KR20210084030A - Skin whitening composition containing lycopi herba extract as an active ingredient - Google Patents

Abstract

According to an embodiment of the present invention, provided is a cosmetic composition for skin whitening. More particularly, it is possible to reduce the production of melanin by providing a cosmetic composition for skin whitening containing an extract of Lycopi herba as an active ingredient.

Description

Composition for skin whitening containing taekran extract as an active ingredient {SKIN WHITENING COMPOSITION CONTAINING LYCOPI HERBA EXTRACT AS AN ACTIVE INGREDIENT}

The present invention relates to a composition for skin whitening, and more particularly, to provide a composition for skin whitening capable of reducing the production of melanin in the skin by containing an extract of taekrane as an active ingredient.

Lycopi Herba is a plant in the family Lamiaceae, also called sagebrush, distributed in Korea and Asia. It is wild in Korea and grows mainly near wetlands such as ponds and waterside. The soft part is used as a vegetable, and the mature part (outpost) is used for medicinal purposes. Glucoside, tannin, flavonoid glycoside, phenol, amino acid, organic acid, saponin, glucose, galactose, lycopose (lycopose), raffinose (raffinose), stachyose (stachyose) and contains fructose (fructose).

Taekran, which shows excellent efficacy in gynecological diseases, is mainly used in oriental medicine. It is used to relieve/treat the symptoms of painful menstrual cramps during menstruation, menstruation without or stopping menstruation, postpartum eohyeol abdominal pain with abdominal pain due to eohyeol left after childbirth, edema of the body, and vaginal tuberculosis damaged by falling or bumping. .

Melanin is a blackish-brown or black pigment, and it is present in the skin/hair/eyes, etc. and gives it color. Cells that make melanin are called melanocytes, and the produced melanin is created/stored in an organelle called melanosome in melanocytes. The color of the skin is determined according to the amount of melanin. The greater the amount of melanin, the darker the skin color. If melanin is not produced or does not exist, diseases such as albinism occur. In addition, excessively generated melanin causes diseases such as melasma, freckle, and melanoma. The main role of melanin is to protect the human body (skin) by absorbing more than a certain amount of ultraviolet rays and blocking harmful ultraviolet rays from penetrating into the human body. Therefore, when the skin turns brown by sunlight (ultraviolet rays), the melanin formed in melanocytes that exist under the keratinocytes forms melanin pigments to protect the skin and moves to the keratinocytes so that the ultraviolet rays penetrate deep into the skin. because it prevents Melanin pigments include eumelanin, which has a large blackish-brown or black color, and pheomelanin, which has a yellow or red color.

Although many studies have been conducted on each of taekran and skin whitening, research related to the skin whitening effect of taekran is insufficient. Therefore, there is a need for research to obtain skin whitening effect using taekran.

The technical problem to be solved by the present invention is to provide a composition for skin whitening that can reduce melanin production in the skin by containing taekran extract as an active ingredient.

In addition, it is to provide a health functional food for skin whitening containing extract of lycopi herba as an active ingredient, a pharmaceutical composition for preventing or treating skin pigmentation disease, and a quasi-drug composition for preventing or improving skin pigmentation disease.

In order to solve the above problems, the cosmetic composition for skin whitening of the present invention may contain an extract of lycopi herba as an active ingredient.

In addition, the extract of lycopi herba may be extracted with at least one solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, ethyl acetate, and acetone.

According to an embodiment of the present invention, there may be provided a cosmetic composition for skin whitening, characterized in that it contains the extract of lycopi herba at a concentration of 100 to 500 μg/ml.

In addition, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray formulation, characterized in that can do.

According to another embodiment of the present invention, a health functional food for skin whitening containing extract of lycopi herba as an active ingredient, a pharmaceutical composition for preventing or treating skin pigmentation disease, and a quasi-drug composition for preventing or improving skin pigmentation disease may be provided.

The cosmetic composition for skin whitening of the present invention has the advantage of reducing melanin production in the skin by containing taekran extract as an active ingredient.

