KR101644753B1 - Cosmetic composition comprising Sargassum serratifolium extract for skin whitening - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening and, more specifically, to a cosmetic composition having skin whitening effects by comprising a Sargassum serratifolium extract or fraction thereof as an active ingredient. According to the present invention, the Sargassum serratifolium extract or a compound isolated therefrom has excellent melanin generation inhibition and tyrosinase inhibition activities, so that the composition comprising the same as the active ingredient can be usefully used as a cosmetic composition for skin whitening. More specifically, the Sargassum serratifolium is a natural material, and thus is edible, so that the cosmetic composition for skin whitening comprising the extract and the compound derived from the Sargassum serratifolium is safe even when being taken for a long period of time.

Description

[0001] The present invention relates to a cosmetic composition comprising Sargassum serratifolium extract for skin whitening as an active ingredient,

TECHNICAL FIELD The present invention relates to a cosmetic composition for skin whitening, and more particularly to a cosmetic composition having a skin whitening effect, which comprises an extract of Tooth mackerel or a fraction thereof as an active ingredient.

The skin color of a person is determined by the amount of melanin, carotene and hemoglobin. Melanin is the most important factor. Melanin is synthesized in the melanocyte, a pigmented cell in the basal layer of the skin (epidermis), and transformed into keratinocyte to show the color of human skin. Unusually low production of melanin causes skin lesions such as Vitiligo. On the contrary, overproduction is one of the major concerns of middle-aged women, forming spots and freckles, and is also closely related to skin cancer.

The whitening effect will be described in more detail as follows. Melanin is produced by enzymes and nonenzymatic oxidation of tyrosine in cells called melanocytes, which are found in the basal layer of skin epidermis, and transformed into keratinocytes constituting the epidermis. Important enzymes involved in the biosynthesis of melanin include tyrosinase, tyrosinase-related protein 1 and dopachrome tautomerase. In addition to tyrosinase, these enzymes attract attention, but the enzyme that plays a crucial role in the synthesis of melanin is tyrosinase. The first step of biosynthesis of melanin is triggered by tyrosinase. Tyrosine is converted into tyrosine by tyrosine, and this tyrosine is converted into Dopa-quinone. The next step in these two reactions is known to be possible by non-enzymatic reactions, and tyrosinase alone is known to be able to produce melanin.

Recently, new theories about the process of melanin formation due to the expression of signaling substances in skin cells have been suggested. When the skin is exposed to ultraviolet light, interleukin-alpha, which causes inflammation, is expressed from keratinocytes in the epidermis, and this interleukin-alpha keratinocyte endothelin expression Promote. This series of processes increases the formation of melanocytes and activates melanocytes. Melanin is formed by tyrosine tyrosinase in melanocytes in melanocytes thus formed.

In addition, NF-κB is activated in response to various stimuli, and enhances the expression of various genes in relation to immunity, cell proliferation, and inflammation. It has been reported that excessive activation of NF-κB is associated with cancer, rheumatism, psoriasis, etc., and nowadays drugs are actively developed to inhibit NF-κB activation worldwide. NF-κB is also present in epidermal keratinocytes and dermal fibroblasts of the skin, and it is confirmed that NF-κB is excessively activated by ultraviolet rays, extracellular stimuli or inflammatory cytokines, thereby increasing transcription of genes involved in inflammation and apoptosis . The overactivation of NF-κB produces inflammatory cytokines in the cells, which further activate NF-κB. In addition, the synthesis of basic fibroblast growth factor (bFGF) or extracellular matrix degrading enzyme MMP-1, which is a cell proliferation factor, has been increased to cause hyperpigmentation of epidermal keratinocytes, epidermal hypertrophy due to keratinization abnormality, pigmentation by melanocyte proliferation, It is also involved in photoaging of the skin by causing reduction of skin elasticity due to decomposition of dermal collagen of MMP-1.

The skin melanocyte cells that make up the pigment, melanocyte of the skin, are caused by excessive stimulation of the melanin pigment in the skin due to ultraviolet rays or chronic inflammation, resulting in blackness, spots and freckles.

NF-κB in epidermal keratinocytes is excessively activated by external stimuli such as inflammation and ultraviolet rays to induce excessive melanin production and promote the production of bFGF which induces cell proliferation of epidermal keratinocytes, To induce pigmentation by proliferation of the pigment. Therefore, a substance that inhibits the action of NF-κB has an effect of preventing pigmentation such as spots or freckles caused by inflammation or ultraviolet rays.

It has already been reported that ascorbic acid (JP 4-9320), hydroquinone (JP 6-192062), kojic acid (JP 56-7710), arbutin (JP 4-9315) and some compounds have tyrosinase- It is used as a raw material for cosmetics, but its use is limited due to its stability in a prescription system, degradation and coloration, generation of bad odor, efficacy at a living body level, unclear effect and safety problem. And the inhibitory effect of tyrosinase has been proven, but in experiments similar to actual levels of biosynthesis, the effect is low, and hydroquinone is prescribed as a carcinogenic substance and its use is prohibited.

Therefore, it is necessary to develop a natural substance having skin whitening activity with excellent stability.

Korean Patent No. 0562348 Korea Patent No. 0850686

Accordingly, an object of the present invention is to provide a cosmetic composition which is stable to human body and has excellent skin whitening activity even if it is used for a long time by using an extract of natural seaweed.

In order to achieve the above-mentioned object of the present invention,

The present invention provides a cosmetic composition for skin whitening comprising an extract of Tooth mackerel as an active ingredient.

In one embodiment of the present invention, the Saw Mab Extract may be extracted with at least one solvent selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, acetone, water and hexane.

