CN117561050A - A composition comprising Saururus chinensis Baill leaf extract for whitening skin - Google Patents
A composition comprising Saururus chinensis Baill leaf extract for whitening skin Download PDFInfo
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- CN117561050A CN117561050A CN202280045631.8A CN202280045631A CN117561050A CN 117561050 A CN117561050 A CN 117561050A CN 202280045631 A CN202280045631 A CN 202280045631A CN 117561050 A CN117561050 A CN 117561050A
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- melanin
- saururus chinensis
- leaf extract
- extract
- composition
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
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Abstract
Relates to a cosmetic composition for skin whitening, a pharmaceutical composition for preventing, treating or improving melanin hyper-pigmentation diseases and a health functional food composition, which comprises saururus chinensis white leaf extract as an active ingredient. The saururus chinensis leaf extract is highly effective in inhibiting melanin generation and secretion and excellent in inhibiting skin pigmentation compared to saururus chinensis green leaf extract, and thus can be used as a cosmetic composition or pharmaceutical composition for skin whitening.
Description
Technical Field
The present application claims priority from korean patent application No. 10-2021-0090492, filed on 7/9 of 2021, and the disclosure of which is incorporated herein by reference in its entirety.
Relates to a cosmetic composition for skin whitening, a pharmaceutical composition for preventing, treating or improving melanin hyper-pigmentation diseases and a health-care functional food composition.
Background
Pigmentation, such as chloasma, freckles, etc., occurring on the skin is caused by an increase in melanin in the epidermis, and an important factor determining the skin color is also melanin.
The skin pigmentation mechanism can be broadly divided into melanin biosynthesis (melanogenesis), melanosome transport (melanosome transport), and melanosome transfer (melanosome transfer). The mechanism of melanin biosynthesis refers to a process of forming melanin in melanosomes, which are small organs of melanocytes (melanocytes), by the action of tyrosinase (tyrosinase) and tyrosinase-related protein 1/2 (TRP-1/2,tyrosinase related protein-1/2). The melanosome delivery mechanism is the process by which melanosomes are linked to the microcellular organelles and actin, thereby migrating to the ends of dendrites. Finally, the melanosome transfer mechanism refers to the process by which melanosomes that migrate to the dendritic end of melanocytes migrate to the surrounding diffuse epidermis. Through the three mechanisms, tyrosinase synthesis in UV-activated melanocytes is promoted, and melanogenesis is promoted to be delivered and excreted out of the epidermis (Jody P. Ebanks, mechanisms Regulating SkinPigmentation: the Rise and Fall of Complexion Coloration, int J Mol Sci. Sep;10 (9): 4066-4087, 2009).
Skin color depends on the content and distribution of melanin and is related to the amount and distribution of melanosomes released outside the cell after being produced in the melanocyte inside the cell. Hyperpigmentation of the skin can be caused by various factors such as hormonal abnormalities in the body after inflammatory reactions of the skin, genetic diseases, ultraviolet irradiation, etc., which are mainly due to abnormal melanin synthesis and distribution. The main function of melanin is to protect skin from damage caused by oxygen free radicals by scavenging oxygen free radicals, and more melanin means that melanin has an effective coping system and can protect skin from physical and chemical toxic substances.
However, melanin hyperpigmentation is a cause of pathological problems such as skin, but is also considered to be a cosmetic problem such as chloasma, freckles, nevi, and senile plaques. Therefore, there is a need to develop a whitening agent that inhibits melanin formation from the source.
Recent studies have shown that the development of substances that inhibit melanin biosynthesis and factors involved in melanosome transport and transfer processes is also a method capable of preventing skin pigmentation (Jong il Park at., inhibitory effect of-methyl-naptho [1,2,3-de ] quinone-8-one on melanosome transport and skin pigmentation, scientific Reports volume 6,Article number:29189, 2016).
Therefore, there is a need to develop a novel whitening raw material that can inhibit melanin biosynthesis and melanosome transfer process-related factors.
Disclosure of Invention
Technical problem
A composition is provided which comprises saururus chinensis white leaf extract as an active ingredient.
A method for preparing Saururus chinensis Baill leaf extract is provided.
A method of preventing or treating a melanin hyper-pigmentation disorder is provided, comprising administering to a subject in need thereof a composition comprising an effective amount of saururus chinensis white leaf extract.
There is provided a use of saururus chinensis white leaf extract for the preparation of a medicament for preventing or treating a melanin hyper-pigmentation disorder.
Technical proposal
In one aspect, a composition is provided that includes saururus chinensis white leaf extract as an active ingredient.
