WO2014088296A1 - Skin whitening composition using xylosma congesta extract - Google Patents

Abstract

The present invention relates to a skin whitening composition using a Xylosma congesta extract which has an inhibitory activity on mushroom tyrosinase, and shows an inhibitory activity on the generation of melanin when being treated in B16F10, which is a melanoma cell stimulated by α-MSH, which is a melanocyte stimulator.

Description

Skin Whitening Composition Using Acid Citrus Extract

The present invention relates to a composition for skin whitening using an extract of Xylosma congesta.

It is well known that skin whitening is directly related to melanin pigment.

Melanin protects sub-dermal skin organs by blocking UV rays and protects the damage of proteins and genes in the skin caused by free radicals generated in the skin. It causes hyperpigmentation such as blemishes and freckles.

Melanin synthesis is achieved by the action of tyrosinase and tyrosinase related proteins present in the melanosomes. Specifically, tyrosine is oxidized by tyrosinase and is converted into dopa (DOPA, 3,4-dihydroxyphenylalanine), followed by oxidization of dopa by the action of dopaoxidase and then dopaquinone. In addition, melanin is produced by the action of tyrosinase-related proteins TRP-1 (5,6-dihydroxy indole-2-carboxylic acid oxidase) and TRP-2 (dopachrome tautomerase). Tyrosinase, TRP-1 and TRP-2 are promoted by the transcription factors microphthalmia-associated transcription factor (MITF), which is known to inhibit the expression of extracellular signal-regulated kinase (ERK) (DS). Kim, et al., J Cell Sci. 2003; 116: 1699-706).

The present invention discloses an extract of citron oil having a tyrosinase inhibitory activity and a melanogenesis inhibitory activity in B16F10, a melanoma cell stimulated with α-MSH, which is a melanogenesis-stimulating agent.

It is an object of the present invention to provide a composition for skin whitening using an acid extract.

Other and specific objects of the present invention will be presented below.

As confirmed in the following examples and experimental examples, the inventors of the present invention showed that 70% ethanol extract of citron oil leaves B16F10, which is a melanoma cell stimulated with α-MSH, which has mushroom tyrosinase inhibitory activity and is also a melanogenesis-stimulating agent. When treated, it was confirmed that it has a melanin production inhibitory activity in a concentration-dependent manner.

The composition for skin whitening of the present invention is provided based on the results of these experiments, and is characterized by including an acid extract as an active ingredient.

In the present specification, "acid oil extract" refers to an acid oil leaf, stem, root, or mixture thereof, which is an extract, alcohol having 1 to 4 carbon atoms such as methanol, ethanol, acetone, ethyl acetate, chloroform, methylene chloride, water, or these It is understood as meaning including the extract obtained by extraction with a mixed solvent of and fractions fractionated with the solvents listed above. Here, the extraction method includes both a method of dipping the extraction target in the extraction solvent and a method of distilling the extraction target with the extraction solvent. In addition, the extraction method by dipping may be applied to any method such as cold soaking, heating, ultrasonic, reflux. Nevertheless, the term "extract" preferably means that the extract is obtained by extraction (cold, hot water extract or distillation) with water, ethanol or a mixed solvent thereof, more preferably 70% to 90% Mean ethanol aqueous solution. In the above,% is a concentration (v / v) expression by volume percentage, and means a volume of solute dissolved in a volume of solution as known in the art. For example, 30% (v / v) means that 30 ml of solute is dissolved in 100 ml of the solution. In addition, the meaning of the "extract" of the present specification includes an extract in a form purified through filtration, a concentrated liquid extract from which some of the extraction solvent is removed, and a powdery extract from which the extraction solvent is completely removed.

In addition, in the present specification, the meaning of "active ingredient" means a component that can exhibit activity alone or in combination with a carrier having no activity in itself.

