WO2006068254A1

WO2006068254A1 - Skin-whitening agent

Info

Publication number: WO2006068254A1
Application number: PCT/JP2005/023680
Authority: WO
Inventors: Yoshihide Suwa; Seiichi Koshimizu; Haruo Nukaya; Kenji Oguchi; Yoshinori Nozawa; Yukihiro Akao
Original assignee: Suntory Limited; Gifu Research And Development Foundation
Priority date: 2004-12-24
Filing date: 2005-12-22
Publication date: 2006-06-29
Also published as: JP2012116847A; TWI369993B; JPWO2006068254A1; CN101102744B; CN101102744A; TW200637588A; JP5095221B2; JP5497809B2

Abstract

A skin-whitening agent comprising a lyoniresinol as an effective ingredient for skin-whitening. This skin-whitening agent is useful as a food or beverage, or cosmetic, or pharmaceutical, etc. for the prevention and treatment of melanin pigment deposit.

Description

Specification

Whitening agent

Technical field

[0001] The present invention relates to a whitening agent, and in particular, to prevent or treat the formation (deposition) of black (melanin) pigment, which is also the cause of the occurrence of skin spots, freckles, etc. The present invention relates to a whitening agent that uses rio-resinol, which is effective in promoting reduction, as a whitening active ingredient. In addition, by containing the whitening active ingredient, the action of tyrosinase is inhibited, and the activity of melanocytes by melanocyte-stimulating hormone produced in the skin by Z or ultraviolet rays is inhibited, thereby producing a black (melanin) pigment. It relates to foods, beverages, cosmetics and pharmaceuticals that have the ability to suppress and promote the reduction of the deposited pigment.

Background art

[0002] Skin Blemish 'Sobacus is regarded as a serious skin problem. In particular, stains and freckles, and black pigmentation on the skin after tanning, occur and increase with age and become disappeared, which is an inevitable problem for middle-aged and elderly people. Yes. Most of the black pigment deposited on the skin is considered to be melanin, which is produced in melanin granules (melanosomes) in melanocytes (melanocytes) between the epidermis and dermis. It is said that it diffuses to neighboring cells by osmotic action.

[0003] Conventionally, biochemical reactions carried out in melanocytes have been explained as follows. That is, tyrosine, an essential amino acid, is subjected to the action of tyrosinase to become dopaquinone, which undergoes an enzymatic or non-enzymatic oxidation action and changes to a red melanin through a red pigment and a colorless pigment. This process force is the main cause of S melanin pigment formation. Therefore, inhibition or suppression of the action of tyrosinase occurring in the first stage of this reaction can lead to suppression of melanin production, so that it has the action of inhibiting or suppressing the action of tyrosinase. The search for a drug is extremely important for preventing or suppressing the production of melanin caused by the action of tyrosinase.

[0004] From this point of view, many compounds have been researched and developed, and a whitening effect can be expected. For example, there are drugs on the market under the name of whitening agents.

Among these, for example, those relating to ellagic acid (see Patent Document 1), ascorbic acid and ascorbic acid derivatives (see Patent Document 2), which mainly reduce blackish melanin by reducing melanin. , Kojic acid (see patent document 3), hydroquinone (see non-patent document 1) and aractin, which acts directly on melanin cells to inhibit tyrosinase activity and inhibit melanin production to suppress darkness Others, such as those that promote metabolism and excrete melanin pigments outside the body, such as placenta extract (placenta extract), are known (see Patent Document 4).

[0005] In recent years, it has been reported that melanocyte stimulating hormone (hereinafter referred to as “HI_MSH”) force S and stains caused by ultraviolet rays are caused in the skin. This one MSH is one of the melanocyte activators, and promotes melanin synthesis through the receptor MC1R (melanocortin-1 receptor) on melanocytes (see Non-patent Documents 2 to 5). In addition, it has been confirmed that α-MSH production and MC1R expression are increased by ultraviolet irradiation (see Non-Patent Documents 6 to 7). From these findings, it is considered that those having the action of inhibiting the activation of melanocytes by a-MSH can work efficiently from the outside of the cell and exhibit a high whitening effect. As this type of whitening agent, canton carrot and the like are known (see Patent Document 5).

[0006] On the other hand, (+)-Lioniresinol and / or (-)-Lioniresinol are known as compounds having antibacterial action, antioxidant action, and aroma enhancing action (see Patent Document 6), and antibacterial makeup. It is used as a fee (see Patent Document 7). Furthermore, since rio-resinol glycoside has an epidermal cell growth promoting action, it is also used as an anti-oxidant effect and a cosmetic for preventing aging (wrinkle, rough skin) using its performance (Patent Document) 8). Furthermore, it is known to have various antioxidant activities such as lipid peroxide production inhibition, radical scavenging activity, oxygen radical scavenging activity. However, nothing was known about the whitening action (the action of suppressing or inhibiting melanin production) of rioniresinol.

