KR102096788B1 - Composition for skin whitening comprising beauvericin or beauvericin derivative - Google Patents

Abstract

The present invention relates to a composition for skin whitening that includes beaverine (Beauvericin) as an active ingredient, and more particularly, the present invention provides a composition for skin whitening that can have an excellent whitening effect without irritating the skin. .

Description

Composition for skin whitening that includes biliverine as an active ingredient {COMPOSITION FOR SKIN WHITENING COMPRISING BEAUVERICIN OR BEAUVERICIN DERIVATIVE}

The present invention relates to a composition for skin whitening comprising bubericin as an active ingredient.

In general, a person's eyes, hair, and skin color are determined by the concentration and distribution of melanin in the skin, and are also influenced by various environmental or physiological factors such as ultraviolet rays, stress, and hormones.

Melanin is a series of enzymatic oxidations, starting from the melanosome of melanocytes present in the basal layer of the epidermis, where tyrosine, an amino acid, is oxidized as a dopa by tyrosinase and converted to dopachrome. It is produced through a process and a non-enzymatic oxidation process. This melanin has the net function of absorbing a certain amount of ultraviolet rays to protect the skin while maintaining body temperature, but the over-synthesized melanin darkens the skin tone and causes skin hyperpigmentation such as spots, freckles, and skin spots. It is known to promote skin aging.

Conventionally, substances that show tyrosinase inhibitory activity, such as hydroquinone, ascorbic acid, glutathione, and cysteine, were applied to improve skin whitening or skin hyperpigmentation. However, hydroquinone has a problem that the skin irritation is high and thus the formulation must be limited to a very small amount, and ascorbic acid has low stability against external stimuli such as heat, light, and oxygen, and thus discoloration or odor occurs in the case of a cosmetic or pharmaceutical composition to which it is applied. There was a problem. In addition, thiol-based compounds such as glutathione and cysteine not only lowered consumer preference due to the peculiar unpleasant odor, but also had low percutaneous absorption and low long-term stability.

Accordingly, there is a need to develop a cosmetic or pharmaceutical composition using a natural material or a compound derived from a natural material that has a whitening effect and is not irritating to the skin.

Republic of Korea Patent Registration No. 10-1776692 discloses a composition for skin whitening, comprising as an active ingredient an extract of Gogol tree having a skin whitening effect by inhibiting the activity of tyrosinase. In addition, Korean Patent Registration No. 10-1069907 discloses a composition for skin whitening that exhibits a higher melanin inhibitory activity than arbutin by containing an extract, fraction, or a cyclopentadione compound purified therefrom as an active ingredient. However, the conventional natural substances or natural substance-derived compounds including the above patents do not have sufficient whitening effect, and there is a problem that stability, discoloration, or odor problems still remain.

On the other hand, beaverin (Beauvericin) is one of the antibiotics, one of the niatin (Enniatin) cyclic depsipeptide (Depsipeptide), has been reported to have anti-cancer, antibacterial, anti-inflammatory effect. However, it is not well known about the use of whitening of the present berry.

Thus, the present inventors have confirmed that vitamincin can be used as an active ingredient in skin whitening compositions by confirming that it can be used for skin whitening purposes, even though it does not irritate the skin, but has melanin production inhibitory activity. The present invention has been completed.

Republic of Korea Registered Patent No. 10-1085059 Republic of Korea Registered Patent No. 10-1776692 Republic of Korea Registered Patent No. 10-1069907

Shivani et al., J Proteomics Bioinform 2017, 10: 1, pp18-23 (see Abstract)

An object of the present invention is to provide a composition for skin whitening, which includes beauvericin as an active ingredient that is capable of realizing an excellent whitening effect without being irritating to the skin.

The object of the present invention is not limited to the technical problems as described above, and another technical problem may be derived from the following description.

In order to achieve the above object, the present invention provides a composition for skin whitening that includes beaverine (Beauvericin) as an active ingredient.

The composition for skin whitening may inhibit the production of melanin pigments.

The skin whitening composition may include 0.0001 to 15% by weight of the birberrycin based on the total weight of the skin whitening composition.

The skin whitening composition may be a cosmetic composition or a pharmaceutical composition.

The cosmetic composition is one selected from the group consisting of softening lotion, nourishing lotion, converging lotion, skin, lotion, essence, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream and cleansing water It may be the above formulation.

