KR100519851B1 - Cosmetic Compositions for Skin-Whitening Comprising Diacetylboldine as Active Ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening comprising diacetylboldine. The cosmetic composition of the present invention has a skin whitening effect of inhibiting melanin production of melanocytes and inhibiting the activity of tyrosinase to inhibit the synthesis of melanin to brighten the color of the face and reduce excessive black spots (smear, freckles, etc.). In addition, there are no side effects such as cytotoxicity.

Description

Cosmetic Compositions for Skin-Whitening Comprising Diacetylboldine as Active Ingredient

The present invention relates to a cosmetic composition for skin whitening comprising diacetylboldine as an active ingredient, and more particularly to a cosmetic composition for skin whitening having a skin whitening effect by inhibiting melanin production.

The pigments that determine human skin color include melanin, hemoglobin, carotenoids, and the like. In addition to the skin color, various colors of hair and eyes are determined by the pigment. The most important pigment that determines skin color is melanin, which depends on the amount, nature and extent of distribution of melanin, which is genetically determined. Melanin production is affected by environmental factors such as the sun's ultraviolet rays and physiological factors such as fatigue or stress.

Melanin is produced through non-enzymatic oxidation after tyrosine is converted to dopa (DOPA) and dopaquinone by the action of an enzyme called tyrosinase present in pigment cells. As such, the melanin synthesis pathway is known to some extent, but the mechanism of inducing melanin synthesis before the action of tyrosinase has not been elucidated in detail.

Melanin is widely distributed in the skin and removes oxygen radicals to protect the skin from damage caused by it. However, the excessive production of melanin is closely related to skin cancer (melanoma), causing skin blackening, producing blemishes, freckles, etc., and the development of cosmetics and pharmaceuticals for the purpose of preventing overproduction of melanin is being actively performed. . Factors that promote the production of melanin include the above-mentioned sunlight (ultraviolet rays) as well as hormones such as estrogen, prostaglandin and bleomycin.

Inhibition of skin blackening can significantly reduce skin pigmentation or prevent excessive pigmentation. In this regard, inhibition of melanocyte activity, reduction of chemical and biological waveguide (L-DOPA), elimination of inflammatory reactions, Research is underway in the direction of reduced synthase activity.

Conventionally, chemicals such as hydroquinone, which whitens the skin by destroying the organized melanin structure, have been used. However, hydroquinone not only has very low anti-tyrosinase activity but also shows high cytotoxicity against normal melanocytes. For this reason, arbutine and hydroquinone derivatives with weak cytotoxicity have been developed and used, but they lack specificity for the pigmentation mechanism. Therefore, kojic acid, ascorbic acid and derivatives thereof, which regulate the melanin production mechanism at the molecular level, have been used targeting tyrosinase. However, kojic acid is rapidly oxidized by other raw materials of cosmetics, exhibits allergic phenomena, and is known as a substance causing cancer, and its use is limited in Korea. On the other hand, ascorbic acid has a disadvantage in that its use is limited due to its poor stability such as oxidation and decomposition in aqueous solution.

In order to solve the problems of the aforementioned conventional skin whitening substances, that is, to develop a whitening agent having an excellent whitening effect by acting specifically on the pigmentation mechanism as well as excellent safety on the skin. As a result, diacetylboldine, which diacetylated boldine, reduced the melanogenesis of melanocytes, inhibited melanin production, and inhibited the synthesis of melanin by inhibiting tyrosinase activity. It has been found that the skin whitening effect is little and that there is little toxicity to the cells, and a cosmetic composition for skin whitening containing the same as an active ingredient was developed.

Accordingly, it is an object of the present invention to provide a cosmetic composition for skin whitening having a skin whitening effect by inhibiting melanin production of melanocytes and inhibiting the activity of tyrosinase.

According to an aspect of the present invention, the present invention provides a cosmetic composition for skin whitening comprising diacetylboldine (2,6-diacetyl-3,5-dimethoxyaporphine) of Formula 1 as an active ingredient:

Diacetylboldine included in the cosmetic composition of the present invention is a compound having a structure of Formula 1, more specifically, 2,6-diacetyl-3,5-dimethoxyamorphine (2,6-diacetyl-3, 5-dimethoxyaporphine), which can be obtained through deacetylation of boldine.

