JP2002370962A - Bleaching preparation and cosmetic for preventing and improving aging of skin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a tyrosinase inhibitor, an elastase inhibitor and a collagenase inhibitor, a bleaching preparation and a cosmetic for preventing and improving aging of the skin. SOLUTION: An extract of Japanese wisteria tea is brought to be included as effective ingredients of a tyrosinase inhibitor, an elastase inhibitor and a collagenase inhibitor. This bleaching preparation is obtained by formulating the aforesaid tyrosinase inhibitor. This cosmetic for preventing and improving aging of the skin is obtained by formulating the aforesaid elastase inhibitor and/or the collagenase inhibitor.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a tyrosinase inhibitor which inhibits tyrosinase activity involved in the production of melanin which causes spots, freckles and skin pigmentation, and an elastase activity which inhibits elastin reduction / denaturation. And a collagenase inhibitor that inhibits collagenase activity involved in the reduction and denaturation of collagen. Further, the present invention relates to a cosmetic for whitening and a cosmetic for preventing / improving skin aging.

[0002]

2. Description of the Related Art Skin spots, freckles and skin pigmentation after sunburn are the result of a marked increase in melanin production due to the activation of pigment cells (melanocytes) present in the skin. It is one of the worries.

In general, melanin changes from tyrosine to dopa and dopa to dopaquinone by the action of an enzyme tyrosinase biosynthesized in pigment cells.
It is supposed to be formed via an intermediate such as 6-dihydroxyindophenol. Therefore, in order to prevent and treat skin darkening (skin pigmentation), it is conceivable to inhibit the melanin production process or to bleach the already produced melanin in pale color.

[0004] Based on such a concept, various whitening components have been conventionally proposed. For example, sulfur compounds typified by colloidal sulfuric acid and glutathione have been used to inhibit tyrosinase activity and suppress melanin production. Ascorbic acids, hydrogen peroxide, hydroquinone, catechol, and the like have been used as light-color bleaching of the produced melanin.

However, regarding the prevention and treatment of skin pigmentation, ascorbic acids are easily oxidized and unstable in a system containing a large amount of water, such as a hydrated cosmetic, and cause discoloration. Further, aqueous hydrogen peroxide has a problem in storage stability and safety in use, and glutathione and colloidal sulfur have a problem in that they emit a noticeable odor. Furthermore, those such as hydroquinone and catechol have problems in use safety such as skin irritation and allergy. Therefore, the use of these whitening components in products such as cosmetics is restricted, and a sufficiently satisfactory whitening component has not yet been obtained.

[0006] As the change in the tissue level of the skin which has lost elasticity due to aging and has formed wrinkles and lumps, the decrease in collagen in the dermis layer is most remarkable. That is, in the skin dermis layer of healthy young people, fibrous collagen and elastin, which is a coiled hard protein existing entangled with the fibrous collagen, keep the skin firm and elastic. Both collagen and elastin are successively synthesized in the fibroblasts and replenished, and metabolism is repeated.However, when the amount of new collagen and elastin replenished decreases with age, or when elastin degradation is accelerated by UV irradiation, However, it becomes difficult to maintain the structures involved in maintaining the elasticity of the skin, and the skin becomes aged.

[0007] As described above, changes accompanying the aging of the skin, ie, wrinkles, dullness, dullness, disappearance of texture, decrease in elasticity, etc., involve reduction and denaturation of extracellular matrix components such as collagen and elastin. are doing.

In recent years, it has been pointed out that the involvement of matrix-based proteases as factors inducing these changes. Among matrix-based proteases, collagenase, that is, MMP-1 (matrix metalloprotease) is known as an enzyme that degrades type I and III collagens, which are the main components of the extracellular matrix of the dermis of the skin. It is thought that it greatly increases due to the irradiation of UV light, causes a decrease or denaturation of collagen by ultraviolet rays, and is a major factor such as formation of skin wrinkles. Therefore, promotion of collagen production and inhibition of collagenase activity are important in preventing and improving skin aging.

Hitherto, as a method for preventing and improving skin aging such as wrinkles and tarmi, protection from ultraviolet rays, elimination of antioxidants and active oxygen has been considered to be useful, and development of various substances has been attempted. However, the effect has not been obtained yet.