The pharmaceutical composition includes granules, lemonades, powders, syrups, liquids, exes, elixirs, fluid extracts, emulsions, suspensions, premise, needles, tablets, tablets, capsules, tinctures, pills, transdermal absorbers, It may be one or more formulations selected from the group consisting of lotions, linen agents, aerosols, ointments, patch, pasta, cataplasma, cream, paste, non-aqueous, lyophilized, suppository and injection.

The composition for skin whitening comprising the berry of the present invention (Beauvericin) as an active ingredient may have an excellent whitening effect without irritating the skin. In particular, the composition for skin whitening comprising the birberrycin of the present invention as an active ingredient may have a whitening effect by inhibiting the activity or reducing the concentration of transcription factors, enzymes, and signal transduction substances related to melanin production.

1 is a diagram showing the results of Experimental Example 1.
2 is a diagram showing the results of Experimental Example 2.
3 is a diagram showing the results of Experimental Example 3.
4 is a diagram showing the results of (1) in Experimental Example 4.
5 is a diagram showing the results of (2) in Experimental Example 4.
6 is a diagram showing the results of Experimental Example 5.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art to which the present invention pertains can easily practice. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. In addition, in order to clearly describe the present invention, parts not related to the description are omitted.

The terms or words used in the specification and claims of the present invention are not to be construed as being limited to a conventional or lexical meaning, and the inventor can appropriately define the concept of terms to describe his or her invention in the best way. Based on the principles, it should be interpreted as meaning and concept consistent with the technical idea of the present invention.

Throughout the specification of the present invention, when a part “includes” a certain component, it means that the component may further include other components, not to exclude other components, unless otherwise specified. .

Throughout the specification of the present invention, "A and / or B" means A or B, or A and B.

Hereinafter, the present invention has been specifically described with reference to the accompanying drawings, but the present invention is not limited thereto.

The present invention provides a composition for skin whitening that includes beauvericin as an active ingredient.

The composition for skin whitening according to an embodiment of the present invention may have a skin whitening effect as it contains beauvericin as an active ingredient. In particular, the composition for skin whitening according to the present embodiment can implement a skin whitening effect by inhibiting the production of melanin, and more specifically, the activity of transcription factors, enzymes, and signal transduction substances related to melanin production. By inhibiting or reducing the concentration, it can have a whitening effect.

Beauvericin is a cyclic depsipeptide, belonging to the Enniatin family of antibiotics, and has been reported to have anti-cancer, anti-bacterial, anti-inflammatory, and anti-cholesterol effects. In addition, it has been reported that bubericin is involved in apoptosis, anti-platelet aggregation, etc. in a biochemical process (Shivani et al., J Proteomics Bioinform 2017, 10: 1, pp18-23).

The name of the IUPAC of Viverycin is (3S, 6R, 9S, 12R, 15S, 18R) -3,9,15-Tribenzyl-6,12,18-triisopropyl-4,10,16-trimethyl-1,7,13-trioxa -4,10,16-triazacyclooctadecane-2,5,8,11,14,17-hexone, and has the structure of Formula 1 below.

 [Formula 1]

The berry of this embodiment may be isolated, extracted, purified, or produced from Cordyceps or Endophytic fungi of medicinal plants. Some of the fungi that produce berry are Beauveria , Paecilomyces , Polyporus , Isaria , and Fusarium . Fusarium Fusarium Proliferatum , Fusarium Semitectum , Fusarium Subglutinans , Fusarium Begoniae, etc., and the Beauveria include Beauveria Bassiana .

The composition for skin whitening of the present embodiment may include 0.0001 to 15% by weight, preferably 0.001 to 10% by weight, and more preferably 0.001 to 5% by weight based on the total weight of the composition for skin whitening.

If the composition for skin whitening of the present embodiment contains less than 0.0001% by weight of berry, based on the total weight of the composition for skin whitening, there may be a problem that the effect of whitening by berry is insufficient. On the other hand, when the composition for skin whitening of the present embodiment contains more than 15% by weight of berry, based on the total weight of the composition for skin whitening, the increase in whitening effect by berry is not economical because it is insignificant, and the formulation of the composition There may be a problem that stability is deteriorated.

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The composition for skin whitening comprising the berry of the present invention may be a cosmetic composition.