"Saururus chinensis (Asian lizard's tail, saururus chinensis)" is called Saururus chinensis because of the white color of the leaves, roots, and flowers. Leaves of saururus chinensis are green before flowering, but become white for insect attraction at the flowering stage and become green again after flowering. The blooming period of saururus chinensis is known to be june to june. That is, saururus chinensis will only appear "white leaf" during a certain period.
Thus, the term "white leaf of saururus chinensis" refers to the white leaf of saururus chinensis harvested during the flowering period of saururus chinensis. The white leaf of the saururus chinensis and the green leaf of the saururus chinensis have differences in color, acquisition period and the like.
The term "extract" includes all substances obtained by extracting components of natural products, and is independent of the extraction method, the extraction solvent, the extraction conditions, the extracted components or the form of the extract, and also includes substances which can be processed or treated by other methods after extracting components of natural products. For example, the processing or treatment may include dilution, concentration, drying, purification, fractionation, filtration, fermentation, enzymatic treatment, and the like. Thus, the extract may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, and a fraction after fractionation.
The above extraction method may use known natural extraction methods such as hot water extraction, ethanol extraction, heat extraction, cold soaking extraction, reflux cooling extraction, ultrasonic extraction, subcritical extraction, and the like, but is not limited thereto. In a specific embodiment, the extraction method may be hot water extraction or ethanol extraction.
The extraction solvent may be selected from water, organic solvents or mixtures thereof. The water may be distilled water or purified water. The organic solvent may include, but is not limited to, at least one of C1 to C6 lower alcohols, acetone, diethyl ether, ethyl acetate, diethyl ether, methyl ethyl ketone, and chloroform.
In a specific embodiment, the extraction solvent of the extract may be water, a C1 to C4 alcohol, or a mixture thereof.
In another embodiment, the extraction solvent of the extract may be a mixture of water and a C1 to C4 alcohol. In another embodiment, the extraction solvent of the extract may be ethanol. In another embodiment, the extract may have an extraction solvent of 10% (v/v) to 90% (v/v), 20% (v/v) to 80%
(v/v), 30% (v/v) to 70% (v/v), 40% (v/v) to 60% (v/v), or 50% (v/v) ethanol. Thus, the saururus chinensis white leaf extract may be a hot water extract of saururus chinensis white leaf or an ethanol extract of saururus chinensis white leaf. In one example, it was confirmed that the ethanol extract of the white leaf of saururus chinensis had excellent melanin generation-inhibiting effect and melanin secretion-inhibiting effect as compared to the hot water extract of the white leaf of saururus chinensis. Thus, the saururus chinensis white leaf extract may be an ethanol extract of saururus chinensis white leaf.
The extraction conditions refer to conditions for extracting natural product components, such as extraction time, extraction temperature, etc. The extraction time is 30 minutes to 120 hours, 30 minutes to 100 hours, 30 minutes to 80 hours, 30 minutes to 60 hours, 30 minutes to 40 hours, 30 minutes to 20 hours, 30 minutes to 10 hours, 2 hours to 120 hours, 2 hours to 100 hours, 2 hours to 80 hours, 2 hours to 60 hours, 2 hours to 40 hours, 2 hours to 20 hours, or 2 hours to 10 hours, but may be appropriately selected according to other conditions such as an extraction solvent, an extraction temperature, and the like. The extraction temperature can be 10 deg.C to 150 deg.C, 10 deg.C to 120 deg.C, 10 deg.C to 100 deg.C, 10 deg.C to 80 deg.C, 2 deg.C
0 ℃ to 150 ℃, 20 ℃ to 120 ℃, 20 ℃ to 100 ℃, 20 ℃ to 80 ℃, 20 ℃ to 60 ℃, 20 ℃ to 4%
The temperature is 0℃or normal temperature, but may be appropriately selected depending on other conditions such as the extraction solvent and the extraction time.
The term "comprising as an active ingredient" means that the saururus chinensis white leaf extract is added to such an extent that the effects mentioned in the present specification can be exhibited, and the meaning thereof includes that various forms are formulated after various ingredients as subcomponents are added for the purpose of delivering and stabilizing a drug.
The composition comprising Saururus chinensis Baill leaf extract as an active ingredient can be used for skin whitening, inhibiting melanin generation, inhibiting melanin secretion, reducing melanin, inhibiting melanin hyperpigmentation, or preventing, treating or ameliorating melanin hyperpigmentation diseases.
The term "skin whitening" may refer to lightening the skin tone by inhibiting melanin synthesis and improving skin hyperpigmentation caused by ultraviolet rays, hormones, or genetics to chloasma or freckles, etc.
