KR20160146406A - Cosmetic composition comprising sea cucumber steaming juice or a fraction thereof as functional component - Google Patents

Abstract

The present invention relates to a cosmetic composition which contains boiled sea cucumber extract or a fraction thereof as an active ingredient and has skin-whitening and wrinkle-ameliorating effects.

Description

A cosmetic composition comprising a sea cucumber juice liquid or a fraction thereof as an active ingredient,

The present invention relates to a cosmetic composition comprising a sea cucumber juice extract or a fraction thereof as an active ingredient and having an effect of improving whitening and wrinkles.

Sea cucumber is a collective term for sea cucumbers belonging to the echinoderms of echinoderms. In food science, 90.5% of the sea cucumber is composed of water, 3.2% of other proteins, 0.2% of fat and 3.5% of minerals. The proteins contained in the sea cucumbers are of high quality. The minerals contain calcium and phosphorus in an appropriate ratio, and chondroitin sulfate contains many chondroitin sulfate. Also, free amino acids, sweetness of alanine and glutamic acid are the most distributed, and the highest content of saponin known as functional ingredient of ginseng is also called ginseng of the sea. The main components of dried sea cucumber ginseng are glycoprotein and chondroitin. Studies on mucopolysaccharide and chondroitin sulfate structure have been carried out.

The sea cucumber juice is a by-product which is produced in large quantities during the process of producing dried sea cucumber by heating and drying the sea cucumber. Most of the sea cucumber juice is economically lost and environmental pollution problem is raised. In addition, although the sea cucumber juice contains a large amount of functional ingredients such as polyphenols, proteins and glycogens, the research on conventional sea cucumbers has mostly resulted in the development of food using sea cucumbers themselves, Research on extracting functional materials from liquids or developing functional products is very limited.

Domestic patent registration 1513892 (April 15, 2015) Domestic patent registration No. 1493158 (Feb. 2, 2015)

In accordance with the present invention, it was confirmed that the sea cucumber juice was fractionated and purified according to the molecular weight to separate the glycoprotein, and the fractions containing the glycoprotein exhibited whitening activity and / or wrinkle improving effect. Accordingly, the present invention provides a sea cucumber juice, which is a by-product produced in the processing of sea cucumber, as a functional material for cosmetics.

 Accordingly, an object of the present invention is to provide a cosmetic composition comprising a sea cucumber juice extract or a fraction thereof as an active ingredient.

Another object of the present invention is to provide a cosmetic comprising the cosmetic composition.

Another object of the present invention is to provide a process for producing the cosmetic composition.

For example, the present invention relates to a cosmetic composition comprising a sea cucumber juice extract or a fraction thereof as an active ingredient.

In one embodiment, the cosmetic composition may be used for whitening or wrinkle-improving purposes.

In another embodiment, the fraction of the sea cucumber juice may be a fraction comprising a glycoprotein having a molecular weight of at least 50 kDa, a concentrate of the fraction, or a lyophilizate of the fraction.

As another example, the present invention provides a cosmetic comprising the above cosmetic composition.

In one embodiment, the cosmetic may comprise one or more formulations selected from the group consisting of a lotion, a milky lotion, a cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation, a powder, Can be.

As another example, the present invention provides a method for preparing a sea cucumber, comprising obtaining a sea cucumber juice by dipping the sea cucumber at 80 to 90 캜, filtering the sea cucumber juice under reduced pressure, and separating the fraction containing the glycoprotein having a molecular weight of 50 kDa or more. And a method for producing a cosmetic composition comprising the sea cucumber juice liquid fraction as an active ingredient.

Hereinafter, the present invention will be described in more detail.

In the present invention, the juvenile juvenile juice was fractionated and purified by molecular size to obtain a juvenile juvenile fraction having a molecular weight of 50 kDa or more or 50 kDa or less, and the fraction exhibited whitening activity and / or wrinkle-reducing activity. Accordingly, there is a need to provide cosmetic compositions and cosmetics comprising the sea cucumber juice extract or fractions thereof.