Boldin is a component of a plant that is also (boldo) view, in Fig. Gwamyeong (boldo) viewed on the monitor is lost three (Monimiaceae) nomenclature is penyu commerce see Saunders (Peumus boldus). Bold is an evergreen tree, commonly called boldo, boldu, boldus, boldoa, boldina, etc., which grows to 6-8 m. The tree is native to Chile and Turkey, and is also grown in the Mediterranean, including Europe. In Chile and Turkey, bold has been used as a folk remedy for a long time, and it is known to have analgesic effect, antiparasitic effect, liver, digestive, sedative, diarrhea, diuretic and gallstones. Voldo leaves are also used in Europe and Canada as health supplements. In addition, 17 kinds of alkaloids are known to be present in Voldo, and among them, the boldin component has been recently reported to promote appetite, is good for the liver, and has an anti-inflammatory effect. Voldin is especially present in the leaves of Volvo and 25% of the alkaloids in Volvo are known as Voldin.

Within the cell there are many biological reactions in which Ca ⁺⁺ ions are involved as signal carriers. In general, changes in the concentration of Ca ⁺⁺ ions inside and outside cells regulate biochemical reactions and enzyme activity in cells.

Ca ⁺⁺ ions play a very important role in the mechanism of melanin production by UV irradiation, and it is known that the concentration of Ca ⁺⁺ ions increases when UV is applied to melanocytes. Ca ⁺⁺ ions in B-16 cells are important factors regulating receptor-α-MSH (melanocyte stimulating hormone) binding and activity of enzymes present in melanosomes. Ca ⁺⁺ is not alone involved in increasing melanogenesis, but Ca ⁺⁺ is involved as an intermediate mechanism of transcription or enzyme activity regulation.

If well differentiated keratinocytes and melanocytes are in close proximity, there are 7000 β-adrenergic receptors and 4000 α-adrenergic receptors expressed on the surface of each melanocytes. Differentiated keratinocytes secrete catecholamines, in particular epinephrine. Catecholamines regulate calcium homeostasis (the number of expressed adrenergic receptors associated with calcium flow) and melanin production, while catecholamines are inositol triphosphate (IP3) and diacylglycerol (DAG). Increase As a result, it activates tyrosinase.

The number of α-adrenergic receptors expressed in melanocytes is related to the influx of Ca ⁺⁺ in cells (influx) and catecholamines secreted by keratinocytes. Recently, studies on inhibiting the production of melanin have been reported by regulating the interrelationship between Ca ⁺⁺ ions, catecholamines secreted from keratinocytes and α-adrenergic receptors of melanocytes. Voldin, on the other hand, is known to interfere with the flow of Ca ⁺⁺ ions and have α-adrenergic antagonist properties.

Therefore, the present inventors confirmed whether or not boldin can exert melanin production inhibitory ability through the above-described action, and finally obtained positive experimental results, and further to further optimize boldin as a melanogenesis inhibitor. Several derivatives were synthesized, and among the synthesized derivatives, diacetyl fluorine was found to show very strong activity in melanocytes and also to substantially reduce skin spots on the skin.

Diacetylboldine contained in the cosmetic composition of the present invention is preferably used by dissolving in capric / caprylic triglyceride (0.1%).

According to a preferred embodiment of the present invention, the composition of the present invention comprises EDTA in addition to diacetylboldine, EDTA is bound to tyrosine by binding to copper ions, which are metallic cofactors required to oxidize tyrosine during melanogenesis in the diacetylboldine of the present invention. It acts to inhibit cinases. EDTA is added in the form of a stable salt, disodium EDTA.

In the whitening cosmetic composition of the present invention, the amount of diacetylboldine is preferably 0.0001-10% by weight based on the total cosmetic composition, and when the amount is less than 0.0001% by weight, the whitening effect is too weak, and when the content exceeds 10% by weight There is a problem that the increase of the effect of the increase is insufficient, the cytotoxicity is increased and the stability of the formulation is not secured.

On the other hand, among derivatives obtained by the addition or substitution reaction of the substituents commonly used in the art to the diacetylboldine of the formula (1) derivatives exhibiting skin whitening effect (by inhibiting melanin production and / or tyrosinase activity) It is apparent to those skilled in the art that the present invention is included in the scope of the present invention in consideration of the technical level in the art.

The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to diacetylboldine as an active ingredient, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes, And carriers.

The whitening composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing It may be formulated as cleansing, oil, powder foundation, 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, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition for skin whitening containing the diacetylboldine of the present invention exhibits a skin whitening effect through inhibition of melanin production and inhibition of tyrosinase activity. Therefore, the cosmetic composition for whitening of the present invention specifically acts on the pigmentation mechanism to achieve an excellent skin whitening effect.

In addition, the whitening cosmetic composition of the present invention has no side effects such as cytotoxicity.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. Will be self-evident.

