが見られ、数種の異性体の存 在が予想された。またそのうちの一つと考えられる、以下のシグナルを有する化合物 が認められた。  [0100] When the obtained fraction was analyzed, the molecular weight was 218 according to the mass spectrum. In proton NMR, several overlapping signals were observed, and the existence of several isomers was expected. In addition, one of the compounds considered to have one of the following signals was recognized.
[0101] プロトン NMR ^ベクトル(500MHz) (CDC13) δ (ppm) : 5.23 (1H, S) , 3.20 (1H, d, J= 17.4Hz) , 2.90 (1H, d, J=17.4Hz) , 4.29 (2H, m) , 1.37 (3H, t, J=7.4Hz) 0 [0101] Proton NMR ^ vector (500MHz) (CDC13) δ (ppm): 5.23 (1H, S), 3.20 (1H, d, J = 17.4Hz), 2.90 (1H, d, J = 17.4Hz), 4.29 (2H, m), 1.37 (3H, t, J = 7.4Hz) 0
[0102] 以上の分析結果と、 Hibiscus sabdariffaし中にハイビスカス酸が多く含まれる事から 、該分画物には、ハイビスカス酸モノェチルエステル及びその異性体が含まれると推 した。  [0102] From the above analysis results and the fact that Hibiscus sabdariffa contained a large amount of hibiscus acid, it was presumed that the fraction contained hibiscus monoethyl ester and isomers thereof.
[0103] 該ノ、ィビスカス酸モノェチルエステル及びその異性体の構造は、下記 (a)及び (b)と 推察される。  [0103] The structures of the nobis, bisbisic acid monoethyl ester and isomers thereof are presumed to be the following (a) and (b).
[0104] [化 4] H o 0 (b) [0104] [Chemical 4] H o 0 (b)
、H C2H50 H , HC 2 H 5 0 H
O  O
[0105] 以下、得られた分画物を「ノ、ィビスカス酸モノェチルエステル分画物」ともいう。 Hereinafter, the obtained fraction is also referred to as “no biscus acid monoethyl ester fraction”.
[0106] ¾告例 3 (ハイビスカス酸ジメチルエステル) [0106] Example 3 (Hibiscus acid dimethyl ester)
製造例 1で得たハイビスカス酸 450mgをメチル -t-ブチルエーテル(MTBE) 1.8mlとメ タノール 1.8mlの混合溶媒に溶解した。この溶液を氷水浴にて冷却しながら攪拌し、ト リメチルシリルジァゾメタン 2.0Mジェチルエーテル溶液 3.5mlを少しずつ添カ卩した。で きた析出物をろ過し、 MTBEで洗浄後、減圧乾燥させ、ノ、ィビスカス酸ジメチルエステ ルを 300mg得た。  450 mg of Hibiscus acid obtained in Production Example 1 was dissolved in a mixed solvent of 1.8 ml of methyl-t-butyl ether (MTBE) and 1.8 ml of methanol. This solution was stirred while cooling in an ice-water bath, and 3.5 ml of trimethylsilyl diazomethane 2.0M jetyl ether solution was added little by little. The resulting precipitate was filtered, washed with MTBE, and dried under reduced pressure to obtain 300 mg of bis (bisbisamic acid) dimethyl ester.
[0107] 確認のため得られた物質を分析したところ、マススペクトルによる分子量は 218であ つた。またプロトン NMRは次のとおりであった。  [0107] When the substance obtained for confirmation was analyzed, the molecular weight by mass spectrum was 218. Proton NMR was as follows.
[0108] プロトン NMR ^ベクトル(400MHz) (クロ口ホルム dl) δ (ppm) : 5.11 (1H, S) , 3.08 ( 1H, d, J=17.5Hz) , 2.88 (1H, d, J=17.2Hz) , 3.80 (1H, S), 3.84 (3H, S), 3.96 (3H, S[0108] Proton NMR ^ vector (400 MHz) (black mouth form dl) δ (ppm): 5.11 (1H, S), 3.08 (1H, d, J = 17.5Hz), 2.88 (1H, d, J = 17.2Hz ), 3.80 (1H, S), 3.84 (3H, S), 3.96 (3H, S
)。 ).
[0109] これらの値は、 IBnusaudら(Tetrahedron,2002年, Vol.58, 4887- 4892頁)によって報 告されたハイビスカス酸ジメチルエステルの値と一致した。  [0109] These values were consistent with those of hibiscus acid dimethyl ester reported by IBnusaud et al. (Tetrahedron, 2002, Vol. 58, 4887-4892).
[0110] 製造例 4レヽィビスカスエキス) [0110] Production Example 4 Lebiscus extract)
比較として用いるために以下の方法で、ノ、ィビスカスエキス (抽出物)を製造した。  For use as a comparison, a biscus extract (extract) was produced by the following method.
[0111] 乾燥した Hibiscus sabdariffa L.の花 100gをミルで粉砕し、これに 30%エタノール水 溶液 500mLをカ卩え、室温で撹拌しながら 3日間抽出処理した。次いで、濾過により固 形分を取り除き、濾液を減圧濃縮した後、凍結乾燥して 15gの HibkmiSsabdariffa L.抽出物を得た。 [0111] 100 g of dried Hibiscus sabdariffa L. flower was pulverized with a mill, and 500 mL of a 30% aqueous ethanol solution was added to this and extracted for 3 days with stirring at room temperature. Next, the solid content was removed by filtration, and the filtrate was concentrated under reduced pressure, and then freeze-dried to obtain 15 g of Hibkmi Ssabdariffa L. extract.
[0112] ¾施{列 ίチロシナーゼ カ の ¾平 ノ、イビスカス酸のチロシナーゼ生成抑制効果を評価するために、以下のようにゥェ スタンプロットを行った。 [0112] ¾ application {line ί tyrosinase ¾ flat In order to evaluate the inhibitory effect of bis and ibiscus acid on the production of tyrosinase, we stamped lots as follows.
[0113] マウス由来の B16メラノーマ細胞(大日本製薬製、 B16F0)を 25cm2フラスコで 8mlの 培養液中で 24時間培養した後に、製造例 1で得られたノヽィビスカス酸の 5重量%水 溶液をそれぞれ 12.5、 25、 50又は 100ml添加し、 72時間培養を行った。 [0113] Mouse-derived B16 melanoma cells (D16, manufactured by Dainippon Pharmaceutical Co., Ltd., B16F0) were cultured for 24 hours in an 8 ml culture medium in a 25 cm 2 flask. Were added at 12.5, 25, 50 or 100 ml, respectively, and cultured for 72 hours.
[0114] 培養終了後、細胞を粉砕処理してタンパク質を抽出し、電気泳動を行った。次に、 ポリビ-リデンジフルオリド膜 (PVDF膜)にタンパク質を転写し、ャギ抗チロシナーゼ 抗体を反応させた後、さらにアルカリフォスファターゼ標識抗ャギ抗体を反応させ、最 後に発色基質をカ卩えることによって、チロシナーゼを検出した。なお、比較として、ハ ィビスカス酸無添カ卩のものを用いて、同様に試験を行った。  [0114] After completion of the culture, the cells were pulverized to extract proteins, and subjected to electrophoresis. Next, the protein is transferred to a polyvinylidene difluoride membrane (PVDF membrane), reacted with a goat anti-tyrosinase antibody, further reacted with an alkaline phosphatase-labeled anti-goat antibody, and finally a chromogenic substrate is obtained. As a result, tyrosinase was detected. For comparison, the same test was carried out using a bismuth acid-free additive.