The skin whitening composition of this embodiment may be a cosmetic composition.

The cosmetic composition for whitening the skin of the present embodiment may include a cosmetically acceptable carrier or additive in addition to the above birberrycin. For example, fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, blowing agents, fragrances, surfactants, emulsifiers, fillers, metal ion blockers, chelating agents, preservatives, Vitamins, blockers, wetting agents, oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles, etc., but are not limited thereto, and may include any known material applicable to cosmetic compositions.

In addition, the cosmetic composition for skin whitening of the present embodiment is a softening lotion, nutrient makeup, convergent makeup, skin, lotion, essence, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream and cleansing It may be formed of one or more formulations selected from the group consisting of water, but is not limited thereto, and may be formed of all known formulations.

On the other hand, when the cosmetic composition for skin whitening of the present embodiment having the above formulation is formed in a paste, cream or gel state, animal oil, vegetable oil, flax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol , Silicon, bentonite, silica, talc, zinc oxide, and the like.

When the cosmetic composition for skin whitening of the present embodiment is formed in the form of a solution or an emulsion, a solvent, a solubilizing agent, and an emulsifying agent may be further included. The solvent, solubilizing agent, and emulsifying agent include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, sorbitan Fatty acid esters, and the like.

When the cosmetic composition for skin whitening of the present embodiment is formed in the form of a suspension, liquid diluents such as water, ethanol, and propylene glycol, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester Agent, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tragacanth gum, and the like.

When the cosmetic composition for skin whitening of the present embodiment is a cleansing foam, cleansing cream or cleansing water, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarco It may further include a sinate-based compound, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, and the like.

The cosmetic composition for skin whitening comprising birberrycin of the present embodiment may include all components of the composition for skin whitening containing birberrycin.

The composition for skin whitening comprising the berry of the present invention may be a pharmaceutical composition.

The skin whitening composition of the present embodiment may be a pharmaceutical composition.

The dosage form of the pharmaceutical composition for skin whitening of this embodiment may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other active pharmaceutical compounds. The salt is not particularly limited as long as it is pharmaceutically acceptable, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methane Sulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid and the like can be used.

The pharmaceutical composition includes granules, lemonades, powders, syrups, liquids, exes, elixirs, fluid extracts, emulsions, suspensions, premise, needles, tablets, tablets, capsules, tinctures, pills, transdermal absorbers, It may be one or more formulations selected from the group consisting of lotions, linen agents, aerosols, ointments, patch, pasta, cataplasma, cream, paste, non-aqueous, lyophilized, suppository and injection.

Accordingly, the pharmaceutical composition for skin whitening of this embodiment may further include a diluent, excipient, carrier, such as filler, extender, binder, wetting agent, disintegrant, surfactant, etc., in addition to the above-described berry.

For example, solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in one or more compounds. ) Or by mixing lactose, gelatin, and the like. In addition, lubricants such as magnesium stearate, talc and the like are used in addition to simple excipients. Liquid preparations for oral administration include suspensions, lemonates, elixirs, emulsions, syrups, etc. In addition to the simple diluents commonly used, water and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, preservatives And the like.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspension solvents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The pharmaceutical composition for skin whitening of this embodiment can be administered to various mammals, such as rats, mice, livestock, and humans. Any mode of administration can be expected, for example, oral, intraperitoneal, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura or intracranial injection. At this time, transdermal administration is preferred as the parenteral route, and topical application may be most preferable.

The preferred dosage of the pharmaceutical composition for skin whitening of the present embodiment varies depending on the patient's condition and body weight, the degree of disease, the drug form, the route and duration of administration, and may be appropriately selected by those skilled in the art. However, for a desired effect, the pharmaceutical composition for skin whitening of the present embodiment may be administered by dividing the amount of 0.1 to 100 mg / kg once to several times a day, but is not limited thereto.

The parenteral route dosage of the pharmaceutical composition for skin whitening of this embodiment varies depending on the patient's condition and body weight, the degree of disease, the drug form, the route of administration and the duration, but for external use, 1.0 to 3.0 ml per day can be used Circuits may be applied separately, but are not limited thereto.

The pharmaceutical composition for skin whitening containing birberrycin of the present embodiment may include all components of the composition for skin whitening containing birberrycin.