The cosmetic composition of the present invention may contain 0.01 to 10% of the sea cucumber juice extract or its fraction based on the total weight of the composition and can be suitably adjusted according to the use of the composition, the application form, the intended use and the desired effect. However, when the amount is less than the above range, substantial whitening or wrinkle-reducing effect can not be obtained, and if it is more than the above range, the stability of the formulation may deteriorate.

 In the present invention, the sea cucumber juice is a liquid by-product generated after boiling the raw sea cucumber. For example, in the representative process of obtaining the sea cucumber juice of the present invention, salt and impurities can be removed by first cutting the collected sea cucumber, removing the viscera, and then washing. At this time, the collection of sea cucumbers is usually done by diving, and it is good to prevent the sea cucumbers from being scratched or deteriorated in commerciality. In addition, the use of the sea cucumber collected on the same day is preferable because the freshness of the sea cucumber or sea cucumber juice can be adjusted and the yield (severity rate) can be increased. The cuts of the sea cucumbers are 2 to 3 cm in length, and they are cleanly cut in a straight line. The smaller the cucumber is, the more clean the sea cucumbers when boiled, and the less the flocculation liquid is. The sea cucumbers can be cleaned with fresh water after they are completely removed, and they can be boiled in boiling water (eg, about 80 to 90 ° C) . After that, all of the sea cucumbers are removed and the remaining liquid phase can be obtained as a sea cucumber juice. The above process is merely an example of realizing the present invention, and it is obvious that a person skilled in the art can appropriately change and modify the sea cucumber juice.

In the present invention, the fraction of the sea cucumber juice liquid refers to a fraction obtained by desalination of seaweed juice and its molecular size. Wherein the sea cucumber juiced liquid fraction is obtained by dipping a sea cucumber in a sea cucumber at 80 to 90 ° C to obtain a sea cucumber juice liquid, filtering the sea cucumber juice under reduced pressure, and separating a fraction containing a glycoprotein having a molecular weight of 50 kDa or more ≪ / RTI >

In the present invention, a typical process for obtaining a fraction of juvenile juvenile juice is described in more detail. First, the juvenile juvenile juice is thawed in a refrigerated state (for example, about 4 to 6 ° C) and then filtered under reduced pressure to remove impurities. In order to obtain the liquid fraction of the sea cucumber juice according to the molecular weight, the juice liquid having a molecular weight of 50 kDa or more and 50 kDa or less can be separated from the juvenile juice by using a desalting concentrator. Alternatively, it may be fractionated using a conventional membrane having a specific pore size, or may be carried out through liquid chromatography. The obtained fractions can be concentrated with a filter having an appropriate pore size or concentrated until a desired glycoprotein concentration is obtained by a conventional method such as concentration under reduced pressure. However, the above process is merely an example of realizing the present invention, and it is obvious that a person skilled in the art can appropriately change and modify the seaweed juice fraction.

The sea cucumber juice liquid fraction can be dried or lyophilized and dissolved in a solvent such as purified water.

In the present invention, the term " comprising as an active ingredient " means a cosmetic composition containing an effective amount of a component exhibiting skin improving effect such as wrinkle improving and / or whitening effect.

In a specific example herein, the glycoprotein was identified by Alcian blue staining and Lectin blotting on the juvenile juvenile fraction having a molecular weight of 50 kDa or greater or 50 kDa or less, and the intracellular tyrosinase (tyrosinase) activity and elastase activity (activity). In addition, an essence formulation containing a sea cucumber juice liquid fraction was prepared, and it was confirmed that it was excellent in the effect of improving wrinkles without irritating the skin by applying it to the clinic. Therefore, the cosmetic composition of the present invention can be applied for improving wrinkles and / or whitening.

Wrinkle improvement includes maintaining or enhancing the ability to associate with the wrinkles and elasticity of the skin. In this regard, elastase activity is known to increase skin aging and reduce skin elasticity, so that an activity inhibitor of elastase can be used to inhibit skin aging and improve wrinkles.

