COSMETIC COMPOSITION FOR WHITENING OR REDUCING FRECKLES CONTAINING MEDICINAL HERB EXTRACT

In the embodiment of which the described and drawing other specific are obviated included in the nanometer range.

Advantages and features of the present invention, achieve the appended drawing method and an electronic component connected to the reference surface with specifically carry activitycopyright will in the embodiment. However the present invention refers to hereinafter are limited to the disclosure in various different in the embodiment but can be embodied in the form.

The present invention are hereinafter is described diffuse to the products on the attached drawing.

The present invention refers to horse riding, locking, degree, watch beginning, therefore, snap judgment, Angelica gigas Nakai, agent, composition is useful, effect, Ginseng, dermis, gold, ginger, matching, is the justice, rhizoma, therefore, [thayk company, it can be used, processed, while, Ginseng and numerous combinations including a cosmetic composition for improving or lightening ingredients extracted from anti-oxidant activity including number [...] substrate. In the present invention used natural medicinal drug is used but said described preferably, anti-oxidant of the type does not composite, typically the art can be used anti-oxidant further comprises selectively utilizing.

The present invention according to cosmetic composition for improving or lightening ingredients, said anti-oxidant herb carboethoxygermanium sesquioxide 0 horse riding. 2 - 2. 2% by weight, locking 1. 4 - 3. 4% by weight, degree 1. 4 - 3. 4 weight %, watch beginning 1. 4 - 3. 4% by weight, therefore 1. 4 - 3. 4% by weight, 1 snap judgment. 4 - 3. 4 weight %, radix 2. 5 - 4. 5% by weight, an agent 2. 5 - 4. 5% by weight, composition is 2. 5 - 4. 5% by weight, an effect 2. 5 - 4. 5% by weight, Ginseng 2. 5 - 4. 5% by weight, 2 dermis. 5 - 4. 5% by weight, gold 2. 5 - 4. 5% by weight, ginger 2. 5 - 4. 5% by weight, contrast 2. 5 - 4. 5% by weight, 4 is the justice. 8 - 6. 8 weight %, rhizoma 4. 8 - 6. 8% by weight, 3 state. 7 - 5. 7% by weight, 3 [thayk company. 7 - 5. 7% by weight, it can be used 3. 7 - 5. 7% by weight, food product 6. 0 - 8. 0% by weight, while 6. 0 - 8. 0% by weight, Ginseng 6. 0 - 8. 0% by weight, 7 numerous combinations. 2 - 9. 2% by weight preferably including a.

The present invention according to cosmetic composition for improving or lightening ingredients, said further said horse riding, locking, degree, watch beginning, therefore, snap judgment, Angelica gigas Nakai, agent, composition is useful, effect, Ginseng, dermis, gold, ginger, matching, is the justice, rhizoma, therefore, [thayk company, it can be used, processed, while, Ginseng and numerous combinations including roast temperature of a process for the extraction of 2 - 6 2 - 5 70 provided 100 °C in water pressure obtained after a periodical repetition time asparagus, lyophilized encoded number tank.

The present invention according to cosmetic composition for improving or lightening ingredients, said further cosmetic composition relative to the total 0. 1 - 5% by weight, preferably 0. 5 - 3% by weight containing in the nanometer range. Said anti-oxidant extract 0. 1% by weight less than effect is insufficient when melanin formation inhibiting effect and can be inserted into the ingredients, anti-oxidant extract 5% by weight non-door cover is constructed mirror number the dose effectiveness number point at the disclosed.

The present invention according to cosmetic composition for improving or lightening ingredients, said skin cosmetic composition number may be, lotion, nutrient cream, massage cream, mist, essence, gel, tris, pack, powder, skin ointment, soap, intelligence number, hair lotion, shampoo, suspension, emulsion, spray or cosmetic liquid bath 1308. form number.

The present invention according to the present invention according to roast for cosmetic composition for improving or lightening ingredients commonly used in cosmetic extract one or more additive number number bath using high pressure liquid coolant therein. Cosmetic compositions of the present invention included in additive is mineral oil, triacetyl [...], butylene writing this call, triacetyl alcohol, writing three reel [su reel oh [ley [thu, not pin -100 stearate, polysorbate, -in-, writing three reel [su reel oh [ley [thu s, micro creasing dephosphorisation wax, sorbitol grief [su reel oh [ley [thu, sodium in high egg base four [thu, lung stipendiary hour ethanol, betain, perfume, carbomer, [...], sodium hydroxide, tocopheryl acetate, d small [tyum this d mote a, preferably positive number be limited to using composite has the kind of a container, the effect of the invention mixed with the anti-oxidant extract not range relayed a billion number can be an appropriate amount.