The term "inhibiting melanin production" may refer to inhibiting melanin regeneration in cells or the like.
The term "inhibiting melanin secretion" may refer to inhibiting melanin secretion in cells or the like.
The term "reducing melanin" may refer to reducing the amount of melanin originally present in a cell or the like.
The term "melanin hyperpigmentation" is a broad concept including all melanin hyperpigmentation caused by various causes, and refers to the fact that a specific area of the skin or fingernail or toenail becomes darker than other areas due to excessive increase in melanin.
The melanin hyper-pigmentation disease may be any one selected from the group consisting of freckle, chloasma, liver spots, senile spots, female spots, solar blackens, melanospora, peutz-jegher's syndrome, gestational chloasma (chloasma gravidarum), hyperpigmentation after the use of a drug, and post-inflammatory pigmentation (postinflammatory hyperpigmentation), but is not limited thereto.
The term "preventing" includes inhibiting the occurrence of a disease. The term "treating" includes inhibiting, reducing or eliminating the progression of a disease. The term "ameliorating" may refer to alleviation of a state or at least alleviation of all behavior of a treatment-related parameter, e.g. at least alleviation of symptoms.
The saururus chinensis green leaf extract has a high skin whitening effect, melanin generation inhibiting ability, melanin secretion inhibiting ability, melanin reducing ability, or melanin hyper-pigmentation inhibiting ability, compared to the saururus chinensis green leaf extract.
The saururus chinensis white leaf extract may include any one or more of quercitrin (quercitin) and afzelin (afzelin). In one embodiment, it was confirmed that the white leaf extract of Saururi herba contained significantly higher levels of quercetin and afugin than the green leaf extract of Saururus chinensis.
The composition can inhibit melanin generation or secretion. In one example, it was confirmed that the melanins production-inhibiting effect and melanin secretion-inhibiting effect of the saururus chinensis white leaf extract were superior to those of saururus chinensis green leaf extract. Further, it was confirmed that quercetin contained in the saururus chinensis white leaf extract has excellent melanin production-inhibiting effect and melanin secretion-inhibiting effect. Further, it was confirmed that the ethanol extract of the saururus chinensis leaf has excellent melanin production-inhibiting effect and melanin secretion-inhibiting effect as compared with the hot water extract of the saururus chinensis leaf.
The composition can inhibit the expression of protease activated receptor-2 (protease-activated receptor-2, PAR-2) involved in melanosome transfer. In one embodiment, it was confirmed that the alfumoside contained in the saururus chinensis white leaf extract inhibits the expression of PAR-2 involved in melanosome transfer, and thus has a skin pigmentation inhibiting effect.
The concentration of the saururus chinensis white leaf extract may be, but is not limited to, 0.0001 μg/ml to 0.1 μg/ml, 0.0001 μg/ml to 0.05 μg/ml, or 0.0001 μg/ml to 0.001 μg/ml based on the whole composition. The concentration of the saururus chinensis white leaf extract may be selected to be within a moderate range that exhibits no cytotoxicity while exhibiting whitening efficacy.
The saururus chinensis white leaf extract contains quercetin at a concentration of 0.0001 μg/ml to 100 μg/ml, 0.0001 μg/ml to 10 μg/ml, 0.0001 μg/ml to 1 μg/ml, 0.001 μg/ml
However, the present invention is not limited thereto, and may be used in combination of/ml to 100. Mu.g/ml, 0.001. Mu.g/ml to 10. Mu.g/ml, 0.001. Mu.g/ml to 1. Mu.g/ml, 0.01. Mu.g/ml to 100. Mu.g/ml, 0.01. Mu.g/ml to 10. Mu.g/ml, or 0.01. Mu.g/ml to 1. Mu.g/ml.
The concentration of quercetin can be selected to be within a moderate range that exhibits no cytotoxicity while exhibiting whitening efficacy.
The saururus chinensis white leaf extract can be 0.0001 wt% to 20.0 wt%, 0.00 wt% based on the whole composition
01 wt% to 10.0 wt%, 0.0001 wt% to 5.0 wt%, 0.0001 wt% to 1.0 wt%, 0.00 wt%
1 to 20.0 wt%, 0.001 to 10.0 wt%, 0.001 to 5.0 wt%, 0.001 to 1.0 wt%, 0.01 to 20.0 wt%, 0.01 to 10.0 wt%, 0.01 wt%
To 5.0 wt%, or 0.01 wt% to 1.0 wt% is included therein, but is not limited thereto. The content of the saururus chinensis white leaf extract may be selected to be a moderate range that exhibits no cytotoxicity while exhibiting whitening efficacy.