In one embodiment of the present invention, the Saw Moth extract may have a skin whitening effect through inhibition of melanin formation and tyrosinase inhibitory activity.

In one embodiment of the present invention, the toothed moth extract may be included in an amount of 0.0001 to 20% by weight based on the total weight of the cosmetic composition.

The present invention provides a cosmetic composition for skin whitening comprising as an active ingredient a compound isolated from a teaspoon mallow extract.

In one embodiment of the present invention, the compound may be sagahydroquinoic acid, sagaquinoic acid or sagachromenol.

In one embodiment of the present invention, the compound may have a skin whitening effect through inhibition of melanin formation and tyrosinase inhibitory activity.

In an embodiment of the present invention, the cosmetic composition may be a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, , Nutrient essence, pack, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, latex, lipstick, makeup base, foundation, press powder and loose powder .

Since the extract of Tooth mothia or the compound isolated therefrom according to the present invention has excellent melanin production inhibitory activity and tyrosinase inhibitory activity, the composition comprising it as an active ingredient can be usefully used as a cosmetic composition for skin whitening. In particular, since the sawtooth moth can be edible as a natural substance, the whitening cosmetic composition of the present invention containing the extract and the compound derived therefrom has a safety advantage for long-term use.

Figure 1 shows four fraction chromatograms obtained from the n-hexane fraction of the toothed mats.
2 is a second fraction chromatogram of fraction 1 of the four fractions obtained from the n-hexane fraction of the toothed mackerel.
FIG. 3 is a chromatogram of sagahydroquinoic acid identified by analyzing NMR by purifying a fraction obtained by fractionating the first fraction of the four fractions obtained from the n-hexane fraction of the toothed mackerel.
FIG. 4 shows the result of confirming three peaks of a second fraction chromatogram of the fraction No. 2 among the four fractions obtained from the n-hexane fraction of the toothed mackerel.
FIG. 5 is a chromatogram of sagachromenol identified by NMR analysis after purifying the second peak in the second fraction of the second fraction of the four fractions obtained from the n-hexane fraction of the toothed mackerel.
FIG. 6 shows the results of confirming four peaks in a second fraction chromatogram of the fraction No. 3 in the four fractions obtained from the n-hexane fraction of the toothed mackerel.
FIG. 7 is a chromatogram of sagaquinoic acid identified by NMR analysis after purifying the fourth peak in the second fraction of the second fraction of the four fractions obtained from the naphtha fraction of the toothed mackerel.
FIG. 8 shows the result of MTS assay analysis to determine whether or not the extract of the present invention has cytotoxicity.
FIG. 9 is a result of MTS assay analysis to determine whether cytotoxicity of hydroxycinnamic acid (SHQ, sagahydroquinoic acid) was detected by the compound sphaeroa extract of the present invention.
FIG. 10 is a result of MTS assay analysis to determine whether cytotoxicity of sQA (sagaquinoic acid) was detected by the compound syrup isolated from the extract of the teaspoon of the present invention.
FIG. 11 is a result of MTS assay analysis to determine whether cytotoxicity of spermine (SCM, sagachromenol acid) from the extract of the extracts of Tooth Moth extract of the present invention is cytotoxic.
FIG. 12 shows the result of confirming the intracellular melanin content in order to examine whether or not there is melanin formation inhibitory activity in the Teaspoon Corn Extract (ESS).
FIG. 13 shows the results of confirming the intracellular melanin content in order to examine whether the compound syrup separated from the extract of the teaspoon of the mother has inhibitory activity on melanin formation by hydroquinone acid (SHQ, sagahydroquinic acid).
FIG. 14 shows the result of confirming the intracellular melanin content in order to examine whether the compound saury extracted from the extract of the toothed mackerel has a melanin formation inhibitory activity on squequinic acid (SQA).
FIG. 15 shows the results of confirming intracellular melanin content in order to examine whether the compound SMA isolated from the extracts of teaspoon mushroom has melanin formation inhibitory activity on chromenol (SCM, sagachromenol).
16 shows the results of measurement of tyrosinase expression in order to examine whether or not whitening activity is exerted on the extract of the teaspoon of the present invention.
FIG. 17 shows the results of measurement of tyrosinase, pERK and ERK expression levels in order to determine whether the present invention has whitening activity on sagahydroquinoic acid (SHQ).
FIG. 18 shows the results of measurement of tyrosinase, pERK and ERK expression levels in order to determine whether the present invention has a whitening activity on SQA (sagaquinoic acid).
FIG. 19 shows the result of measuring the phosphorylation of ERK in order to determine whether the sagachromenol (SCM) of the present invention has a whitening activity.

As a result of screening for substances which are effective for skin whitening among various natural substances of natural ones, it was confirmed that the extract of Sawtooth mackerel has an excellent skin whitening effect.

Particularly, the present inventors have searched for a material derived from a natural substance which is stable to human body and exhibited excellent whitening effect, and focused on extracts of Sawtooth mildew.

That is, according to an embodiment of the present invention, B16F10 cells were treated with the extracts of Sawtooth mildew at a concentration of 100% or more, showing no cytotoxicity and stable to the human body (see FIGS. 8 to 11).

On the other hand, melanin is a pigment present in the skin and plays an important role in protecting the body from external stimuli such as ultraviolet rays. However, when it is overproduced and deposited, it forms spots and freckles, which can cause problems in the appearance of the skin. Further, since it can induce skin cancer by promoting skin aging, a substance which can inhibit melanin excess production can be used as a whitening product It is possible.