[0115] 得られた結果を図 1に示す。図 1は、ノヽィビスカス酸の存在下又は非存在下で培養 した B16メラノーマ細胞のチロシナーゼの発現量を示す PVDF膜像を示す。  [0115] Fig. 1 shows the obtained results. FIG. 1 shows a PVDF membrane image showing the expression level of tyrosinase in B16 melanoma cells cultured in the presence or absence of neubiscus acid.
[0116] 図中レーン Iはハイビスカス酸溶液無添加、レーン IIは 5%ハイビスカス酸溶液を 100 μ 1添加、レーン IIIは 5%ハイビスカス酸溶液 50 μ 1添加、レーン IVは 5%ハイビスカス酸 溶液 25 μ 1添加、レーン Vは 5%ハイビスカス酸 12.5 μ 1添カ卩したサンプルの結果を示 す。  [0116] In the figure, lane I shows no addition of hibiscus acid solution, lane II adds 100 μ1 of 5% hibiscus acid solution, lane III adds 50 μ1 of 5% hibiscus acid solution, lane IV shows 5% hibiscus acid solution 25 Lane 1 shows the results for a sample with 1 μl of 5% bismuth acid added.
[0117] 図 1に示されるバンドの太さから明らかなように、ノ、ィビスカス酸無添加のものとくら ベ、ノ、ィビスカス酸を添加して培養したものは、その添加濃度の上昇に伴って、チロ シナーゼの発現が抑制される傾向があることが確認された。  [0117] As is apparent from the thickness of the band shown in Fig. 1, the cultures added with no, ibiscus acid and those added with no, ibiscus acid increased with the increase of the concentration. Thus, it was confirmed that the expression of tyrosinase tends to be suppressed.
[0118] この結果、ノ、イビスカス酸は、チロシナーゼ生成抑制作用を有することが明らかとな つた ο  As a result, it has been clarified that bis and ibiscus acid have a tyrosinase production inhibitory effect.
[0119] 実施例 2チロシナーゼ活件阳.害効 の評価 (ハイビスカス酸)  Example 2 Tyrosinase Activity V. Evaluation of Harmful Effect (Hibiscus Acid)
ノ、ィビスカス酸のチロシナーゼの活性阻害効果を評価するために、以下の試験を 行った。  In order to evaluate the activity inhibitory effect of tyrosinase of bis and biscus acids, the following tests were conducted.
[0120] マウス由来 B16メラノーマ細胞(大日本製薬製、 B16F0)を 96ウェルマイク口プレート に 5 X 104cells/wellとなるように播種してー晚培養し、細胞を定着させた。次に、培養 プレートの培養液をァスピレーターで取り除き、 1%トライトン X-100を含む PBS (リン酸 緩衝食塩水)を lZwell添加し、細胞の膜構造を破壊して、粗チロシナーゼ溶液 とした。 [0120] Mouse-derived B16 melanoma cells (D16, manufactured by Dainippon Pharmaceutical Co., Ltd., B16F0) were seeded in a 96-well microphone mouth plate at 5 × 10 4 cells / well, and cultured in sputum to establish the cells. Next, remove the culture solution from the culture plate with an aspirator, add 1Z Triton X-100-containing PBS (phosphate buffered saline) to lZwell, destroy the cell membrane structure, and remove the crude tyrosinase solution. It was.
次に、ゥエル中のハイビスカス酸の最終濃度が 0.3〜2.5mg/mlとなるように、製造例 1 で得られたノヽィビスカス酸を PBSで希釈した溶液(下式数 1にお 、てハイビスカス酸添 加サンプルともいう。)を 50 lZwell添カ卩した。その後、 PBSで希釈した 0.1%ドーパ溶 液を各ウエノレ〖こ 50 /z L添加し、直ちに 37°Cで 4時間インキュベート後、マイクロプレート リーダーを用いて、 405nmの吸光度を測定した。  Next, a solution obtained by diluting the nobiscus acid obtained in Production Example 1 with PBS so that the final concentration of hibiscus acid in the well becomes 0.3 to 2.5 mg / ml (in the following formula 1, the hibiscus acid is used). (Also referred to as an added sample) was added at 50 lZwell. Thereafter, 50 / zL of 0.1% dopa solution diluted with PBS was added and immediately incubated at 37 ° C for 4 hours, and then the absorbance at 405 nm was measured using a microplate reader.
[0121] なお、コントロールとして、下記の条件についても同様の方法で測定した。  [0121] As a control, the following conditions were also measured in the same manner.
[0122] 測定した吸光度の値力も下式数 1に基づ 、てチロシナーゼ活性阻害率 (%)を算出 した。 [0122] Based on the measured absorbance value, the tyrosinase activity inhibition rate (%) was calculated based on Equation 1 below.
•ブランク: B 16メラノーマ細胞無播種:ハイビスカス酸溶液無添カロ  • Blank: B 16 melanoma cells unseeded: Hibiscus acid solution-free caro
•ハイビスカス酸添加サンプル: B 16メラノーマ細胞播種 ·ハイビスカス酸溶液添カロ • Hibiscus acid added sample: B 16 melanoma cell seeding · Hibiscus acid solution added caro
•コントロール 1 : B16メラノーマ細胞播種'ハイビスカス酸溶液無添カ卩(代わりに PBSを 添加) • Control 1: B16 melanoma cell seeding 'Hibiscus acid solution free supplemented solution (PBS added instead)
•コントロール 2 : B 16メラノーマ細胞無播種 'ハイビスカス酸溶液無添カ卩(代わりに PB Sを添加)  • Control 2: B 16 melanoma cell-free seeding 'Hibiscus acid solution free additive (added PBS instead)
[0123] [数 1]  [0123] [Equation 1]
(ハイビスカス酸添加サンプルの吸光度  (Absorbance of hibiscus acid added sample
チロシナーゼ活性阻害率 (X): 一ブランクの吸光ほ)  Tyrosinase activity inhibition rate (X): Absorbance of one blank)
X 100 X 100
(コントロール 1の吸光度一コントロール 2の吸光度) (Control 1 absorbance vs. Control 2 absorbance)
[0124] 又、比較のため、ハイビスカス酸の代わりに、製造例 4で得られたノヽィビスカス (Hibi scus sabdariffa L.)エキスを用いる以外は上記と同様に試験を行い、ノヽィビスカスェ キスのチロシナーゼ活性阻害率(%)も算出した。 [0124] For comparison, a test was carried out in the same manner as above except that the biscus acid (Hibi scus sabdariffa L.) extract obtained in Production Example 4 was used instead of hibiscus acid, and the tyrosinase activity of The inhibition rate (%) was also calculated.
[0125] 得られた結果を図 2に示す。図 2には、ノ、ィビスカス酸およびハイビスカスエキスの 添加濃度とチロシナーゼ活性の阻害率(%)の関係を図示して!/ヽる。  [0125] The obtained results are shown in FIG. Figure 2 illustrates the relationship between the added concentration of bismuth, ibiscus acid and hibiscus extract and the inhibition rate (%) of tyrosinase activity! / Speak.
[0126] この結果から、ノ、ィビスカス酸には、ハイビスカスエキスよりも高いチロシナーゼ活性 阻害作用があること、そしてその効果は濃度依存的に増強することが確認された。  [0126] From these results, it was confirmed that bis and biscus acid have a higher tyrosinase activity inhibitory action than hibiscus extract, and that the effect is enhanced in a concentration-dependent manner.