Patent Document 1: Japanese Patent Application Laid-Open No. 64-79103

Patent Document 2: JP-A-2-45408 Patent Document 3: JP-A-53-3538

Patent Document 4: Japanese Patent Laid-Open No. 8-104616

Patent Document 5: Republished WO2004 / 017980

Patent Document 6: Japanese Unexamined Patent Publication No. 2003-128568

Patent Document 7: JP-A-10-139601

Patent Document 8: Japanese Patent Laid-Open No. 10-236940

Patent Document 9: Japanese Patent Laid-Open No. 11-2555639

Non-Patent Document 1 JAMA., No. 194, p. 965-967, 1965

Non-Patent Document 2: J. Cell Sci., 105, p. 1079-1084, 1993

Non-Patent Document 3: J. Cell Sci., 107, p. 205-211, 1994

Non-Patent Document 4: Anal. Biochem., 159, p. 191-197, 1986

Non-Patent Document 5: Pro Natl. Acad. Sci. USA, 92nd, p. 1792-1793, 1995

Non-Patent Document 6: Biochem. Biophys. Acta., Pp. 1313, p. 130-138, 1996 Non-Patent Document 7: British J. Darmatol, p. 139, p. 216-224, 1998 Disclosure of the Invention

Problems to be solved by the invention

As mentioned above, various drugs have been proposed as whitening agents, but many of the whitening cosmetics that only contain these whitening agents have weak expected effects. In terms of sexuality, there were insufficient points, which was not satisfactory. For example, ascorbic acid is known to have poor stability and induce dermatitis, and kojic acid and its derivatives are known to be easily decomposed by light and heat, although they have a strong whitening effect. Hydone quinone also has a strong whitening effect but is unstable and has a problem that it tends to be discolored in the manufacturing methods of micelles and emulsions, and also induces allergic contact dermatitis even at low concentrations. H. Ind. Med., 45 (6) IV, ρ · 376 — 80, 1988). In addition, crude drugs such as canton carrot have a problem that the crude drug itself is expensive and difficult to provide to consumers as an inexpensive whitening agent. [0008] The present invention provides a whitening agent having an excellent whitening effect even at a low blending amount, which provides a compound having excellent safety such as skin irritation and excellent stability. It provides food, drink, cosmetics and pharmaceuticals to facilitate ingestion or administration.

Means for solving the problem

[0009] As a result of intensive studies to solve the above problems, the present inventors have incorporated an extract obtained by extracting a beech family Quercus plant such as oak with an aqueous solution of a lower alcohol such as ethanol. It has been discovered that a whitening composition can be obtained, and a patent application has been filed (International Publication W005 / 28126). As a result of further diligent examination of this composition, it was found that the active ingredient of whitening was rioniresinol and its isomer. As a result of further investigations, (+) rio diresinol has a high inhibitory action on melanocyte activation by a-MSH, and (1) rio diresinol has a tyrosinase activity inhibitory action. It has been found that a mixture of (+)-rioniresinol and (1) rioniresinol, which is a whitening material having different mechanisms of action, is useful as a whitening agent, suppressing or preventing melanin formation. The invention has been completed.

[0010] That is, the present invention provides:

(1) The following formula (Α) to (Η)

[Chemical 1]

(Α) (Β) (C) (D)

(E) (F) (G) and (H) (In the formula, Me represents a methyl group.)

A whitening agent characterized by containing at least one lyoniresinol represented by the formula:

(2) Whitening active ingredient is formula (A)

[Chemical 2]

(E)

(In the formula, Me represents a methyl group.)

The whitening agent according to the above (1), characterized in that it is rioniresinol represented by

(3) The whitening agent according to the above (1) or (2), wherein the whitening active ingredient has a tyrosinase activity inhibitory action,

(4) The active ingredient that inhibits tyrosinase activity is represented by the formula (E)

[Chemical 4]

(E)

(In the formula, Me represents a methyl group.)

(1) Whitening agent according to the above (3), characterized in that it is rio diresinol,

(5) Whitening active ingredient strength Whitening agent according to (1) or (2), characterized by having an inhibitory action on S-melanin production,

(6) The active ingredient that inhibits melanogenesis is expressed by the formula (A)

[Chemical 5]

(A)

(In the formula, Me represents a methyl group.)

(+)-Lio diresinol represented by

(7) A cosmetic, food or drink, or pharmaceutical comprising at least one compound described in (1) or (2) as a whitening active ingredient,

(8) The cosmetic or pharmaceutical product according to (7) for external use,

(9) The cosmetic or pharmaceutical product according to (7), which is for oral use,

(10) The food and drink according to the above (7), which is labeled for whitening action, tyrosinase activity inhibitory action and / or melanin production inhibitory action,

(E) (F) (G) and (H)

(In the formula, Me represents a methyl group.)

A method of whitening the skin, which comprises administering to a mammal a whitening agent containing at least one of rio-resinols represented by the formula:

(12) Whitening active ingredient is formula (A)

[Chemical 7]

(A)

(In the formula, Me represents a methyl group.)

And / or formula (E)

(E)

(In the formula, Me represents a methyl group.)

The whitening method according to (11), wherein the whitening method is represented by

(13) The whitening method according to (11) or (12), wherein the whitening active ingredient has a tyrosinase activity inhibitory action,

(14) The active ingredient for inhibiting tyrosinase activity is represented by the formula (E)

[Chemical 9]

(E)

(In the formula, Me represents a methyl group.)

(1) Whitening method according to (13), characterized in that it is represented by

(15) The whitening method according to the above (11) or (12), wherein the whitening active ingredient has an inhibitory action on melanin production,

(16) The active ingredient that inhibits melanogenesis is expressed by the formula (A)

[Chemical 10]

(A)

(In the formula, Me represents a methyl group.)

The whitening method according to the above (15), wherein the whitening method is (+)-rio diresinol represented by:

(17) The whitening method according to the above (11) or (12), wherein the whitening agent is a cosmetic, a food or drink, or a medicine.

