JP2016041668A - Skin whitening composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel skin-whitening agent that has excellent safety and skin functionality, and also has high tyrosinase and/or melanin production inhibitory activity.MEANS FOR SOLVING THE PROBLEM: The present invention provides a tyrosinase inhibitor and a melanin production inhibitor comprising Myrtaceae Myrcia plants or extracts thereof as an active ingredient.SELECTED DRAWING: None

Description

  The present invention relates to a whitening agent. More specifically, a melanin production inhibitor and / or a tyrosinase inhibitor, which is a plant of Myrtaceae Myrcia, particularly preferably Myrcia breachata, or an extract thereof, and a whitening agent containing the same The present invention relates to a pharmaceutical composition, a cosmetic composition, and a food.

  Skin pigmentation due to aging, skin freckles and sunburn are caused as a result of markedly increased melanin production due to activation of pigment cells (melanocytes) present in the skin. It has become one of

  In general, melanin is converted from tyrosine to dopa and from dopa to dopaquinone by the action of tyrosinase, an enzyme biosynthesized in pigment cells, and then formed through intermediates such as 5,6-dihydroxyindophenol. It is said that. Therefore, in order to prevent and treat skin pigmentation etc., it is effective to inhibit the melanin production process.

  Based on such an idea, various whitening components have been conventionally proposed. For example, as compounds that inhibit tyrosinase activity and suppress melanin production, sulfur compounds such as colloidal sulfate and glutathione, kojic acid, arbutin, and the like are known.

  Of these, kojic acid and its derivatives have been used in whitening cosmetics as tyrosinase inhibitors and melanin production inhibitors (see Patent Documents 1-3). However, in recent years, the possibility that kojic acid has carcinogenicity and genotoxicity has been pointed out, and a safe tyrosinase inhibitor and a melanin production inhibitor are demanded instead.

  Furthermore, in whitening cosmetics, an attempt has been made to find a plant extract having an inhibitory effect on melanin production, reflecting the demand for low skin irritation and natural orientation (Patent Document 4). See -8).

Japanese Examined Patent Publication No. 56-18869 Japanese Patent Publication No. 60-9722 Japanese Patent Publication No. 60-10005 Japanese Examined Patent Publication No. 61-10442 Japanese Patent Publication No. 5-27605 Japanese Patent Publication No. 5-87482 Japanese Patent No. 5149548 Patent No. 4975069

  In view of the present situation as described above, the main object of the present invention is to provide a novel whitening agent that is excellent in safety and skin functionality and has a high tyrosinase inhibitory action and / or melanin production inhibitory action.

  In order to solve the above problems, the present inventor has searched for various natural products and found for the first time that plants of the genus Myrchiaceae have a strong tyrosinase inhibitory action and a melanin production inhibitory action, thereby completing the present invention. It was.

The present invention provides the following tyrosinase inhibitors and / or melanin production inhibitors.
1. A tyrosinase inhibitor and / or a melanin production inhibitor comprising as an active ingredient a plant of the genus Myrtia spp. Or an extract thereof.
A tyrosinase inhibitor and / or a melanin production inhibitor, wherein the plant of 2.1 is Myrcia bracteata.
3. The tyrosinase inhibitor and / or melanin production inhibitor according to 1-2, wherein the extract is an extract of water, alcohol, or a mixed solvent thereof.
4). The tyrosinase inhibitor and / or melanin production inhibitor according to 1 to 3, wherein the alcohol described in the previous period is ethanol.
5. The tyrosinase inhibitor and / or melanin production inhibitor according to 1-4, wherein the ethanol concentration of the solvent described in the previous period is 10 to 100% by volume.
A whitening agent comprising 6.1 to 5.
The skin external preparation formed by mix | blending 7.1-6.

Melanin pigments that cause skin coloring are produced by pigment cells (melanocytes) in the skin tissue. Pigmentation such as skin spots and buckwheat is caused by abnormal deposition of melanin as a result of activation of pigment cells due to stimulation by ultraviolet exposure from sunlight, hormonal abnormalities, or genetic factors. It is thought to do. Normally, melanin is metabolized by metabolism (turnover) of skin tissue, but a decrease in metabolism of skin tissue with aging also causes pigmentation.

  The plant of the genus Myrchiaceae used in the present invention or an extract thereof, such as Murta Caberuda or an extract thereof, is a highly safe material used as a decoction in the country of origin. As shown in the experimental examples to be described later, Murta Caberuda has an inhibitory activity superior to arbutin, which has an inhibitory effect on the enzyme tyrosinase involved in melanin synthesis, and has an action of suppressing melanin production. To whitening (improves pigmentation and dullness). Therefore, Murta Caberuda is useful as an active ingredient of a tyrosinase inhibitor, a melanin production inhibitor, and a whitening agent, and is also useful as an active ingredient of a skin external preparation (whitening external preparation) for the purpose of whitening effect.