[0127] 実施例 3チロシナーゼ活件阳.害効 の評価 (ハイビスカス酸  Example 3 Tyrosinase Activity V. Evaluation of Harmful Effect (Hibiscus Acid
実施例 2にお 、て、ノ、ィビスカス酸に代えて製造例 2で得られたハイビスカス酸モノ ェチルエステル分画物を用い、ハイビスカス酸モノェチルエステルの最終濃度を 0.25 〜5mg/mlとなるように調製した以外は、実施例 2と同様にして評価を行った。 In Example 2, the final concentration of bismuth acid monoethyl ester was set to 0.25 by using the bismuth acid monoethyl ester fraction obtained in Production Example 2 instead of bis and biscus acid. Evaluation was performed in the same manner as in Example 2 except that the concentration was adjusted to ˜5 mg / ml.
[0128] 又、実施例 2と同様に、比較のため、ハイビスカス酸モノェチルエステル分画物の 代わりに、製造例 4で得られたハイビスカス(Hibiscus sabdarilfa [0128] As in Example 2, for comparison, instead of the hibiscus monoethyl ester fraction, the hibiscus (Hibiscus sabdarilfa) obtained in Production Example 4 was used.
し)エキスを用いる以外は同様に試験を行い、ハイビスカスエキスのチロシナーゼ活 性阻害率 (%)も算出した。  The same test was conducted except that the extract was used, and the tyrosinase activity inhibition rate (%) of the hibiscus extract was also calculated.
[0129] 得られた結果を図 3に示す。図 3には、ハイビスカス酸モノェチルエステル分画物お よびノヽィビスカスエキスの添加濃度とチロシナーゼ活性の阻害率(%)の関係を図示 している。 [0129] Fig. 3 shows the obtained results. FIG. 3 shows the relationship between the added concentration of the hibiscus monoethyl ester fraction and the nobiscus extract and the inhibition rate (%) of tyrosinase activity.
[0130] この結果から、ハイビスカス酸モノェチルエステル分画物には、ノ、イビスカスエキス よりも高いチロシナーゼ活性阻害作用があること、そしてその効果は濃度依存的に増 強することが確認された。  [0130] From this result, it was confirmed that the hibiscus acid monoethyl ester fraction had a higher tyrosinase activity inhibitory effect than that of biscus ibis extract, and that the effect increased in a concentration-dependent manner. It was.
[0131] uチロシナーゼ活件阳.害効奥の評 (ハイビスカス酸ジメチルヱステル)  [0131] u tyrosinase activity 評. Evaluation of harmful effects (Dimethyl Hibiscusate ヱ)
実施例 2にお 、て、ノ、ィビスカス酸に代えて製造例 3で得られたハイビスカス酸ジメ チルステル分画物を用い、ハイビスカス酸ジメチルエステルの最終濃度を 0.25〜5mg /mlとなるように調製した以外は、実施例 2と同様にして評価を行った。  In Example 2, instead of bis and biscus acids, the bismuth acid dimethyl ester fraction obtained in Production Example 3 was used, and the final concentration of bismuth acid dimethyl ester was adjusted to 0.25 to 5 mg / ml. Evaluation was performed in the same manner as in Example 2 except that.
[0132] 又、実施例 2と同様に、比較のため、ハイビスカス酸ジメチルエステルの代わりに、 製造例 4で得られたハイビスカス(Hibiscus sabdarilfa L.)エキスを用いる以外は同様 に試験を行い、ハイビスカスエキスのチロシナーゼ活性阻害率(%)も算出した。  [0132] For comparison, in the same manner as in Example 2, a test was conducted in the same manner except that the Hibiscus sabdarilfa L. extract obtained in Production Example 4 was used instead of Hibiscus acid dimethyl ester. The tyrosinase activity inhibition rate (%) of the extract was also calculated.
[0133] 得られた結果を図 4に示す。図 4には、ハイビスカス酸ジメチルエステルおよびハイ ビスカスエキスの添加濃度とチロシナーゼ活性の阻害率(%)の関係を図示して 、る  [0133] The obtained results are shown in FIG. Fig. 4 illustrates the relationship between the concentration of bismuth acid dimethyl ester and hibiscus extract and the inhibition rate (%) of tyrosinase activity.
[0134] この結果から、ハイビスカス酸ジメチルエステルには、ノ、イビスカスエキスと同等に チロシナーゼ活性阻害作用があること、そしてその効果は濃度依存的に増強すること が確認された。 [0134] From these results, it was confirmed that bismuth acid dimethyl ester has a tyrosinase activity inhibitory action equivalent to that of biscus ibis extract and that the effect is enhanced in a concentration-dependent manner.
[0135] 実施例 5 チロシナーゼ活件阳.害効果の持続件の評価 (ハイビスカス酸)  [0135] Example 5 Tyrosinase activity 評 価. Evaluation of sustained effects of harmful effects (Hibiscus acid)
ノ、ィビスカス酸のメラニン生成抑制効果の持続性を評価するために、以下の方法に 従って、チロシナーゼ活性阻害効果の持続性確認試験を行った。  In order to evaluate the persistence of the inhibitory effect of bismuth acid and bismuth acid on melanin production, a test for confirming the persistence of the tyrosinase activity inhibitory effect was performed according to the following method.
[0136] マウス由来 B16メラノーマ細胞(大日本製薬製、 B16F0)を 96ウェルマイク口プレート に 5 X 104cells/wellとなるように播種してー晚培養し、細胞を定着させた。次に、培養 プレートの培養液をァスピレーターで取り除き、 1%トライトン X-100を含む PBS (リン酸 緩衝食塩水)を 50 μ lZweU添加し、細胞の膜構造を破壊して粗チロシナーゼ溶液と した。 [0136] Mouse-derived B16 melanoma cells (Dainippon Pharmaceutical, B16F0) in a 96-well microphone mouthplate The cells were seeded at 5 × 10 4 cells / well and cultured in a sputum to establish the cells. Next, the culture solution on the culture plate was removed with an aspirator, and 50 μl ZweU of PBS (phosphate buffered saline) containing 1% Triton X-100 was added to disrupt the cell membrane structure to obtain a crude tyrosinase solution.
次に、ゥエル中のハイビスカス酸の最終濃度が 0.3〜10mg/mLとなるように製造例 1で 得られたノヽィビスカス酸を PBSで希釈した溶液を、 50 lZwell添カ卩した(以下、ハイビ スカス酸添加サンプルともいう。 ) oその後、 PBSで希釈した 0.1%ドーパ溶液を各ゥェ ノレ〖こ 50 μ L添加し、直ちに 37°Cでインキュベートした。  Next, a solution obtained by diluting the nobiscus acid obtained in Production Example 1 with PBS so that the final concentration of hibiscus acid in the well was 0.3 to 10 mg / mL was added with 50 lZwell (hereinafter referred to as Hibiscus). (It is also referred to as acid-added sample.) O After that, 50 μL of 0.1% dopa solution diluted with PBS was added and incubated at 37 ° C immediately.
[0137] インキュベート開始力 20時間後及び 108時間後に、マイクロプレートリーダーを用 いて、 405nmの吸光度を測定した。 [0137] The absorbance at 405 nm was measured using a microplate reader 20 hours and 108 hours after the initiation of incubation.
[0138] なお、コントロールとして、下記の条件についても同様の方法で測定した。 [0138] As a control, the following conditions were also measured in the same manner.
[0139] 測定した吸光度の値から下式数 2に基づいてチロシナーゼ活性阻害率 (%)を算出 した。 [0139] The tyrosinase activity inhibition rate (%) was calculated from the measured absorbance value based on the following equation (2).