Hereinafter, a composition for skin whitening including the berry of the present invention as an active ingredient will be described in detail through experimental examples. Since these examples are only for illustrating the present invention, it should not be construed that the scope of the present invention is limited by these examples.

Experimental example

Experimental example  1: Cytotoxicity test

After inoculating murine melanoma (B16 F10) cells into 6-well plates (using DMEM medium containing 10% fetal bovine serum (FBS)) at 1 × 10 ⁵ per well, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more.

After incubation, B16 F10 cells were treated with Vibericin [purchase: Sigma-Aldrich, ≥97% (HPLC)] according to concentration (1 μM, 10 μM, 100 μM), respectively, and 5% CO _{2 and} 37 ° C. conditions Incubated for about 1 day. Subsequently, cytotoxicity was measured using the MTT (3- (4,5-dimetylthiazol-2-yl) 2,5-diphenyltetrazolium bromide) assay method, and the cell viability of the control group was compared for comparison. After measurement, the results are shown in FIG. 1.

On the other hand, MTT is a light yellow substrate, which is cleaved by the respiratory chain enzyme in the mitochondria of living cells to produce dark blue formazan, which does not react in dead cells, so the number of live cells can be measured using the amount of formazan produced. You can.

As shown in FIG. 1, according to the MTT assay method, it can be confirmed that virberine has little cytotoxicity at a treatment concentration of about 1 to 100 μM. In other words, it means that berry is hypoallergenic to the skin.

Experimental example  2: Melanin production Inhibitory ability  evaluation

After inoculating murine melanoma (B16 F10) cells into 6-well plates (using DMEM medium containing 10% fetal bovine serum (FBS)) at 1 × 10 ⁵ per well, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more.

After incubation, the medium was removed, and B16 F10 cells were treated with Vibericin [purchase: Sigma-Aldrich, ≥97% (HPLC)] according to concentration (1 μM, 5 μM, 10 μM), respectively, and melanin as a control thereof. H89 (N- [2- (p-bromocinnamylamino) ethyl] -5-isoquinolinesulfonamide), known to inhibit production, was treated with 10 μM, and incubated for about 1 day in 5% CO _{2 and} 37 ° C. conditions in diluted medium. .

The cells from which the medium was removed were washed with PBS (phosphated buffer saline), and treated with trypsin to recover the cells. After measuring the cell number of the recovered cells using a hematocytometer, the number of cells for each treatment group is divided into 2 mL tubes with equal number of cells, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant is removed to remove pellet cells. (Cell precipitate) was obtained.

The obtained cell precipitate was placed in 300 µl of a 1M sodium hydroxide solution containing 10% DMSO, and the melanin pigment was dissolved while leaving it in a constant temperature bath at 65 ° C for a certain period of time. Subsequently, the amount of melanin per cell was determined by measuring the absorbance at 405 nm using a microplate reader for the solution in which the melanin pigment was dissolved, and the results are shown in FIG. 2.

Referring to FIG. 2, it can be seen that melanin production was inhibited as the treatment concentration of bubericin increased. Particularly, as a result of treating the control groups H89 and bubericin at the same concentration, it can be seen that bubericin has a significantly superior melanin production inhibitory effect compared to H89, and thus it can have a whitening effect.

Experimental example  3: Expression of melanin-related genes Inhibitory ability  evaluation

After inoculating murine melanoma (B16 F10) cells in DMEM medium containing 10% fetal bovine serum (FBS) with a 60φ dish, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more.

After incubation, H16 F10 cells were treated with Vibericin [purchase: Sigma-Aldrich, ≥97% (HPLC)] according to concentration (1 μM, 5 μM, 10 μM), respectively, and 5% CO _{2 and} 37 ° C. conditions Incubated for about 1 day.

After removing the medium, the cells were washed with phosphated buffer saline (PBS), and then the cells were recovered using a scraper. Subsequently, the recovered cells were lysed using a RIPA (Radioimmunoprecipitation assay) lysis buffer to recover the protein. The recovered protein was separated by size using 8% to 10% dodecyl sulfate sodium polyacrylamide gel electrophoresis, and then transferred to a polyvinylidene fluoride membrane. Thereafter, the membrane was blocked with 5% skim milk for 1 hour, and the primary antibody was incubated at 4 ° C for overnight.