A whitening external preparation containing a tyrosinase inhibitor, a melanin production inhibitor, and a whitening agent of the present invention is safe and has an excellent whitening action, and is capable of reducing pigmentation, blemishes, freckles, erythema and the like after sunburn. Since it has a beautifying effect such as making it inconspicuous and can maintain healthy skin, it is particularly useful as a whitening cosmetic, a beautiful skin cosmetic, a skin care cosmetic, and the like.

  In the present invention, the active ingredient having a tyrosinase inhibitory action and / or a melanin production inhibitory action is derived from a plant belonging to the genus Myrchiaceae, and extracts of these plants are preferably used. As for the plant of the genus Myrtia, Myrcia cabruda, Myrcia lanceola, Myrcia longipes, Pedra, Umia, Mircia, and M. Preferred examples include Murta Caberuda.

  Murta Caberuda (also known as Myrcia bracteta, Multi-cabeluda, Myrtus bacteta, Goabinha, etc.) belongs to the Myrtus genus Myrtus bractata and is distributed in South America.

The plants of the genus Mirkia are well known and are widely distributed mainly in South America. Particularly well known is Myrcia multiflora, which is an HMG-CoA reductase inhibitory action (JP 2008-297230), a 5α reductase inhibitory action (JP 2006-257060), a lipase inhibitory action (special 2006-257058), serine protease activity inhibitory action (Japanese Patent Laid-Open No. 2001-240551) and the like have been reported. Japanese Patent Application Laid-Open No. 2003-055246 proposes use as a melanin production inhibitor, but there is no report on the effect.

Among the species belonging to the genus Mirkia, there are few reports on Murta Caberuda. All of the proposed biological activities have been established in folk remedies, and are expected to be effective against indigestion and stomach weakness and to prevent diarrhea in children. As a result of searching for various natural products, it has been surprisingly found that Murta Caberuda has a strong tyrosinase inhibitory action and melanin production inhibitory action over arbutin.

The extract of the genus Myrchiaceae (for example, Murta Caberuda) used in the present invention is a part of the plant body such as leaves, bark (stem or trunk), flower, fruit, root, or the whole plant. It is obtained by extracting from the body. When the plant is Murta Caberuda, extracts from leaves, bark and flowers are preferably exemplified. The method for producing the extract is not particularly limited, and examples thereof include those extracted at normal temperature or heat using an appropriate extraction solvent. Examples of the extraction solvent include water, hydrous alcohol (eg, hydrous ethanol), lower alcohols having 1 to 4 carbon atoms (eg, methanol, ethanol, n-propanol, isopropanol), ketones such as acetone and methyl ethyl ketone, and ethyl acetate. Examples thereof include esters, polyhydric alcohols (ethylene glycol, 1,3-butylene glycol, propylene glycol, glycerin and the like), ethers such as tetrahydrofuran and diethyl ether, and organic solvents such as hexane. Water, a water-containing solvent, or a water-miscible solvent is preferable, and water, ethanol, and water-containing ethanol are particularly preferable. In the case of water-containing ethanol, although the ethanol concentration is not limited, 20 to 90% by volume, particularly 40 to 80% by volume can be preferably mentioned. These solvents may be used alone or in combination of two or more.
The plant extract may be used as it is in the extracted solution, and may be used after treatment such as concentration, dilution, filtration, etc. if necessary, and the extract may be evaporated to dryness, spray dried, freeze dried, etc. It may be processed and used as a dry product (powder).

  In the tyrosinase inhibitor and / or melanin production inhibitor of the present invention, the blending amount of the extract of the plant belonging to the genus Myrtia is not particularly limited as long as it has a melanin production inhibitory action, but in terms of dryness, 0.0001 to 100 % By weight, preferably 0.001 to 50% by weight, more preferably 0.01 to 25% by weight.

  The tyrosinase inhibitor and / or melanin production inhibitor of the present invention has an action of suppressing the production of melanin, and is suitably used as an active ingredient of a whitening agent and as an active ingredient of a cosmetic for the purpose of a whitening effect. Can be used.

  The whitening agent of the present invention is characterized by containing the aforementioned tyrosinase and / or melanin production inhibitor as an active ingredient. The ratio of the melanin production inhibitor to be added to the whitening agent is not particularly limited as long as it has a whitening action based on the tyrosinase and / or melanin production inhibitory action of tyrosinase and / or melanin production. In terms of the amount of the plant or the extract thereof (in terms of dry matter), it is 0.0005 to 20% by weight, preferably 0.001 to 10% by weight, more preferably 0.01 to 1% by weight. .

  The form of the whitening agent of the present invention is not particularly limited. For example, the whitening agent may have a solid preparation form such as powder, granule, tablet, capsule or the like, and may be liquid, suspension, emulsion. You may have liquid formulation forms, such as a shape.