The whitening function includes a function of preventing the deposition of excessive melanin pigment on the skin or thinning the color of the melanin pigment previously deposited to suppress the generation of spots and freckles, thereby helping to whiten the skin. Melanin pigments in human skin are a major mechanism to protect against UV damage, but they cause undesirable problems such as melasma, freckles, senile lentigines, or abnormal pigmentation such as excessive pigmentation , Tyrosinase, causes melanin biosynthesis in human skin. Thus, tyrosinase inhibitors are known to be important materials used in cosmetics for whitening effects.

The cosmetic composition of the present invention is a functional cosmetic composition for whitening or wrinkle improvement, which can be applied to the skin, transdermally or subcutaneously, depending on the route of administration, and is preferably a composition that can be administered externally or transdermally, more preferably, Lt; / RTI >

The cosmetic composition may be percutaneously applied to the skin, scalp or hair, and may be applied percutaneously to the skin, scalp or hair, and may be applied to skin, scalp or hair such as foundation cosmetics, makeup cosmetics, body products (body cleanser, body lotion, etc.), shaving products, hair products (shampoo, Means a composition which can be used in the production of all cosmetic products and may be formulated in the form of a warning agent, a spray, a suspension, an emulsion, a cream, a gel or a foam, but there is no particular limitation on its form. For example, it may have formulations such as lotion, milky lotion, cream, essence, cosmetic lotion, spray, gel, pack, sunscreen, makeup base, foundation, powder, makeup remover and detergent.

The cosmetic composition may contain, in addition to the active ingredient, all kinds of components that can be used for ordinary commercialization or formulation such as a perfume, a pigment, a bactericide, an antioxidant, an antiseptic, a moisturizer, a thickening agent, an excipient, a diluent, And the kind and content thereof can be appropriately adjusted depending on the use of the final product and the intended use.

In addition, the cosmetic composition may contain a solvent ordinarily included in the application form, and examples thereof include ethanol, glycerin, butylene glycol, propylene glycol, polyethylene glycol, 1,2,4-butanetriol, sorbitol ester , 1,2,6-hexanetriol, benzyl alcohol, isopropanol, butanediol, diethylene glycol monoethyl ether, dimethylisosorbide, N-methyl-2-pyrrolidone, propylene carbonate, glycereth- At least one member selected from the group consisting of isosetyl myristate, isocetyl octanoate, octyldodecyl myristate, octyldodecanol, isostearyl diacetate, cetyl octanoate, neopentyl glycol dicaprate, etc. . ≪ / RTI > Those skilled in the art will appreciate that the type and amount of solvent may be suitably selected according to the characteristics of the product.

In addition, the cosmetic composition may contain various substances for enhancing transdermal permeation during transdermal administration. For example, there may be mentioned, for example, laurolactam derivatives and oleic acid, ester derivatives of monooleate derivatives, aderapalene, tritinoin, retinaldehyde, tazotrotin, salicylic acid, azelaic acid, glycolic acid, ethoxydiglycol, 80, lecithin olganogel, and the like. In order to impart additional functions, the cosmetic composition of the present invention may contain a co-surfactant, a surfactant, an anti-dandruff agent, a keratin softener, a blood circulation accelerator, a cytostatic agent, , A moisturizer, an antioxidant, a pH adjuster, a purified water, and the like, and may contain additives such as suitable fragrances, pigments, preservatives and excipients, depending on the application.

The present invention provides functional cosmetics having wrinkle improving and whitening effect without skin cell toxicity using a sea cucumber juice liquid or a composition thereof, thereby reducing the cost of processing by-products produced in sea cucumber culture, reducing environmental pollution and creating new industrial demand Can make an important contribution.