In a specific embodiment, the composition may be a cosmetic composition.
The cosmetic composition can be used for whitening skin, inhibiting melanin generation, inhibiting melanin secretion, reducing melanin, or inhibiting melanin hyperpigmentation.
In addition to the active ingredients disclosed in the present specification, the cosmetic composition may further include ingredients conventionally used in cosmetic compositions, functional additives, and the like, such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, thickeners, preservatives, vitamins, pigments, fragrances, and the like, and cosmetically acceptable carriers.
The cosmetic composition may be prepared in any dosage form conventionally prepared. For example, the number of the cells to be processed, the cosmetic composition can be made into toner, face cream, essence, face cream, cleansing water, facial mask, ampoule, skin lotion, skin oil, etc,
Skin-moistening gel, shampoo, hair conditioner, hair cream, hair gel, foundation, lipstick, mascara or make-up cosmetic dosage form.
In a specific embodiment, the composition may be a skin external composition.
The skin external composition can be used for whitening skin, inhibiting melanin generation, inhibiting melanin secretion, reducing melanin, inhibiting melanin hyperpigmentation, or preventing, treating or ameliorating melanin hyperpigmentation diseases.
The skin external agent may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug impregnated bandage, lotion, or a combination thereof. The external preparation for skin can be appropriately mixed with components conventionally used for external preparations for skin such as cosmetics or medicines, for example, aqueous components, oily components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof, as required.
In a specific embodiment, the composition may be a pharmaceutical composition.
The pharmaceutical composition can be used for skin whitening, inhibiting melanin production, inhibiting melanin secretion, reducing melanin, inhibiting melanin hyperpigmentation, or preventing or treating melanin hyperpigmentation diseases.
The pharmaceutical composition may include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose or mannitol and the lubricant may be magnesium stearate, talc or a combination thereof. The carrier may be an excipient, a disintegrant, a binder lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxy cellulose, or a combination thereof. The disintegrant may be calcium carboxymethyl cellulose, sodium starch glycolate, anhydrous dibasic calcium phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low substituted hydroxypropyl cellulose, or a combination thereof. The lubricant may be magnesium stearate, silica, talc, or a combination thereof.
The pharmaceutical compositions may be formulated for oral or parenteral administration. The oral dosage form may be a granule, powder, liquid, tablet, capsule, dry syrup, or a combination thereof. The parenteral dosage form may be an injection.
In a specific embodiment, the composition may be a health functional food composition.
It can be used for whitening skin, inhibiting melanin generation, inhibiting melanin secretion, reducing melanin, inhibiting melanin hyperpigmentation, or preventing or improving melanin hyperpigmentation diseases.
The health functional food composition may use the extract alone or may use the extract together with other foods or food ingredients, and may be moderately used according to conventional methods. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use (prophylactic, healthy or therapeutic treatment). The kind of the health functional food is not particularly limited. The beverage composition in the category of health functional foods may contain various flavors or natural carbohydrates, etc. as additional ingredients as in the conventional beverage. The natural carbohydrate is monosaccharide such as glucose and fructose, disaccharide such as maltose and sucrose, polysaccharide such as dextrin and cyclodextrin, and sugar alcohol such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The health functional food composition may contain nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents for carbonated beverages, or combinations thereof. The health functional food composition may contain pulp for producing natural juice, juice beverage, vegetable beverage, or a combination thereof.
In another aspect, a method of preparing a saururus chinensis white leaf extract is provided.
The method comprises obtaining an extract by incubating saururus chinensis white leaves with an extraction solvent.
The detailed description about the saururus chinensis white leaf, the extraction solvent and the extract is as described above.
The incubation may be performed under conventional extraction conditions. The detailed description about the extraction conditions is as described above.
The method may further comprise filtering and/or drying the obtained extract. The filtration and drying may be carried out by known conventional methods.
In another aspect, a method is provided for preventing or treating a melanin hyper-pigmentation disorder comprising administering to a subject in need thereof a composition comprising an effective amount of saururus chinensis white leaf extract.
The term "effective amount" refers to an amount effective to exhibit the effects described above.
The terms "administering," "introducing," and "transplanting" are used interchangeably and may refer to administering a composition according to one embodiment to an individual by a method or route that at least partially localizes the composition according to the one embodiment to a desired site.
The application may be performed according to methods known in the art. The administration to the individual may be direct in any manner by a route such as intravenous, intramuscular, oral, transdermal (transmucosal), intranasal (intranasal), intratracheal or subcutaneous administration. The administration may be systemic or local.
The application may be by coating on the skin.