  The whitening agent of the present invention has an action of suppressing the production of melanin based on the tyrosinase inhibitory activity of an extract of the genus Myrtia spp., Which is effective for whitening (including prevention and reduction of spots and dullness). It can be suitably used as an active ingredient in skin external preparations (whitening external preparations), particularly whitening cosmetics.

  You may mix | blend a conventionally well-known whitening agent with the whitening agent of this invention in combination with the plant of the Myrtaceae mirukia genus or these extracts. In addition, components usually used in cosmetics, such as humectants, film agents, antioxidants, oily components, ultraviolet absorbers, surfactants, thickeners, alcohols, powder components, color materials, aqueous components, Water, various skin nutrients, and the like can be appropriately blended as necessary.

The dosage forms of the whitening agent of the present invention are aqueous solution system, solubilization system, emulsification system, powder system, oil liquid system, gel system, ointment system, aerosol system, water-oil two-layer system, and water-oil-powder 3 A wide range of dosage forms such as layer systems can be adopted. For example, the cosmetic liquid, milky lotion, cream, gel, cosmetic liquid, pack, mask, etc. can be mentioned as the form.

  The whitening agent of the present invention can also be used as an external preparation for skin for the purpose of whitening, particularly as an active ingredient in cosmetics. In this case, the blending ratio of the whitening agent to the external preparation for skin (preferably cosmetic) is not particularly limited as long as the external preparation for skin exhibits a whitening effect. In terms of dry matter (in terms of dry matter), 0.0005 to 20% by weight, preferably 0.001 to 10% by weight, and more preferably 0.01 to 1% by weight. The form of the external preparation for skin is not particularly limited. For example, when the external preparation for skin is a basic cosmetic, examples of the form include lotion, milky lotion, cream, gel, essence, cosmetic liquid, pack, and mask. In addition, if the external preparation for skin is a makeup cosmetic, examples of the form include foundation, lipstick, blusher, eye shadow, eyeliner, mascara and the like. Other forms include facial cleansing agents, massage agents, cleansing agents, after-shave lotions, pre-shave lotions, shaving creams, body soaps, body creams, soaps and the like.

  EXAMPLES Hereinafter, although an Example, a test example, and a formulation example demonstrate this invention in detail, this invention is not limited at all by these Examples.

(Example 1) Preparation of water-containing ethanol extract of Murta Caberuda 50 ml of 80% by volume water-containing ethanol (ethanol: purified water = 8: 2 (v / v)) was added to 5 g of the dried plant body, and 1 at room temperature. Mixed with shaking overnight. Thereafter, the mixture was centrifuged at 3,000 rpm for 15 minutes to obtain a supernatant. The obtained supernatant was subjected to a rotary evaporator to remove ethanol to some extent and then freeze-dried to obtain a hydrous ethanol extract (586 mg).

(Example 2) Preparation of water-containing ethanol extract of pedra, ume, and kaa 50 ml of 80 volume% water-containing ethanol (ethanol: purified water = 8: 2 (v / v)) was added to 5 g of the dried plant body at room temperature. And mixed with shaking overnight. Thereafter, the mixture was centrifuged at 3,000 rpm for 15 minutes to obtain a supernatant. The obtained supernatant was subjected to a rotary evaporator to remove ethanol to some extent and then freeze-dried to obtain a hydrous ethanol extract (590 mg).

(Test Example 1) Measurement of Tyrosinase Inhibitory Activity Tyrosinase inhibitory activity was measured by the following method using the Murta Caberuda and Pedra Ume Kaa extracts prepared in Examples 1 and 2 as test samples.
(1) Test method Two plates were prepared for measuring tyrosinase inhibitory activity and confirming cytotoxicity. In a 96-well plate, mouse melanoma B16 cells suspended in 20 ng / ml MSH / DMEM medium (Gibco) were seeded at a rate of 5 × 10 ³ cells / well, under conditions of 37 ° C. and 5% CO ₂ . Incubated for 24 hours. Thereafter, Murta Caberuda extract, Pedra Ume kaa extract, arbutin as a test sample, and solvent (DMSO) as a control were added, and the cells were cultured under conditions of 37 ° C. and 5% CO ₂ for 3 days. After incubation, the supernatant of one plate was removed, the wells were washed with PBS, 1% TRITON / PBS solution, 1.97 mg / ml L-DOPA was added at a ratio of 9: 1, and addition 0, Abs492 after 1 hour was measured and used to calculate tyrosinase inhibitory activity. The remaining one plate was used for viable cell measurement using CellTiter-Glo kit (Promega).