FIG. 1 shows a manufacturing process chart according to the method of manufacturing a sea cucumber juice liquid fraction (each having a molecular weight of 50 kDa or more and 50 kDa or less) according to the present invention.
2 is a graph showing the results of confirming N-glycan by treating lectin (RCA-120) and confirming the staining with Alcian blue (AB) in order to confirm the glycoprotein in the sea cucumber juice fraction (not less than 50 kDa and not more than 50 kDa) And the result of checking 0-glycan is shown. M is a marker, B is a fraction of 50 kDa or less, A is a fraction of 50 kDa or more, C is a control mucin (Bovine submaxillary glands), AB is an Alcian blue stain, and RCA-120 is a lectin derived from Ricinus communis (β-Gal).
FIG. 3 shows the cytotoxic effect of various concentrations (0.1, 1, 2, 5 mg / ml) of the sea cucumber juice fraction (over 50 kDa, 50 kDa or less) according to the present invention. PC represents a positive control (100% Ethanol).
FIG. 4 is a graph showing the amino acid analysis values of the sea cucumber juice, sea cucumber and sea cucumber tablets (50 kDa or more) according to the present invention. The sea cucumber juice is one of its own freeze-dried powder, the sea cucumber is a dried sea cucumber powder, and the sea cucumber purified water is a fraction of 50 kDa or more.
FIG. 5 is a graph showing the tyrosinase inhibitory effect of the sea cucumber juice fraction (50 kDa or more, 50 kDa or less) according to the present invention. Ascorbic acid and arbutin were used as positive control.
FIG. 6 is a graph showing the wrinkle-reducing effect of the sea cucumber juice liquid fraction (50 kDa or more, 50 kDa or less) according to the present invention. Vitamin C (ascorbic acid) was used as a positive control.
FIG. 7 is a graph showing the degree of improvement of eye wrinkles after use of the cosmetic composition containing the sea cucumber juice fraction (over 50 kDa) according to the present invention.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  1. Sea cucumbers Homemade  Produce

The size of the sea cucumber was 200g or more and the domestic sea cucumber collected on the day was used. When the sea cucumbers were removed, they were incised up to 3 cm from the anus to the center of the back. The sea cucumbers were cleaned with fresh water to remove primary salts and impurities. The sea cucumbers that had been washed with fresh water were boiled in fresh water. For this purpose, about 50 kg of sea cucumbers with internal organs removed from the boiling water were poured into the pot bottom with a spatula or the like. When the sea cucumber was boiled, it was poured with a spatula and stirred to keep it from burning on the floor. When the sea cucumber started to breathe more and began to shrink more than the first one, it added an additional sea cucumber with the intestines removed. The above procedure was repeated repeatedly while boiling the sea cucumber. When the sea cucumbers swell like a ball, they rescued and continued to receive sea cucumber. The heating temperature was maintained at 80 to 90 占 폚.

After the above steps, all the solids (sea cucumbers) were removed and the liquid phase was the sea cucumber juice. The juvenile juice was frozen at -20 ° C after collection and stored until the juice fraction was prepared.

Example  2. Sea cucumbers Dipping solution Fraction  Preparation and identification of glycoproteins

2-1. Sea cucumber Dipping solution Fraction  Separation and purification

The sea cucumber juice prepared in Example 1 was thawed in a refrigerated state (4 to 6 ° C), and subjected to a first vacuum filtration using a SUPRAcap depth filter K500P grade. Using a SUPRAcap depth filter 10 inch capsule, To remove impurities. Next, the fraction of the mature liquid having a molecular weight of 50 kDa or more and 50 kDa or less was separated by using a desalting and concentration apparatus (UF SYSTEM, manufacturer, Vitis) and was used as a cosmetic composition by freeze-drying. FIG. 1 is a diagram showing a method of separating and purifying a sea cucumber juice liquid fraction.

2-2. Sea cucumber Dipping solution Fraction  Trypsin treatment

Tryptine (Porcine, Sigma-Aldrich) was added in an amount of 0.2% to the fraction of the mature food liquid having a molecular weight of 50 kDa or more and 50 kDa or less and cultured at 37 ° C for 3 hours.