Skin pigmentation is caused by stimuli such as ultraviolet rays or inflammation, and it is also known to be caused by factors such as genetic factors, hormones, foods, and chemicals. The process of biosynthesis of melanin involves melanocytes in the skin. In other words, the biosynthesis of melanin occurs in a special form of melanocyte subcellular organ called melanosome. In this melanocyte, an enzyme called tyrosinase plays the most important role in the production of melanin. Tyrosine is converted to dopa by the action of this enzyme, which is converted to dopachrome by the formation of dopaquinone through a series of oxidation processes and ultimately forms melanin, a black brown color. do. Protein genes involved in melanin pigment formation include tyrosinase, tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and extracellular signal-regulated kinases (ERK) Expression of MITF (Microphthalmia-associated transcription factor), a major transcription factor that regulates expression of synthetic factors, plays an important role in melanin synthesis.

According to one embodiment of the present invention, it was found that the sawtooth moth extract of the present invention has an excellent melanin synthesis inhibitory effect (see Figs. 12 to 15).

Furthermore, in order to determine whether the extract of the present invention affects the phosphorylation of ERK protein important for the synthesis of tyrosinase, which is an important enzyme for melanogenesis, Western blotting analysis confirmed the inhibition of ERK protein phosphorylation. As a result, Showed that tyrosinase protein expression was decreased compared with the control group (see Figs. 16 to 19).

From these results, the inventors of the present invention have experimentally proved that the teaspoon mallow extract of the present invention has a skin whitening effect.

Therefore, the present invention can provide a cosmetic composition for skin whitening containing an extract of Tooth mackerel as an active ingredient.

In order to prove the superiority of the extract of the present invention on the inhibition of the melanin production of the Korean mushroom repellent moth, it is possible to use other kinds of moths such as the staffs, the horns, the moths, the moths, And the extracts of Solanum tuberosum, and by measuring the EC50 values thereof, it was confirmed that the sawtooth moth extract of the present invention had much higher whitening effect than the other hat repellent extracts.

As a result of EC50 value calculation of the sawtooth moth extract of the present invention, a value of 7.2 ug / mL was obtained, and it was confirmed that the efficacy was at least 13 times higher than that of other hat repellents.

'EC50' means a concentration that inhibits 50% of the amount of melanin produced when the amount of melanin produced by the α-MSH treatment according to the present invention is 100%, and is referred to as 50% effective concentration or 50% efficacy concentration .

On the other hand, the present inventors have further tested whether any of the above-mentioned Sawmill extracts have a skin whitening effect.

As a result, it was found that SHQ, sagahydroquinoic acid (SQA), sagaquinoic acid (SQA), sagachromenol (SCM), and sagachromenol And it was confirmed that all of the above three compounds had excellent tyrosinase synthesis inhibiting ability.

Accordingly, the present invention provides a skin whitening cosmetic composition comprising, as an active ingredient, not only an extract of Tooth mackerel, but also a combination of sachet hydroquinic acid (SHQ), sagequinoic acid (SQA), and sagachromenol .

The toothed mothball extract according to the present invention can be obtained by extracting and isolating from nature using an extraction and separation method known in the art, and the extract defined in the present invention can be obtained by extracting from a sawtooth moth with an appropriate solvent And includes, for example, a crude extract of a saw moth, a polar solvent-soluble extract or a non-polar solvent-soluble extract.

As an appropriate solvent for extracting the extract from the sawtooth mackerel, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used. Examples of the solvent include, but are not limited to, purified water, methanol alcohol having 1 to 4 carbon atoms, acetone, ether, benzene, chloroform (including methanol, ethanol, propanol, isopropanol, and butanol) various solvents such as chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination. Preferably, ethanol (alcohol), butanol or ethyl acetate can be used.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the production method of the sawtooth extract of the present invention, and any known method may be used.

For example, in the composition of the present invention, the first extract extracted with the hot water extraction method or the solvent extraction method can be prepared into a powder state by an additional process such as vacuum distillation, freeze drying or spray drying have. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

In the present invention, the teaspoon mushroom extract is a concept that includes all the extracts, fractions and tablets obtained in each step of extraction, fractionation or purification, their diluted solutions, concentrates or dried products.

Methods for preparing extracts and fractions of teaspoon mine according to one embodiment of the present invention will be described in more detail as follows.

In the present invention, 1.5 kg of the powder obtained by natural drying and grinding of the saw blades in a sunny place was put in 6 L of a syrup, and the mixture was refluxed and cooled twice at 70 캜 for about 3 hours, and then the foreign substances were removed by an ultrafilter. The crude oil extract was obtained.

The crude extract of the sawtooth moths was suspended in a mixed solvent of water: ethanol (9: 1, v / v), and n-hexane soluble fraction was obtained by using n-hexane.

The toothed moth extract or the fractions thereof according to an embodiment of the present invention may be contained in the cosmetic composition at a concentration of 1 ppm to 1000 ppm.

When the extract or the compound isolated therefrom is prepared from a cosmetic composition as described above, the composition of the present invention may contain components commonly used in the cosmetic composition as well as the above-mentioned Sawmill extract, Antioxidants, stabilizers, solubilizers, vitamins, customary adjuvants such as pigments and flavoring agents, and carriers.

In addition, the composition of the present invention may be mixed with an organic UV blocking agent which has been used in the past so long as it does not deteriorate the skin protecting effect by reacting with the sawtooth moth extract.