The individual may be a mammal such as a human, cow, horse, pig, dog, sheep, goat, or cat. The individual may be an individual in need of prevention or treatment of a melanin hyper-pigmentation disorder.
The dosage may be varied depending on the formulation method, the administration mode, the age, weight, sex, the degree of illness, diet, administration time, administration route, excretion rate, and response sensitivity of the patient, and the dosage may be appropriately adjusted by those skilled in the art in consideration of these factors. The number of administrations may be 1 time per day or two or more times per day within the scope of clinically acceptable side effects, the site of administration may also be 1 site or 2 sites or more, and the total administration date may be 1 day to 30 days per day or every 1 day to 5 days, 1 time of treatment. The same treatment can be repeated after an appropriate period of time if necessary. For animals other than humans, the dose per kilogram may be the same as the human, or the dose may be administered after calculation, for example, in terms of the volume ratio (e.g., average value) of the target animal and human organs (heart, etc.), etc.
Another aspect provides a use of an extract of saururus chinensis white leaf for the preparation of a medicament for preventing or treating a melanin hyper-pigmentation disorder.
In view of the complexity of the present description, duplicate matters are omitted, and terms not defined in the present description have meanings conventionally used in the art to which the present invention pertains.
Advantageous effects
The composition according to an aspect comprises saururus chinensis white leaf extract, and thus can have effects of skin whitening, inhibiting melanin generation and secretion, and inhibiting melanin hyper-pigmentation. The saururus chinensis leaf extract is highly effective in inhibiting melanin generation and secretion and excellent in inhibiting skin pigmentation compared with saururus chinensis green leaf extract, and thus can be used as a cosmetic composition or pharmaceutical composition for skin whitening.
Drawings
FIG. 1a shows the results of LC-QTOF-MS analysis of the Saururus chinensis white leaf extract in example 1.
FIG. 1b shows the results of LC-QTOF-MS analysis of the Saururi herba green leaf extract in comparative example 1.
FIGS. 2a and 2b show MS fragmentation patterns of quercetin contained in Saururus chinensis Bai Shezhong.
Fig. 3a and 3b show the MS fragmentation pattern of african-containing african-herb Bai Shezhong.
Fig. 4 shows cytotoxicity test results of example 1, comparative example 1, example 2, and comparative example 2.
FIG. 5 shows the cytotoxicity test results of quercetin contained in Saururus chinensis Baill leaf extract.
Fig. 6 shows the cytotoxicity test results of african-white grass leaf extract containing african-bean glycoside.
Fig. 7 shows melanin biosynthesis results of example 1, comparative example 1, example 2, and comparative example 2.
FIG. 8 shows melanin biosynthesis results of quercetin contained in Saururus chinensis Baill leaf extract.
Fig. 9 shows melanin secretion rate results of example 1 and comparative example 1.
FIG. 10 shows the melanin secretion rate results of quercetin contained in Saururus chinensis Baill leaf extract.
FIG. 11 shows the PAR-2 expression inhibiting effect of african-containing african-white leaf extract.
Detailed Description
Hereinafter, the present invention will be described in detail by way of examples. However, these examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.
EXAMPLE 1 preparation of Saururus chinensis Baill leaf extract
Only white leaves were separated from Saururus chinensis, and extraction was performed with 50% ethanol at room temperature for 3 days to prevent decomposition of components. The filtrate was concentrated after filtration and dried in vacuo.
Comparative example 1 preparation of Saururus chinensis green leaf extract
Green leaves were separated from saururus chinensis only, and extracted with 50% ethanol at room temperature for 3 days to prevent decomposition of components. The filtrate was concentrated after filtration and dried in vacuo.
EXAMPLE 2 preparation of Hot Water extract of Saururus chinensis Baill leaf
White leaves were separated from only Saururi herba, distilled water was added thereto in an amount 10 times the weight of the raw material of Saurururus chinensis white leaves and mixed, and then extracted at a temperature of 90℃for 4.5 hours. The resulting extract was filtered through filter paper, and then the filtrate was concentrated under reduced pressure using a rotary evaporation dryer. The powdery saururus chinensis white leaf hot water extract is obtained through freeze-drying so as to remove the residual solvent in the solution after reduced pressure concentration.
Comparative example 2 preparation of Hot Water extract of Saururus chinensis green leaves
Green leaves were separated from only saururus chinensis, distilled water was added thereto in an amount 10 times by weight as much as the weight of the saururus chinensis green leaf raw material and mixed, and then extracted at a temperature of 90 ℃ for 4.5 hours. The resulting extract was filtered through filter paper, and then the filtrate was concentrated under reduced pressure using a rotary evaporation dryer. The powdery saururus chinensis green leaf hot water extract is obtained through freeze-drying, so as to remove the residual solvent in the solution after reduced pressure concentration.