2-3. Identification of glycoproteins

(1) Dyeing of Alcian blue

20 [mu] l of a 4X sample buffer was added to each 60 [mu] i of each of the samples of the fraction of the matured liquor of 50 kDa or more or 50 kDa or less prepared in Example 2-1 and left for 10 minutes at 100 [deg.] C. SDS-PAGE was performed. SDS-PAGE gels were washed three times for 5 minutes each with tertiary distilled water. The O-glycan glycoprotein was stained with Alcian blue (AB) staining solution (0.1% Alcian blue, 3% acetic acid) For 20 minutes and then dyed in tertiary distilled water to confirm the band.

As a result, as shown in Fig. 2, the presence of the glycoprotein in the sea cucumber juice could be confirmed through Alcian blue staining. Specifically, it was difficult to identify the band below 50 kDa. In the case of 50 kDa or more, two band - type proteins were identified in all the cyanoblue staining. As a result, the presence of O-glycan glycoprotein in the sea cucumber juice could be confirmed.

(2) Lectin blotting

Next, for analysis of N-glycan glycoprotein, β-Gal saccharide of N-glycan was confirmed through a lectin antigen-antibody reaction using RCA120. For this, the proteins of SDS-PAGE gels were transferred to PVDF membranes. Membranes were washed twice in TBST (Tris-buffered saline, 0.1% Tween 20) solution for 5 min each. The TBST solution was reacted with RCA-120 diluted to 1000: 1 for 1 hour, and then washed three times for 10 minutes in the TBST solution. Streptavidin-HRP (Thermo) was reacted with the TBST solution diluted 1: 5000 for 1 hour and then washed 3 times for 10 minutes in the TBST solution. The reacted membrane was reacted in a stable peroxide substrate buffer of DAP substrate kit (Thermo) until the band was clearly visible in the 1X diluted DAP solution, followed by washing with distilled water to confirm the band.

As a result, as shown in FIG. 2, a band of about 52 kDa of the fermented juice after trypsin treatment was confirmed as RCA120 lectin (FIG. 2).

Example  3. Sea cucumbers Dipping solution Fraction  Stability evaluation

B16-F10 melanocytes (manufacturer ATCC LOT. 60508145) were used to evaluate the stability of the sea cucumber juice liquid fraction. 100 μl of cell suspension (5000 cells / well) was dispensed into a 96-well plate and incubated for 24 hours at 37 ° C and 5% CO ₂ The cells were pre-incubated and attached to well plates. In the blank well, 100 μl of cell-free medium DMEM (Dulbecco's Modified Eagle Medium, manufacturer ATCC) alone was dispensed.

10 μl of the test sample (fraction of 50 kDa or more of the sea cucumber juice extract and fraction of less than 50 kDa) was treated with 10 μl of each of the wells and the negative control was treated with 10 μl of the medium. CO ₂ conditions. 10 μl of CCK-8 solution was added to each well, incubated for 1 hour, and absorbance was measured at 450 nm. The blank was measured with the sample in the absence of cells.

Survival rate (%) = (A _sample -A _b ) / (A _c -A _b ) × 100

A _sample : sample absorbance

A _b : blank absorbance

A _c : Negative control absorbance (measured with distilled water in cell)

As shown in FIG. 3, the cell viability at 0.1, 1, 2, and 5 mg / ml concentrations of each of the molecular size fractions (50 kDa or more and 50 kDa or less) The stability of the sea cucumber juice liquid fraction was confirmed. In contrast, the positive control (100% Ethanol) showed cell viability below 10%.

Example  4. Sea cucumbers Dipping solution Fraction  And amino acids in sea cucumbers

Sea cucumber juice, sea cucumber, and sea cucumber were added to the test tube with about 2-5g of the sample. The sea cucumber juice samples were prepared by lyophilization, and the sea cucumber samples were analyzed by using the fractions over 50 kDa. 6N hydrochloric acid (10 mL) was added to the flask, and the flask was filled with nitrogen gas for 1 minute. Then, the flask was closed and hydrolyzed at 110 DEG C for 22 hours. The hydrolyzed specimen was filtered by DW in a 50 mL flask and diluted 10-fold. This was filtered with a 0.45 μm syringe filter.