Organic UV protectors include, but are not limited to, glyceryl paraben, drometrizol trisiloxane, drometrizole, dipaloyyl trioleate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexylbutamidotriazone, Benzoylhexyl benzoate, di-methoxy cinnamate, a mixture of Rawson and dihydroxyacetone, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidene camphor, menthyl anthranilate, benzophenone- Benzophenone-3 (oxybenzone), benzophenone-4, benzophenone-8 (dioxyphenylbenzene), butylmethoxydibenzoylmethane, bishexylhexyloxyphenol methoxyphenyltriazine, synoxate, Octyl-p-methoxycinnamate, polysilicon-15 (dimethicone-ethylbenzylphthalate, ethylhexylmethoxycinnamate, ethylhexylmethoxycinnamate, ethylhexylsalicylate, ethylhexyltriazone, isoamyl- Yeah Bit), terephthalic ylidene dikaem peoseol sulphonic Acid and their salts, tea yieyi - may be used salicylate and at least one member selected from the group consisting of amino-benzo ripening Acid (paba).

In addition, the cosmetic composition of the present invention may be prepared in any form conventionally produced in the art, and examples thereof include solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactants- Containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, the cosmetic composition of the present invention can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

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 as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

The present invention also relates to a cosmetic composition comprising, as an active ingredient, a cosmetic composition comprising an extract of Tortola extract or sagahydroquinoic acid, sagaquinoic acid and sagachromenol as an active ingredient, And a cosmetic method comprising the steps of:

The cosmetic process of the present invention refers to all cosmetic processes using the cosmetic composition of the present invention. That is, all methods known in the art using a cosmetic composition belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in combination, or may be used in combination with other cosmetic compositions other than the present invention. The cosmetic composition according to the present invention may be used according to a conventional method of use, and may be used in a number of times depending on the skin condition or taste of the user.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, washed or rinsed with water. The surfactant-containing cleansing formulation may be a cleansing foam, a cleansing water, a cleansing towel and a cleansing pack. The surfactant-free cleansing formulation may be a cleansing cream, , Cleansing lotion, cleansing water and cleansing gel, but is not limited thereto.

According to the present invention, the content of the active ingredient (trehalose extract or the compound isolated therefrom) of the present invention is 0.0001-40% by weight, preferably 0.0005-40%, more preferably 0.0005-40% -10% by weight. If the content of the active ingredient (the extract of the sawtooth mackerel or the compound separated therefrom) is less than 0.0001% by weight, the skin whitening effect is greatly reduced. If the content is more than 40% by weight, skin irritation may be caused. Lt; / RTI >

On the other hand, the cosmetic composition according to the present invention may be formulated by incorporating an effective ingredient (a saphenic extract or a compound isolated therefrom) into the nanoliposome and stabilizing it. When the above active ingredient (the extract of Tooth mackerel or the compound isolated therefrom) is contained in the nanoliposome, the active ingredient is stabilized, thereby solving problems such as formation of precipitate, discoloration and detachment during formulation, and solubility and transdermal absorption So that the expected efficacy from the extract can be maximized.

In the present invention, nanoliposome refers to a liposome having a typical liposome form and having an average particle diameter of 10 to 500 nm. According to a preferred embodiment of the present invention, the average particle diameter of the nanoliposome is 50 to 300 nm, more preferably 100 to 200 nm. When the average particle diameter of the nanoliposome exceeds 500 nm, improvement of skin penetration and improvement of formulation stability is very weak among technical effects to be achieved in the present invention.

The nanoliposomes used to stabilize the extract according to the present invention may be prepared by a mixture comprising a polyol, an oily component, a surfactant, a phospholipid, a fatty acid and water.

The polyol used in the nanoliposome of the present invention is not particularly limited and is preferably a polyol such as propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, methylpropanediol, isopropylene glycol, pentylene glycol, erythritol, , Sorbitol, and mixtures thereof. The amount thereof is 10 to 80% by weight, preferably 30 to 70% by weight, based on the total weight of the nanoliposome.

The oil component used in the preparation of the nanoliposome of the present invention may be selected from a variety of oils known in the art and is preferably a hydrocarbon oil such as hexadecane and paraffin oil, Vegetable oils such as silicon oil such as dimethicone and cyclomethicone, sunflower oil, corn oil, soybean oil, avocado oil, sesame oil and fish oil, ethoxylated alkyl ether oils, propoxylated alkyl ether oils , Sphingoid lipids such as phytosphingosine, sphingosine and sphinganine, celecrose cholesterol, sitosterol cholesteryl sulfate, cetostearyl sulfate, C10-40 fatty alcohols and mixtures thereof. The amount thereof may be 1.0 to 30.0% by weight, and preferably 3.0 to 20.0% by weight based on the total weight of the nanoliposome.

Any surfactant known in the art may be used as the surfactant used in the preparation of the nanoliposome of the present invention. For example, anionic surfactants, cationic surfactants, amphoteric surfactants and nonionic surfactants can be used. Preferred are anionic surfactants and nonionic surfactants. Specific examples of anionic surfactants include alkyl acyl glutamates, alkyl phosphates, alkyl lactylates, dialkyl phosphates and trialkyl phosphates. Specific examples of nonionic surfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters, alkyl polyglycosides, polyglyceryl esters and sugar esters. Particularly preferred surfactants are polysorbates belonging to nonionic surfactants. The amount thereof may be 0.1 to 10% by weight, preferably 0.5 to 5.0% by weight, based on the total weight of the nanoliposome.

The phospholipid, another component used in the preparation of the nanoliposomes of the present invention, is a lipophilic lipid that is used with both affinity lipids and is composed of natural phospholipids (e.g., egg yolk lecithin or soy lecithin, sphingomyelin) Dipalmitoyl phosphatidylcholine or hydrogenated lecithin), preferably lecithin. In particular, unsaturated lecithin or saturated lecithin derived from natural origin extracted from soybean or egg yolk is preferable. Normally, the amount of phosphatidylcholine is 23 to 95% and the amount of phosphatidylethanolamine is 20% or less in natural derived lecithin. In the production of the nanoliposome of the present invention, the amount of the phospholipid to be used is 0.5 to 20.0% by weight, preferably 2.0 to 8.0% by weight based on the total weight of the nanoliposome.