(18) The whitening method as described in (11) or (12) above, wherein the whitening agent is for external use

(19) The whitening method according to the above (11) or (12), wherein the whitening agent is for oral use,

(20) The following formulas (A) to (H) for producing a whitening agent

[Chemical 11]

(A) (B) (C) (D)

(E) (F) (G) and (H) (In the formula, Me represents a methyl group.)

Use of at least one of the rioniresinols represented by

(21) Rio diresinol is represented by the formula (A)

[Chemical 12]

(E)

(In the formula, Me represents a methyl group.)

The use according to (20) above, characterized in that it is rioniresinol represented by

(22) The use according to the above (20) or (21), wherein the whitening agent has a tyrosinase activity inhibitory action,

(23) Rio diresinol power formula)

[Chemical 14]

(E)

(In the formula, Me represents a methyl group.)

(1) The use according to the above (22), characterized by being rio diresinol,

(24) The use according to the above (20) or (21), wherein the whitening agent has a melanin production inhibitory action,

(25) Rio diresinol is represented by the formula (A)

[Chemical 15]

(A)

(In the formula, Me represents a methyl group.)

The use according to (24) above, characterized in that it is (+)-rio diresinol represented by:

(26) The use according to the above (20) or (21), wherein the whitening agent is a cosmetic, a food or drink, or a medicine.

(27) The use according to (20) or (21) above, wherein the whitening agent is for external use, and

(28) The use according to (20) or (21) above, wherein the whitening agent is for oral use. The invention's effect

[0011] The whitening agent provided by the present invention inhibits or suppresses the action of tyrosinase, which is the first step in the above-mentioned cause of melanogenesis, and melanocyte-stimulating hormone (hypertensive hormone) produced in the skin by Z or ultraviolet rays. Inhibiting the activity of melanocytes by (MSH) suppresses the production of the melanin pigment itself and at the same time reduces the amount of the pigment deposited. In particular, a whitening material with a different mechanism of action, that is, having a tyrosinase activity inhibitory action (1) a mixture of lio diresinol and melanocyte activity inhibitory action (+)-lio diresinol is formulated at a low concentration. However, it is possible to obtain an excellent whitening effect. In addition, the whitening agent provided by the present invention contains rio diresinol and its isomers contained in whiskey, ume vinegar and the like that have been ingested for food and drink as active whitening ingredients, and only skin irritation. Or when taken orally, it is safe. Further, the whitening agent provided by the present invention has a feature that it is excellent in stability. Furthermore, since the whitening agent provided by the present invention can be produced by chemical synthesis, it has the advantage that it can be provided to consumers as an inexpensive whitening agent.

Brief Description of Drawings

[0012] FIG. 1 is a graph showing the lyoniresinol content of extracts obtained by extracting American white oak and Spanish oak (new wood, old wood) with aqueous ethanol solutions of various concentrations. In the figure, △ indicates 40% ethanol, ▽ indicates 60% ethanol, mouth indicates 70% ethanol, and ○ indicates 96% ethanol.

[FIG. 2] FIG. 2 shows a process for identifying an active ingredient for inhibiting tyrosinase activity from an ethanol extract (whiskey) of a beech family Quercus.

[FIG. 3] FIG. 3 shows the tyrosinase inhibitory activity of (+) rio-niresinol and (1) rioniresinol. The vertical axis represents the inhibition rate (%).

[FIG. 4] FIG. 4 is a graph showing the amount of melanin produced in melanoma cells in mouse cells of lyoniresinol (racemate), (+)-ryoniresinol and (1) mono-lioresinol.

[FIG. 5] FIG. 5 shows the transition of lightness of guinea pig skin with respect to test sample application and ultraviolet irradiation. The vertical axis shows the UV value when the L value immediately before application of the test sample is 0. The difference in L value (AL) force from the start of application of the test sample to before irradiation, the number of days is shown on the horizontal axis.

BEST MODE FOR CARRYING OUT THE INVENTION

[0013] The following equations (A) to (H)

[Chemical 16]

(A) (B) (C) (D)

(E) (F) (G) and (H)

(In the formula, Me represents a methyl group.)

A whitening agent characterized by containing at least one lioresinol of a compound represented by the formula as a whitening active ingredient. The above formulas (A) to (H) are stereoisomers, and in the above formula, the compound represented by (A) is represented as (+) _ Lioniresinol ((+) _ Lyoniresinol) and represented by (E). The resulting compound is known as (1) 1 lyone diresinol ((1) 1 Lyoniresinol). Accordingly, lio-resinol is a compound represented by the formulas (A) and (E).

[0014] As a tyrosinase activity inhibitor, at least one of the two benzene rings having a carbon skeleton of lignans or norlignans is a 4-substituted resorcinol skeleton, followed by a benzylic skeleton. There has been proposed a tyrosinase activity inhibitor (see Patent Document 9) containing a lignan derivative and / or a kalelignan derivative as a whitening active ingredient in which the carbon atom has no substituent (see Patent Document 9). Since the compounds according to the present invention represented by A) to (H) are lignans having an arylene tetralin skeleton, they are different from the above lignans having a resorcinol skeleton. [0015] Rio diresinols used in the present invention, that is, rio diresinol and its isomers are naturally occurring, but can also be obtained by chemical synthesis, and can be obtained by any means. Even if it exists, it can be used without trouble in the present invention. When using a natural product, it is not limited to purified rioniresinol, but a raw material extract or crude product containing rioniresinol can also be used as a whitening agent of the present invention, for example, beechaceae (Fagaceae) Quercus plants, aqueous extracts of lower alcohols of these plants, or purified products thereof can be used. In addition, ume vinegar that is produced as a by-product in the production of umeshu and umeboshi, extracts thereof, or purified products from ume vinegar extract can be used.