AccQ-Tag Kit. In the derivatization step, 70 μl of borate buffer, 10 μl of sample and standard, and 20 μl of the prepared AccQ-Tag Reagent were added to the PCR tube and reacted in a heating block at 55 ° C. for 10 minutes. After cooling sufficiently, it was transferred to an LC vial and analyzed by HPLC. The analysis conditions are shown in Table 1 and Table 2 below

Column AccQ-Tag column 3.9x150nm Detector fluorescence lex = 250 nm len = 395 nm Flow rate 1 mL / min

Mobile phase (A: acetate phosphate buffer B: 60% ACN) Time A (%) B (%) 0 100 0 0.5 98 2 15 93 7 19 90 10 32 67 33 33 67 33 34 0 100 37 0 100 38 100 0 45 100 0

As shown in FIG. 4, the total amino acid content of the sea cucumber juice was 16.59 g / 100 g and the sea cucumber was 54.21 g / 100 g. It can be seen that the sea cucumber juice fraction (over 50 kDa) contains about 4.21 g / 100 g of sea cucumber amino acid.

Example  5. Sea cucumbers Dipping solution Fraction  Whitening efficacy verification

5-1. Intracellular tyrosinase ( tyrosinase ) Inhibition activity measurement

B16-F10 melanocytes (manufacturer ATCC LOT. 60508145) were used to measure intracellular tyrosinase inhibitory activity of the sea cucumber juvenile fraction. Cells were pre-incubated for 24 h at 37 ° C and 5% CO ₂ in a 24-well plate (B16-F10, manufacturer ATCC LOT.60508145) at a rate of 500 μl / .

50 ㎕ or less of 50 kDa fractions and 50 ㎕ of vitamin C and arbutin as positive control were added to each well and incubated at 37 캜 and 5% CO ₂ for 24 hours. The pellet obtained by trypsinization was dissolved in 0.5 ml of 1% triton X-100 PBS, mixed with 0.5 ml of 0.2% L-DOPA 0.1 M sodium phosphate buffer, incubated at 37 ° C for 2 hours and absorbance at 490 nm Respectively.

Inhibition rate (%) = (absorbance of control group - absorbance of addition group) / absorbance of control group * 100

As a result, as shown in FIG. 5, tyrosinase activity was measured at 0.1, 1, 2, and 5 mg / ml concentrations of the sea cucumber matured liquid according to their molecular sizes (50 kDa or more and 50 kDa or less) The fractions over 50 kDa showed higher tyrosinase inhibitory activity than the fractions below 50 kDa and showed high whitening activity and activity in a concentration - dependent manner. Vitamin C (ascorbic acid) and arbutin were used as positive controls.

5-2. Elastase ( Elastase ) Inhibitory activity assay

In order to measure the inhibitory activity of Elastase, 10 쨉 l of each fraction was subjected to 50 쨉 l of a 50 kDa or higher fraction and 50 D or less of a fraction of 50 kDa or less, and a positive control group of vitamin C and arbutin. N-succinyl- (L-Ala) ₃ - p- nitroanilide (0.5 mg / mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) as a substrate was added to 50 μl of porcine pancreas elastase (10 μg / mL) was added and the reaction was carried out for 20 minutes. The inhibitory activity of elastase was expressed by the absorbance reduction rate of the sample solution and the non-added sample.

As a result, as shown in FIG. 6, when the fractions of the molecular sizes derived from the sea cucumber juice (50 kDa or more and 50 kDa or less) were measured at 0.1, 1, 5 and 10 mg / ml, the fractions of 50 kDa or more The activity of inhibition of elastase inhibition was higher than that of fractions. The fractions over 50 kDa showed a concentration - dependent activity. Positive control vitamin C (ascorbic acid) was used.

Example  6. Sea cucumbers Dipping solution The fraction  Clinical trial of cosmetics containing active ingredient

6-1. Cosmetic formulation of essence formulation

A cosmetic product of an essence formulation containing the sea cucumber juice extract fraction of the present invention as an active ingredient was prepared. The specific composition is shown in Table 3.