Example 1: Diacetylboldine Synthesis

Boldine (1 mmol, purchased from Sigma) was dissolved in a chloroform (10 mL) solution and then stirred at 0 ° C. for 10 min. Triethylamine (1.2 mmol) was slowly added to the mixed solution as a base, followed by stirring for 10 minutes, followed by addition of acetyl chloride (1.1 mmol). After tracing to TLC (TLC), the reaction was terminated, neutralized with 1N HCl, extracted with chloroform solution and water, dried over MgSO ₄ , filtered and distilled under reduced pressure to obtain diacetylboldine.

Experimental Example 1: Influence on melanin production in melanocytes

Melanosite was used by purchasing a mouse-derived B-16 melanoma (ATCC CRL 6323) cell line. Melanoma cell lines were inoculated in DMEM medium containing 4.5 g / l glucose, 10% serum and 1% antibiotic and incubated at 37 ° C. in a 50 ml T-flask. After incubation for 24 hours under 5% CO ₂ condition, cells were isolated by treatment with 0.05% trypsin containing 0.02% EDTA, and then incubated in 50 ml T-flask for 48 hours. At this time the cell number was 4.57 × 10 ⁶ cells / flask. Samples of appropriate concentration were diluted in DMEM medium, treated with cultured melanoma cells, and incubated at 37 ° C. for 5 days. After incubation, all of the medium was removed, cells were separated by treatment with 1 ml of phosphate buffered saline (PBS) containing 0.02% EDTA and 0.05% trypsin, followed by centrifugation for 5 minutes to collect only cells. Then, 5% trichloroacetate (TCA) was treated, stirred, and centrifuged to precipitate precipitated melanin with PBS. 1N NaOH was dissolved and dissolved in the precipitated melanin, and the absorbance was measured at 475 nm. Melanin concentrations were determined from standard concentration curves of synthetic melanin (Sigma). The experimental results are shown in Table 1.

Sample concentration Melanin production inhibition rate Kojic acid Diacetylboldine 0.0012% - 27.2 ± 17.4% 0.0025% - 60.2 ± 10.4% 0.0037% - 69.3 ± 3.3% 0.03% 21 ± 4.0% - 0.1% 49.2 ± 2.3% -

As can be seen in Table 1, it can be seen that about 40 ppm of diacetylboldine shows a higher melanogenesis inhibitory effect than about 1000 ppm of kojic acid. It can be determined that diacetylboldine has a melanin production inhibitory effect about 30 times greater than kojic acid, and therefore, the diacetylboldine of the present invention significantly inhibits melanin production of melanocytes.

Experimental Example 2: Tyrosinase Inhibition Activity Test

To 500 μl of 0.1 mg / ml tyrosine (Sigma), phytolite, kojic acid, ascorbic acid and hydroquinone, which act as inhibitors, were added at various concentrations to prepare a reaction solution. Sodium phosphate buffer (pH 6.8) was used as the buffer solution. 500 μl of tyrosinase (Sigma, 250 U / mL) was added to the reaction solution, and the resultant was reacted at 37 ° C. for 10 minutes, and then the absorbance was measured at 475 nm. No inhibitor was added as a control. Inhibition was calculated according to the following equation (1). The experimental results are shown in Table 2.

% Inhibition =

{1- [Compare absorbance (ODsub ₄₅₇ )] / [Control absorbance (ODsub ₄₅₇ )]} x100

Sample (wt%) Inhibition (%) Control (0) 0 Diacetylboldine (0.0012) 10 ± 19.4 Diacetylboldine (0.0025) 44.7 ± 9.8 Diacetylboldine (0.0037) 53.7 ± 5.1 Phytolite (10) 95 ± 2.9 Kojic acid (0.03) 33 ± 4.0 Kojic acid (0.1) 60 ± 2.8 Ascorbic acid (0.1) 66 ± 2.2 Hydroquinone (0.4) 0 Hydroquinone (0.8) 0 Hydroquinone (1.6) 0

As can be seen in Table 2, it can be seen that the diacetylboldine of the present invention greatly inhibits the activity of tyrosinase, and moreover, about 27-fold inhibition compared to kojic acid and ascorbic acid, which are known as tyrosinase inhibitors. Effect.

Experimental Example 3: Cytotoxicity Test

Incubate for 45 min by adding 1% of kojic acid, ascorbic acid and diacetylboldine to NHFB cells (Korea Cell Line Bank) incubated in a 96-well plate for 24 hours, remove the supernatant and inject 200 μl of medium again. Incubated for 48 hours. After incubating for 4 hours by adding 50 µl of 5 mg / ml MTT reagent to the culture, the absorbance was measured at 540 nm with an ELISA reader. Cytotoxicity was expressed in% by the following equation. Cells cultured without any material were used as controls. The experimental results are shown in Table 3.