The cosmetic composition for skin whitening of the present invention has the advantage that side effects are not high by using a natural extract.

According to another embodiment of the present invention, there may be provided a health functional food for skin whitening, a pharmaceutical composition for preventing or treating skin pigmentation disease, and a quasi-drug composition for preventing or improving skin pigmentation disease.

Figure 1 A is melanoma (B16F1) cells treated with taekran extract and kojic acid by concentration, Figure 1 B is melanoma (B16F1) cells by concentration taekran extract, kojic acid and α- It is a graph of the cytotoxicity test result measured by MTT assay after treatment with melanocyte stimulating hormone (α-MSH).
2A and 2C are melanoma (B16F1) mouse melanoma cells treated with taekran extract, kojic acid, and α-melanocyte stimulating hormone (α-MSH) by concentration at the same time, and absorbance using a spectrophotometer was measured to indicate the content of melanin, and B and D are images showing the color change of the cell culture medium and the cell pellet after treatment with the taekran extract.
3 is an image showing the effect of inhibiting protein expression related to melanin synthesis of taekran extract in mouse melanoma cells induced by α-melanocyte stimulating hormone (α-MSH).

Hereinafter, preferred embodiments of the present invention will be described in detail. Advantages and features of the present invention, as well as embodiments for achieving them, will become apparent with reference to the following embodiments. However, the present invention is not limited to the embodiments published below, but may be implemented in various different forms, and only these embodiments allow the publication of the present invention to be complete, and common knowledge in the art to which the present invention pertains. It is provided to fully inform the possessor of the scope of the invention, and the present invention is only defined by the scope of the claims. Like reference numerals refer to like elements throughout.

Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used with the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs. In addition, terms defined in a commonly used dictionary are not to be interpreted ideally or excessively unless clearly defined in particular. The terminology used herein is for the purpose of describing the embodiments and is not intended to limit the present invention. As used herein, the singular also includes the plural unless specifically stated otherwise in the phrase.

As used herein, lycopi herba extract means obtained by extracting from various organs or parts (eg, leaves, flowers, roots, stems, branches, bark and seeds, etc.) of taekran. Preferably, it means an extract obtained from the outpost of taekran.

In addition, taekran (lycopi herba) extract is glucoside (glucoside), tannin (tannin), flavonoid (flavonoide) glycosides, phenols (phenol), amino acids, organic acids, saponin (aponin), glucose (glucose), galactose (galactose) , lycopose, raffinose, stachyose and fructose.

The extract of the present invention includes not only the extract by the above-mentioned extraction solvent, but also the extract that has undergone a conventional purification process. For example, reduced pressure by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by extraction using ultrasound, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity Active fractions obtained through various additional purification methods, such as separation by sex), are also included in the extract of the present invention.

According to an embodiment of the present invention, a cosmetic composition for skin whitening containing extract of lycopi herba as an active ingredient may be provided.

The extract of lycopi herba of the present invention may be prepared according to a conventional method known in the art, that is, under conditions of ordinary temperature and pressure, using a conventional solvent, preferably purified water (or water). ), lower alcohols having 1 to 4 carbon atoms, glycerin, ethyl acetate, acetone, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol and hexane using at least one solvent selected from the group consisting of extracted, more preferably using ethanol. However, the present invention is not limited thereto.

According to an embodiment of the present invention, the cosmetic composition for skin whitening may include the extract of lycopi herba at a concentration of 100 to 500 μg/ml. When the concentration of the extract is less than 100 μg / ml, the whitening effect is extremely weak, and when it exceeds 500 μg / ml, the degree of increase in the effect of increasing the content is insignificant, there is a problem in safety and stability of the formulation, and it is not economical .

According to another embodiment of the present invention, the cosmetic composition for skin whitening of the present invention is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion It may be in the form of foundation, wax foundation, or spray, but is not limited thereto.

According to another embodiment of the present invention, a health functional food for skin whitening containing extract of lycopi herba as an active ingredient may be provided.