[0010]

Accordingly, the present invention provides a substance which is easily available and has a tyrosinase inhibitory action, an elastase inhibitory action or a collagenase inhibitory action from natural products whose safety has been confirmed. Headings,
An object of the present invention is to provide a tyrosinase inhibitor, an elastase inhibitor or a collagenase inhibitor containing the same as an active ingredient.

Another object of the present invention is to find a substance having a tyrosinase inhibitory effect from natural products whose safety has been confirmed, and to provide a whitening cosmetic containing the same. With the goal.

Further, the present invention has found a substance which is easily available and has an elastase inhibitory action or a collagenase inhibitory action from natural products which have been confirmed to be safe. It is intended to provide cosmetics for improvement.

[0013]

In order to achieve the above object, the tyrosinase inhibitor, elastase inhibitor and collagenase inhibitor of the present invention are characterized by containing a wisteria tea extract as an active ingredient. The whitening cosmetics and the skin aging preventing / improving cosmetics are characterized by containing a wisteria tea extract.

The whitening cosmetic of the present invention preferably further contains a whitening agent. The whitening agent is a group consisting of ascorbic acid and its derivatives, placenta extract, kojic acid, rucinol, ellagic acid and chamomile extract. It is preferable to use one or more whitening agents selected from the group consisting of:

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, "wisteria tea extract"
Means an extract obtained using wisteria tea as an extraction raw material,
"Wisteria tea extract" includes an extract obtained using wisteria tea as an extraction raw material, a diluent or a concentrate of the extract, a dried product obtained by drying the extract, or a crude product or a purified product thereof. Any of the things are included.

Fuji tea (scientific name: Ampe) used as an extraction raw material
lopsis grossedentata) is a perennial plant belonging to the grape family. The leaves of wisteria tea are used as beverages, and the roots or whole plants of wisteria tea are used as folk medicine for treatment of jaundice hepatitis, colds, sore throat, acute conjunctival inflammation and the like. Wisteria is native to a wide area from central to southern China, and is also cultivated in Taiwan. Therefore, wisteria tea is readily available from these regions. As the extraction raw material, whole plant, branch, leaf, branch and leaf of wisteria tea can be used, but it is preferable to use branch and leaf.

The wisteria tea used as an extraction raw material is suitably dried and pulverized immediately after collection. Drying may be performed in the sun or using a commonly used dryer. Wisteria tea may be used as an extraction raw material after pretreatment such as degreasing with a nonpolar solvent such as hexane or benzene. By performing a pretreatment such as degreasing, the extraction treatment of wisteria tea with a polar solvent can be performed efficiently.

At the time of the extraction treatment, it is preferable to use a polar solvent as the extraction solvent. The components having a tyrosinase inhibitory action, an elastase inhibitory action or a collagenase inhibitory action contained in wisteria tea can be easily extracted by an extraction treatment using a polar solvent as an extraction solvent.

Specific examples of suitable extraction solvents include water, lower aliphatic alcohols, and water-containing lower aliphatic alcohols, and these can be used alone or as a mixture of two or more thereof. Specific examples of suitable lower aliphatic alcohols include methanol, ethanol, propanol, 1,3-butylene glycol, glycerin,
Propylene glycol and the like can be exemplified.

The water that can be used as the extraction solvent includes pure water,
Tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, etc., as well as those subjected to various treatments are included. As a treatment to be applied to water, for example, purification, heating, sterilization, sterilization,
Includes filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

When a mixture of two or more polar solvents is used as the extraction solvent, the mixing ratio can be adjusted appropriately. For example, when a mixture of water and a lower aliphatic alcohol is used, the mixing ratio between water and the lower aliphatic alcohol can be freely adjusted, but 7: 3 to 2: 8.
(Weight ratio) is preferable.

The extraction treatment is not particularly limited as long as the soluble components contained in the wisteria tea can be eluted into the extraction solvent, and can be carried out according to a conventional method. In the extraction treatment, it is not necessary to employ a special extraction method, and any device can be used at room temperature or under reflux.

For example, an extraction raw material is charged into a treatment tank filled with an extraction solvent, and a soluble component is eluted with occasional stirring. At this time, the extraction conditions can be appropriately adjusted according to the extraction raw material and the like, but the amount of the extraction solvent is usually 5 to 15 times (weight ratio) the extraction raw material, and the extraction time is usually 1 to 3 hours.
The extraction temperature is usually from room temperature to 95 ° C.