[0016] Examples of the above Quercus genus plants include, for example, Q. mongolica Fisch. ), White oak (Quercus alba L.), common oak (Quercus robur L .; also called Limousin oak, French oak or Spanish oak), cecil oak (Quercus oetraea L.), cork oak (Quercus suber L.), etc. Can be mentioned. Different forces depending on the plant's production area, harvest time, extraction conditions, etc. Among the above Quercus plants, the plant group (plants called oaks) used as raw materials for the production and storage of whiskey and brandy etc. In particular, common oak and Mizunara are preferred because they contain rio-niresinol at a high concentration.

[0017] Examples of the extraction solvent used for the production of the lower alcohol aqueous extract of the Quercus genus plant include aqueous solutions of lower alcohols having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol, butanol, etc.). be able to. It is important that the concentration of the lower alcohol in the aqueous solution is a concentration at which rioniresinol can be efficiently extracted. Specifically, the lower alcohol concentration in the lower alcohol aqueous solution is about 10 to 100 volumes. %, Preferably about 30-70% by volume, more preferably about 40-60% by volume. Among the above lower alcohols, considering that it can be finally incorporated into foods and drinks, an aqueous ethanol solution is preferably used as the extraction solvent from the viewpoint of safety. In addition, here, the extraction solvent is not limited to lower alcohol, and other solvents may be used as long as the extraction efficiency is not greatly impaired. Components, for example, water-soluble components such as sugars, salts, acids, alkalis or amino acids, and various other solvents such as ethyl acetate and acetone may be contained. The extraction time may be set as appropriate, but in general, the longer the extraction time, the more rioniresinol can be extracted.

[0018] Although there is no particular limitation on the method for purifying rio-niresinol from the above Quercus genus plant extract or ume vinegar extract, it is preferable to purify by column chromatography, particularly by gel filtration chromatography. Purification is efficient and preferable. As a resin carrier used for purification, a resin commonly used such as Sephadex (registered trademark) or polyacrylamide gel (Bio-Gel) manufactured by Pharmacia can be selected and used according to the desired purity. When purifying using these resins, it is preferable to use a solvent such as acetonitrile, ethanol, methanol, acetone or benzene and an aqueous solution thereof as a developing solution.

[0019] Furthermore, the rioniresinols according to the present invention can be produced by chemical synthesis, and the synthesis can be carried out generally as follows. That is, using 4-hydroxy-1,3,5-dimethyoxyphenylpropionic acid as a raw material, Indian Journal Chemistry, 1976, 14-8, 128 (INDIAN J. CHEM., VOL. 14B, FEBRUARY, 19 (P. 128, p. 128).

[0020] It should be noted that, from the above natural product extract or lyoniresinol by chemical synthesis,

[Chemical 17]

(A)

(In the formula, Me represents a methyl group.)

Represented by (+)-Lio-resinol and formula (E) [Chemical 18]

(E)

(In the formula, Me represents a methyl group.)

Isolation and purification of (1) mono- and di-resinol represented by (1) can be performed by high performance liquid chromatography (HPLC) using a chiral column. The above (+)-Lioniresinol and (I) Mono-Lioresinol are both useful materials as whitening agents, and their action mechanisms are different. Specifically, (+) _ Lionisinol has a high inhibitory effect on melanocyte activation by melanocyte-stimulating hormone (a-MSH), and effectively exerts a whitening effect from outside the cell. ) Irioniresinol works directly on melanocytes to inhibit tyrosinase activity and inhibit melanin production. In the whitening agent of the present invention, these purified (+) _ rioniresinol and (1)-rioniresinol are used at an arbitrary ratio [about 1:99 to about 99: 1 (WZW)] depending on the purpose. It can be used by mixing.

[0021] The whitening agent of the present invention is produced by blending lio-resinols as a whitening active ingredient. For example, foods, drinks, cosmetics and pharmaceuticals having an excellent whitening effect can be obtained. The amount of rioniresinols to be added to the food, beverage, cosmetics and pharmaceuticals of the present invention varies depending on the form and use. Usually, about 0.0001 to 10% by weight is preferred. Weight percent is particularly preferred.

[0022] Examples of the food and drink containing the whitening active ingredient in the present invention include sports drinks, carbonated drinks, various drinks including fruit juices, soft drinks such as tea drinks, cakes, biscuits, breads, strawberries, and ice creams. Sweets such as cream, udon, soba, ramen, nosta, noodles such as somen, miso, soy sauce, vinegar, salad oil, sesame oil, butter, cheese, soy milk or What can be ingested as food, regardless of the type and form, such as milk. These can be produced by dissolving, mixing, etc., the whitening active ingredient of the present invention in food.