Experimental example 1. Secondary metabolites of white leaf and green leaf extracts were confirmed and compared
Experiments were performed to confirm secondary metabolites of the saururus chinensis white leaf extract of example 1 and the saururus chinensis green leaf extract of comparative example 1 and analyze the contents thereof.
First, the saururus chinensis white leaf extract and saururus chinensis green leaf extract were dissolved in 50% methanol, respectively, so that the concentration thereof reached 1mg/mL. Then, filtration was performed with a 0.45 μmPVDF (Polyamide Fluoride) filter and analysis was performed with LC-QTOF-MS (liquid chromatography-quaterpole-time of flight mass spectrometry). The LC-QTOF-MS analysis results are shown in fig. 1a to 1b.
As shown in fig. 1a and 1b, it was confirmed that the content of the white leaf of saururus chinensis appeared to be higher than the content of 4 peaks (peaks) in the green leaf. The MS fragmentation pattern was analyzed for 4 peaks (fragmentation pattern) and all the results were predicted to be flavonol glycosides (flavonoid glycoside).
Among the 4 flavonol glycosides, MS fragmentation patterns for 2 peaks mainly showing differences are shown in fig. 2a to 2b and fig. 3a to 3b. In addition, the MS fragment deduplication (duplicate) results of the two peaks are shown in table 1.
[ Table 1 ]
As shown in fig. 2a, 2b, 3a, 3b and table 1,2 major peaks in white leaves that appear to be more than in green leaves were predicted to be quercetin and afugin. The corresponding ingredients were confirmed after measurement with standards of quercetin and afugin, and the content in white leaves was confirmed to be higher than that in green leaves.
Experimental example 2 evaluation of cytotoxicity against B16 melanoma cells
The white leaf extract and green leaf extract of Saururus chinensis and the cytotoxicity of the indicator component quercetin on B16 melanoma cells were evaluated.
Specifically, after inoculating melanoma cells into a 6-well plate for culturing for 24 hours, the cells were replaced with a new DMEM medium containing α -MSH and 10% bovine serum, and then each sample was treated at a different concentration and cultured for 72 hours. Replacement of cells which have been terminated in culture 2.5mg/ml of MTT (3- (4, 5-dimethylazol-2-yl) -2,5-diphenyltetrazolium bromide) solution was diluted 10-fold with the medium and allowed to react. Then, the supernatant was removed, 2ml of DMSO (dimethyl sulfoxide) was added to dissolve the MTT-formazan crystals thus formed, and the absorbance was measured at 570nm by ELISA reader. Samples treated with alpha-MSH were used as control. Unlike the control group, cytotoxicity was expressed as a percentage of absorbance, and the results are shown in tables 2,3, 4 and 5.
[ Table 2 ]
[ Table 3 ]
As shown in table 2, table 3, fig. 4 and fig. 5, the saururus chinensis white leaf extract of example 1, the saururus chinensis green leaf extract of comparative example 1, the saururus chinensis white leaf hot water extract of example 2 and the saururus chinensis green leaf hot water extract of comparative example 2 showed no toxicity up to a concentration of 0.1 μg/ml in melanoma cells. Furthermore, the index component quercetin did not show toxicity up to a concentration of 100. Mu.g/ml.
Experimental example 3 evaluation of cytotoxicity on human immortalized keratinocytes (HaCaT)
Cytotoxicity of the index component african bean glycoside on human immortalized keratinocytes (HaCaT) was evaluated.
Specifically, human immortalized keratinocytes were inoculated into a 24-well plate and cultured for 24 hours, and then the medium of the cultured cells was replaced with serum-free medium, and the samples were treated at different concentrations and cultured for 24 hours. Cells that have been terminated in culture are replaced with a medium in which 2.5mg/ml of MTT solution is diluted 10-fold, and then allowed to react. Then, the supernatant was removed, 1ml of DMSO was added to dissolve the MTT-formazan crystals thus produced, and absorbance was measured at 570nm by ELISA reader. Untreated groups were used as control groups. Unlike the control group, cytotoxicity was expressed as a percentage of absorbance, and the results thereof are shown in table 4 and fig. 6.
[ Table 4 ]
As shown in Table 4 and FIG. 6, the index component, alfoside, did not show toxicity to human immortalized keratinocytes up to a concentration of 100. Mu.g/ml.
Experimental example 4 detection of whitening efficacy per unit cell inhibiting melanin biosynthesis in melanoma cells
Whitening efficacy of the saururus chinensis white leaf extract and the green leaf extract in inhibiting melanin biosynthesis per unit cell of the index component quercetin in melanoma cells was examined.