After the soluble component is eluted by the extraction treatment, an extraction liquid can be obtained by filtering to remove the extraction residue. The obtained extract is diluted according to a conventional method to obtain a diluent or concentrate of the extract, a dried product of the extract, or a crude product or a purified product thereof.
Treatment such as concentration, drying, and purification may be performed.

The obtained extract can be used as it is as a tyrosinase inhibitor, an elastase inhibitor or a collagenase inhibitor, but a concentrated solution or a dried product thereof is easier to use. In order to obtain a dried extract, a carrier such as dextrin or cyclodextrin may be added to improve the hygroscopicity.

Further, since the wisteria tea extract has a peculiar smell and taste, it can be purified for the purpose of decolorization, deodorization, etc., as long as the physiological activity is not reduced. Since it is not used in large quantities when added to cosmetics, there is no practical problem even if it is not purified. The purification can be specifically performed by activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like.

Although the wisteria tea extract can be used as it is as a tyrosinase inhibitor, an elastase inhibitor or a collagenase inhibitor, it can also be formulated into a conventional method and used. In the case of formulation, a pharmaceutically acceptable carrier such as dextrin and cyclodextrin and other optional auxiliaries can be added to facilitate storage and handling. The wisteria tea extract can be made into any dosage form such as powder, granule, and tablet by formulation.

The tyrosinase inhibitor of the present invention can inhibit the activity of tyrosinase. It is thought that melanin is converted from tyrosine to dopa and dopa to dopaquinone by the action of the enzyme tyrosinase biosynthesized in pigment cells, and then formed via intermediates such as 5,6-dihydroxyindophenol. ing. Therefore, according to the tyrosinase inhibitor of the present invention, melanin production can be suppressed by inhibiting tyrosinase activity involved in melanin production. Further, according to the tyrosinase inhibitor of the present invention, a whitening effect can be exerted through suppression of melanin production.

The elastase inhibitor of the present invention can inhibit the activity of elastase. According to the elastase inhibitor of the present invention, through the elastase inhibitory action, elastin decreases elastin, suppresses denaturation, etc.,
It can prevent and / or improve skin aging caused by elastin decrease, degeneration, and the like.

The collagenase inhibitor of the present invention can inhibit the activity of collagenase. According to the collagenase inhibitor of the present invention, through the collagenase inhibitory action, the reduction of collagen by collagenase, suppression of denaturation, etc.,
It is possible to prevent and / or improve skin aging caused by a decrease or denaturation of collagen.

The tyrosinase inhibitor of the present invention can exhibit a whitening effect through tyrosinase inhibitory action and is excellent in use feeling and safety when applied to the skin. It is suitable for blending into ingredients.

The whitening cosmetic is a skin cosmetic used for the purpose of whitening, and the type thereof is not particularly limited. Examples thereof include ointments, creams, emulsions, lotions, and the like.
Packs, bath additives and the like can be exemplified.

It is preferable that a whitening agent is added to the whitening cosmetic in addition to the tyrosinase inhibitor of the present invention. The type of whitening agent is not particularly limited, but may be one or more whitening agents selected from the group consisting of ascorbic acid and its derivatives, placenta extract, kojic acid, rucinol, ellagic acid and chamomile extract. Preferably, there is. As long as the whitening effect does not hinder the whitening effect, various main ingredients and auxiliaries usually used in the manufacture of skin cosmetics and other optional auxiliaries can be blended. The tyrosinase inhibitor of the present invention is not limited to the main agent alone.

The amount of the tyrosinase inhibitor of the present invention in the whitening cosmetic can be appropriately adjusted according to the kind of the skin cosmetic and the physiological activity of the extract. About 0.001 to 1 in terms of extract
0% by weight, preferably 0.05 to 5% by weight.

The elastase inhibitor or collagenase inhibitor of the present invention can prevent and / or improve skin aging through an elastase inhibitory action or a collagenase inhibitory action, and can provide a feeling of use and safety when applied to the skin. It is suitable for blending into skin cosmetics, especially cosmetics for preventing and improving skin aging.

The cosmetic for preventing / improving skin aging is a skin cosmetic used for the purpose of preventing / preventing skin aging, and its type is not particularly limited. Prevents skin aging
Specific examples of the preventive cosmetics include ointments, creams, emulsions, lotions, packs, bath salts and the like.