Cell survival rate (%)

{1- [Compare absorbance (ODsub ₅₄₀ )] / [Control absorbance (ODsub ₅₄₀ )]} x100

Sample (wt%) Cell survival rate (%) Control 100 Kojic acid (1%) 44 ± 2.2 Ascorbic acid (1%) 15 ± 1.8 Diacetylboldine (1%) 85.8 ± 1.5

As can be seen in Table 3, while the diacetylboldine of the present invention has little cytotoxicity, it can be seen that the conventional whitening cosmetics, kojic acid and ascorbic acid, greatly reduce the survival rate of cells.

Hereinafter, in the formulation example, the cosmetic formulation containing the diacetylboldine of the present invention is exemplified as a flexible lotion, lotion, cream and pack, but is not limited thereto.

Formulation Example 1 Flexible Lotion (Skin)

ingredient Content (% by weight) Diacetylboldine 1.0 1,3-butylene glycol 5.2 Oleyl alcohol 1.5 ethanol 3.2 Polysorbate 20 3.2 Benzophenone-9 2.0 Carboxyl Vinyl Polymer 1.0 glycerin 3.5 incense a very small amount antiseptic a very small amount Purified water Remaining amount Sum 100

Formulation Example 2: Emulsion (Lotion)

ingredient Content (% by weight) Diacetylboldine 1.0 glycerin 5.1 Propyl glycol 4.2 Tocopheryl Acetate 3.0 Floating paraffin 4.6 Triethanolamine 1.0 Squalane 3.1 Macadamia Nut Oil 2.5 Polysorbate 60 1.6 Sorbitan sesquirrolate 1.6 Propylparaben 0.6 Carboxyl Vinyl Polymer 1.5 incense a very small amount antiseptic a very small amount Purified water Remaining amount Sum 100

Formulation Example 3: Cream

ingredient Content (% by weight) Diacetylboldine 1.0 glycerin 4.0 vaseline 3.5 Triethanolamine 0.5 Liquid paraffin 24.0 Squalane 3.0 Beeswax 2.1 Tocopheryl Acetate 0.1 Polysorbate 60 2.4 Carboxyl Vinyl Polymer 1.0 Sorbitan sesquioleate 2.3 incense a very small amount antiseptic a very small amount Purified water Remaining amount Sum 100

Formulation Example 4: Pack

ingredient Content (% by weight) Diacetylboldine 1.0 Ethyl alcohol 3.0 EDTA-2Na 0.02 Propylene glycol 5.1 glycerin 4.5 Cabopol 1.0 Polyoxide 0.1 antiseptic a very small amount incense a very small amount Purified water Remaining amount Sum 100

Experimental Example 4: Measurement of the whitening effect

Thirty Korean female volunteers aged 19-40 years old were selected, especially those with dark skin. Minolta CR 300 was used as a measuring instrument and measured at 2 week intervals over a 2 month period. The five arms of the arm were marked by 1 cm in width as the measurement site, and four different prescriptions (Formulation Examples 1 and 2 and Comparative Examples 1 and 2) were applied to four locations and one was placed as a control value for comparison. The difference from the control is shown in Table 5. Comparative Examples 1 and 2 are formulations without addition of diacetylboldine in Formulation Examples 1 and 2.

Prescription Formulation Example 1 Formulation Example 2 Comparative Example 1 Comparative Example 2 Control ΔL value 1.95 2.15 0.80 0.65 0

As can be seen from Table 8, the ΔL value after 8 weeks is higher in Formulations 1 and 2 of the present invention in which Diacetylboldine was added than in Comparative Examples 1 and 2 without addition of diacetylboldine, and thus whitening to diacetylboldine. It can be seen that there is an effect.

As described in detail above, the present invention provides a cosmetic composition for skin whitening comprising diacetylboldine as an active ingredient. The cosmetic composition for skin whitening of the present invention exhibits excellent skin whitening effect by inhibiting melanin production of melanocytes and inhibiting melanin synthesis by inhibiting tyrosinase activity. In addition, there are no side effects such as cytotoxicity.

Claims (4)

A cosmetic composition for skin whitening comprising diacetylboldine of formula 1 as an active ingredient: Formula 1 The method of claim 1, The composition is a cosmetic composition for skin whitening, characterized by having a skin whitening effect by inhibiting melanin production of melanocytes and inhibiting the activity of tyrosinase. The method according to claim 1 and 2, The composition is a cosmetic composition for skin whitening, characterized in that containing 0.0001-10% by weight of diacetylboldine relative to the total composition. The composition of claim 1 or 2, wherein the composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and Cosmetic composition for skin whitening, characterized in that it has a formulation selected from the group consisting of a spray.