Specifically, after inoculating melanoma cells into 6-well plates for 24 hours of culture, the cells were replaced with cells containing alpha-MSH and 10%
The new DMEM medium of bovine serum was then treated with samples at different concentrations and incubated for 72 hours. After culturing for 72 hours, to detect the amount of melanin secreted into the medium, the medium was transferred to a 96-well plate, and then absorbance was measured at 450nm, thereby obtaining the amount of melanin secreted per unit cell. Then, cells of all media were washed with 2ml of PBS (phosphate-buffered saline), then treated with 0.25ml of 1N NaOH, and the cells were collected into 1.5ml tubes, thereby obtaining intracellular melanin. After the collected cell lysate was incubated at 60℃for 30 minutes, vortexing was performed, and absorbance at 450nm was measured after transfer to a 96-well plate, thereby obtaining the melanin production per unit cell. The control group was treated with α -MSH, and the positive control group was administered with arbutin (manufactured by HyundaiiBioland). The melanin biosynthesis results per unit cell are shown in tables 5, 6, 7 and 8, and the melanin secretion rate results per unit cell are shown in tables 7, 8, 9 and 10.
[ Table 5 ]
[ Table 6 ]
[ Table 7 ]
/>
[ Table 8 ]
As shown in tables 5 to 8 and fig. 7 to 10, the saururus chinensis white leaf ethanol extract and the green leaf ethanol extract reduced the melanin production rate and secretion rate per unit cell increased by α -MSH in a concentration-dependent manner. In particular, it was confirmed that the melaninogenesis and secretion-inhibiting effect per unit cell of the saururus chinensis leaf ethanol extract was superior to that of the green leaf ethanol extract. In addition, the indicator component quercetin is considered to exhibit whitening efficacy by inhibiting melanin production rate and secretion rate per unit cell, due to inhibition of melanin biosynthesis and secretion.
Further, it was confirmed that the ethanol extracts of the white leaf and the green leaf of saururus chinensis were excellent in the efficiency of reducing the melanin production rate per unit cell in a concentration-dependent manner, as compared with the hot water extract.
Experimental example 5 detection of whitening efficacy to inhibit skin pigmentation by blocking melanosomes transferred to epidermis
The skin pigmentation inhibiting effect caused by melanosome inhibition of the index component afugin was examined.
Melanosome transfer (melanosome transfer) is the process of transferring melanosomes from melanocytes (melanocytes) to the epidermis, and there are various mechanisms, well known among which phagocytosis (phagocytosis) is caused by PAR-2 (protease-activated receptor-2) as a key mediator. The whitening efficacy of inhibiting melanosomes transferred to the epidermis and thus skin pigmentation by blocking the expression of such PAR-2 was confirmed (Eun-Jung CHOI at al, macelignan Inhibits Melanosome Transfer Mediated by Protease-Activated Receptor-2in Keratinocytes,Biol.Pharm.Bull.34 (5) 748-754, 2011).
Inoculating human immortalized keratinocytes (HaCaT) in a 60 mm dish (corning), culturing for 24 hr, washing with HBSS (Hank's Buffered Salt Solution) 2 times, adding HBSS and irradiating UVB 30mJ/cm 2 . Then, each sample was treated at a different concentration and replaced with serum-free mediumAfter 8 hours of culture, the cells were collected to QIAzol TM Lysis Reagent (Qiagen) and RNA was isolated according to the method of manufacturing company. After the isolated RNA was quantified, cDNA was synthesized using 1. Mu.g of the RNA, and real-time fluorescent quantitative PCR was performed. PAR-2, beta-action primers for PCR were synthesized by cosmagenetech company (Korea) and the sequences of the primers are shown in Table 9. The UVB-irradiated group was used as a control group, and nicotinamide (manufactured by Sigma) was used as a positive control group. The PAR-2 expression results are shown in Table 10 and FIG. 11.
[ Table 9 ]
[ Table 10 ]
As shown in table 10 and fig. 11, inhibition of PAR-2 expression due to UVB increase was confirmed after treatment of human immortalized keratinocytes with the index component african. Thus, it was confirmed that the index ingredient afugin exhibits a whitening effect of inhibiting skin pigmentation by inhibiting melanosomes transferred to the epidermis.