Raw material name content Purified water to 100g Seaweed juice fraction 50kDa or more 0.01 Herbal extract 0.01 Hexanediol 2g glycerin 3g Centipede extract 0.1 g Niacinamide 2g Marine Collagen 0.5 g Hyaluronic acid 2g Adenosine 0.5 g Allantoin 0.1 g Panthenol 0.5 g Solvite 80 1g Xanthan gum 0.05 g Hydroxyethylcellulose 0.05 g

6-2. Skin irritation test

The subjects' basic information is shown in Table 4 below

Total number of subjects 30 people gender M: 2 people W: 28 Average age 41 years old

A 24-hour patch test was carried out using an essence-type cosmetic product containing the sea cucumber juice extract fraction prepared in Example 6-1 as an active ingredient. The patches were applied to the skin for 24 hours, and the dermatologist judged whether or not primary skin irritation had occurred respectively at 30 minutes, 24 hours (i.e. 48 hours after patch application) and 48 hours (i.e. 72 hours after patch application) Respectively. The skin response was determined according to the ICDRG criteria and the US Cosmetics Association (PCPC) guidelines, and the irritation index was calculated using the skin response scores of each subject. The evaluation criteria for the 24-hour adhesive test are shown in Table 5 below. As a result, as shown in Table 6, a skin irritation index of 0 was obtained and the cosmetic formulation was judged to be a non-irritant substance.

score Criteria 0(-) No signs of inflammation, normal skin
(No signs of inflammation, normal skin) 0.5 (±) Uncertain or faint reaction
(Doubtful or slight reaction) 1 (+) Slight erythema 2 (++) Common erythema with or without partial swelling or palsy
(Moderate erythema with or without partial edema or papules) 3 (+++) Normal erythema with diffuse edema
(Moderate erythema with diffuse edema) 4 (++++) Strong erythema with blisters and diffuse edema
Intense erythema with diffuse edema with vesicles

Subject
ID The cosmetic formulation of Example 6-1 BLANK 24h * Reaction 48h 72h 24h * Reaction 48h 72h 1720 0 0 0 0 0 0 1721 0 0 0 0 0 0 1293 0 0 0 0 0 0 1722 0 0 0 0 0.5 0 1018 0 0 0 0 0 0 1349 0 0 0 0 0 0 1724 0 0 0 0 0 0 1526 0 0 0 0 0 0 222 0 0 0 0 0 0 904 0 0 0 0 0 0 1713 0 0 0 0 0 0 1725 0 0 0 0 0 0 1430 0 0 0 0 0 0 1712 0 0 0 0 0 0 1719 0 0 0 0 0 0 1726 0 0 0 0 0 0 1385 0 0 0 0 0 0 674 0 0 0 0 0 0 1072 0 0 0 0 0 0 1482 0 0 0 0 0 0 157 0 0 0 0 0 0 1127 0 0 0 0 0 0 1684 0 0 0 0 0 0 856 0 0 0 0 0 0 1686 0 0 0 0 0 0 892 0 0 0 0 0 0 1309 0 0 0 0 0 0 940 0 0 0 0 0 0 1061 0 0 0 0 0 0 624 0 0 0 0 0 0 Stimulation index 0 0.006 Judgment No stimulation -

In the above table, "24h *" is the reading after 30 minutes from the patch removal, and "48h "and" 72h " represents the reading after 48 hours and 72 hours after the patch application. Blank Represents the data that has not been sampled.

6-3. Wrinkle improvement efficacy evaluation experiment

A cosmetic formulation containing the sea cucumber juice extract fraction prepared in Example 6-1 as an active ingredient was used for a total of 22 subjects and a total of 4 weeks of the experiment was performed to evaluate the effect of improving the eye wrinkles. Twenty-two subjects completed the test without dropouts.

(1) Equipment used

ANTERA 3D ^TM (Miravex, Ireland): Antera 3D ^TM Is a device that can measure the state of epidermis and dermis by using multi wavelength LED light source. It is possible to reconstruct 2-D and 3-D images using built-in programs.