The skin aging prevention / prevention cosmetic contains various main ingredients and auxiliaries usually used in the manufacture of skin cosmetology, as well as any other auxiliaries, as long as they do not hinder the skin aging prevention / prevention effect. With regard to the effect of preventing and preventing skin aging, it is not limited that only the elastase inhibitor or the collagenase inhibitor of the present invention is used as the main agent.

The amount of the elastase inhibitor or collagenase inhibitor of the present invention in the cosmetic for preventing and improving skin aging can be appropriately adjusted depending on the kind of the skin cosmetic, the physiological activity of the extract, and the like. The compounding ratio is about 0.001 to 10% by weight, preferably 0.05 to 5% by weight in terms of a standard wisteria tea extract.

Specific examples of cosmetics for whitening or cosmetics for preventing / improving skin aging which can be used as constituents together with the tyrosinase inhibitor, elastase inhibitor or collagenase inhibitor of the present invention are as follows. When the following components are used in combination with the wisteria tea extract, the synergistic action between the wisteria tea extract and the components used in combination may bring about superior use effects that are generally expected.

Examples of the astringent include citric acid or salts thereof, tartaric acid or salts thereof, lactic acid or salts thereof, aluminum chloride, aluminum potassium sulfate, allantoindihydroxyaluminum, zinc sulfate, proanthocyanidins, sycamore extract, rhubarb extract, and horse chestnut. Extract, cucumber extract, melissa extract and the like.

Examples of the bactericidal and antibacterial agents include benzoic acid, sodium benzoate, p-hydroxybenzoic acid ester, distearylmethylammonium chloride, orthophenylphenol, photosensitive element 101, photosensitive element 201, salicylic acid, sodium salicylate, sorbin Acid, resorcinol, phenoxyethanol, bisabolol, hinokitiol, oil-soluble licorice extract (licorice hydrophobic flavonoid,
Gravlidine, Gravrene, Ricochalcone A) and the like.

Examples of the ultraviolet absorber include β-isopropylfuranone derivative, urocanic acid, ethyl urocanate, oxybenzone, octyl paradimethylbenzoate, octyl paraaminobenzoate, paraaminobenzoic acid, ethyl paraaminobenzoate, and butylmethoxydibenzoyl. Methane, titanium oxide, β-carotene and the like can be mentioned.

Examples of humectants include serine, glycine, threonine, alanine, collagen, hydrolyzed collagen, hydronectin, fibronectin, keratin, elastin, royal jelly, chondroitin heparin, glycerophospholipid, lactic acid fermentation, yeast extract,
And soybean phospholipids, γ-oryzanol, below oil extract, yoquinin extract and the like.

Examples of the cell activator include riboflavin or a derivative thereof, pyridoxine or a derivative thereof, nicotinic acid or a derivative thereof, pantothenic acid or a derivative thereof,
α-Tocopherol or a derivative thereof, saxifrage extract, garlic extract, mannen wax extract and the like can be mentioned.

Examples of anti-inflammatory and anti-allergic agents include:
Azulene, allantoin, aminocaproic acid, indomethacin, lysozyme chloride, glycyrrhizic acid or its derivative, glycyrrhetinic acid or its derivative, photosensitizer 301
No. 401, diphenhydramine hydrochloride, adenosine phosphate, estradiol, esulone, ethinyl estradiol, spinach extract, perilla extract, pentagon extract and the like.

Examples of the antioxidant / active oxygen scavenger include dibutylhydroxytoluene, butylhydroxyanisole, propyl gallate, baicalin, baicalein, superoxide dismutase, catalase,
Rosemary extract, nutmeg extract, mace extract,
Laurel extract, turmeric extract and the like.

Examples of fats and oils include soybean oil, linseed oil, sesame oil, rapeseed oil, safflower oil, olive oil, camellia oil, almond oil, castor oil, coconut oil, tallow, jojoba oil, evening primrose oil and the like. .

The waxes include, for example, carnauba wax, candelilla wax, beeswax, salami beeswax, whale wax, Celax, lanolins and the like.

Examples of the hydrocarbons include liquid paraffin, petrolatum, microcrystalline wax, ceresin, squalane, polyethylene powder and the like.