In addition, in the present specification, "skin whitening" is understood as a result of the inhibition of the production of melanin, specifically, means the prevention, delayed expression or treatment of symptoms caused by the excessive production of melanin, such as blemishes, freckles, blotch, etc. .

In addition to the active ingredient, the skin whitening composition of the present invention may further include other known compounds or plant extracts that are known to have a skin whitening effect so as to enhance or enhance the skin whitening effect.

Herein, the other compounds or plant extracts include, for example, mercaptosuccinic acid, mercaptodextran, tefrenone, dihydroxy-isoquinoline, 3-hydroxymanul, vitamin K, thiazolidone, kynurenine, lemon extract, cucumber Extracts, mulberry extracts, licorice extracts, rosemary extracts, acerola cherry extracts, ginkgo extracts, carob (carob) extracts, geranium extracts, herbal extracts, and the like, but is not limited thereto. Such compounds or plant extracts may be included in the composition for skin whitening of the present invention with one or more active ingredients thereof.

In the composition for skin whitening of the present invention, the active ingredient may be included in any amount within a range that does not toxic to the human body depending on the use, formulation (cosmetic, soap, ointment, etc.), specifically, the whole composition It may be included in an amount of 0.01% to 99.99% by weight based on the weight.

The skin whitening composition of the present invention can be regarded as a cosmetic composition in a specific embodiment.

When the composition for skin whitening of the present invention is identified as a cosmetic composition, the cosmetic composition may be prepared in various formulations, such as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants- Contains cleansing, oils, powder found

Emulsion, emulsion foundation, wax foundation, spray, and the like, but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), nutrition lotion (milk lotion), nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .

The cosmetic composition of the present invention is prepared by including a suitable carrier component according to the dosage form, and in addition to the active ingredient, conventionally used ingredients such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. It may be prepared including an adjuvant.

Auxiliaries such as stabilizers, solubilizers, vitamins, fragrances, carrier components, etc. according to the formulation are known in the art, it can be selected and used appropriate materials.

When the formulation of the cosmetic composition is a cream or a paste, animal fats such as larch, sunny, and fish oils, vegetable oils such as palm oil, palm oil, soybean oil, perm oil, olive oil, wax, paraffin, starch and the like may be used as carrier components. Even when the formulation of the composition is a soap, the vegetable or animal fats and oils exemplified above can be used as the carrier component. If the formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, or the like may be used as a carrier component, and in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether. have.

The skin lightening composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.

When the composition for skin whitening of the present invention is identified as a pharmaceutical composition, the pharmacological effect may be understood as a therapeutic or prophylactic effect for a disease resulting from an increase in melanin, and the meaning of the skin whitening is the treatment or It can be understood as the meaning of prevention. Examples of diseases resulting from an increase in melanin include skin hyperpigmentation such as blemishes, freckles and blotch, and skin aging.

Pharmaceutical compositions of the present invention, in addition to its active ingredients, include pharmaceutically acceptable carriers, excipients, and the like, oral formulations (tablets, suspensions, granules, emulsions, capsules, syrups, etc.), parenteral formulations (sterile injectable aqueous solutions). Or oily suspensions), topical formulations (solutions, creams, ointments, gels, lotions, patches) and the like.

As used herein, "pharmaceutically acceptable" means that the subject of application (prescription) is not as toxic as applicable, without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (such as corn starch, potato starch, etc.), cellulose, derivatives thereof (such as sodium carboxymethyl cellulose, ethylcellulose, etc.), malt, gelatin, talc, solids Lubricants (such as stearic acid, magnesium stearate, etc.), calcium sulfate, vegetable oils (such as peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, etc.), alginic acid, and the like. May include emulsifiers (such as TWEENS), wetting agents (such as sodium lauryl sulfate), colorants, flavors, tablets, stabilizers, antioxidants, preservatives, water, saline, phosphate buffer solutions, and the like. These ingredients may be used one or more selected as appropriate depending on the formulation of the pharmaceutical composition of the present invention.