A health functional food for skin whitening containing lycopi herba extract as an active ingredient, in addition to containing a mixture of the taekran extract as an active ingredient, contains various flavoring agents or natural carbohydrates as an additional ingredient like a conventional food composition. may contain.

Formulations of the health functional food may be tablets, capsules, powders, granules, liquids, pills, and the like. For example, for formulation in the form of a tablet or capsule, the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or required, suitable binders, lubricants, disintegrants and color-developers may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as needed.

According to another embodiment of the present invention, there may be provided a pharmaceutical composition for preventing or treating skin pigmentation diseases containing extract of lycopi herba as an active ingredient.

In the present invention, the skin pigmentation disease may include at least one selected from the group consisting of melasma, freckles, and melanoma, but is not limited thereto.

The extract of the pharmaceutical composition for preventing or treating skin pigmentation disease according to the present invention further comprises at least one of a carrier, a diluent and an excipient, such as granules, lemonades, powders, syrups, liquids, extracts, elixirs, fluid extracts, Emulsions, suspensions, preparations, saliva, tablets, alcohol tablets, capsules, tinctures, pills, transdermal absorbents, lotions, liniments, aerosols, ointments, patches, pastas, cataplasmas, creams, One selected from the group consisting of emulsions, pastes, non-aqueous solvents, freeze-dried preparations, suppositories and injections may be used as a formulation.

In addition, carriers, excipients and diluents that may be further included in the present invention include sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, which are generally used in formulation. , methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, but is not limited thereto.

When formulating the pharmaceutical composition for preventing or treating skin pigmentation disease according to the present invention, it is prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose or lactose. , gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. In addition, formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used. The content of the extract according to the present invention as an active ingredient in the above-mentioned formulation can be appropriately adjusted depending on the type and purpose of use, the patient's condition, the type and severity of symptoms, etc.

The dosage of the extract-containing composition of the present invention may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and may be administered once to several times a day at regular time intervals according to the judgment of a doctor or pharmacist It can also be administered in divided doses.

According to another embodiment of the present invention, there may be provided a quasi-drug composition for preventing or improving skin pigmentation disease containing extract of lycopi herba as an active ingredient.

The term "quasi-drug" used in the present invention refers to articles with a milder action than pharmaceuticals among articles used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs exclude products used for medicinal purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products with minor or no direct action on the human body.

The quasi-drug composition of the present invention is used for the purpose of improving skin pigmentation, and is not particularly limited in its formulation, for example, softening lotion, nutrient lotion, massage cream, nourishing cream, pack, mask pack, mask sheet , may be a cosmetic composition having a formulation of a gel or skin adhesion type cosmetic, and may also be a transdermal formulation such as a lotion, ointment, gel, cream, patch or spray.

In addition, in each formulation, the quasi-drug composition can be formulated by selecting other components arbitrarily according to the formulation or purpose of use of other quasi-drugs. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use (suppression or alleviation). For example, it may contain conventional adjuvants such as thickeners, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers and the like.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

In order to confirm the effect of taekran extract on the biosynthesis of melanin, an experiment was conducted using B16F1 mouse melanoma cells. First, it was evaluated whether the taekran extract showed cytotoxicity in melanoma (B16F1) cells.

Cytotoxicity was measured by MTT assay. The result is shown in FIG. 1 . 1 shows the cytotoxicity of taekran extract in B16f1 mouse melanoma cells. 1A is the result of treating melanoma (B16F1) cells with taekran extract (100, 250, 500 μg/ml) and standard material kojic acid (1 mM) for 48 hours. 1B is a melanoma (B16F1) cell concentration-specific extract (100, 250, 500 μg/ml), kojic acid (1 mM), a standard material, and α-melanocyte stimulating hormone (α-MSH, 0.2 μM) is the result of simultaneous treatment. The control group used a medium not treated with the sample. After standing for 48 hours, MTT reagent (80 μg/ml, dark condition) was treated, and the absorbance was measured to indicate the cell viability. As a result of the experiment, it was confirmed that when the taekran extract was treated by concentration and when α-melanocyte stimulating hormone (α-MSH) was simultaneously treated, it did not show any particular toxicity compared to the control group.