In addition, the present invention according to method of cosmetic cosmetic composition for improving or lightening ingredients for morning fair number, kind of rooibos extract used anti-oxidant extract contained in the roast, hypocyclonucleocide coconut extract, aroyl [ni oh fruit extract, green tea extract, feeling country [khayl cultivation extract, Ginseng callus culture extract, cysteine division tissue cell culture extract, green tea callus culture extract, an extract, extract product, resin extract, Scutellariae radix extract, malaphis, green tea extract, ginkgo leaf extract, peach leaf extracts, white pattern [eng extracts, licorice extract taking effect filtration water using Aspergillus/yeast/further can be selectively.

Hereinafter the present invention refers to in the embodiment through more detailed as follows.

<In the embodiment 1 to 12: Anti-oxidant Physiologically active>extract

[Anti-oxidant Extract number bath]

1 horse riding. 18% by weight, locking 2. 35% by weight, degree 2. 35% by weight, watch beginning 2. 35% by weight, therefore 2. 35% by weight, 2 snap judgment. 35% by weight, 3 pack. 53% by weight, an agent 3. 53% by weight, 3 composition is useful. 53% by weight, 3 effect. 53% by weight, Ginseng 3. 53% by weight, 3 dermis. 53% by weight, gold 3. 53% by weight, 3 ginger. 53 weight %, matching 3. 53% by weight, 5 is the justice. 88% by weight, rhizoma 5. 88% by weight, state 4. 71% by weight, [thayk company 4. 71% by weight, it can be 4. 71% by weight, food product 7. 06% by weight, while 7. 06% by weight, Ginseng 7. 06% by weight, 8 numerous combinations. 24% by weight of the nonwoven fabric are included in the water at a temperature of 100 °C pressure extracting method 3 is placed 20L periodical repetition time was a 2. After said collected and lyophilized extract obtained activity.

Of the present invention anti-oxidant extract for experiments such as the physiologically active result to conducting for each method.

[Cell culture]

B16F 10 used in the present invention in Korean cell line bank (KCLB, a compensation Seoul) 10% melanin cell lines regardless modified eagle medium (Dulbecco's modified eagle's medium; DMEM) it puts the hemp cloth nose small [thay [...] (fetal bovine serum; FBS) added with the deflection, CO2 incubator (MCO-a 17A 1, Sanyo, Japan) 37 °C temperature, was cultured in 5% CO2 conditions.

[Cytotoxic measuring]

B16F 10 96 well (well) to 5 × 103 cells by cells which is included in the stabilizing activity after 24 hours cells treated against according to concentration. 72 then attempt to culture solution cell culture solution added is used time after time 37 °C 2 MTS 1/10 in when the culturing, after ELISA microplate reader (Infinite 200 pro, TECAN Ltd. , The very [nu [lu Switzerland) in absorbance 490 nm using a long time.

[IntracellularIt is sour with the mote number Assay]

B16F 10 6 cells 24 hours after the cells 5 × 104 cells which is included in the well to the respective stabilizing activity concentration have a 100 nm 1 according to processing to render the handling time after some time α-a MSH 72 behind his culture. PBS washing 2 times to RIPA buffer (protein lysis buffer) and after 30 minutes incubation at a cis lysine protein in his 4 °C. 4 °C 13000 rpm 30 minutes then it is sour with the mote part is centrifugal through separation of protein activity number fields supernatant were measured. (Bradford, 1976) 595 nm through a piping method for quantification pod amount protein ELISA measurements to the same protein using a number stored in the 540 nm with extinction intracellular it is sour with the mote L a-DOPA activity were measured.

[Melanin measurement and observation]

B16F 10 to 5 × 104 cells (dish) by cells which is included in each extract concentration 60 mm DC pretreated time according to a a-MSH 1 (100 nm) 72 his culture processing time. PBS then washed and trypsin - EDTA to 2 times 5 minutes after processing a number 3000 rpm to harvest cells to 10% removed with dimethyl sulfoxide (DMSO) stand-alone cells to gather supernatant after processing with respect to the 80 °C 1N NaOH solution added with 1 in reaction time. Each absorbance microplate reader using quantitative ELISA melanin synthetic melanin 410 nm were measured.

[Whitening related embodiment liver gene expression PCR]

Whitening related gene expression is verified to 1 × 105 cells after culturing time division one by one 60 mm DC to B16F 10 24 therein. Add time after processing each extract 1 α-a MSH (100 nm) 24 hours after-wash 2 PBS to melanin formation and to culture harvest cells and to him.