[0140] ·ブランク: B16メラノーマ細胞無播種:ハイビスカス酸溶液無添カロ  [0140] · Blank: B16 melanoma cell non-seeding: Hibiscus acid solution-free caro
•ハイビスカス酸添加サンプル: B16メラノーマ細胞播種 ·ハイビスカス酸溶液添カロ •コントロール 1 : B16メラノーマ細胞播種'ハイビスカス酸溶液無添カ卩(代わりに PBS を添加)  • Hibiscus acid added sample: B16 melanoma cell seeding • Hibiscus acid solution added caro • Control 1: B16 melanoma cell seeded 'Hibiscus acid solution not added (PBS added instead)
•コントロール 2: B 16メラノーマ細胞無播種 'ハイビスカス酸溶液無添カ卩(代わりに PB Sを添加)  • Control 2: B 16 melanoma cell-free seeding 'Hibiscus acid solution-free potassium (added PBS instead)
[0141] [数 2] [0141] [Equation 2]
「 (ハイビスカス酸添加サンブルの吸光度一ブランクの吸光度) "(Absorbance of Hibiscus acid added sample-Absorbance of blank)
チロシナ―ゼ活性阻害率 1 - (コントロール,の吸光度 -コントロール 2の吸光度) ^ J 00 Tyrosinase activity inhibition rate 1-(Absorbance of control, Absorbance of control 2 ) ^ J 00
[0142] 又、比較のため、ハイビスカス酸の代わりにアルブチンを用いる以外は同様に試験 を行い、アルブチンのチロシナーゼ活性阻害率(%)を算出した(以下、アルブチン 添加サンプルという。)。 [0142] For comparison, a test was conducted in the same manner except that arbutin was used in place of hibiscus acid, and the tyrosinase activity inhibition rate (%) of arbutin was calculated (hereinafter referred to as an arbutin-added sample).
[0143] 得られた結果を図 5に示す。図 5において、〇及び參はハイビスカス酸添加サンプ ルのチロシナーゼ活性阻害率(%)、△及び▲はアルブチン添加サンプルのチロシ ナーゼ活性阻害率(%)を示す。また、点線は 20時間後の測定結果、実線は 108時間 後の測定結果を [0143] Fig. 5 shows the obtained results. In FIG. 5, 〇 and 參 indicate the tyrosinase activity inhibition rate (%) of the sample added with hibiscus acid, and △ and ▲ indicate the tyrosinase activity inhibition rate (%) of the sample added with arbutin. The dotted line is the measurement result after 20 hours, and the solid line is 108 hours. Later measurement results
示す。  Show.
[0144] 図 5から明らかなように、 20時間後の測定結果において、ノ、イビスカス酸はアルブチ ンより高いチロシナーゼ阻害効果を示していた。さらに 108時間後の測定結果におい てノ、ィビスカス酸添加サンプルはその高 、チロシナーゼ阻害効果を維持して 、た。  As is clear from FIG. 5, in the measurement results after 20 hours, biscus acid and biscus acid showed a higher tyrosinase inhibitory effect than arbutin. Further, in the measurement result after 108 hours, the sample added with biscus acid maintained a high tyrosinase inhibitory effect.
[0145] それに対し、アルブチン添加サンプルでは、 20時間後に比べ 108時間後ではチロシ ナーゼ活性阻害率の顕著な減少が認められた。  [0145] In contrast, in the arbutin-added sample, a significant decrease in the inhibition rate of tyrosinase activity was observed after 108 hours compared to 20 hours later.
[0146] この結果から、ノ、ィビスカス酸は高く且つ持続的なチロシナーゼ活性阻害効果を有 し、高い美白効果を長時間にわたって持続できることが示唆された。  [0146] From these results, it was suggested that bis and biscus acid have a high and sustained tyrosinase activity inhibitory effect and can maintain a high whitening effect over a long period of time.
[0147] meメラニン牛成 ¾制効 (ハイビスカス酴)  [0147] me melanin cattle ¾ effect (Hibiscus coffee)
上記の結果からハイビスカス酸は、メラニンの生成に関与するチロシナーゼ生成抑 制作用、及びチロシナーゼ活性阻害作用を有していることが明らかとなったが、更に 、ノ、ィビスカス酸のメラニン自体に対する生成抑制効果にっ ヽても評価した。  From the above results, it has been clarified that hibiscus acid has an inhibitory effect on the production of tyrosinase involved in the production of melanin and an inhibitory effect on tyrosinase activity. Even the effect was evaluated.
[0148] 評価試験は、以下の方法に従って行った。  [0148] The evaluation test was performed according to the following method.
[0149] マウス由来 B16メラノーマ細胞(大日本製薬製、 B16F0)を、 6ゥエルプレートに約 1 X  [0149] Mouse-derived B16 melanoma cells (Dainippon Pharmaceutical Co., Ltd., B16F0) were placed on a 6-well plate at about 1 X
105cellsZwellとなるように播種し、一晩培養して、細胞を定着させた。次に、最終的 にハイビスカス酸のゥエル内濃度が 0.2〜2mgZmlとなるように製造例 1で得られたノヽ ィビスカス酸を PBSで希釈した溶液(下式数 3にお 、て目的サンプルとも ヽぅ。)を添 加し、 4日培養後、 0.25%トリプシン液を用いて細胞を剥がし、遠心操作により細胞を 回収した。回収した細胞の色差を目視で確認した後、 5NNaOHにて細胞の膜構造を 破壊しメラニンを溶出させて、 405nmの吸光度を測定した。 The cells were seeded to 10 5 cellsZwell and cultured overnight to fix the cells. Next, a solution obtained by diluting the nobiscus acid obtained in Production Example 1 with PBS so that the final concentration of the hibiscus acid in the well is 0.2 to 2 mgZml (in the following formula 3, the target sample is also included). ) Was added, and after 4 days of culture, the cells were detached using a 0.25% trypsin solution, and the cells were collected by centrifugation. After visually confirming the color difference of the collected cells, the membrane structure of the cells was destroyed with 5N NaOH to elute melanin, and the absorbance at 405 nm was measured.
[0150] なお、コントロールとして、ハイビスカス酸を添カ卩しない系(以下、下式数 3において 、無添加サンプルという。)について同様に測定した。  [0150] As a control, the same measurement was performed on a system not added with hibiscus acid (hereinafter referred to as an additive-free sample in Formula 3 below).
[0151] さらに、溶液調製時の誤差を補正する為、サンプル中のタンパク量を、定量キットを 用いて測定し、単位タンパク量で補正し、メラニン生成率(%)を下式数 3に基づいて 算出した。  [0151] Furthermore, in order to correct errors during preparation of the solution, the amount of protein in the sample was measured using a quantification kit, corrected with the amount of unit protein, and the melanin production rate (%) was based on Equation 3 below. Calculated.
[0152] [数 3] ニン生成率 (¾) = (目的サンプルの吸光度) ノ(目的サンプルのタンパク量) X  [0152] [Equation 3] Nin production rate (¾) = (Absorbance of target sample) No (Amount of protein in target sample) X
(無添加サンプルの吸光度) Z (無添加サンプルのタンパク量) [0153] 得られた結果を図 6に示す。図 6は、ノ、ィビスカス酸の各添加濃度におけるメラニン 生成率 (%)を示す。 (Absorbance of additive-free sample) Z (protein amount of additive-free sample) [0153] The obtained results are shown in FIG. Figure 6 shows the melanin production rate (%) at each additive concentration of bis and biscus acids.
[0154] この結果、ハイビスカス酸の添カ卩によって、メラニンの生成が有意に抑制されている ことが確認され、ハイビスカス酸力 ラニン生成抑制剤として有用であることが明ら力と なった。  [0154] As a result, it was confirmed that the addition of hibiscus acid significantly suppressed the production of melanin, and it became clear that it was useful as a hibiscus acid lanine production inhibitor.