Whether or not the taekran extract inhibits the production of melanin, the final product of the melanin biosynthesis process, was confirmed by treating α-melanocyte stimulating hormone (α-MSH)-induced B16F1 mouse melanoma cells with the taekran extract and the standard substance kojic acid. did. In Figure 2, B16F1 mouse melanoma cells by concentration extract (100, 250, 500 μg / ml), standard material kojic acid (kojic acid, 1 mM) and α- melanocyte stimulating hormone (α-MSH, 0.2 μM) were simultaneously treated, and the absorbance was measured at 405 nm using a spectrophotometer after 72 hours to indicate the content of melanin (FIG. 2A and FIG. 2C). The content of melanin was expressed as a percentage of the control group. The images in which the color of the cell pellet and the cell culture medium changed after treatment with the Taekran extract are B of FIG. 2 and D of FIG. 2 .

Referring to Figure 2, as a result of the experiment, the production of melanin was induced in the α-melanocyte stimulating hormone (α-MSH) alone treatment group. It can be seen that the reduction was reduced to a degree similar to that of the control group. It can be seen that the color of the cell pellet (Fig. 2D) and the cell culture medium (Fig. 2B) is also reduced in a concentration-dependent manner when the taekran extract is treated. Therefore, it is shown that taekran extract is effective in reducing the production of melanin, the final product of the melanin biosynthesis process.

MITF is a microphthalmia-associated transcription factor, a transcription factor known to regulate the expression of various genes essential for melanin synthesis in melanocytes. In addition, it is involved in the regulation of melanocyte differentiation, pigmentation, proliferation and survival. The signaling pathway related to MITF regulates the production of melanin, and when melanocyte-stimulating hormone (α-MSH) produced from ultraviolet light binds to the Melanocortin 1 receptor (MC1R), it is essential for melanin synthesis through a series of signal transduction processes. It induces the synthesis of melanin by increasing the expression of genes tyrosinase, tyrosinase-related protein 1, and tyrosinase-related protein 2. Among them, tyrosinase is the main enzyme that regulates the production of melanin, ① oxidizes L-tyrosine to dopaquinone ② oxidizes L-tyrosine to L-Dopa ③ oxidizes 5,6-dihydroxyindole (DHI) to indole-2-carboxtlic acid-5, It is involved in oxidation to 6-quinone (IQ). Therefore, the expression of the protein was confirmed through western blot analysis in order to confirm whether the taekran extract regulates the expression of these enzymes (proteins) (FIG. 3).

3A shows that α-MSH-induced B16F1 mouse melanoma cells were treated with taekran extract (100, 250, 500 μg/ml) and standard material kojic acid (1 mM) by concentration, α-melanocyte-stimulating hormone. Measured after treatment for 72 hours in the presence or absence of (α-MSH, 0.2 μM) (anti-tyrosinase antibody), B of FIG. 3 shows taekran extract (100, 250, 500 μg/ml) and standard It was measured after treatment with the substance kojic acid (1 mM) for 3 hours in the presence or absence of α-melanocyte stimulating hormone (α-MSH, 0.2 μM) (anti-MITF antibody).

Referring to FIG. 3, as a result of the experiment, the expression of tyrosinase and MITF was induced by α-melanocyte stimulating hormone (α-MSH), and the expression amount was decreased in a concentration-dependent manner by treating the taekran extract for each concentration. Therefore, the extract of taekran shows that it is effective in reducing the production of melanin by suppressing the expression of enzymes essential for melanin synthesis.

[Experimental Example 1]

<1-1> Cell culture

As the cells, melanoma (B16F1) cells, a mouse melanoma cell line, were used, and these cells were purchased from Korean cell line bank (Korea Cell Line Bank). Using DMEM medium containing 10% fetal bovin serum (FBS) and 1% penicillin streptomycin, it was maintained in an incubator at 37° C. containing _{5% CO 2 .} For maintenance, cells were subcultured once every two days.