The fatty acids include, for example, stearic acid, linoleic acid, lauric acid, myristic acid, palmitic acid, heveninic acid, lanolinic acid, oleic acid and the like.

Examples of alcohols include lauryl alcohol, cetyl alcohol, stearyl alcohol, lanolin alcohol, oleyl alcohol, hexadecyl alcohol, propylene glycol, 1,3-butylene glycol, ethylene glycol or a polymer thereof,
Cetostearyl alcohol and the like.

Examples of the esters include decyl oleate, butyl stearate, myristyl myristate, glyceryl monostearate, glyceryl trimyristate, lanolin acetate, and cetyl lactate.

Examples of the surfactant include an anionic surfactant, a cationic surfactant, an amphoteric surfactant, a nonionic surfactant and the like.

Examples of the flavor include menthol, carvone, eugenol, anethole, peppermint oil, spearmint oil, peppermint oil, eucalyptus oil, anise oil and other oil-like flavors from various animals and plants.

[0055]

The present invention will be described in more detail with reference to the following examples. [Preparation Example 1] 500 g each of wisteria tea leaves was extracted with 5 liters of water, 50% ethanol and ethanol at 80 ° C for 4 hours each. Thereafter, the extract obtained by filtration was concentrated under reduced pressure, and the extract from the wisteria tea leaves was separated by 97 times.
g, 130 g and 79 g were obtained.

[Test Example 1] Tyrosinase inhibitory activity test Mcclvaine buffer (pH 6.8) 1
mL, tyrosine solution (0.3 mg / ml) 0.3 mL,
And 5 mg of the wisteria tea extract of Production Example 1 in 25 (v / v)% DM
0.9 ml of a sample solution obtained by dissolving in 5 ml of an SO solution was added to a test tube, and pre-incubated at 37 ° C. for 10 minutes. Thereafter, 0.1 mL of a tyrosinase solution (5000 units / mL derived from mushrooms manufactured by Sigma) was added, and the mixture was incubated at 37 ° C. for 15 minutes. After the completion of the reaction, the absorbance at a wavelength of 475 nm was measured (hereinafter, this absorbance is referred to as "absorbance at the time of addition of the enzyme solution / sample solution").

The same operation and absorbance measurement were carried out by adding Macklepine buffer (pH 6.8) instead of the tyrosinase solution (hereinafter, this absorbance was referred to as "absorbance when no enzyme solution was added / when a sample solution was added"). ). In addition, the same operation and absorbance measurement were performed by adding a solvent that dissolves the sample instead of the sample solution (hereinafter, this absorbance is referred to as “absorbance when the enzyme solution is added and no sample solution is added”). Further, the same operation and absorbance measurement were performed by adding Macclepine buffer (pH 6.8) instead of the tyrosinase solution and adding a solvent that dissolves the sample instead of the sample solution (hereinafter, this absorbance was measured). `` No enzyme solution added
Absorbance when no sample solution is added ”).

Since the absorbance is proportional to the concentration of a group of coloring components such as melanin generated from tyrosine, the tyrosinase activity inhibition rate (%) was calculated from the obtained absorbance using the following equation.

[0059]

Formula 1: Tyrosinase activity inhibition rate (%) = ％ 1- (A−
B) / (CD)｝ × 100

In Equation 1, A is “absorbance when adding enzyme solution / sample solution”, B is “absorbance when adding enzyme solution / no sample solution”, and C is “absorbance when adding enzyme solution / no sample solution”. Absorbance at the time of addition "and D represent" absorbance at the time of no addition of the enzyme solution and no addition of the sample solution ".

The tyrosinase activity inhibition rate is measured by gradually decreasing the sample concentration, and the sample concentration IC ₅₀ (ppm; μg / mL) at which the tyrosinase activity inhibition rate becomes 50%.
Was determined by interpolation. Table 1 shows the results.

[0062]

[Table 1]

From the results shown in Table 1, it was confirmed that the extract from the wisteria tea leaves had tyrosinase inhibitory activity. In particular, it was confirmed that the 50% ethanol extract and the ethanol extract from the wisteria tea leaves had excellent tyrosinase inhibitory activity.