The membrane was washed with TBS (tris-buffered saline) containing Tween20, and then the secondary antibody was incubated for 1 hour at room temperature. The finally obtained membrane protein was detected by ECL (Enchanced chemiluminescence), and the results are shown in FIG. 3.

Referring to FIG. 3, it can be seen that as a result of treatment with bubericin, the expression of the enzyme or transfer factor tyrosinase, TRP1, TRP2, and MITF involved in the melanin production step is suppressed. Particularly, it was confirmed that the expression of tyrosinase, TRP1, TRP2, and MITF is suppressed as the treatment concentration of birberrycin increases, and among them, the expression of tyrosinase, TRP1, TRP2, and MITF is significantly suppressed at a concentration of 10 μM of berryberry. Did.

That is, through the above results, it can be seen that the treatment with bubericin can inhibit melanin production and have a whitening effect.

Experimental example  4

(One) cAMP  Production inhibition measurement test

After inoculating murine melanoma (B16 F10) cells into 6-well plates (DMEM medium containing 10% fetal bovine serum (FBS)) at 1 × 10 ⁵ per well, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more.

After incubation, the medium was removed, and H16 F10 cells were treated with Vibericin [purchase: Sigma-Aldrich, ≥97% (HPLC)] according to concentration (1 μM, 5 μM, 10 μM), respectively, and diluted medium In 5% CO _2, and cultured at 37 ° C. for about 1 day.

After removing the medium, the cells were washed with phosphated buffer saline (PBS), and then the cells were recovered using a scraper. The recovered cells were then lysed through incubation at room temperature for about 30 minutes using 0.1 M hydrogen chloride, and then the supernatant was added to a 96-well plate with rabbit polyclonal antibody.

Subsequently, it was incubated at 4 ° C. for 18 hours, and then the amount of cAMP was confirmed using Ellman's solution, and the absorbance was measured at 405 nm with a microplate reader, and the results are shown in FIG. 4 below. At this time, H89 (10 μM) and Fk (Forskolin, 20 μM) were treated as controls.

Referring to FIG. 4, it can be seen that buberrycin inhibits the production of cAMP that induces the expression of MITF and tyrosinase. Particularly, as a result of treating both berry and H89 at the same concentration, it can be confirmed that the effect of inhibiting cAMP production of berry is superior. Therefore, it can be seen that bubericin can suppress the expression of MITF and tyrosinase by inhibiting the production of cAMP, thereby inhibiting the production of melanin, and thus has a whitening effect.

(2) CREB  Expression inhibition measurement test

After inoculating murine melanoma (B16 F10) cells in DMEM medium containing 10% fetal bovine serum (FBS) with a 60φ dish, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more. After incubation, B16 F10 cells were treated with 10 μM of Vivericin [purchased: Sigma-Aldrich, ≥97% (HPLC)] at 5 μm CO _{2 at} 37 ° C. for 0 to 90 min, approximately 30 min intervals The degree of phosphorylation of CREB protein was measured.

In more detail, after culturing, treatment with Buribercin on B16 F10 cells and partial separation of each B16 F10 cell and Burberrycin mixture at about 0 minutes, 30 minutes, 60 minutes, and 90 minutes, followed by CREB The expression level was evaluated.

After removing the medium, the cells were washed with phosphated buffer saline (PBS), and then the cells were recovered using a scraper. Subsequently, the recovered cells were lysed using a RIPA (Radioimmunoprecipitation assay) lysis buffer to recover the protein. The recovered protein was separated by size according to 8% to 10% dodecyl sulfate sodium polyacrylamide gel electrophoresis, and then transferred to a polyvinylidene fluoride membrane. Thereafter, the membrane to which the protein was transferred was blocked with 5% skim milk for 1 hour, and the primary antibody was incubated at 4 ° C for overnight.

The membrane was washed with TBS (tris-buffered saline) containing Tween20, and then the secondary antibody was incubated for 1 hour at room temperature. The membrane protein finally obtained was detected by ECL (Enchanced chemiluminescence), and the results are shown in FIG. 5.

Referring to FIG. 5, it can be seen that after treatment with Viberin at 10 μM, phosphorylation of CREB decreases over time. In other words, it means that the production of melanin can be inhibited by suppressing the activity of CREB.

That is, according to FIGS. 4 and 5, it means that birberrycin can suppress melanin production and thereby have a whitening effect by inhibiting cAMP production or phosphorylation of CREB.