<1-2> Cytotoxicity test (Cell viability - MTT assay)

In order to confirm the cell viability of Taekran extract against melanoma (B16F1) cells, a mouse melanoma cell line, MTT assay was performed. Melanoma (B16F1) cells were spread in a 24-well cell culture plate at a density of ^{2.5Х10 4 .} These cells were treated with taekran extract (100, 250, 500 μg/ml) and a standard material, kojic acid (1 mM), at each concentration, and α-melanocyte stimulating hormone (α-MSH, 0.2 μM) in the presence or absence. After 48 hours, 20 μL of MTT reagent (80 μg/ml) was put into each well and reacted at 37° C. under dark conditions for 1 hour. The medium containing the MTT reagent was removed, and 400 μL of dimethyl sulfoxide (DMSO) was added to dissolve intracellular formazan. For absorbance measurement, 200 μL of each was transferred to a 96 well plate, and absorbance was measured at 552 nm.

<1-3> Melanin content measurement

In order to measure the melanin content in the cells, it was spread on a 60 mm cell culture dish melanoma (B16F1) to the cell ⁴ 5Х10 density. These cells were treated with taekran extract (100, 250, 500 μg/ml) and a standard material, kojic acid (1 mM), at each concentration, and α-melanocyte stimulating hormone (α-MSH, 0.2 μM). After 72 hours, the cell pellet was collected and dissolved by reacting at 80° C. for 1 hour with 1M sodium hydroxide (NaOH) containing 10% dimethyl sulfoxide (DMSO). The content of melanin was shown by measuring the absorbance at 405 nm.

To measure the extracellular melanin content, a cell culture medium was used. After treating melanoma (B16F1) cells with α-melanocyte-stimulating hormone (α-MSH, 0.2 μM), treated with taekran extract and standard kojic acid (1 mM) at 37° C. for 72 hours, After transferring 150 μL of cell culture medium to a 96-well plate, absorbance at 405 nm was measured and indicated.

<1-4> Protein expression analysis (Western blot analysis)

Melanoma (B16F1) cells ^{were spread on a 60 mm cell culture dish at a density of 5Х10 4} , and α-melanin extract (100, 250, 500 μg/ml) and kojic acid (1 mM), a standard material, were added to each concentration. It was reacted with cell-stimulating hormone (α-MSH, 0.2 μM) at 37° C. for 72 hours. After 72 hours, the remaining medium and other residues were washed twice with phosphate buffered saline (PBS). Cell pellets were collected and proteins were extracted with a lysis buffer (RIPA buffer) containing a protein inhibitor. The extracted protein was quantified by DC protein assay, and the quantified protein was separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes. The membrane was blocked with 5% non-fat dry milk dissolved in Tris-buffered saline-Tween 20 solutions (TBS-T), and reacted with a primary antibody at 4°C for 24 hours. After 24 hours, the reaction was carried out at room temperature for 1 hour, rinsed with TBS-T, and then reacted with horseradish peroxidase secondary antibody (anti-mouse and anti-rabbit IgG) at room temperature for 2 hours. Protein bands were visualized using western HRP substrate and ImageQuant LAS 500 (GE Healthcare Life Science, USA).

Although embodiments of the present invention have been described above, those of ordinary skill in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without changing the technical spirit or essential features thereof. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (7)

A cosmetic composition for skin whitening containing extract of lycopi herba as an active ingredient. According to claim 1, wherein the extract of taekran (lycopi herba),
A cosmetic composition for skin whitening, characterized in that it is extracted with at least one solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, ethyl acetate, and acetone. According to claim 1,
A cosmetic composition for skin whitening, characterized in that it contains the extract of lycopi herba at a concentration of 100 to 500 μg/ml. According to claim 1,
The cosmetic composition is in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray formulation. A cosmetic composition for whitening. A health functional food for skin whitening containing lycopi herba extract as an active ingredient. A pharmaceutical composition for preventing or treating skin pigmentation diseases containing extract of lycopi herba as an active ingredient. A quasi-drug composition for preventing or improving skin pigmentation diseases containing extract of lycopi herba as an active ingredient.

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