Test Example 2 Elastase Inhibition Test A 96-well plate was prepared. To one well, 8 mg of the wisteria tea extract of Production Example 1 was added to a 0.2 mol / L Tris-HCl buffer (containing 2% DMSO, pH 8). ) 50 µL of a sample solution obtained by dissolving in 5 ml and 50 µL of an elastase solution were added, and 100 µL of a substrate solution was further added and mixed. 2
After incubation at 5 ° C for 15 minutes, a wavelength of 41
The absorbance at 5 nm was measured (hereinafter, this absorbance is referred to as “absorbance at the time of addition of an enzyme solution / sample solution”). In addition,
As elastase solution, Sigma Elastase T
5 mg of ypeIII was dissolved in 1 mL of the above-mentioned Tris-HCl buffer, and this was diluted 250-fold with the above-mentioned Tris-HCl buffer and used. In addition, as the substrate solution, Sigma's NS
UCCINYL-ALA-ALA-ALA-p-NIT
ROANILIDE was dissolved in DMSO to a concentration of 45.1.
A 4 mg / mL solution was prepared, and this was diluted 100-fold with the above Tris-HCl buffer and used.

The same enzyme reaction and absorbance measurement were carried out by adding only the same amount of solvent as the sample solution instead of the sample solution (hereinafter, this absorbance is referred to as “absorbance when enzyme solution is added / no sample solution is added”). ). The same enzyme reaction and absorbance measurement were performed by adding the above-mentioned Tris-HCl buffer instead of the elastase solution (hereinafter, this absorbance is referred to as "absorbance when no enzyme solution is added / when a sample solution is added"). Further, the same enzyme reaction and absorbance measurement were carried out by adding only the same amount of solvent as the sample solution instead of the sample solution, and adding the above Tris-HCl buffer instead of the elastase solution (hereinafter, this absorbance was used). Is referred to as “absorbance when no enzyme solution is added and no sample solution is added”). The elastase activity inhibition rate (%) was determined by the following equation.

[0066]

Formula 2: Elastase activity inhibition rate (%) = {1- (A-
B) / (CD)} × 100

In Equation 2, A is “absorbance when adding enzyme solution / sample solution”, B is “absorbance when adding enzyme solution / no sample solution”, and C is “absorbance when adding enzyme solution / no sample solution”. Absorbance at the time of addition "and D represent" absorbance at the time of no addition of the enzyme solution and no addition of the sample solution ".

The elastase activity inhibition rate was measured by gradually decreasing the sample concentration, and the elastase activity was reduced by 50%.
The inhibitory sample concentration IC ₅₀ (ppm; μg / mL) was determined by interpolation. The results are shown in Table 2.

[0069]

[Table 2]

From the results shown in Table 2, it was confirmed that the extract from the wisteria tea leaves had elastase inhibitory activity. In particular, it was confirmed that the water extract from the wisteria tea leaves had excellent elastase inhibitory activity.

Test Example 3 Collagenase Inhibition Test 20 mg of the wisteria tea extract of Production Example 1 was dissolved in 5 ml of 0.1 mol / L Tris-HCl buffer (containing 20 mmol / L calcium chloride and 5% DMSO) (pH 7.1). The obtained sample solution (50 μL), the collagenase solution (50 μL) and the substrate solution (400 μL) were mixed, and incubated at 37 ° C. for 30 minutes. Then, the reaction was stopped with 1 mL of a 25 mmol / L citric acid solution, and extracted with 5 mL of ethyl acetate. The absorbance of the obtained extract at a wavelength of 320 nm (control solution: ethyl acetate) was measured (hereinafter, this absorbance is referred to as "absorbance at the time of addition of an enzyme solution / addition of a sample solution"). As the collagenase solution, a solution prepared by dissolving 5 mg of Sigma Collagenase Type IV in 1 mL of the above Tris-HCl buffer and diluting it 50-fold was used. As the substrate solution, BAC was added to the above Tris-HCl buffer.
HEM Fenichemikalieen AG Pz-
The peptide was used after being dissolved to a concentration of 0.5 mol / L.