Next, cosmetics include, for example, lotions, jewels, lotions, creams, pack si IJs, emulsions, emulsions or cream-like foundations, lipsticks, powders, facial cleansers, hair tonics, solids, etc. , Sol-like, paste-like, suitable for external use, soft capsules that break in the mouth, or those sprayed in the mouth, etc. are mentioned as cosmetics of the present invention. These can be produced by a method known per se using other commonly used materials in addition to the whitening active ingredient according to the present invention. Other commonly used materials include oils and fats (e.g. waxes such as beeswax and carnavalou, jojoba oil, mink oil, cacao butter, palm oil, palm kernel oil, camellia oil, sesame oil, castor Oil, olive oil, etc.), surfactant (for example, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene alkyl ether, polyoxyethylene cetyl ether, sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyglycerin) Fatty acid esters), lower or higher alcohols (for example, cetanol, isostearyl alcohol, lauryl alcohol, hexadecyl alcohol, behenyl alcohol, otatildodecanol, etc.), fatty acids (for example, lauric acid, myristic acid) , Palmitic acid, Aric acid, undecylenic acid, oleic acid, etc.), water-soluble polymers (eg, carboxyvinyl polymer, alkyl-modified carboxybier polymer, cellulose, calcium alginate, etc.), polysaccharides (eg, hyaluronic acid, chondroitin sulfate, etc.) ), Peptides (such as collagen), preservatives (such as benzoic acid and its salts, isopropylmethylphenol, chlorhexidine hydrochloride, orthofelf enoreno, chronolehexidine gnoleconate, chronorecrezo monole, chronolephenesin) , Chlorobutanol, sorbic acid and its salts, dehydroacetic acid and its salts, paraoxyethylene benzoate, halocarban, etc.), thickener (for example, sodium carboxymethylcellulose, calcium alginate, polysaccharides, etc.) Humectants (such as glycerol, xylitol Honoré, Sonorebitonore, dipropylene glycol Honoré, butylene glycol Honoré, propylene glycol Honoré

Polyethylene glycolol 200 to 600, polyoxyethylene methyl darcoside, multitol, mannitol, etc.), pigments, fragrances, water or pH adjusters. [0024] Further, as pharmaceuticals, for example, externally used liniments, patches, ointments, sol coatings, etc., granules to be taken orally, fine granules, tablets, capsules, syrups Or a liquid agent etc. are mentioned. These can be produced by a method known per se using other commonly used materials in addition to the whitening active ingredient according to the present invention. Other commonly used materials include various additives such as excipients (eg, lactose, sucrose, glucose, starch, crystalline cellulose, etc.), binders (eg, starch paste, hydroxypropyl cellulose solution). , Carmellose solution, gum arabic solution, gelatin solution, sodium alginate solution, etc.), disintegrant (eg starch, carmellose sodium, calcium carbonate etc.), lubricant (eg magnesium stearate, talc, stearic acid, stearin) Acid calcium, etc.), surfactants (eg polysorbate 80, polyoxyethylene hydrogenated castor oil, etc.) or thickeners (eg hydroxyethyl cellulose, hydroxypropyl cellulose, polybutyl alcohol, polyethylene glycol etc.) It is done. Examples of dosage forms for oral medicines and cosmetics include chewable tablets and troches.

[0025] Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not construed as being limited thereto.

Example 1

[0026] Manufacture of extract containing lio-resinol (1)

As a plant material, oak (American white oak and Spanish oak, old wood) was used. Prepare 40, 60, 70, 96 volume% ethanol aqueous solution by mixing industrial ethanol with water, add 40 g of the above oak chips (1 XIX 2 cm) 2 to this ethanol aqueous solution, and 85 ° C For 5 minutes, then left at room temperature for 24 hours and again heated at 85 ° C for 5 minutes. The lyoniresinol content of the obtained extract was measured. The measurement conditions are as follows.

Column: Cosmosil 5C18AR-II

Column size: 3 X 250mm

Mobile phase: acetonitrile / water / formic acid = 20/80 / 0.1 (v / v / v)

Flow rate: 0.5mL / min

Measurement wavelength: 280nm The results are shown in Figure 1. Rio diresinol was found to be most extracted with 40-60% ethanol, especially 60% ethanol. Example 2

[0027] Manufacture of lyoniresinol-containing extract (2)

A new pot was prepared as a whiskey storage raw liquor. That is, the germinated barley (malt) is pulverized, mixed with warm water, saccharified, yeast is added to the filtered sugar solution and fermented to obtain moromi having an alcohol content of 7.0 to 7.5% by volume. It was. The koji was placed in a copper pot still (single distiller) and distilled twice to obtain a composition (new pot) having an alcohol concentration of 60% by volume. Next, whiskey production barrels (white oak, spanish oak, mizunara new barrels) are used, the above new pots are put in these barrels, stoppered, and stored in the storage for 5 years to obtain barrel extract. It was. After 5 years, the rio-resinol content of the obtained barrel extract was measured.

The lyoniresinol content of the various barrel extracts was 4.9 mg / L for white oak, 11 · 39 mg / L for Spanish oak, and 10 · 7 mg / L for Mizunara.

Example 3

[0028] Manufacture of rio-resinol-containing extract (3)

A barrel extract of whiskey was made by using American white oak barrels (old wood) for whiskey production and a new pot produced in the same manner as in Example 2. Barrel extracts were collected over time (0, 4, 8, 12 years), and the lio-resinol content of the extract was measured.

The rio-resinol content of the barrel extract collected over time is 0 years: Omg / L, 4 years: 0.668 mg / L, 8 years: 1. 13 mg / L, 12 years: 2. 15 mg / L Met.