Experimental Example 5: Evaluation of the association of MAPK and NF-kB in the melanin synthesis inhibition process

After inoculating murine melanoma (B16 F10) cells in DMEM medium containing 10% fetal bovine serum (FBS) with a 60φ dish, 5% CO ₂ and Cells were cultured at 37 ° C. until the cells were attached to the bottom of the well plate by about 80% or more.

After incubation, B16 F10 cells were treated with 10 μM of Vivericin [purchased: Sigma-Aldrich, ≥97% (HPLC)] at 5 μm CO _{2 at} 37 ° C. for 0 to 90 min, approximately 30 min intervals The phosphorylation of MAPK protein and NF-kB protein was measured.

In more detail, after incubation, B16 F10 cells were treated with Vivericin, and each B16 F10 cell and Viverin mixture was partially separated at approximately 0 minutes, 30 minutes, 60 minutes, and 90 minutes, followed by MAPK as follows. And NF-kB phosphorylation.

After removing the medium, the cells were washed with phosphated buffer saline (PBS), and then the cells were recovered using a scraper. Subsequently, the recovered cells were lysed using a RIPA (Radioimmunoprecipitation assay) lysis buffer to recover the protein. The recovered protein was separated by size using 8% to 10% dodecyl sulfate sodium polyacrylamide gel electrophoresis, and then transferred to a polyvinylidene fluoride membrane. Thereafter, the membrane to which the protein was transferred was blocked with 5% skim milk for 1 hour, and the primary antibody was incubated at 4 ° C for overnight.

The membrane was washed with TBS (tris-buffered saline) containing Tween20, and then the secondary antibody was incubated for 1 hour at room temperature. The finally obtained membrane protein was detected by ECL (Enchanced chemiluminescence), and the results are shown in FIG. 6.

Referring to FIG. 6, it can be confirmed that while vivericin does not affect phosphorylation of ERK, JNK and NF-kB proteins, phosphorylation of p38 MAPK protein is inhibited.

According to the prior art, phosphorylation of p38 MAPK is a major mechanism for increasing melanin biosynthesis (see Kor. J. Aesthet. Cosmetol., Vol. 11 No. 3, 417-426, June 2013). That is, in the present invention, birberrycin can inhibit the phosphorylation of the p38 MAPK protein, thereby inhibiting the phosphorylation of the CREB protein, thereby inhibiting melanin production. Therefore, it can be seen that it can have a whitening effect.

Formulation example

Formulation example  1: Production of flexible cosmetics

The composition of the softening cosmetic composition containing the above-mentioned virberine as an active ingredient is shown in Table 1 below.

Specifically, sodium hyaluronate was dispersed in purified water with a propeller mixer (3000 rpm) to prepare a 1% solution. Raw materials 1 to 8, except sodium hyaluronate, were homogenized at 500 rpm in a water phase dissolution tank using a propeller mixer, heated at 75 ° C. to complete dissolution, and then cooled to room temperature. Subsequently, after completely dissolving the raw materials 9 to 11 in a separate dissolution tank, the mixture was added to the aqueous phase dissolution tank and stirred and mixed. Here, the berry juice was added and mixed thoroughly with stirring to prepare a flexible cosmetic. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Flexible cosmetic production ingredient Content (% by weight) One Burberry 0.01 2 Polyoxyethylene cured castor oil 0.5 3 Glycine 3.3 4 Dipotassium glyceride 0.1 5 1,3-butylene glycol 3.0 6 Sodium hyaluronate 0.1 7 ethanol 5.0 8 Antioxidant 0.1 9 Triethanolamine 0.1 10 EDTA 0.1 11 antiseptic Proper 12 Purified water Balance

Formulation example  2. Production of nutrient makeup

The composition of the nutrient cosmetic composition containing the above-described berry is as shown in Table 2.