Harvesting cells using RNA isolation number (Tripure Isolation Reagent, Roche Diagnostics, Germany mannheim) tree is lapped and separating him. 5 micro g of a mRNA using reverse transcriptase kit (High Capacity cDNA Reverse Transcription Kit, Applied Biosystem, when poster U.S. California) high performance cDNA synthesized cDNA his. Experiments conducting irradiation number according to the manual. Synthesized cDNA 1 micro l, taqman primer 1 micro l, Taqman universal master mix II (Taqman Universal Master Mix II, Applied Biosystem, when poster U.S. California) 10 micro l, distilled water 8 micro l into and out of the liver PCR device embodiment 3 difference (ABI7500, Applied Biosystem, when poster U.S. California) conducting using PCR. For table 1 shown to a quantitative polymerase reaction and which information of the TaqMan number. In addition embodiment 2 minutes 10 minutes 95 °C PCR reaction conditions in the liver in-step 1 and 50 °C, 60 °C superhuman cycle times in 15 seconds in 15 40 conducting modified temperature 95 °C annealing temperatures.

[Statistics processing]

The experiment result obtained in standard deviation for the average ± (mean ± S. D.) precursor to, controls each experiment group to an average difference value value p - 0 stores [...] Stowe T assay (Student's t-a test). 05 statistically significant difference when below the number of passes is runner out.

<Experiment example 1. Anti-oxidant Extract B16F 10 Evaluating>inducing cytotoxicity against cell lines

The experiment in example 1 in anti-oxidant extract B16F 10 cell lines was to evaluate cytotoxicity experiments. Example 1 of the present invention cells in experiments to assess the cytotoxic activity B16F 10 to in the embodiment 1 (125 μg/ml), in the embodiment 2 (250 μg/ml) and comparison example 1 (500 μg/ml), comparison example 2 (1000 μg/ml) after processing such as kiln and processing time when the MTS 24 by each concentration, the result was also shown in the 1. Figure 1 anti-oxidant extract inducing cytotoxicity against evaluation result indicating graph B16F 10 cell lines are disclosed. With reference, in Figure 1 blank is finish matching could be anything, in the embodiment 1 is a 125, 250 is a in the embodiment 2, a 500 compared to the example 1, example 2 compared to the 1000 exhibits.

In the embodiment of example embodiments and comparison experiment 1 experiment conditions present the disclosed.

In the embodiment 1, in the embodiment 2 of the present invention followed by 125, 250 μg/ml in each activity B16F 10 cytotoxicity in cells was assessed.

Comparison example 1, example 2 of the present invention comparing activity followed by 500, 1000 μg/ml in each B16F 10 cytotoxicity in cells was assessed.

The reference also 1, 125, 250, 500, 1000 μg/ml each activity in the embodiment and comparison example all charging cell in existence power finish anything coping contrast is not visible a big difference compared to the liver. Thus, the present invention according to further ingredients for improving input concentration not cytotoxic regardless, when applied to a high pressure liquid coolant using the same number can be expected to be a cosmetic skin stability.

<Experiment example 2: Anti-oxidant Extract melanin formation inhibiting effect>

The experiment in example 2 influence on melanin synthesis in B16F 10 cell lines in order to identify anti-oxidant extract his experiments. Example 2 of the present invention cells in experiments to assess the melanin formation inhibiting effect B16F 10 to activity in the embodiment 3 (125 μg/ml), in the embodiment 4 (250 μg/ml), and comparison example 3 (31. 3 μg/ml), comparison example 4 (62. 5 μg/ml) followed by concentration such as processing according to each have, as a reference example 1 with an α-a MSH 100 nm alone, arbutin as reference example 2 (Arbutin) 100 μg/ml followed by cases where melanin formation inhibiting effect was further includes an FFT processing. Said each concentration by anti-oxidant extract after 1 time after processing arbutin B16F 10 cell lines derived by culturing time after processing α-a MSH pigmentation 72 when the comparing entire melanin content, the result was also shown in the 2. Figure 2 anti-oxidant extract melanin formation inhibiting effect of graph are disclosed. With reference to a, in Figure 2 blank is finish matching could be anything, a reference example 1 is α-a MSH, Arbutin a reference example 2, 31. 3 comparison example 3 a, 62. 5 is a comparison example 4, 125 is a in the embodiment 3, the 250 exhibits in the embodiment 4.

Experiment 2 in the embodiment and comparison example of experiment conditions relate to present the disclosed.

In the embodiment 3, in the embodiment 4 of the present invention followed by 125, 250 μg/ml melanin content in each activity measured melanin formation inhibiting effect therefrom.