Example 4

[0029] Purification of rioniresinol and its isomers

Commercially available whiskey as a lower alcohol aqueous solution extract from the beech family Quercus (Yamazaki 18 years, Suntory Co., Ltd., alcohol concentration 60./.) Using U. Evaporation of 400 mL of whiskey, followed by freeze-drying to obtain a dried product (whiskey congener) 1. Omg was obtained, pure water was added to this dried product, and the resulting pure water fraction was mixed with n— With hexane, ethyl acetate or n-butanol The tyrosinase inhibitory activity was measured by the following method (Figure 2-1). Pure water was added to the highly active ethyl acetate fraction and subjected to gel filtration chromatography. For gel filtration chromatography, a Sephadex (registered trademark) LH-20 force ram (Φ1.3 x 90cm) manufactured by Pharmacia was used, and it was separated with methanol at a flow rate of lmL / lOmin. . : Fractions of! ~ 3 [fractional self-coefficient (Kd) = 0—0.5, 0.5-1.0, 1.0—1.5] were fractionated. Among those fractions, the fractions with the highest activity were further divided into 1 to 5 fractions (Kd = 0.0.4-0.5, 0.5-5.0.6, 0.6-6.0.7, 0 7—0.8 and 0.8—0.9) were fractionated, and tyrosinase inhibitory activity was measured (FIG. 2 _ 2). As a result of the measurement, the tyrosinase inhibitory activity was highest at Kd = 0.6 -0.7.

[0030] The active fractions at the above Kd = 0.6 -0.7 were fractionated by HPLC. For the column, YMC-Pack ODS-AM (10 X 300 mm) was used, fractionated at a moving bed: 38% (v / v) methanol aqueous solution, flow rate: 2. OmL / min, detection wavelength: 280 nm. As a result, tyrosinase inhibitory activity was highest at peak No. 2 (Figure 2-3). This fraction was purified to a single peak on HPLC. The retention time of the fraction under the above HPLC conditions was about 17.5 minutes.

[0031] (Method for measuring tyrosinase inhibitory activity)

For B16 mouse melanoma cells, use DMEM (Dulbecco's Modified Eagle) medium containing 10% (W / W) urine fetal serum, 5% (v / v) CO at 37 ° C. In culture. Measurement

2

The sample and crude tyrosinase enzyme solution prepared from B16 mouse melanoma cells were mixed, and L-dopa was added as a substrate to a concentration of 0 · 05% (W / W). The mixture was reacted at 37 ° C for 20 minutes, and absorbance A at 492 nm (proportional to the amount of dopachrome) was measured. As a control, the same procedure was carried out without adding the test sample to the same reaction system, the absorbance B at 492 nm was measured, and the tyrosinase inhibition rate was calculated from the following formula.

Inhibition rate (%) = (1 Absorbance AZ Absorbance B) X 100

Example 5

[0032] Determination of structure of active substance

The active substance (fraction) isolated and purified in Example 4 was subjected to mass spectrometry (FAB MASS) and nuclear magnetic resonance NMR, ¹³ C-NMR) according to conventional methods, and spectral analysis was performed. went. As a result of dissolving FAB-MASS after dissolving the active substance in DMSO-D,

6

When turned on, the mass / charge (m / z) was found to be 443 (M + Na) ⁺ and the molecular weight to be 420. Next, nuclear magnetic resonance (^ H-NM ^ ^13C- NMR) was performed and a spectrum analysis was performed. Bmker Biospin DMX-750 ('H-NMR) or DMX-500 ( ¹³ C-NMR) was used for the measurement. As a result of the measurement, the chemical shift (ppm) and molecular weight of the carbon atom of the isolated and purified active material were completely the same as those described in Magn. Reson. Chem., 1985, No. 23, ρ369. Matched.

[0033] Therefore, as a result of mass spectrometry (FAB-MASS) and nuclear magnetic resonance spectra (ΝMR, ^13C- NMR) analysis, the chemical structure of the active substance showing inhibition of tyrosinase activity is represented by the following formulas (A) to ( It was found to be one or a mixture of two or more of H).

[Chemical 19]

(A) (B) (C) (D)

(E) (F) (G) and (H)

(In the formula, Me represents a methyl group.)

Example 6

[0034] Synthesis of lyoniresinol

Rioniresinol was produced according to a method known per se described in INDIAN J. CHEM., VOL.14B, FEBRUARY, 1976, page 128 (hereinafter abbreviated as “Document A”). More specifically, rioniresinol used in the following Examples 7 to 13 was prepared according to the following reaction formula. That is, [Chemical 20]

(In the formula, Ac represents a acetyl group, and Me represents a methyl group.)

In the same manner as in Reference A except that the above formula (1 (2 (3 (4)) was used instead of the formula (1 (11 (111 (IV)) in Reference A, respectively. The nuclear magnetic resonance and mass spectrometry of this rioniresinol were completely consistent with those of Example 5. As a result of circular dichroism (CD) spectral analysis, the synthesized product was found to be (+ It was found to be a mixture (racemate) of) -Rioniresinol and (1) Mono-Rioresinol.

Purification of (+) —Lioniresinol and (1) —Lioniresinol

Separation of rioniresinol (racemate) synthesized in Example 6 by high performance liquid chromatography (HPLC) was performed using a chiral column. The HPLC analysis conditions are as follows.

Column: CHIRALCEL AD— H

Column size: 0.461. D X 25cm

Mobile phase: Methanol / ethanol Z acetic acid = 50Z50Z0.1 (ν / ν / ν

Flow rate: 1. OmL / min Measurement wavelength: 254nm

The optical purity of the obtained fractions (+)-rioniresinol and (1) rioniresinol was 98% ee or higher, respectively. In addition, the ratio of (+)-lio diresinol and (1) -lio diresinol obtained was about 1: 1 (w / w).

Example 8

[0036] Measurement of Tyrosinase Inhibitory Activity (IC)

50

As the test substance, (+)-lio diresinol and (1) lio diresinol collected in Example 7 were used. 5 mg of the test substance was dissolved in DMSO (dimethyl sulfoxide), and tyrosinase inhibitory activity was measured in the same manner as in Example 4. As a positive control, arbutin, which is widely used in the market as having tyrosinase activity inhibition, was prepared in the same manner as described above, and tyrosinase inhibitory activity was measured.