[0155] 実施例 7メラニン生成抑制効果レ、ィビスカス酸モノェチルエステル)  Example 7 Inhibitory Effect of Melanin Formation (Ibiscus acid monoethyl ester)
実施例 6において、ノ、ィビスカス酸に代えて、製造例 2で得られたノヽィビスカス酸モ ノエチルエステル分画物を用いる以外は、実施例 6と同様に試験し、評価を行った。  In Example 6, tests and evaluations were performed in the same manner as in Example 6 except that the bisbiscus monoethyl ester fraction obtained in Production Example 2 was used in place of bis and biscus acids.
[0156] 得られた結果を図 7に示す。図 7は、ハイビスカス酸モノェチルエステル分画物の各 添加濃度におけるメラニン生成率(%)を示す。 [0156] The obtained results are shown in FIG. FIG. 7 shows the melanin production rate (%) at each addition concentration of the hibiscus monoethyl ester fraction.
[0157] この結果、ハイビスカス酸モノェチルエステル分画物の添カ卩によって、メラニンの生 成が有意に抑制されて 、ることが確認され、ハイビスカス酸モノェチルエステルカ^ラ ニン生成抑制剤として有用であることが明らかとなった。 [0157] As a result, it was confirmed that the addition of the bismuth acid monoethyl ester fraction significantly suppressed the production of melanin, and the bismuth acid monoethyl ester carbonate production was suppressed. It became clear that it was useful as an agent.
[0158] msメラユン牛成¾制効 (ハイビスカス酸ジメチルエステル)  [0158] ms Melayun cattle growth control effect (Hibiscus acid dimethyl ester)
実施例 6において、ノ、ィビスカス酸に代えて、製造例 3で得られたノヽィビスカス酸ジ メチルエステルを用いる以外は、実施例 6と同様に試験し、評価を行った。  In Example 6, tests and evaluations were carried out in the same manner as in Example 6 except that the bis-biscus acid dimethyl ester obtained in Production Example 3 was used instead of bis-biscus acid.
[0159] 得られた結果を図 8に示す。図 8は、ハイビスカス酸ジメチルエステルの各添加濃度 におけるメラニン生成率(%)を示す。 [0159] Fig. 8 shows the obtained results. FIG. 8 shows the melanin production rate (%) at each addition concentration of hibiscus dimethyl ester.
[0160] この結果、ハイビスカス酸ジメチルエステルの添カ卩によって、メラニンの生成が有意 に抑制されて 、ることが確認され、ハイビスカス酸ジメチルエステル力 ラニン生成抑 制剤として有用であることが明らかとなった。 [0160] As a result, it was confirmed that the addition of bismuth acid dimethyl ester significantly suppressed the production of melanin, and it was clear that it was useful as an inhibitor of bismuth acid dimethyl ester power lanine production. became.
[0161] 試験例 [0161] Test example
ノ、イビスカス酸には光学異性体が存在するため、植物力ら得られる抽出物にはノ、ィ ビスカス酸の光学異性体も含有されて ヽることが考えられる。下式(2)で表されるハイ ビスカス酸の光学異性体のうち、天然に存在するものとしては、下式(3)で表されるガ ルシニア酸が考えられる。  Since the optical isomers of bis and ibiscus acids exist, the extract obtained from plant power may contain the isomers of bis and biscus acids. Among the optical isomers of hibiscus acid represented by the following formula (2), galsinic acid represented by the following formula (3) is considered as a naturally occurring one.
[0162] [化 5] [0162] [Chemical 5]
Figure imgf000023_0001
Figure imgf000023_0001
[0163] そのため、美白作用がハイビスカス酸の立体構造に特徴的なものであるかを確認 するために、ハイビスカス酸とガルシニア酸について、メラニン生成抑制作用に関す る試験を以下の手順に従って行った。また、併せて、ハイビスカス酸の安全性を確認 するために、細胞毒性に関しても評価した。  [0163] Therefore, in order to confirm whether the whitening action was characteristic of the three-dimensional structure of hibiscus acid, a test on the melanin production inhibitory action of hibiscus acid and garcinia acid was performed according to the following procedure. In addition, cytotoxicity was also evaluated to confirm the safety of hibiscus acid.
[0164] メラニン生成抑制作用については、次のように試験した。  [0164] The melanin production inhibitory action was tested as follows.
[0165] マウス由来 B16メラノーマ細胞(大日本製薬製、 B16F0)を、 6ゥエルプレートに約 I X  [0165] Mouse-derived B16 melanoma cells (B16F0, manufactured by Dainippon Pharmaceutical Co., Ltd.)
105cellsZwellとなるように播種し、ー晚培養して、細胞を定着させた。次に、ノ、ィビス カス酸又はガルシニア酸の最終的なゥエル内濃度が 0.5mg/mlとなるように PBSで希釈 した溶液(下式数 4にお ヽて「目的サンプル」と ヽぅ。)を添加し、 4日培養後、 0.25%ト リブシン液を用いて細胞を剥がし、遠心操作により細胞を回収した。回収した細胞の 色差を目視で確認した後、 5NNaOHにて細胞の膜構造を破壊しメラニンを溶出させ て、 405nmの吸光度を測定した。 The cells were seeded to 10 5 cellsZwell and cultured in anther culture to establish the cells. Next, a solution diluted with PBS so that the final concentration of noci, biscus acid or garcinia acid in the well is 0.5 mg / ml (the “target sample” in Equation 4 below). After culturing for 4 days, the cells were detached using a 0.25% tribsin solution, and the cells were collected by centrifugation. After visually confirming the color difference of the collected cells, the membrane structure of the cells was destroyed with 5N NaOH to elute melanin, and the absorbance at 405 nm was measured.
[0166] なお、コントロールとして、ハイビスカス酸又はガルシニア酸を添カ卩しない系(以下、 下式数 4において「無添加サンプル」という。 )について同様に測定した。  [0166] As a control, the measurement was carried out in the same manner with respect to a system not added with hibiscus acid or garcinia acid (hereinafter referred to as "non-added sample" in formula 4 below).
[0167] さらに、溶液調製時の誤差を補正する為、サンプル中のタンパク量を定量キットを 用いて測定し、単位タンパク量で補正し、メラニン生成率 (%)を次式に基づいて算出 した。  [0167] Furthermore, in order to correct errors during solution preparation, the amount of protein in the sample was measured using a quantification kit, corrected with the amount of unit protein, and the melanin production rate (%) was calculated based on the following equation. .
[0168] [数 4] メラー、'生成率 = (目的サンプルの吸光度) z (目的サンプルのタンパク量) X 1 00 [0168] [Equation 4] Meller, 'Production rate = (Absorbance of target sample) z (Protein amount of target sample) X 1 00
(無添加サンプルの吸光度) / (無添加サンプルのタンパク量)  (Absorbance of additive-free sample) / (protein amount of additive-free sample)
[0169] また細胞毒性については、回収後の培養液をトリパンブルーにて染色し、生細胞数 を血球計算盤にて計測した。 [0170] 得られた結果を図 9に示す。図 9は、濃度 0.5%におけるガルシニア酸及びノヽィビス カス酸のメラニン生成率(%)及び細胞数を示す。また、 BKはブランク(無添加サンプ ル)を示す。 [0169] Regarding cytotoxicity, the collected culture was stained with trypan blue, and the number of viable cells was measured with a hemocytometer. [0170] Fig. 9 shows the obtained results. FIG. 9 shows the melanin production rate (%) and the number of cells of garcinia acid and neurobiscus acid at a concentration of 0.5%. BK indicates a blank (no additive sample).