The same enzyme reaction and absorbance measurement were carried out by adding an equal amount of a Tris-HCl buffer solution to the sample solution instead of the sample solution (hereinafter, this absorbance was measured when the enzyme solution was added and the sample solution was not added). Absorbance "). The same enzyme reaction and absorbance measurement were performed by adding a Tris-HCl buffer instead of the collagenase solution (hereinafter, this absorbance is referred to as "absorbance when no enzyme solution is added / when a sample solution is added"). Further, the same enzymatic reaction and absorbance measurement were carried out by adding only a solvent equivalent to the sample solution in place of the sample solution and adding a Tris-HCl buffer in place of the collagenase solution (hereinafter, this absorbance was measured). This is referred to as “absorbance when no enzyme solution is added and no sample solution is added”). The collagenase activity inhibition rate (%) was determined by the following equation.

[0073]

Formula 3: Collagenase activity inhibition rate (%) = {1- (A-
B) / (CD)} × 100

In Equation 3, A is “absorbance when adding enzyme solution / sample solution”, B is “absorbance when adding enzyme solution / no sample solution”, and C is “absorbance when adding enzyme solution / no sample solution”. Absorbance at the time of addition "and D represent" absorbance at the time of no addition of the enzyme solution and no addition of the sample solution ".

The collagenase activity inhibition rate was measured by gradually decreasing the sample concentration, and the collagenase activity was reduced by 50%.
The inhibitory sample concentration IC ₅₀ (ppm; μg / mL) was determined by interpolation. Table 3 shows the results.

[0076]

[Table 3]

From the results shown in Table 3, it was confirmed that the extract from the leaves of the wisteria tea leaves had collagenase inhibitory activity. In particular, it was confirmed that a 50% ethanol extract from wisteria tea leaves had excellent elastase inhibitory activity.

Test Example 4 The lotions of Examples 1 to 5 and Comparative Example were produced by a conventional method and subjected to a feel test during use. The test method consisted of 6 groups of 24 women in their 20s and 40s.
After washing the face twice a day for two weeks in the morning and evening, the example product was used on the right face and the comparative example product was used on the left face. that time,
Evaluation was made on four items: feel during use, stickiness after use, sustainability of moisturizing, and effect of improving skin roughness.

The evaluation was carried out on a five-point scale from 1 to 5, with “5 points” being “very good” (very good feel at the time of use, no stickiness after use, no moisture retention, "Very good", "4 points" are "Good" (Good feel at use, almost non-sticky after use, high moisture retention, high skin roughness improvement effect) , "3 points" are "normal" (the feel when using is normal,
There is not much stickiness after use, persistence of moisturizing is a little, skin roughness improvement effect is a little), "two points" is "somewhat bad" (feel at the time of use is slightly bad, there is some stickiness after use, Moisture retention is not very good, skin roughening is not so effective. "One point" is "poor" (feeling bad when used, stickiness after use is considerable, no moisturizing persistence, skin roughness improvement No effect).
Table 4 shows the average of the evaluation points.

[0080]

[Table 4]

From the results shown in Table 4, the wisteria tea extract, wisteria tea
Lotion containing 50% ethanol extract or wisteria tea ethanol extract (containing ascorbic acid derivative, kojic acid, rucinol, ellagic acid or chamomile extract as a whitening agent) has a good feel when used, and after use It was confirmed that there was little stickiness, and it was excellent in the moisturizing persistence and the skin roughness improving effect.

Test Example 5 The back of a colored guinea pig was shaved with a shaver, and four skin sections were set to be irradiated with ultraviolet rays. One section at that time was 1.5 × 1.5 cm. A lotion (Examples 6 to 7 and Comparative Examples 2 to 3) of the formulation shown in Table 5 was applied to each designated section at a daily dose of 50 μl.
It was applied for one day, and the lightness (L value) before ultraviolet irradiation was measured by a color pigment meter (manufactured by Minolta). After the measurement is completed, the back is irradiated with 0.2 J / cm ² of UV-B under the same conditions as above,
The same amount of UV-B was irradiated on the second day, and the degree of pigmentation was observed 4 days and 7 days after the irradiation. The degree of skin pigmentation was compared between the lightness before irradiation (Lb: L before) and the lightness 4 days and 7 days after irradiation (La: L after). Table 6 shows the results. In Table 6, the lightness (L
Value) is larger, the degree of pigmentation is smaller,
The less the difference in brightness before and after ultraviolet irradiation (that is, the smaller the Lb value-La value), the greater the whitening effect (pigmentation suppression effect) of the lotion applied to the skin.