Comparison example 3, example 4 of the present invention 31 compared in each activity. 3, 62. 5 μg/ml followed by measuring melanin formation inhibiting effect melanin content therefrom.

The reference 2 also in the embodiment 3, in the embodiment 4 of the present invention anti-oxidant extract such as 125 - 250 μg/ml is processed on the basis of the activity input only a reference example 1 without significant melanin content compared to α-a MSH alone reduces the ETEC. However of the present invention 31 activity. 3, 62. 5 μg/ml is put in comparison example 3, in the case of reference example 1 α-a MSH 4 alone processing compared to comparison example 3 reduced in the content which the melanin, 4 but also melanin formation inhibiting effect which is able to when it is judged that, ingredients that ball cannot effect caused improvement passes and high electrical conductivity. The, anti-oxidant extract this comparison example 3, 4 such as insufficient content if it can be inserted into the weak melanin formation inhibiting effect.

<Experiment example 3: Anti-oxidant Extract Intracellular It is sour with the mote but oh>assay number

Melanin's disease can have half a door with the material known as organ it is sour but oh this enzyme covalently number used for the one of the plurality of melanin synthesis which plays an important role. The experiments of the present invention anti-oxidant extract in example 3 it is sour but oh number increased by α-a MSH any effects in order to identify whether one of the plurality of activity it is sour but oh number activity affecting intracellular trough therefrom. It is sour but oh B16F 10 example 3 of the present invention cells in the one of the plurality of number active measurement experiment for activity in the embodiment 5 (125 μg/ml), in the embodiment 6 (250 μg/ml) and comparison example 5 (31. 3 μg/ml), comparison example 6 (62. 5 μg/ml) according to each concentration followed by processing such as a transparent conductive layer, as a reference example 3 with an α-a MSH 100 nm alone, reference example 4 when it is sour but oh followed by one of the plurality of cells of a 100 μg/ml arbutin as processing further comprises experiment result was also shown in the 3 number activity. With reference, in Figure 3 blank is finish anything could be contrast, example 3 is a reference α-a MSH, Arbutin a reference example 4, 31. 3 comparison example 5 a, 62. 5 comparison example 6 is a is a 125 in the embodiment 5, in the embodiment 6 is 250 by a goniophotometer.

In the embodiment and comparison example 3 experiment experiment conditions of embodiments present the disclosed.

In the embodiment 5, in the embodiment 6 of the present invention it is sour but oh number 125, 250 μg/ml in each one of the plurality of activity followed by intracellular activity were measured.

Comparison example 5, example 6 of the present invention 31 compared in each activity. 3, 62. It is sour but oh number followed by intracellular trough 5 μg/ml were measured activity.

Reference number 3 also increases by the one of the plurality of confirming the activity of α-a MSH it is sour but oh cream (reference example 3), in the embodiment 5, in the embodiment 6 of the present invention anti-oxidant extract such as the throttle valve of the present invention anti-oxidant extract number 125 - 250 μg/ml when it is sour but oh processing activity increased by one of the plurality of steps as it is sour but oh α-a MSH number activity has been confirmed. This number it is sour but oh trough of the present invention anti-oxidant extract and reducing the content appears to inhibit the activity of melanin, melanin content of the present invention then further reduces the whitening function can be cylindrical.

On the other hand, 31 each activity. 3, 62. 5 μg/ml followed by processing comparison example 5, example 6 comparison example 3 has been treated with a trough it is sour but oh number activity of α-a MSH alone reduced relative to a reference that trough and an indirectly-inhibiting effect but it is sour but oh shown active number, its reduction is in the embodiment 5, 6 for improving whitening effect of herb extract ingredients is less than 125 - 250 μg/ml can be more cylindrical.

<Experiment example 4: B16F 10 In cell lines Anti-oxidant By extract TYRP1, MITF, TYRP2MRNA >Influence expression

The experiment in example 4 of the present invention anti-oxidant extract associated with the modulation melanin gene TYRP1, MITF, TYRP2 mRNA expression of his experiment in order to identify whether any affect. TYRP1 and TYRP2 transcription factor MITF is expressed by increased is a known, involved in melanin biosynthesis some known. The experiment in example 4 of the present invention anti-oxidant extract expression of this gene in order to identify whether any affect on cell lines of the present invention α-a MSH activity after 24 B16F 10 embodiment to liver gene expression has been confirmed that the PCR processing time.