SEQUENCE LISTING
<110> Ke Si Mei poetry corporation
<120> a composition for skin whitening comprising Saururi herba white leaf extract
<130> PX068311PCT
<150> KR 10-2021-0090492
<151> 2021-07-09
<160> 4
<170> PatentIn version 3.2
<210> 1
<211> 20
<212> DNA
<213> Artificial work
<220>
<223> PAR-2 Forward primer
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<210> 2
<211> 20
<212> DNA
<213> Artificial work
<220>
<223> PAR-2 reverse primer
<400> 2
ccagtgagga cagatgcaga 20
<210> 3
<211> 19
<212> DNA
<213> Artificial work
<220>
<223> beta-action Forward primer
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Claims (12)
1. A cosmetic composition for skin whitening comprises Saururi herba white leaf extract as active ingredient.
2. The cosmetic composition for skin whitening according to claim 1, wherein,
the extraction solvent of the extract is water, C1-C4 alcohol, or mixture thereof.
3. The cosmetic composition for skin whitening according to claim 1, wherein,
the extraction solvent of the extract is 10% (v/v) to 90% (v/v) ethanol.
4. The cosmetic composition for skin whitening according to claim 1, wherein,
the saururus chinensis white leaf extract has better skin whitening effect than saururus chinensis green leaf extract.
5. The cosmetic composition for skin whitening according to claim 1, wherein,
the extract comprises any one or more of quercetin and afugin.
6. The cosmetic composition for skin whitening according to claim 1, wherein,
it inhibits melanin production or secretion.
7. The cosmetic composition for skin whitening according to claim 1, wherein,
which inhibits the expression of the protease activated receptor-2 involved in melanosome transfer.
8. A pharmaceutical composition for preventing or treating a melanin hyper-pigmentation disease, comprising saururus chinensis white leaf extract as an active ingredient.
9. The pharmaceutical composition for preventing or treating melanin pigmentation disorders according to claim 8, wherein,
the melanin hyper-pigmentation disease is any one selected from freckle, chloasma, liver spots, senile plaque, sunlight blackness, boitz-Yeger syndrome, gestational chloasma, hyperpigmentation after using medicine and pigmentation after inflammation.
10. A health functional food composition for preventing or improving melanin hyper-pigmentation diseases comprises Saururus chinensis Baill leaf extract as active ingredient.
11. A method of preventing or treating a melanin hyper-pigmentation disorder comprising administering to a subject in need thereof a composition comprising an effective amount of saururus chinensis white leaf extract.
12. Use of saururus chinensis white leaf extract for the preparation of a medicament for the prevention or treatment of a melanin hyper-pigmentation disorder.
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|---|---|---|---|
| KR10-2021-0090492 | 2021-07-09 | ||
| KR1020210090492A KR102560191B1 (en) | 2021-07-09 | 2021-07-09 | Composition for skin whitening comprising Saururus chinensis white leaf extract |
| PCT/KR2022/009314 WO2023282528A1 (en) | 2021-07-09 | 2022-06-29 | Skin whitening composition comprising extract from white leaf of saururus chinensis (lour.) baill |
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| CN117561050A true CN117561050A (en) | 2024-02-13 |
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| CN202280045631.8A Pending CN117561050A (en) | 2021-07-09 | 2022-06-29 | A composition comprising Saururus chinensis Baill leaf extract for whitening skin |
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| KR (1) | KR102560191B1 (en) |
| CN (1) | CN117561050A (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4530832B2 (en) | 2004-12-17 | 2010-08-25 | 株式会社資生堂 | Skin external preparation for whitening, whitening agent, whitening method, and method for producing skin external preparation for whitening |
| KR100865071B1 (en) | 2007-04-27 | 2008-10-23 | 영남대학교 산학협력단 | Composition comprising the saururus chinensis extract or compound isolated therefrom for skin whitening |
| KR20100086728A (en) * | 2009-01-23 | 2010-08-02 | (주)아모레퍼시픽 | Cosmetic composition for skin whitening |
| KR20150023601A (en) * | 2015-02-03 | 2015-03-05 | (주)아모레퍼시픽 | Composition Containing Extracts of Saururus chinensis |
| KR101993699B1 (en) * | 2017-04-19 | 2019-06-28 | 주식회사 진켐 | Composition for improving skin |
| KR102202554B1 (en) * | 2019-07-23 | 2021-01-14 | 주식회사 현대바이오랜드 | Skin whitening cosmetic composition with the extract of Saururus chinensis leaf, the extract of Grape seed, the extract of Portulaca oleracea and rice fermented product |
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2021
- 2021-07-09 KR KR1020210090492A patent/KR102560191B1/en active IP Right Grant
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- 2022-06-29 CN CN202280045631.8A patent/CN117561050A/en active Pending
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| WO2023282528A1 (en) | 2023-01-12 |
| KR20230009735A (en) | 2023-01-17 |
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