Excipients may be selected and used according to the formulation of the pharmaceutical composition of the present invention, for example, when the pharmaceutical composition of the present invention is prepared with an aqueous suspending agent, suitable excipients are sodium carboxymethyl cellulose, methyl cellulose, hydropropylmethylcellulose And sodium alginate, polyvinylpyrrolidone and the like. Suitable excipients when prepared as injections include Ringer's solution, isotonic sodium chloride, and the like.

The pharmaceutical compositions of the present invention may be administered orally or parenterally, but will typically be administered directly to the skin topically in a topical formulation.

The daily dosage of the pharmaceutical composition of the present invention is usually 0.001 ~ 150 mg / kg body weight range, it can be administered once or divided into several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage limits the scope of the present invention in any aspect. It should not be understood.

The composition of this invention can be grasped | ascertained as a food composition in a specific aspect.

The food composition of the present invention may be prepared as a dietary supplement, a special nutritional supplement, a functional beverage, and the like.

The food composition of the present invention may include sweeteners, flavoring agents, bioactive ingredients, minerals, etc. in addition to the active ingredients.

Sweeteners may be used in amounts that give the food a suitable sweet taste, and may be natural or synthetic. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavoring agents can be used to enhance the taste or aroma, both natural and synthetic. It is the case of using a natural thing preferably. In addition to flavors, the use of natural ones can be combined with nutritional purposes. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. In addition, ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo and the like can be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavoring agents may be used, and synthetic flavoring agents may include esters, alcohols, aldehydes, terpenes, and the like.

As the physiologically active substance, catechins such as catechin, epicatechin, gallocatechin, epigallocatechin, vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine, riboflavin, and the like can be used.

As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium, zinc and the like can be used.

In addition, the food composition of the present invention may contain a preservative, an emulsifier, an acidulant, a thickener, and the like, in addition to the sweetener.

Such preservatives, emulsifiers and the like are preferably added and used in very small amounts as long as the use to which they are added can be achieved. By trace amount is meant numerically expressed in the range of 0.0005% to about 0.5% by weight based on the total weight of the food composition.

Examples of preservatives that can be used include sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), and the like.

Emulsifiers that can be used include acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like.

Examples of acidulants that may be used include lead acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Such acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing taste.

Thickeners that can be used include suspending implements, sedimenters, gel formers, swelling agents and the like.

In addition, the food composition of the present invention may include powders or extracts derived from natural products in order to enhance flavor and palatability and to enhance and enhance skin related functions such as wrinkle improvement, sun powder or extracts, bean sprout powder or extracts, shellfish powder or Extract, oyster powder or extract, hawthorn powder or extract, radish juice or extract, cucumber juice or extract, leek juice or extract, spinach juice or extract, lotus root juice or extract, sesame juice or extract, pine needle juice or extract, ginseng juice Or extracts, baekryechosa powder or extracts, licorice powder or extracts, browning powder or extracts, reed powder or extracts, sine powder or extracts, gourd powder or extracts, ginger powder or extracts, jujube powder or extracts, phosphorus powder or extracts, strips Purple Powder or Extract, Numerical Powder or Extract, White Extract Horse or extract, age powder or extract, tangerine seed (peel of tangerine) powder or extract, wolfberry powder or extract, green tea powder or extract, schizandra powder or extract, holly tree powder or extract, borage powder or extract, albino powder or extract , Cinnamon powder or extract, decosinol and the like can be exemplified. Such extracts may be obtained by mixing the extraction objects.

As described above, according to the present invention can provide a composition for skin whitening using an acid extract. The skin whitening composition of the present invention may be commercialized into cosmetics, drugs, foods and the like.