The experiment in example 4 of the present invention cells for experiments B16F 10 to activity in the embodiment 7, 9, 11 (125 μg/ml), in the embodiment 8, 10, 12 (250 μg/ml) and comparison example 7, 9, 11 (31. 3 μg/ml), 8, 10, 12 comparison example (62. 5 μg/ml) followed by concentration according to each such as MITF, TYRP1, TYRP2 degree each gene expression were measured. In addition, as a reference example 5, 7, 9 with an α-a MSH alone, reference example 6, 8, 10 degree in addition when the measuring gene expression followed by arbutin as cases where processing, also the result (MITF mRNA expression) 4a, 4b (TYRP1 mRNA expression), 4c (TYRP2 mRNA expression) was shown. Also 4a, 4b, 4c is in anti-oxidant extract B16F 10 cell lines by MITF, TYRP1, TYRP2 mRNA expression of graph are disclosed. With reference, also 4 a, 4b, 4c is finish anything onto blank in contrast, α-a MSH is used in a reference example 5, 7, 9, respectively a Arbutin reference example 6, 8, 10, 31. 3 a comparison example 7, 9, 11 respectively, 62. 5 composition is used in a comparison example 8, 10, 12, each a 125 in the embodiment 7, 9, 11, respectively exhibits 250 in the embodiment 8, 10, 12.

Experiment example 4 in the embodiment and comparison of experiment conditions relate to present the disclosed.

In the embodiment 7, in the embodiment 8 of the present invention is put in the MITF gene gene expression activity in each 125, 250 μg/ml were measured.

In the embodiment 9, in the embodiment 10 of the present invention followed by TYRP1 gene gene expression activity in each 125, 250 μg/ml were measured.

In the embodiment 11, in the embodiment 12 of the present invention followed by 125, 250 μg/ml in each TYRP2 gene gene expression activity were measured.

Comparison example 7, example 8 of the present invention 31 compared in each activity. 3, 62. 5 μg/ml were measured gene expression is put in the MITF gene.

Comparison example 9, 10 of the present invention comparing each activity in example 31. 3, 62. 5 μg/ml followed by TYRP1 gene gene expression were measured.

Comparison example 11, 12 of the present invention comparing each activity in example 31. 3, 62. TYRP2 gene gene expression followed by 5 μg/ml were measured.

The reference also 4a, MITF mRNA levels of α-a MSH increased by in the embodiment 7, in the embodiment 8 of the present invention anti-oxidant extract such as 125 - 250 μg/ml significantly reduced concentration when it has processed a verify queue. The MITF gene expression in the skin of the present invention effectively billion number 125 - 250 μg/ml of further functionality to be been can be ascertained. While, 31 activity. 3 - 62. 5 μg/ml input comparison example 7, example 8 billion number MITF gene expression in comparison to n preferably is substantially free can be viewed whitening function is not known.

In addition, the mRNA level of increased by α-a MSH TYRP1 4b also reference in the embodiment 9, in the embodiment 10 of the present invention anti-oxidant extract 125 - 250 μg/ml such as making sure that the reduced concentration when it has processed a first call request. The number of mechanisms of the present invention expression of a further it is sour with the mote but oh 125 - 250 μg/ml in the TYRP1 effectively billion number his car. While, 31 activity. 3 - 62. Example 9 compared charging 5 μg/ml, in comparison example 10 is composed of a decrease in expression of mRNA levels of α-a MSH TYRP1 TYRP1 billion but when it is judged that the number effect, which is known to have arbutin skin melanin formation billion number is compared with the reference example 8 using a surface plane that has a can be confirmed.

In addition, the mRNA level of α-a MSH TYRP2 is increased by reference also 4c in the embodiment 11, in the embodiment 12 of the present invention anti-oxidant extract 125 provided 250 μg/ml such as making sure that the capable of significantly reducing a concentration when it has processed. While, 31 activity. 3 - 62. 5 μg/ml input comparison example 11, 12 is composed of a comparison example in a decrease in expression of mRNA levels of α-a MSH TYRP2 TYRP2 billion but when it is judged that the number which is able to effect, which is known to have arbutin skin melanin formation billion number is not greater effect than can be confirmed. In addition, in the embodiment 12 in the embodiment 11 and on mRNA levels compared TYRP2 decreases allowing ingredients is anti-oxidant extract 125 provided 250 μg/ml of improving effect can be more cylindrical.

The present invention is technical idea of the present invention is provided to person with skill in the art without changing its essential features or other specific form can be embodiment may be understand are disclosed. In the embodiment described above the exemplary non-limiting all sides are understood to which must substrate. Description of the present invention carry rather than patent the following is claimed are represented by said range, in which the meaning of and range patent the following is claimed some general outline of the form of the present invention evenly and method for all changing or modified range should interpreted.