[0083]

[Table 5]

[0084]

[Table 6]

From the results shown in Table 6, it was confirmed that the wisteria tea 50% ethanol extract had a whitening effect (a pigmentation inhibiting effect). Also, the lotion containing both the wisteria tea 50% ethanol extract and magnesium ascorbate (whitening agent) (Example 6) shows a particularly excellent whitening effect (pigmentation inhibitory effect) due to a synergistic effect. Was confirmed.

[Formulation Example 1] An emulsion having the following composition was produced by a conventional method. Jojoba oil 4.0 g Placenta extract 0.1 g Olive oil 2.0 g Squalane 2.0 g Cetanol 2.0 g Glyceryl monostearate 2.0 g Polyoxyethylene cetyl ether (20EO) 2.5 g Polyoxyethylene sorbitan oleate ( 20EO) 2.0 g 1.3-butylene glycol 3.0 g hinokitiol 0.15 g Fragrance 0.05 g Wisteria tea extract 5.0 g Purified water The remainder (total amount is 100 g)

[Formulation Example 2] A cream having the following composition was produced by a conventional method. Liquid paraffin 5.0 g Salami beeswax 4.0 g Cetanol 3.0 g Squalane 10.0 g Lanolin 2.0 g Stearic acid 1.0 g Polyoxyethylene sorbitan oleate (20EO) 1.5 g Glyceryl monostearate 3.0 g 1. 3-butylene glycol 6.0 g Methyl parahydroxybenzoate 1.5 g Fragrance 0.1 g Fujicha 50% ethanol extract 1.0 g Purified water The balance (total amount is 100 g)

[Formulation Example 3] A pack having the following composition was produced by a conventional method. Polyvinyl alcohol 15.0 g Polyethylene glycol 3.0 g Propylene glycol 7.0 g Ethanol 10.0 g Ethyl parahydroxybenzoate 0.05 g Fragrance 0.05 g Fujicha ethanol extract 1.0 g Purified water The remainder (total amount is 100 g)

[0089]

According to the present invention, a tyrosinase inhibitor, an elastase inhibitor and a collagenase inhibitor are provided. The tyrosinase inhibitor of the present invention can exert a whitening effect by inhibiting a tyrosinase activity involved in the production of melanin which causes spots, freckles and skin pigmentation. Therefore, the tyrosinase inhibitor of the present invention is useful as a component of a whitening cosmetic, and a whitening cosmetic containing the tyrosinase inhibitor of the present invention as an active ingredient is also provided by the present invention.

The elastase inhibitor of the present invention can exert the effect of preventing and improving skin aging by inhibiting elastase activity involved in elastin reduction / denaturation. In addition, the collagen inhibitor of the present invention can exert the action of preventing and improving skin aging by inhibiting the collagenase activity involved in collagen reduction / denaturation. Therefore, the elastase inhibitor or the collagenase inhibitor of the present invention is useful as a component of a cosmetic composition for preventing or improving skin aging, and contains the elastase inhibitor or the collagenase inhibitor of the present invention as an active ingredient. Improvement cosmetics are also provided by the present invention.

 ──────────────────────────────────────────────────続 き Continued on the front page F-term (reference) 4C083 AA071 AA072 AA111 AA112 AA122 AC022 AC072 AC082 AC122 AC182 AC242 AC302 AC422 AC442 AC482 AC841 AC842 AD042 AD112 AD512 AD552 AD641 AD642 CC05 CC07 DD31 EE16 4C088 AB56 MA08 MA02 MA03

Claims (7)

[Claims] 1. A tyrosinase inhibitor comprising wisteria tea extract as an active ingredient. 2. An elastase inhibitor comprising a wisteria tea extract as an active ingredient. 3. A collagenase inhibitor comprising a wisteria tea extract as an active ingredient. 4. A whitening cosmetic comprising the tyrosinase inhibitor according to claim 1. 5. The whitening cosmetic according to claim 4, further comprising a whitening agent. 6. The whitening agent comprises ascorbic acid and derivatives thereof, placenta extract, kojic acid, lucinol,
The whitening cosmetic according to claim 5, which is one or more whitening agents selected from the group consisting of ellagic acid and chamomile extract. 7. A cosmetic composition for preventing and improving skin aging, comprising the elastase inhibitor according to claim 2 and / or the collagenase inhibitor according to claim 3.