Specifically, the carbomer was dispersed at 4000 rpm using a propeller mixer to prepare a 2% solution. After adding the raw materials 1 to 6 to the aqueous phase melting tank and stirring and dispersing with a homomixer (2000 rpm), the mixture was heated to 75 ° C. Raw materials 7 to 14 were added to the oil phase dissolution tank and heated and dissolved to 75 ° C. Then, the oil phase dissolved in the aqueous phase dissolution tank was added to emulsify (3000 rpm / 5 minutes), and then cooled to room temperature. Here, the berry juice was added and mixed thoroughly with stirring to prepare nutrient makeup. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Nutritional makeup ingredient Content (% by weight) One Burberry 0.01 2 glycerin 7.0 3 Sorbitan stearate sucrose cocoate 2.0 4 Mineral oil 4.0 5 Trioctanoin 1.0 6 Stearic Acid 1.0 7 Glyceryl stearate 0.5 8 Sorbitan monostearate 1.0 9 Dimethicone 0.5 10 Antioxidant 0.3 11 Triethanolamine 0.1 12 Carbomer 0.2 13 EDTA 0.1 14 antiseptic Proper 15 Purified water Balance

Formulation example  3. Preparation of Essence

The composition of the essence containing the virberine as an active ingredient is shown in Table 3 below.

Specifically, sodium hyaluronate and hydroxyethyl cellulose were each dispersed in purified water with a propeller mixer (2000 rpm) to prepare a 1% by weight solution. In addition, the carbomer was dispersed in purified water with a propeller mixer (4000 rpm) to prepare a 2% by weight containing solution. On the other hand, raw materials 1 to 12 were added to the aqueous phase dissolution tank, stirred and dispersed with a homomixer (2000 rpm), heated to 75 ° C., and the heated aqueous phase was cooled to room temperature again. After completely dissolving the raw materials 13 to 15 in a separate dissolving tank, the mixture was stirred and mixed into the water dissolving tank. Thus, an essence containing bubericin as an active ingredient was prepared. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Essence production ingredient Content (% by weight) One Burberry 0.01 2 glycerin 5.0 3 1,3-butylene glycol 2.0 4 Polyethylene glycol 2.0 5 Carbomer 1.0 6 Sodium hyaluronate 0.1 7 Glycine 3.0 8 Polyacrylamide 2.0 9 Hydroxyethylcellulose 0.2 10 ethanol 3.0 11 Antioxidant 0.3 12 Triethanolamine 0.1 13 Polyoxyethylene cured castor oil 1.0 14 EDTA 0.1 15 antiseptic Proper 16 Purified water Balance

Formulation example  4. Preparation of cream

The composition of the cream containing the above-mentioned berberrycin as an active ingredient is shown in Table 4 below.

Specifically, raw materials 1 to 8 were added to the aqueous phase dissolution tank, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 ° C. The raw materials 9 to 16 were added to a separate oil phase dissolution tank to warmly dissolve to 80 ° C., and then the oil phase dissolved in the water phase dissolution tank was added to emulsify (3000 rpm / 10 min), and then cooled to room temperature. Here, the berry juice was added and mixed thoroughly with stirring to prepare a cream. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Cream composition ingredient Content (% by weight) One Burberry 0.01 2 1,3-butylene glycol 3.0 3 glycerin 3.0 4 Hydrogenated lecithin 1.0 5 Octyldodecanol 3.0 6 Trioctanoin 2.0 7 Stearic Acid 1.5 8 Cetostearyl alcohol 2.0 9 Polysorbate 60 1.5 10 Sorbitan sesquioleate 2.0 11 Dimethicone 3.0 12 Antioxidant 0.3 13 Shotgun 0.2 14 Triethanolamine 0.1 15 EDTA 0.1 16 antiseptic Proper 17 Purified water Balance

Formulation example  5. Manufacture of pack

The composition of the pack containing the birberrycin as an active ingredient is shown in Table 5 below. Specifically, the raw materials 1 to 8 were completely dissolved in a separate dissolving tank, stirred and mixed, and then vorberrycin was added thereto, followed by sufficiently stirring and mixing to prepare a pack. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

The composition of the pack ingredient Content (% by weight) One Burberry 0.01 2 glycerin 7.0 3 1,3-butylene glycol 3.0 4 Squalene 3.0 5 Dimethicone 3.0 6 Sodium hyaluronate 0.1 7 Glycine 2.0 8 Polyacrylamide 5.0 9 Antioxidant 0.3 10 Triethanolamine 0.1 11 EDTA 0.1 12 antiseptic Proper 13 Purified water Balance