Cell viability (%) was derived from the following formula.
(Equation 1)
Cell viability (%) = ([measured value] B− [measured value] S) / [measured value] B × 100
[Measured value] B: Control measured value [Measured value] S: Test sample measured value

  Tyrosinase inhibitory activity (%) was derived from the following formula. Moreover, IC50 of each test sample was calculated | required from obtained inhibitory activity (%).

(Equation 2)
Tyrosinase inhibitory activity (%) = ([measured value] _B− [measured value] _S ) / [measured value] _B × 100
[Measured value] _B : Control measured value (1 hour)-Control measured value (0 hour)
[Measured value] _S : Test sample measurement value (1 hour)-Test sample measurement value (0 hour)

(2) Test results

Table 1 shows the tyrosinase inhibitory activity and cell viability when the sample was added at a final concentration of 300 μg / ml.

The IC50 value of each test sample is shown in Table 1.

From the above results, the Murta Caberuda extract of the present invention does not exhibit strong cytotoxicity like the Pedra Ume Kaa extract, and has a stronger tyrosinase inhibitory activity than arbutin generally used as a whitening agent. It became clear to have.

(Test Example 2) Inhibitory Action on Cell Melanin Production The melanin production inhibitory action was measured using the Murta Caberuda extract prepared in Example 1 as a test sample.
(1) Test method Mouse melanoma B16 cells suspended in DMEM medium (Gibco) were seeded in 10 cm dishes for cell culture at 3 × 10 ⁵ cells and incubated at 37 ° C. under 5% CO ₂ for 24 hours. . Thereafter, Murta Caberuda extract and arbutin were added as test samples, and solvent (DMSO) was added as a control, followed by culturing at 37 ° C. and 5% CO ₂ for 3 days. After culturing, the supernatant was removed, the wells were washed with PBS, trypsin-EDTA solution (Gibco) was added to detach the cells, and the resulting cell suspension was collected in a tube. 20 μl of the obtained cell suspension was subjected to viable cell measurement using CellTiter-Glo kit (Promega).
The tube from which the cell suspension was collected was centrifuged at 15,000 rpm for 10 minutes to accumulate the cells at the bottom of the tube, and then the supernatant was removed. Add 1N NaOH in an amount corresponding to the number of cells, incubate at 95 ° C. for 60 minutes to extract melanin, measure the absorbance at 405 nm after taking a photograph, and measure values (test sample measured values, control measured values) did. Simultaneously, the absorbance of 1N NaOH was measured and used as a background value.

From the measured values obtained, the melanin production inhibition rate (%) was determined for each test sample and control using the following formula.

(Equation 3)
Melanin production inhibition rate (%) =
{(Absorbance of control−absorbance of test sample) / absorbance of control} × 100
Absorbance of test sample: measured value of test sample-background value
Absorbance of control: Control measurement value-background value

(2) The test results are shown in Table 2.

  From the above results, it was revealed that the Murta Caberuda extract of the present invention has a melanin production inhibitory action. From these facts, it is considered that these multa caberuda extracts are useful as a melanin production inhibitor, and as a whitening agent and an active ingredient of a cosmetic having a whitening effect, based on the melanin production inhibitory action.

Formulation Example 1: Whitening agent having a lotion form Using each of the components shown in Table 4 below, a whitening agent having a lotion form was prepared according to a conventional method.

Formulation Example 2: Whitening agent having a cosmetic liquid form Using each of the components shown in Table 5 below, a whitening agent having a cosmetic liquid form was produced according to a conventional method.

Formulation Example 3: Whitening Agent Having an Emulsion Form Using each component shown in Table 6 below, an emulsion was produced according to a conventional method.

Formulation Example 4: Whitening Agent having Cream Form A cream was produced according to a conventional method using each component shown in Table 7 below.

Formulation example 5: Whitening agent which has bath salt form Bath salt was manufactured according to the conventional method using each component shown in the following Table 8.

Formulation Example 6: After sterilizing tea leaves, multa and caberuda leaves as necessary, pulverized into strips, chops, powders or granules, then filled into large bags, tea packs, sticks or three-way seals and packaged . Moreover, you may add a roasting process after a sterilization process.
Formulation Example 7: Tablet Tablets were produced according to a conventional method using the components shown in Table 9 below.

Formulation Example 8: Capsule According to a conventional method, each component shown in Table 10 was mixed and filled into a capsule to obtain a capsule preparation.

Claims (5)

A tyrosinase inhibitor and / or a melanin production inhibitor comprising a plant of Myrtaceae Myrcia or an extract thereof as an active ingredient. A tyrosinase inhibitor and / or a melanin production inhibitor, wherein the plant of claim 1 is Myrcia bracteata. The tyrosinase inhibitor and / or melanin production inhibitor according to claim 1 or 2, wherein the extract is an extract of water, alcohol or a mixed solvent thereof. The whitening agent formed by mix | blending Claims 1-3. The skin external preparation formed by mix | blending Claims 1-4.