[0171] この結果ノヽィビスカス酸にはメラニン生成抑制効果があり、細胞毒性も低いことがわ かった。  [0171] As a result, it was found that neutral biscus acid had an inhibitory effect on melanin production and low cytotoxicity.
[0172] 一方、ガルシニア酸にはメラニン生成抑制効果がほとんどなぐ細胞毒性も高いこと 明らかとなった。  [0172] On the other hand, it has been clarified that garcinia acid is highly cytotoxic with almost no inhibitory effect on melanin production.
[0173] このことから、ハイビスカス酸の立体構造力 メラニン生成抑制効果に関して重要で あることが示唆された。  [0173] From this, it was suggested that the steric strength of hibiscus acid is important for the melanin production inhibitory effect.
[0174] 処方例 [0174] Formulation example
以下に、本発明の処方例を示して、本発明をより詳細に説明するが、本発明はこれ らに限定されるものではない。  Hereinafter, the present invention will be described in more detail with reference to formulation examples of the present invention, but the present invention is not limited thereto.
[0175] <処方例 1 > 化粧水 [0175] <Prescription Example 1> Lotion
下記表に示す配合割合で原料を配合し、常法に従って、化粧水を製造した。  The raw materials were blended at the blending ratios shown in the following table, and a lotion was produced according to a conventional method.
(原料) (重量%)  (Raw material) (wt%)
ノヽィビスカス酸 0. 5  Neubiscus acid 0.5
ァスコルビン酸リン酸エステルマグネシウム 3. 0  Ascorbic acid magnesium phosphate 3.0
ポリプロピレングリコール 2. 0  Polypropylene glycol 2.0
ォキシベンゾン 3. 0  Oxybenzone 3.0
パラォキシ安息香酸エステル 0. 5  Paraxybenzoate ester 0.5
クェン酸 適量  Quenic acid
香料 適量  Perfume
精製水 mm _  Purified water mm _
合計 100. 0  Total 100. 0
[0176] <処方例 2> 化粧用クリーム  [0176] <Prescription Example 2> Cosmetic cream
下記表に示す配合割合で原料を配合し、常法に従って、化粧用クリームを製造し ハイビスカス酸モノメチルエステル 0. Ingredients are blended in the proportions shown in the table below, and cosmetic creams are produced according to conventional methods. Hibiscus acid monomethyl ester 0.
ァスコルビン酸モノステアリン酸エステル 3 Ascorbic acid monostearate 3
1. 0  Ten
ステアリルアルコ 5. 0 Stearyl Arco 5.0
ステアリン酸 8. 0 Stearic acid 8.0
スクヮラン 10. 0 Screen 10.0
自己乳化型グリセリルモノステアレート 3. 0 Self-emulsifying glyceryl monostearate 3.0
ポリオキシエチレンセチルエーテル(20E. 0) 1. 0 Polyoxyethylene cetyl ether (20E. 0) 1.0
プロピレングリコ 5. 0 Propylene glycol 5.0
水酸化カリウム 0. 3 Potassium hydroxide 0.3
香料 Fragrance
防腐剤Preservative
Figure imgf000025_0001
Figure imgf000025_0001
合計 100. 0 Total 100. 0
<処方例 3 >乳液  <Formulation example 3> Emulsion
下記表に示す配合割合で原料を配合し、常法に従って、乳液を製造した (原料) (重量%)  The raw materials were blended at the blending ratios shown in the table below, and the emulsion was produced according to a conventional method (raw materials) (wt%)
ノヽィビスカス酸モノメチノレエステノレ 1. 0 Monobiscusic acid monomethylenoestenole 1. 0
ノヽィビスカス酸モノェチノレエステノレ 1. 0 Nobisci acid monoethino estenole 1. 0
スクヮラン 8. 0 Skulun 8.0
ワセリン 2. 0 Vaseline 2.0
ミツロウ 0. 5 Beeswax 0.5
ソノレビタンセスキォレエート 0. 8 Sonorebitan Sesquitoate 0.8
ポリオキシエチレンォレイルエーテル(20E. 0) 1. 2 Polyoxyethylene oleyl ether (20E. 0) 1.2
カノレボキシビニノレポリマー 0. 2 Canoleboxyvininole polymer 0.2
プロピレングリコール 0. 5 Propylene glycol 0.5
水酸化カリウム 0. 1 Potassium hydroxide 0.1
エタノール 7. 0 香料 Ethanol 7.0 Fragrance
防腐剤  Preservative
纏 7k  Summary 7k
合計 100. 0  Total 100. 0
[0178] <処方例 4>パン  [0178] <Prescription Example 4> Bread
下記表に示す配合割合で原料を配合し、常法に従って、パンを製造した c The raw materials were blended in proportions shown in the following table according to a conventional method, to produce bread c
(原料) (重量%) (Raw material) (wt%)
小麦粉
Figure imgf000026_0001
flour
Figure imgf000026_0001
砂糖 3. 0  Sugar 3.0
食塩 1. 0 〇  Salt 1. 0 〇
ノ、ィビスカス酸 0. 5  No, biscus acid 0.5
脱脂粉乳 1. 0  Nonfat dry milk 1. 0
油脂 3. 1  Oils and fats 3.1
イーストフード 0. 1  East Food 0.1
イースト 1. 0  East 1.0
残部  The rest
A  A
O斗B I 100. 0  O Doo B I 100. 0
[0179] <処方例 5〉タブレット  [0179] <Prescription Example 5> Tablet
下記表に示す配合割合で原料を配合し、常法に従って、タブレットを製造した (原料) (重量%)  Ingredients were blended in the proportions shown in the table below, and tablets were produced according to conventional methods (raw materials) (wt%)
ノヽィビスカス酸 5  Neubiscus acid 5
ノヽィビスカス酸モノェチノレエステノレ 5  Nobiscus acid monoethinore estenore 5
ァスコルビン酸リン酸エステルマグネシウム 10  Ascorbic acid phosphate magnesium 10
プラセンタエキス 10  Placenta extract 10
コラーゲン 20  Collagen 20
結晶セノレロース 20  Crystalline cellulose 20
デキストリン 14  Dextrin 14
グリセリン脂肪酸エステル 6 陚形剤 10 Glycerin fatty acid ester 6 Vaginal form 10
合計 100  Total 100
[0180] <処方例 6 > ドリンク剤  [0180] <Prescription Example 6> Drink
下記表に示す配合割合で原料を配合し、常法に従って、ドリンク剤を製造した The ingredients were blended at the blending ratios shown in the table below, and a drink was produced according to a conventional method.