The fatty acid used in the production of the nanoliposome of the present invention is a higher fatty acid, preferably saturated or unsaturated fatty acids of C12-22 alkyl chain, such as lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid and linoleic acid . The amount thereof may be 0.05 to 3.0% by weight, preferably 0.1 to 1.0% by weight, based on the total weight of the nanoliposome.

The water used in the preparation of the nanoliposome of the present invention is generally deionized distilled water, and the amount thereof may be 5.0-40 wt% based on the total weight of the nanoliposome.

The preparation of nanoliposomes can be accomplished through a variety of methods known in the art, but most preferably is made by applying a mixture comprising the ingredients to a high pressure homogenizer. The preparation of a nanoliposome by a high pressure homogenizer can be carried out under various conditions (for example, pressure, number of times, etc.) depending on a desired particle size, and preferably at a high pressure homogenizer So that the nanoliposome can be produced.

The cosmetic composition of the present invention may be used alone or in combination, or it may be used in combination with other cosmetic compositions other than the present invention. The cosmetic composition having an excellent skin whitening effect according to the present invention can be used according to a conventional method of use, and may be used in a number of times depending on the skin condition or taste of the user.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.

≪ Example 1 >

Saw blade  Manufacture of alcohol extracts

The sawtooth moths used in the present invention were obtained by scuba diving using natural scavengers from Gyujang-gun and Gyeongnam Tongyeong-si in Busan. Professor Chang-geun Choi of Ecological Engineering Department of Pukyong National University and Professor of Department of Marine Life Science, Gyeongsang National University Professor Kim Nam-gil performed this. The toothed mushroom was naturally dried in sunshine, and 1.5 kg of the obtained powder was put into 6 L of a syrup, and the mixture was refluxed and cooled twice at 70 캜 for about 3 hours, and then dehydrated by an ultrafilter (MWCO 50 kDa). The filtrate was extracted with a vacuum rotary condenser at 40 ° C, and 160 g of a crude extract was obtained as an extracted residue (hereinafter, abbreviated as "ESS" in the present invention). The thus obtained Saw Moth extract was used as a sample in the following experiment while being stored at 4 ° C in a refrigerator.

≪ Example 2 >

Separation of Saga Hydroquinone, Sagacromenol and Sagaquinoxaline as Effective Ingredients from Extracts of Sawtooth Matter

<2-1> Isolation of Saga Hydroquinoxaline

Sawtooth mung bean extract 15 g was dissolved in 1 liter of a mixed solvent of water: ethanol (9: 1, v / v), placed in a separatory funnel, equilibrated with the same amount of n-hexane, and the upper n-hexane- After dehydration with anhydrous sodium sulfate, the n-hexane fraction was isolated by concentration. The separated n-hexane fraction (12.2 g) was dissolved in 120 mL of methanol, filtered through a membrane filter, and then 200 μL was injected and separated into four fractions by HPLC equipped with Phenomenex C18 (2) (15 μm, 21.2 mm × 250 mm) column (See Fig. 1). As a mobile phase of the chromatography, a mixed solvent of methanol (A) and water (B) was used and the flow rate was set to 7 mL. The chromatographic conditions were A / B (90/10) to A / B (94/6) for 33 minutes, A / B (94/6) to A / B / B (100/0) for 10 minutes and the column was equilibrated for 10 minutes at A / B (90:10) concentration. Chromatography was performed 50-100 times by automatic sample injection with Autosampler to obtain a large fraction of the first fraction of fraction No. 1.

The same instrument and column were used to increase the purity of the first fraction obtained in large quantities. The chromatographic conditions were A / B (85/15) to A / B (87/13) for 26 min, A / B (87/13) to A / B / B (100/0) for 6 minutes and the column was equilibrated for 10 minutes at A / B (87:13) concentration. Chromatography was performed by automatically injecting the sample into an autosampler to obtain a second fraction of fraction 1 having a higher purity (see FIG. 2).

One of the second fractions of fraction No. 1 was purified by rechromatography (523 mg) under the same conditions and analyzed with an analytical column (Phenomenex C18 (2), 3 μm, 3 mm × 150 mm) , Which was analyzed by NMR and found to be sargahydroquinoic acid (hereinafter referred to as "SHQ") (see FIG. 3).

(1) Sagahydroquinoic acid

<2-2> Isolation of Sagar Chromenol

The fraction 2 of FIG. 1 was re-chromatographed using the same HPLC apparatus and column as the fraction 1 of Example 2-1. The chromatographic conditions were A / B (92.5 / 8.5) to A / B (92.2 / 7.8) for 26 min and A / B (92.2 / / B (100/0) for 6 minutes and the column was equilibrated for 10 minutes at A / B (91.5: 8.5) concentration. Chromatography was performed 50-100 times by automatic sample injection with Autosampler to obtain a large fraction of the second fraction of fraction No. 2 (see FIG. 4). As shown in FIG. 4, fraction No. 2 showed three peaks. The second peak (Frc2-2) was further purified to obtain a chromatogram as shown in FIG. 5 (126 mg). The separated product was analyzed by NMR and found to be sargachromenol (SCA) (see FIG. 5).