[0037] The results are shown in FIG. It was found that both (+)-Lioniresinol and (I) LioniResinol have tyrosinase inhibitory activity.

In addition, IC of tyrosinase inhibitory activity of the test substance was calculated. The results are shown in Table 1. (

50

―) — Lioniresinol's IC is lower than that of arbutin, (−) — Lioniresinol

50 50

It showed higher tyrosinase inhibitory activity than force arbutin.

[0038] [Table 1]

Example 9

[0039] Measurement of melanin production

As test substances, the whiskey congeners containing (+)-Lioniresinol and (I) -LioniResinol prepared in Example 7 and rioniresinol prepared in Example 4 were used, and DMSO ( Each was prepared to have a final concentration force of S100 μg / mL. 4 × 10 ⁴ mouse B 16 melanoma cells were seeded in a 60 mm plastic dish and precultured. After 24 hours of pre-culture, the test substance was added and mixed, and further cultured at 37 ° C for 3 days. The cells were washed with PBS, lysed by adding 1 mL of 1M NaOH to 1 × 10 ⁶ centrifugal residues, and the absorbance at 470 nm was measured. In addition, after pre-culture, 0.1 ImM Nle α-Phe-a-melanocyte stimulating hormone (NDP-a_MSH) and the test substance were added and mixed, and further cultured at 37 ° C for 3 days. Similarly, the absorbance at 470 nm was measured. The absorbance at 470 nm of the sample to which neither the test substance nor NDP-a-MSH was added was defined as an intracellular melanin amount of 100%, and the ratio of each sample was calculated. In addition, saplinole was prepared in the same manner as described above using alpinin as a positive control, and the amount of intracellular melanin was measured.

The results are shown in Fig. 4. In (+)-Lioniresinol and a whiskey congener containing rioniresinols, melanin production was remarkably suppressed regardless of the addition of NDP-a-MSH. This effect was higher than that of arbutin.

Example 10

[0040] Measurement of melanin production inhibition using guinea pigs

(1) Test animals and breeding methods

Weiser maples (brown male guinea pig 4 weeks old) was purchased at room temperature 23.5. C, relative humidity 50 ± 10%, ventilation rate 10-15 times / hour, lighting time 7:00 to 19: 0 In the breeding room set at 09:00, polycarbonate gauge (width 29.2cm, height 20cm, Preliminary breeding for 1 week at a depth of 44 cm). Commercial chow and water (public tap water) were freely consumed. During the study period, body weight was measured once a week.

[0041] (¾ Tested Sampnore

Was dissolved chemically synthesized Rio two ricinoleic the (racemic) 60% (W / W) ethanol aqueous solution _10/0 (W / W) concentration as described in Example 6, containing the Rio two ricinoleic The test sample was used as a test sample. In addition, arbutin was used as a positive control, dissolved in 60% (W / W) ethanol aqueous solution at a concentration of 7% (W / W), and the lyoniresinol (racemate) of the present invention obtained above was used. Compared to the test sampnore containing. For control, a 60% (W / W) ethanol aqueous solution was used as a test sample. [0042] (3) Pigmentation by UV irradiation

A group of 12 guinea pigs preliminarily raised was used, and each back was depilated with an electric clipper and an electric shever. The guinea pig was placed on the back with a pattern, fixed on a compensator made of Sumipec 010 (UV transparent acrylic) and irradiated with ultraviolet rays. For UV irradiation, use UV irradiation device (CS & TOREX DERM ARAY medical UV irradiation device M— DMR 80 type) and UV-B lamp (TOREX FL20S -E-30 / DMR 20WAT TOSHIBA MEDICAL SUPPLY). Irradiated for 6 minutes at an intensity of 46 mWZcm2 (0.526j / cm2) ₀ UV intensity was measured using a UV intensity meter (DERMARAY UVR-3036ZS2). From the first day of the test, the pigment was deposited by irradiating with ultraviolet rays once a day for 3 consecutive days.

[0043] (4) Application of test sample

Immediately after the completion of UV irradiation on the third day of the test, the application of the test sample was started. Once a day, 40 / L was applied to each site.

[0044] (5) Evaluation of inhibitory effect on melanin pigmentation

The measurement site was measured five times using a color difference meter (Koni force Minolta COLOR READER CR-10), the skin color was displayed in the Lab color system, and the L value (lightness) was used for evaluation. The average value of L values (AL value) was calculated and used as an index.

[0045] (6) Evaluation result of L value

Fig. 5 shows the transition of skin color (AL value) when the test guinea pig was irradiated with ultraviolet rays (UV-B). A decrease in L value due to pigmentation was observed after the start of UV irradiation. In order to avoid measurement due to fluctuations in the amount of UV irradiation, the analysis was performed with the L value immediately before the start of application of the test sample as 0.

The group to which the test sample containing rioniresinol according to the present invention was applied showed the same melanin pigmentation inhibitory effect as the test sample containing arbutin. Considering that the concentration of the test sample containing rioniresinol according to the present invention is one-seventh the concentration of the test sample containing arbutin, rioniresinol according to the present invention has an excellent melamine pigmentation inhibitory effect. I found out.

In addition, the test sample containing rio diresinol according to the present invention is apparently dermatitis, etc. No symptoms were observed.