(原料) (material)
ノ、ィビスカス酸 0. 5  No, biscus acid 0.5
果糖ブドウ糖 20. 0  Fructose glucose 20. 0
クェン酸 0. 50  Chenic acid 0.50
クェン酸ナトリウム 0. 10  Sodium quenate 0. 10
ァスコノレビン酸 0. 20  Asconolevic acid 0.20
香料  Fragrance
 Spirit
合計 100. 0  Total 100. 0
[0181] <処方例 7〉練り歯磨き A  [0181] <Prescription Example 7> Toothpaste A
下記表に示す配合割合で原料を配合し、常法に従って、練り歯磨きを製造した (原料) (重量%)  Ingredients were blended in the proportions shown in the table below, and toothpastes were produced according to conventional methods (raw materials) (wt%)
第 2リン酸カルシウム 30. 00  Dicalcium phosphate 30. 00
グ!;セジン 10. 00  Sejin 10. 00
ソノレビトール 20. 00  Sonorbitol 20.00
カルボキシメチルセルロースナトリウム 1. 00  Sodium carboxymethylcellulose 1.00
ラウリル硫酸ナトリウム 1. 50  Sodium lauryl sulfate 1. 50
カラギーナン 0. 50  Carrageenan 0.50
サッカリンナトリウム 0. 10  Saccharin sodium 0. 10
香料 1. 00  Fragrance 1.00
安息香酸ナトリウム 0. 30  Sodium benzoate 0. 30
/、ィビスカス酸 0. 10  /, Biscus acid 0. 10
精製水 翻  Purified water
A斗 100. 0 [0182] <処方例 8 >練り歯磨き B A DOO 100. 0 [0182] <Prescription Example 8> Toothpaste B
下記表に示す配合割合で原料を配合し、常法に従って、練り歯磨きを製造した The raw materials were blended at the blending ratios shown in the table below, and toothpastes were produced according to conventional methods.
(原料) (重量%) (Raw material) (wt%)
ラウリル硫酸ナトリウム  Sodium lauryl sulfate
ラウリル酸ジエタノールアミド 1. 00  Lauric acid diethanolamide 1. 00
プロピレングリコーノレ  Propylene glycol
60% ソルビット液 35. 00  60% sorbite 35. 00
メチルパラベン  Methylparaben
ブチルパラベン 〇 〇 〇  Butylparaben 〇 〇 〇
〇 〇 ο οO C0 0〇〇O C  〇 〇 ο οO C0 00
サッカリンナトリウム 〇 ο 〇 〇 〇  Saccharin sodium 〇 ο 〇 〇 〇
〇 〇 〇 〇 〇1—  ○ ○ ○ ○ ○ 1—
ゼラチン  Gelatin
香料  Fragrance
アルギン酸ナトリウム  Sodium alginate
安息香酸ナトリウム  Sodium benzoate
水酸化アルミニウム 45. 00  Aluminum hydroxide 45. 00
モノフルォリン酸ナトリウム  Sodium monofluorate
ノ、ィビスカス酸  No, biscus acid
合計 100. 0 Total 100. 0
[0183] <処方例 9 >洗口液  [0183] <Prescription Example 9> Mouthwash
下記表に示す配合割合で原料を配合し、常法に従って、洗口液を製造した The raw materials were blended at the blending ratios shown in the table below, and a mouthwash was produced according to a conventional method.
(原料) (重量%) (Raw material) (wt%)
エタノーノレ 10. 00  Ethanor 10.00
グリセリン 5. 00  Glycerin 5.00
クェン酸 0. 01  Quenic acid 0. 01
クェン酸ナトリウム 0. 10  Sodium quenate 0. 10
ポリオキシエチレン硬化ひまし油 0. 50  Polyoxyethylene hydrogenated castor oil 0.50
パラォキシ安息香酸メチル 0. 10 香料 0. 20 Methyl paraoxybenzoate 0. 10 Fragrance 0. 20
ノ、ィビスカス酸 0. 1 No, biscus acid 0.1
精製水 翻 Purified water
合計 100. 0 Total 100. 0
<処方例 10>口腔用パスタ  <Prescription Example 10> Oral Pasta
下記表に示す配合割合で原料を配合し、常法に従って、 口腔用パスタを製造した  Ingredients were blended at the blending ratios shown in the table below, and oral pasta was produced according to conventional methods.
(原料) (重量%) (Raw material) (wt%)
流動パラフィン 13. 00 Liquid paraffin 13.00
セタノーノレ 10. 00 Cetanore 10.00
グリセリン 25. 00 Glycerin 25. 00
ソルビタンモノパルミテート 0. 60 Sorbitan monopalmitate 0.60
ポリオキシエチレンソルビタンモノステアレート 5. 00 Polyoxyethylene sorbitan monostearate 5. 00
ラウリル硫酸ナトリウム 0. 10 Sodium lauryl sulfate 0. 10
塩化べンゾトニゥム 0. 10 Benzotonium chloride 0. 10
サリチル酸メチル 0. 10 Methyl salicylate 0. 10
サッカリン 0. 20 Saccharin 0.20
香料 0. 25 Fragrance 0. 25
ノ、ィビスカス酸 0. 50 No, biscus acid 0.50
精製水 翻 Purified water
合計 100. 0 Total 100. 0
図面の簡単な説明 Brief Description of Drawings
[図 1]実施例 1においてハイビスカス酸の存在下又は非存在下で培養した B16メラノ 一マ細胞のチロシナーゼ発現量を示す PVDF像である。図中レーン Iはハイビスカス 酸無添加;レーン IIは 5%ハイビスカス酸溶液 100 μ 1添カ卩;レーン IIIは 5%ハイビスカス 酸溶液 50 μ 1添カ卩;レーン IVは 5%ハイビスカス酸溶液 25 μ 1添カ卩;レーン Vは 5%ハイ ビスカス酸溶液 12.5 μ 1添カ卩のサンプルを示す図である。 FIG. 1 is a PVDF image showing the expression level of tyrosinase in B16 melanoma cells cultured in Example 1 in the presence or absence of hibiscus acid. In the figure, Lane I is without hibiscus acid; Lane II is with 5% hibiscus acid solution 100 μ1; Lane III is with 5% hibiscus acid solution 50 μ1; Lane IV is 5% hibiscus acid solution 25 μm Lane 1 is a diagram showing a sample of 12.5 μl-added 5% biscus acid solution.
[図 2]実施例 2における、ハイビスカス酸又はハイビスカスエキスの添加濃度とチロシ ナーゼ活性阻害率 (%)の関係を示す図である。 FIG. 2 shows the concentration of hibiscus acid or hibiscus extract and tyrosin in Example 2. It is a figure which shows the relationship of the enzyme activity inhibition rate (%).
[図 3]実施例 3における、ハイビスカス酸モノェチルエステル分画物又はハイビスカス エキスの添加濃度とチロシナーゼ活性阻害率(%)の関係を示す図である。  FIG. 3 is a graph showing the relationship between the added concentration of hibiscus monoethyl ester fraction or hibiscus extract and the tyrosinase activity inhibition rate (%) in Example 3.
[図 4]実施例 4における、ハイビスカス酸ジメチルエステル又はハイビスカスエキスの 添加濃度とチロシナーゼ活性の阻害率(%)の関係を示す。 FIG. 4 shows the relationship between the concentration of hibiscus dimethyl ester or hibiscus extract and the inhibition rate (%) of tyrosinase activity in Example 4.
[図 5]実施例 5における、ハイビスカス酸添加群及びアルブチン添加群のインキュべ ート 20時間後と 108時間後におけるチロシナーゼ活性阻害率(%)を示す図である。  FIG. 5 is a graph showing the tyrosinase activity inhibition rate (%) in Example 5 after 20 hours and 108 hours of incubation in the hibiscus acid added group and arbutin added group.
[図 6]実施例 6における、ハイビスカス酸の添加濃度とメラニン生成率(%)の関係を示 す図である。 FIG. 6 is a graph showing the relationship between the concentration of hibiscus acid added and the melanin production rate (%) in Example 6.
[図 7]実施例 7における、ハイビスカス酸モノェチルエステル分画物の添加濃度とメラ ニン生成率 (%)の関係を示す図である。  FIG. 7 is a graph showing the relationship between the added concentration of the hibiscus monoethyl ester fraction and the melanin production rate (%) in Example 7.