&Lt; Formula 2 > Sagachromenol

<2-3> Isolation of Saga cuneoic acid

The fraction No. 3 in Fig. 1 was re-chromatographed using the same HPLC apparatus and column as the fraction No. 1 in Example 2-1. The chromatographic conditions were A / B (93.4 / 6.6) to A / B (93.8 / 6.2) for 20 min, A / B (93.8 / / B (100/0) for 6 minutes and the column was equilibrated for 10 minutes at A / B (93.8 / 6.2) concentration. Chromatography was performed 50-100 times by automatic sample injection with Autosampler to obtain a large fraction of the second fraction of the fraction No. 3 (see FIG. 6). As shown in FIG. 6, the fraction No. 3 showed four peaks. The peak No. 4 (Frc 3-4) was further purified to obtain a chromatogram as shown in FIG. 7 (171 mg). The separated product was analyzed by NMR and found to be sargaquinoic acid (hereinafter referred to as SQA) (see FIG. 7).

&Lt; Formula 3 > Sagaquinoic acid

<Experimental Example 1>

Evaluation of Cytotoxicity of Extract of Sawtooth Moth

The inventors of the present invention have found that a toothed mucilage extract (ESS), a saccharide hydroquinic acid (hereinafter referred to as "SHQ" in the present invention), a sagaquinoxanic acid (hereinafter referred to as "SQA" (Hereinafter referred to as &quot; SCM &quot;) and sagarpromenol (hereinafter referred to as &quot; SCM &quot;

In detail, B16F10 cells were treated with the extracts and compounds of teaspoon mushroom prepared in Examples 1 and 2, and the cell viability was measured by MTS kit (ellTiter 96 AQueous One Solution Cell Proliferation Assay Kit) B16F10 cells per well were cultured at 6 × 10 ³ cells / well in the presence of 1 mM Tris-HCl After 24 hours of incubation, each extract was pretreated for 1 hour at each concentration, treated with 1 μM α-MSH for 24 hours and cultured, and then 95 μl of medium and 5 μl of MTS solution were added After 3 hours, absorbance was measured at 490 nm.

&Lt; 1-1 > Cytotoxicity of Extract of Spearmill

The present inventors measured the cell viability by the MTS assay by treating the extract of B16F10 with 5, 7.5, 15, and 20 μg / ml of the extract of B16F10 cells to determine the cytotoxicity of the extracts of Echinochloa crus-galli (ESS) prepared by the method of Example 1 Respectively.

As a result of the cytotoxicity measurement of the extracts of teaspoon mushroom, it was confirmed that the extract of teaspoon mushroom did not show cytotoxicity on the B16F10 cell line in the range of 20 / / ml or less (see FIG. 8).

<1-2> Cytotoxicity of Compound Saga Hydroquinoxaline Derived from Spearman's Extract

The present inventors measured the cell survival rate of B16F10 cells at a concentration of 2, 2.5, and 3 μm by measuring the cytotoxicity of hydroquinone acid (SHQ) produced by the method of Example 2, assay.

As a result of the cytotoxicity measurement of the compound saga hydroquinoxalic acid (SHQ) derived from the teaspoon of Chenopodium album filtrate, SHQ showed no cytotoxicity in the B16F10 cell line in the range of 3 μM or less (see FIG. 9).

&Lt; 1-3 > Cytotoxicity of Saga cuneoic acid derived from Tephriton sp. Extract

The present inventors measured SQA at a concentration of 2, 2.5, and 3 μm on B16F10 cells by measuring the cell survival rate using an MTS assay .

As a result of the cytotoxicity measurement of the compound sagaquinoxanic acid (SQA) derived from the teaspoon of Chenopodium albumaline, SQA showed no cytotoxicity in the B16F10 cell line in the range of 3 μM or less (see FIG. 10).

&Lt; 1-4 > Cytotoxicity of sagarchromenol derived from extracts of teaspoon mine extract

The present inventors measured the cell viability of B16F10 cells at a concentration of 1, 2, 4, and 6 μm by measuring the cytotoxicity of chromenol (SMC) MTS assay.

As a result of the cytotoxicity measurement of the compound SCM derived from the teaspoon of spore extract, SCM showed no cytotoxicity on the B16F10 cell line in the range of 6 μM or less (see FIG. 11).

Overall, the cell survival rate was 100% or higher in the experimental group treated with the extracts of the extracts of the Tohmae mushroom extract and the extract of the mushroom extract. Thus, it was confirmed that the extracts of the teaspoon mushroom extract of the present invention, saga hydroquinone acid, sagaquinoxy acid and sagacromenol were safe for skin Respectively.

<Experimental Example 2>

Inhibitory effect on melanin biosynthesis of B16F10 cells in extracts of or from leaves

The present inventors examined the effect of extracts on the melanin biosynthesis of Bombyx mori extract on B16F10 cells at various concentrations and measured intracellular melanin content. Specifically, B16F10 cells were plated at 8 × 10 ⁴ cells / well in a 12-well plate. After 24 hours, the extract or compound of the alcohol was pretreated for 1 hour by concentration, treated with 1 μM of α-MSH for 72 hours, After 2 hours, wash with PBS, dissolve the cells with 1N NaOH, heat in an eppendorf tube and heat at 60 ° C for 30 min. Cell lysates were centrifuged at 5000 xg for 5 min and the absorbance of the supernatant was measured at 405 nm.

<2-1> Determination of Melanin Content in Cells Treated with Extract of Teapot

To investigate the effect of ESS on the melanin biosynthesis, B16F10 cells were treated with 15, 20, and 25 ㎍ / ml Sawtooth milt seed extract and 1 hour after treatment with 1 μM a-MSH. And the melanin content in the cells was measured. As shown in Fig. 12, the extracts of teaspoon mushrooms inhibited melanin biosynthesis in melanoma cells in a concentration-dependent manner.