Example 11

[0046] Preparation of gel for skin

In 80 g of purified water, 0.5 g of poly (sodium acrylate) was dissolved, and rio diresinol (racemic) lg obtained in Example 6 was mixed with 18.5 g of ethyl alcohol, and a small amount of citrus essence was added. In preparation, a gel was prepared.

Example 12

[0047] Preparation of jelly-like peel-off pack

To 65 g of purified water, add 5 g of carboxymethyl cellulose and 10 g of polyvinyl alcohol, dissolve with heating, and add riouresinol (racemic) lg purified from the whiskey obtained in Example 4 to ethino-reanolol record. A jelly peel-off pack was prepared by adding 19g of Nore and adding a small amount of citrus essence.

Industrial applicability

[0048] The present invention relates to a whitening treatment that contains liodiresinols that inhibit melanin production by inhibiting tyrosinase activity involved in melanin pigment production and / or inhibiting melanocyte activity by a-MSH. An agent is provided. Therefore, the present invention is used in various products such as foods and drinks, cosmetics, and pharmaceuticals provided for the purpose of whitening.

Claims

The scope of the claims

The following formula (A) to (H)

[Chemical 1]

(A) (B) (C) (D)

(E) (F) (G) and (H)

(In the formula, Me represents a methyl group.)

A whitening agent characterized in that it contains at least one rio-resinol represented by the formula:

Whitening active ingredient is formula (A)

[Chemical 2]

(A)

(In the formula, Me represents a methyl group.)

And / or formula (E)

[Chemical 3]

(E)

(In the formula, Me represents a methyl group.)

2. The whitening agent according to claim 1, wherein the whitening agent is rioniresinol.

[3] The whitening agent according to claim 1 or 2, wherein the whitening active ingredient has a tyrosinase activity inhibitory action.

[4] The active ingredient that inhibits tyrosinase activity is the formula (E)

[Chemical 4]

(E)

(In the formula, Me represents a methyl group.)

The whitening agent according to claim 3, which is represented by (1) mono-ri-resinol.

[5] The whitening agent according to claim 1 or 2, wherein the whitening active ingredient has a melanin production inhibitory action.

[6] The active ingredient that inhibits melanin production is the formula (A)

[Chemical 5]

(A)

(In the formula, Me represents a methyl group.)

The whitening agent according to claim 5, wherein the whitening agent is (+)-rionidoresinol represented by the following formula.

[7] A cosmetic, food or drink, or pharmaceutical comprising at least one compound described in claim 1 or 2 as a whitening active ingredient.

[8] The cosmetic or pharmaceutical product according to claim 7, which is for external use.

[9] The cosmetic or pharmaceutical product according to claim 7, which is for oral use.

[10] The food or drink according to claim 7, which is labeled as being used for whitening action, tyrosinase activity inhibitory action and / or melanin production inhibitory action.

[11] The following formula (A;) to (H)

[Chemical 6]

(A) (B) (C) (D)

(In the formula, Me represents a methyl group.)

A method of whitening the skin, which comprises administering to a mammal a whitening agent containing at least one rioresinol represented by the formula:

Whitening active ingredient is formula (A)

[Chemical 7]

(E)

(In the formula, Me represents a methyl group.)

The whitening method according to claim 11, wherein the whitening method is represented by

[13] The whitening method according to claim 11 or 12, wherein the whitening active ingredient has a tyrosinase activity inhibitory action.

[14] The active ingredient that inhibits tyrosinase activity is the formula (E)

(E)

(In the formula, Me represents a methyl group.)

The whitening method according to claim 13, wherein the whitening method is represented by:

[15] The whitening active ingredient has an action of inhibiting lanine production.

The whitening method according to item 1 or item 12.

[16] The active ingredient that inhibits melanogenesis is the formula (A)

[Chemical 10]

(A)

(In the formula, Me represents a methyl group.)

The whitening method according to claim 15, wherein the whitening method is (+) -rio-resinol represented by the formula:

[17] The whitening method according to claim 11 or 12, wherein the whitening agent is a cosmetic, a food or drink, or a pharmaceutical.

[18] The whitening agent according to claim 11 or 12, wherein the whitening agent is for external use. Whitening method.

[19] The whitening method according to claim 11 or 12, wherein the whitening agent is for oral use.

[20] The following formulas (A) to (H) for producing a whitening agent

[Chemical 11]

(A) (B) (C) (D)

(E) (F) (G) and (H)

(In the formula, Me represents a methyl group.)

Use of at least one of the rioniresinols represented by

[21] Rio diresinol is represented by formula (A)

[Chemical 12]

(A)

(In the formula, Me represents a methyl group.)

And / or formula (E) [Chemical 13]

(E)

(In the formula, Me represents a methyl group.)

The use according to claim 20, wherein the whitening agent has a tyrosinase activity inhibitory action. Use of.

Rio Niresinol Power S, Formula)

[Chemical 14]

(E)

(In the formula, Me represents a methyl group.)

(1) The use according to claim 22, characterized in that it is rio diresinol.

[24] Use according to claim 20 or 21, wherein the whitening agent has an inhibitory action on melanin production.

[25] Rio diresinol is represented by formula (A) [Chemical 15]

(A)

(In the formula, Me represents a methyl group.)

25. Use according to claim 24, characterized in that it is (+)-Lio diresinol represented by

[26] Use according to claim 20 or 21, wherein the whitening agent is a cosmetic, a food or drink, or a medicine.

[27] The use according to claim 20 or 21, wherein the whitening agent is for external use.

[28] The use according to claim 20 or 21, wherein the whitening agent is for oral use.