[図 8]実施例 8における、ハイビスカス酸ジメチルエステルの添加濃度とメラニン生成 率 (%)の関係を示す。  FIG. 8 shows the relationship between the concentration of bismuth acid dimethyl ester added and the melanin production rate (%) in Example 8.
[図 9]試験例における、ハイビスカス酸とガルシニア酸のゥエル中濃度が 0.5mgZmlで あるときの細胞数とメラニン生成率(%)を示す図である。  FIG. 9 is a graph showing the number of cells and melanin production rate (%) when the concentration of hibiscus acid and garcinic acid in the well is 0.5 mgZml in a test example.

Claims

請求の範囲 [1] 下式 (1) Claim [1] The following formula (1)
[化 1]  [Chemical 1]
Figure imgf000031_0001
Figure imgf000031_0001
(但し、式中の R1及び R2は、それぞれ水素原子、置換又は非置換のアルキル基又は ベンジル基を示す。 )で表されるハイビスカス酸類及びその塩力 なる群力 選ばれ る 1又は 2種以上の化合物を有効成分とするメラニン生成抑制剤。 (In the formula, R 1 and R 2 each represent a hydrogen atom, a substituted or unsubstituted alkyl group or a benzyl group.) Hibiscus acids represented by A melanin production inhibitor containing at least one kind of compound as an active ingredient.
[2] 式(1)で表されるハイビスカス酸類の R1及び R2の少なくとも 1つが置換又は非置換の アルキル基である請求項 1に記載のメラニン生成抑制剤。 [2] The melanin production inhibitor according to claim 1, wherein at least one of R 1 and R 2 of the hibiscus acids represented by the formula (1) is a substituted or unsubstituted alkyl group.
[3] 請求項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む美白剤  [3] A whitening agent comprising the melanin production inhibitor according to any one of claims 1 to 2 as an active ingredient
[4] 請求項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む美白用 化粧料。 [4] A whitening cosmetic comprising the melanin production inhibitor according to any one of claims 1 to 2 as an active ingredient.
[5] 請求項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む美白用  [5] For whitening comprising the melanin production inhibitor according to any one of claims 1 to 2 as an active ingredient
TOo  TOo
[6] 請求項 1〜2のいずれかに記載のメラニン生成抑制剤を有効成分として含む口腔用 組成物。  [6] An oral composition comprising the melanin production inhibitor according to any one of claims 1 to 2 as an active ingredient.
PCT/JP2006/306976 2005-03-31 2006-03-31 Melanin production inhibitory agent WO2006106992A1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016504910A (en) * 2012-12-24 2016-02-18 エルブイエムエイチ レシェルシェ Aptamers that inhibit the enzyme activity of tyrosinase
WO2017019793A1 (en) * 2015-07-27 2017-02-02 Coty Inc. Microdelivery triple acid

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* Cited by examiner, † Cited by third party
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JP2009019010A (en) * 2007-07-12 2009-01-29 Sosin:Kk Composition for oral cavity
KR101007468B1 (en) 2008-11-14 2011-01-12 강원대학교산학협력단 External preparation composition for skin whitening containing sheep extract
EP3006016A1 (en) * 2014-10-06 2016-04-13 Medical Brands Research B.V. Dermatological kit comprising compositions based on Hibiscus flower and Buriti oil
KR102126840B1 (en) * 2018-07-09 2020-06-25 대한민국 method for producing petal of rose of Sharon extract, cosmetic composition for whitening skin comprising petal of rose of Sharon extract and pharmaceutical composition for prevention or treatment of melanin hyperpigmentation disease containing thereof
JP7388748B2 (en) * 2020-11-10 2023-11-29 株式会社デジマ Use of lactic acid bacteria as an additive for improving the taste of Hamabou extract

Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5888305A (en) * 1981-11-19 1983-05-26 Nikko Kemikaruzu Kk Cosmetic
EP0425398A1 (en) * 1989-10-23 1991-05-02 Camille Isnard Aloe solution, preparation process and compositions containing it
JPH0624937A (en) * 1992-06-29 1994-02-01 Narisu Keshohin:Kk Mucopolysaccharide fragmentation-inhibiting agent, active oxygen-scavenging agent, antioxidative agent and cosmetic
JPH09295928A (en) * 1996-05-01 1997-11-18 Narisu Keshohin:Kk Cosmetic material for prevention of aging
JPH11349469A (en) * 1998-06-08 1999-12-21 Kanebo Ltd Ultraviolet-shielding cosmetic
JP2000095663A (en) * 1998-09-24 2000-04-04 Kose Corp Agent for external use containing plant extract
JP2000154134A (en) * 1998-11-17 2000-06-06 Naris Cosmetics Co Ltd Cosmetic
JP2000239164A (en) * 1999-02-19 2000-09-05 Kikkoman Corp Glycosidase inhibitor
US6127553A (en) * 1998-08-03 2000-10-03 Department Of Science And Technology, Goverment Of India Convenient method for large-scale isolation of hibiscus acid
US20030088110A1 (en) * 2000-10-03 2003-05-08 Saud Ibrahim Ibnu Novel chiral derivatives of hibiscus acid bearing lactone ring moiety, process for preparing the same and a convenient method for the large-scale isolation of hibiscus acid
JP2003171300A (en) * 2001-09-28 2003-06-17 Kobayashi Pharmaceut Co Ltd Melanization inhibitor
JP2004346006A (en) * 2003-05-22 2004-12-09 Kyoei Kogyosho:Kk Skin lotion and method for producing the same

Patent Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5888305A (en) * 1981-11-19 1983-05-26 Nikko Kemikaruzu Kk Cosmetic
EP0425398A1 (en) * 1989-10-23 1991-05-02 Camille Isnard Aloe solution, preparation process and compositions containing it
JPH0624937A (en) * 1992-06-29 1994-02-01 Narisu Keshohin:Kk Mucopolysaccharide fragmentation-inhibiting agent, active oxygen-scavenging agent, antioxidative agent and cosmetic
JPH09295928A (en) * 1996-05-01 1997-11-18 Narisu Keshohin:Kk Cosmetic material for prevention of aging
JPH11349469A (en) * 1998-06-08 1999-12-21 Kanebo Ltd Ultraviolet-shielding cosmetic
US6127553A (en) * 1998-08-03 2000-10-03 Department Of Science And Technology, Goverment Of India Convenient method for large-scale isolation of hibiscus acid
JP2000095663A (en) * 1998-09-24 2000-04-04 Kose Corp Agent for external use containing plant extract
JP2000154134A (en) * 1998-11-17 2000-06-06 Naris Cosmetics Co Ltd Cosmetic
JP2000239164A (en) * 1999-02-19 2000-09-05 Kikkoman Corp Glycosidase inhibitor
US20030088110A1 (en) * 2000-10-03 2003-05-08 Saud Ibrahim Ibnu Novel chiral derivatives of hibiscus acid bearing lactone ring moiety, process for preparing the same and a convenient method for the large-scale isolation of hibiscus acid
JP2003171300A (en) * 2001-09-28 2003-06-17 Kobayashi Pharmaceut Co Ltd Melanization inhibitor
JP2004346006A (en) * 2003-05-22 2004-12-09 Kyoei Kogyosho:Kk Skin lotion and method for producing the same

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016504910A (en) * 2012-12-24 2016-02-18 エルブイエムエイチ レシェルシェ Aptamers that inhibit the enzyme activity of tyrosinase
WO2017019793A1 (en) * 2015-07-27 2017-02-02 Coty Inc. Microdelivery triple acid
US10470988B2 (en) 2015-07-27 2019-11-12 Coty Inc. Skin improvement formulations

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