Formulation example  6. Manufacturing of cleansing foam

Table 6 below shows the composition of the cleansing foam containing the above-mentioned bubericin as an active ingredient. After dissolving and dissolving the aqueous phase and the oil phase, respectively, and mixing and saponifying, the mixture was cooled to room temperature to prepare a cleansing foam. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Composition of cleansing foam ingredient Content (% by weight) One Burberry 0.01 2 Stearic acid 6.5 3 Myristic acid 28.0 4 Self emulsifying monostearic acid glycerin 3.0 5 Propylene glycol 5.0 6 Concentrated glycerin 10.0 7 Sodium hydroxide 7.0 8 Sodium ethylenediaminetetraacetate 0.1 9 Spices Proper 10 Purified water Balance

Formulation example  7: Wild  Produce

The powder containing the above-described berry was mixed with 50 mg of the berry and 2 g of crystalline cellulose, and then prepared by a conventional method. Meanwhile, for the preparation, in addition to the above-mentioned ingredients, known ingredients required for manufacturing may be further included.

Formulation example  8: Preparation of tablets

Tablets containing the above-described berberrycin as an active ingredient were prepared by mixing the birberrycin with 50 mg, crystalline cellulose 400 mg, and magnesium stearate 5 mg. In addition to the above components, it may further include known components required for manufacturing.

Formulation example  9: Preparation of capsules

The capsule containing the buberincin as an active ingredient was prepared by mixing the buberincin 30 mg, whey protein 100 mg, crystalline cellulose 400 mg, and magnesium stearate 6 mg. Meanwhile, for the preparation, in addition to the above-mentioned ingredients, known ingredients required for manufacturing may be further included.

Formulation example  10: Preparation of injection

The injectable agent containing the above-described viverin was prepared by a conventional method of filling and sterilizing a 2 ml ampoule by mixing 100 mg of viverin, distilled water for injection, and a pH adjusting agent. Meanwhile, for the preparation, in addition to the above-mentioned ingredients, known ingredients required for manufacturing may be further included.

Formulation example  11. Preparation of ointments

Table 7 below shows the composition of the ointment containing the above-mentioned birberrycin as an active ingredient. On the other hand, for the preparation, in addition to the ingredients disclosed in the table, it may further include known ingredients required for manufacturing.

Ointment composition ingredient Content (% by weight) One Burberry 0.01 2 glycerin 8.0 3 1,3-butylene glycol 4.0 4 Floating paraffin 15.0 5 Beta glucan 7.0 6 Carbomer 0.1 7 Caprylic / Capric Triglyceride 3.0 8 Squalane 1.0 9 Cetylaryl glucoside 1.5 10 Sorbitan stearate 0.4 11 Cetearyl alcohol 1.0 12 antiseptic Proper 13 incense Proper 14 Coloring Proper 15 Purified water Balance

As described above, the description of the present invention is for illustration only, and those having ordinary skill in the art to which the present invention pertains can easily be modified into other specific forms without changing the technical spirit or essential features of the present invention. You will understand. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present invention is indicated by the following claims rather than the above detailed description, and it should be interpreted that all changes or modified forms derived from the meaning and scope of the claims and equivalent concepts thereof are included in the scope of the present invention. do.

Claims (6)

A composition for skin whitening, which includes beauvericin as an active ingredient.
According to claim 1,
The skin whitening composition
Melanin (Melanin) skin whitening composition, characterized in that to inhibit the production of pigments.
According to claim 1,
The skin whitening composition
The composition for skin whitening, wherein the composition for whitening the skin comprises 0.0001 to 15% by weight of the birberrycin.
According to claim 1,
The skin whitening composition
A composition for skin whitening, which is a cosmetic composition or a pharmaceutical composition.
The method of claim 4,
The cosmetic composition
It is one or more formulations selected from the group consisting of flexible cosmetic, nutrient cosmetic, convergent cosmetic, skin, lotion, essence, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream, and cleansing water A composition for skin whitening, characterized by.
The method of claim 4,
The pharmaceutical composition
Granules, lemonade, powder, syrup, liquid, ex, elixir, fluid extract, emulsion, suspension, premise, acupuncture, tablets, tablets, capsules, tinctures, pills, transdermal absorbers, lotions, linens For skin whitening, characterized in that it is at least one formulation selected from the group consisting of cement, aerosol, ointment, patch, pasta, cataplasma, cream, paste, non-aqueous, lyophilized, suppository and injection. Composition.

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