<2-2> Determination of melanin content in cells treated with a compound of sardine hydroquinone acid derived from extracts of Tephritidae

In order to investigate the effect of the extract from the extracts of melanin on the melanin biosynthesis, B16F10 cells were treated with 1, 2, and 3 μM saghydroquinoxanic acid (SHQ) compounds and treated with 1 μM a-MSH For 72 hours to measure intracellular melanin content. As shown in Fig. 13, hydroquinone acid (SHQ), which is a compound derived from a teaspoon of alcohol extract, inhibits intracellular melanin biosynthesis in a concentration-dependent manner.

<2-3> Determination of Melanin Content of Saga cuneoic Acid Derived from Extract of Teapot Matter

In order to examine the effect of the extract from the extract of Sawtooth Bombyx mori extract on melanin biosynthesis, B16F10 cells were treated with 1, 2, 2.5 and 3 μg of SAGA and 1 μM of a-MSH The cells were incubated for 72 hours to measure the intracellular melanin content. As shown in Fig. 14, it was shown that the compound of squeeze mung bean extract-derived compound, quinoxalic acid (SQA), inhibited intracellular melanin biosynthesis in a concentration-dependent manner.

<2-4> Determination of Melanin Contents of Sagar Chromenol Derived from Extract of Teapot

To investigate the effect of sagarchromenol (SCM) on the biosynthesis of melanin, the B16F10 cell line was treated with 3, 4, and 5 μm sagarchromenol (SCM) compound and after 1 hour, 1 μM a -MSH and cultured for 72 hours to measure intracellular melanin content. As shown in FIG. 15, it was found that chromanol (SCM), which is a compound derived from a toothed mung bean extract, inhibits melanin biosynthesis in melanoma cells.

<Experimental Example 3>

Assessment of Inhibition of Activity of Tyrosinase Enzymes of Extract of Sawtooth moth or Extracted Compounds Thereof

It was tried to determine whether the extract of Tooth mothball of the present invention has a whitening effect through regulation of phosphorylation of ERK which is important for tyrosinase and melanin signal transduction mechanism which are important enzymes for melanin production. In the present invention, the degree of phosphorylation of ERK was measured, and the inhibitory effect of the Sawtooth mildew extract on B16F10 cells was confirmed by Western blotting to determine whether the extract of Sawtooth moth was inhibiting the activation of MITF and exhibiting a whitening effect. That is, the cells treated with the sample were incubated for a certain period of time, and the protein of the cells was recovered and the amount of protein expression was analyzed by Western blot analysis.

To investigate the effect of sagarchromenol (SCM) on the melanin biosynthesis, the extracts of Tohmae mushroom extract and hexane fraction of the present invention prepared in Example 1 were added to the B16F10 cell line, After 1 hour, 1 μM of a-MSH was treated and cultured for 72 hours. Proteins were extracted with RIPA buffer containing 1% protease inhibitor, followed by centrifugation at 12,000 rpm for 25 minutes at 4 ° C. to obtain supernatant . Protein content was quantified by using bovine serum albumin as a standard substance using a BCA protein assay kit (Pierce, Rockford, Ill., USA). 20-40 μg of the protein was put into a 5 × SDS sample buffer and denatured by heating at 100 ° C. for 10 minutes and then subjected to electrophoresis. After electrophoresis in a 10% SDS polyacrylamide gel at 120 V for 2 hours, the membrane was transferred to nitrocellulose membrane and blocked in a blocking solution containing 5% skim milk. Lt; / RTI &gt; The primary antibody was diluted 1: 1000, reacted at room temperature for 1 hour, and washed with Tris-Buffered Saline Tween-20 (TBS-T) three times for 10 minutes each. Then, the reaction was performed with a secondary antibody diluted at a ratio of 1: 5000 at room temperature for 1 hour and then washed with TBS-T four times for 10 minutes each. Differences in protein expression were observed using a luminoimager (ChemiDoc ™ XRS +, BIO-RAD) using an ECL western blotting substrate (Thermo Scientific).

As a result, it was confirmed that EGF, SQH, SQA and SCM effectively inhibited the phosphorylation of ERK. Thus, it was confirmed that tyrosinase (SQA) and sagaproic acid And the amount of expression was also decreased, thereby confirming the whitening effect of the teaspoon extract (see Figs. 16 to 19).

<Comparative Example>

Saw blade  Extracts or compounds isolated therefrom and domestic Yan An Mountain  Inhibitory effect of capillary reversal on melanin production

In order to prove the superiority of the extract of the present invention on the inhibition of the melanin production of the Korean mulberry harvest moth, it was found that the other mulberry cultivars such as the chief serpent, horny moth moth moth, moth moth moth moth moth moth moth moth moth moth moth moth moth moth moth, The EC50 value of these extracts was compared with that of the seedless moth, and it was confirmed that the sawtooth moth extract of the present invention had a much higher whitening effect than the other moth repulsive extracts.

Namba name (scientific name) EC ₅₀ Sargassum serratifolium 7.2 ppm S. fulvellum 95 ppm S. horneri 121 ppm Why yezoense ( S. yezoense) > 200 ppm S. confusum 92 ppm S. coreanum = S. ringgoldianum > 200 ppm S. macrocarpum > 200 ppm S. yendoi > 200 ppm Myagropsis myagroides > 200 ppm

As a result of calculation of the EC50 value of the sawtooth mildew extract of the present invention, a value of 7.2 ug / mL was obtained, and it was confirmed that the efficacy was at least 13 times higher than that of the other hatchbacks (see Table 1).

Statistical processing

Each experiment was repeated three times or more, and the results were expressed as mean ± standard deviation for each sample concentration. Significant difference test for each sample concentration group was considered as statistically significant result after Student t test compared with control group.