1 is a result showing the melanin production inhibitory activity of the oleander leaf extract in B16F10, a melanoma cell stimulated with a melanogenesis-stimulating agent α-MSH.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

<Example> Preparation of Citron Leaf Extract

Three times weight of 70% ethanol was added to the citron leaf powder, followed by extraction with stirring at room temperature for 48 hours. The filtrate was concentrated under reduced pressure and lyophilized to obtain a powdery citron extract.

Experimental Example Whitening activity experiment

Experimental Example 1 Tyrosinase Inhibitory Activity Experiment

After adding 20 μl of diluted sample to 220 μl of 0.1M potassium phosphate buffer (pH 6.5), add 20 μl of 1250 unit / ml mushroom tyrosinase. Substrate 1.5 mM L-tyrosine 40 ㎕ was added to mix well by measuring the absorbance using an ELISA reader at 37 ℃ After 10 minutes the reaction 490nm and calculated the inhibition rate according to the treatment concentration of the sample according to the following equation, and then IC ₅₀ value Obtained is shown in Table 1 below. Arbutin was used as a positive control.

Inhibition% = 100- (A-B) / (C-D) x100

A: absorbance after the reaction of the sample liquid

B: Absorbance of sample solution measured by replacing buffer with tyrosinase

C: absorbance after reaction of blank sample liquid

D: absorbance of blank sample measured by buffer instead of tyrosinase

Table 1 Tyrosinase inhibition rate

Sample	IC 50 (μg / ml)
Citron	594
Arbutin	236.93

As shown in Table 1 above, by measuring the absorbance of melanin oxidized by the substrate L-tyrosine and the enzyme mushroom tyrosinase, measurement of the inhibitory activity of tyrosinase, an enzyme that plays an important role in melanogenesis, was carried out. The example of citron oil showed lower activity than Arbutin but tyrosinase inhibitory activity.

<Experiment 2> Melanin production inhibitory activity experiment

Experimental Example 2-1 Cell Culture

B16 / F10 cells were obtained from Korean Cell Line Bank (KCLB) as a melanoma cell line in rats. Incubated in 37 ° C., 5% CO ₂ incubator using RPMI 1640 or DMEM medium (GIBCO, Grand Island, NY, USA) containing Grand Island, NY, USA, subcultures every 3-4 days Was implemented.

Experimental Example 2-2 Evaluation of Melanin Inhibition Activity

B16F10 melanoma cell line was dispensed into 6 well plates at a cell number of 1 × 10 ⁵ / well, and after 24 hours, the medium was removed, and 100 nM of α-melanocyte stimulating hormone (α-MSH) and the positive control group arbutin and samples were treated by concentration. Incubate 72 hours later. After washing twice with PBS, lysis buffer (100 mM sodium phosphate (pH6.8), 0.1 mM PMSF, 1% Triton X-100) was added and dissolved at -80 ° C for 30 minutes. After cell collection, pellet was dissolved by reacting with 1N NaOH containing 10% DMSO at 65 ° C. for 1 hour and absorbance was measured at 405 nm.

The results are shown in [FIG. 1].

Referring to FIG. 1, it can be seen that the acid oil sample of the example showed melanin production inhibitory activity in proportion to the treatment concentration, and melanin production inhibitory activity was much superior to that of the positive control group Arbutin.

Claims (6)

1. Skin whitening composition comprising an acid extract as an active ingredient.
2. The method of claim 1,

   The extract is a skin whitening composition, characterized in that obtained by extracting the citron leaves with water, ethanol or a mixed solvent thereof.
3. The method according to claim 1 or 2,

   The composition is a composition for skin whitening, characterized in that the cosmetic composition.
4. The method according to claim 1 or 2,

   The composition is a composition for skin whitening, characterized in that the food composition.
5. Pharmaceutical composition for the prevention or treatment of hyperpigmentation, comprising an acid oil extract as an active ingredient.
6. The method of claim 5,

   The extract is a pharmaceutical composition for the prevention or treatment of skin hyperpigmentation, characterized in that obtained by extracting the citron leaves with water, ethanol or a mixed solvent thereof.

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