&Lt; Preparation Example 1 &

The whitening cosmetic composition preparation of the present invention

<1-1> Softened skin (skin)

The softened skin containing the teaspoon mine extract of the present invention, sago hydroquinone acid, sagaquinoxanic acid or sagacromenol of the present invention was prepared in a conventional manner at the following proportions.

Examples of the softening longevity formulation of the present invention ingredient Content (% by weight) Sawdust mugwort extract, Saga hydroquinone acid, Saga cuunaic acid or Sagar Chromenol 0.1 to 3 glycerin 5.0 1,3-butylene glycol 3.0 REG 1500 1.0 Allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium hyaruronate 5.0 ethanol 10.0 Octyldodec-16 0.2 Polysorbate 20 0.2 Unicide - Yu 13 0.01 incense a very small amount Distilled water Balance Sum 100

<1-2> Nourishing lotion

Nutritional lotions containing the teaspoon juice extract of the present invention, saga hydroquinone acid, sagaquinoxanic acid or sagacromenol were prepared in a conventional manner at the following proportions.

The nutritional lotion formulation of the present invention ingredient Content (% by weight) Sawdust mugwort extract, Saga hydroquinone acid, Saga cuunaic acid or Sagar Chromenol 0.1 to 3 Glyceryl stearate SE 1.5 Cetearyl alcohol 1.5 lanolin 1.5 Polysorbate 60 1.3 Sorbitastarate 0.5 Hardened vegetable oil 4.0 Mineral oil 5.0 Trioctanoin 2.0 Dimethicone 0.8 Tocopheryl acetate 0.5 Carboxyvinyl polymer 0.12 glycerin 5.0 1,3-butylene glycol 3.0 Sodium hyaruronate 5.0 Triethanolamine 0.12 Unicide - Yu 13 0.02 incense a very small amount Distilled water Balance Sum 100

<1-3> Nourishing cream

The nutritional cream containing the teaspoon juice extract of the present invention, saga hydroquinone acid, sagaquinoxanic acid or sagacromenol was prepared in a conventional manner at the following proportions.

The nutritional cream formulation of the present invention ingredient Content (% by weight) Sawdust mugwort extract, Saga hydroquinone acid, Saga cuunaic acid or Sagar Chromenol 0.1 to 3 Pro-type glycerin monostearate 1.5 Cetearyl alcohol 1.5 Stearic acid 1.0 Polysorbate 60 1.5 Sorbitastarate 0.6 Isostearyl isoserate 5.0 Squalane 5.0 Mineral oil 35.0 Dimethicone 0.5 Hydroxyethylcellulose 0.12 glycerin 6.0 Triethanolamine 0.7 Unicide - Yu 13 0.02 incense a very small amount Distilled water Balance Sum 100

<1-4> Massage Cream

Massage creams containing the teaspoon of the present invention extract of the present invention, saga hydroquinone acid, sagaquinoxanic acid or sagacromenol were prepared in a conventional manner at the following proportions.

Massage Cream Formulation Example of the Present Invention ingredient Content (% by weight) Sawdust mugwort extract, Saga hydroquinone acid, Saga cuunaic acid or Sagar Chromenol 0.1 to 3 Propylene glycol 6 glycerin 4.0 Triethanolamine 0.5 Wax 2.0 Tocopheryl acetate 0.1 Polysorbate 60 3.0 Sorbitan sesquioleate 2.5 Cetearyl alcohol 2.0 Liquid paraffin 30.0 Carboxyvinyl polymer 0.5 incense a very small amount Distilled water Balance Sum 100

<1-5> Pack

A pack containing the sawtooth extract of the present invention, saga hydroquinone acid, sagaquinoxanic acid or sagacromenol of the present invention was prepared in a conventional manner at the following proportions.

Pack Formulation Example of the Present Invention ingredient Content (% by weight) Sawdust mugwort extract, Saga hydroquinone acid, Saga cuunaic acid or Sagar Chromenol 0.1 to 3 glycerin 10.0 Betaine 5.0 REG 1500 2.0 Allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethylcellulose 0.1 Sodium hyaruronate 80. Carboxyvinyl polymer 0.2 Triethanolamine 0.18 Octyldodecanol 0.3 Octyldodec-16 0.4 ethanol 6.0 Unicide - Yu 13 0.01 incense a very small amount Distilled water Balance Sum 100

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (8)

A cosmetic composition for whitening skin, comprising as an active ingredient,
The toothed mugwort extract comprises: a) natural drying of the sawtooth moth in sunny place, followed by grinding and pulverizing; b) adding the alcohol to the powder, refluxing and cooling at 70 ° C, and removing foreign matter through an ultrafilter using a 50 kDa filtration membrane; And c) removing the alcohol from the filtrate extract obtained through step b). &Lt; Desc / Clms Page number 20 &gt; delete The method according to claim 1,
The cosmetic composition for whitening skin according to claim 1, wherein the toothed mugwort extract has melanin production inhibitory activity and tyrosinase inhibitory activity. The method according to claim 1,
The cosmetic composition for whitening skin according to claim 1, wherein the toothed pan extract is contained in an amount of 0.0001 to 20% by weight based on the total weight of the cosmetic composition. A cosmetic composition for skin whitening comprising a sagahydroquinoic acid compound as an active ingredient. delete 6. The method of claim 5,
Wherein the compound has a melanin production inhibitory activity and a tyrosinase inhibitory activity. 6. The method according to any one of claims 1 to 5,
The cosmetic composition of the present invention can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritive cream, a moisturizing cream, a hand cream, Wherein the composition is formulated as one selected from the group consisting of cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, lipstick, makeup base, foundation, press powder and loose powder .

Images