(2) Test procedure

The test procedure was divided into two weeks and four weeks before use, and the first test before use prohibited the use of the base product 12 hours before the visit. After washing with a standard cleanser provided by the tester, it was patted with a paper towel to remove moisture, and then stabilized under constant temperature and humidity conditions (20 to 24 ° C, 40 to 60% RH) for 30 minutes. An image measurement using Antera 3D was performed. The Antera 3D imaging zone was set up in a square area of 6 cm x 6 cm including the orbits and both eyes. After the 2nd and 4th weeks of use, the device was evaluated at the same site in the same manner as the first visit.

(3) Evaluation and analysis methods

Evaluation of eye wrinkle improvement using Antera 3D: Statistical significance between measured eye wrinkles before product use, 2 weeks after use, and 4 weeks after use was analyzed and verified using Wilcoxon sign rank test.

(4) Test results

As shown in FIG. 7, Table 8 and Table 9, the rate of improvement of the wrinkles of the eyes at the test site was found to be about 0.96% in both the left and right eyes after 2 weeks of use and about 2.21% and about 1.45% , And it was confirmed that after two weeks from the use of the product, the improvement of the eye wrinkles was statistically significant as compared with that before use. No adverse events were observed in all subjects during the 4-week trial period. In conclusion, statistically significant improvement of eye wrinkles was observed from the 2nd week of use.

The subjects' basic information is as follows

all Number of subjects 22 people gender M: 0 people W: 22 people Average age 48 years Age distribution 30s 4 people 40s 8 people 50s 10 people

Antera 3D measurement result: Indentation index (Wrinkles: small) Subject
ID Left eye Right eye Before use use after 2 weeks use Four weeks Before use use after 2 weeks use Four weeks 349 69.565 68.950 68.602 68.609 67.899 67.459 1645 70.177 68.855 68.044 69.407 67.718 67.459 1430 66.630 66.627 66.152 67.718 67.661 67.512 1628 67.371 67.290 65.901 68.408 68.368 68.327 1472 71.070 70.995 69.179 69.498 68.920 68.894 1630 68.693 67.888 66.978 68.097 68.163 67.117 1670 69.961 68.927 68.871 69.287 68.891 68.782 1042 73.425 72.455 71.598 65.544 65.064 65.035 1074 73.829 72.637 72.317 69.678 68.836 68.740 1671 71.435 71.242 70.930 69.658 69.040 69.062 1297 68.407 68.422 68.325 68.537 68.463 68.447 1298 68.602 65.852 65.580 67.674 67.463 66.664 1304 73.871 73.766 72.832 70.384 69.238 69.415 1667 76.069 75.765 74.085 75.013 74.788 74.778 1541 72.585 72.336 71.017 72.007 71.940 71.378 892 73.947 72.503 70.608 79.211 77.789 77.681 1637 71.726 71.710 71.383 71.555 71.448 71.364 1292 72.449 72.314 71.192 72.115 70.749 69.692 1370 78.270 77.645 74.248 75.911 71.309 70.590 1672 74.143 73.464 73.007 74.550 74.386 73.289 624 81.529 78.848 77.835 76.145 75.857 74.704 1483 80.724 80.652 80.512 76.303 76.276 76.170

Mean eye wrinkle improvement degree Before use After 2 weeks of use After 4 weeks of use Left eye Measures
Improvement rate
P- value 72.476
0%
- 71.779 ***
0.96%
<0.001 * 70.873 ***
2.21%
<0.001 Right eye Measures
Improvement rate
P- value 71.150
0%
- 70.467 ***
0.96%
<0.001 * 70.116 ***
1.45%
<0.001

*: P < 00.05, **: P < 00.01, ***: P <

Claims (6)

A cosmetic composition for whitening or wrinkle improvement, comprising a sea cucumber juice liquid or a fraction thereof as an active ingredient.
The cosmetic composition according to claim 1, wherein the sea cucumber juice liquid fraction is a fraction containing a glycoprotein having a molecular weight of 50 kDa or more, a concentrate of the fraction or a lyophilizate of the fraction.
The method according to claim 1, wherein the sea cucumber juice-
Dipping the sea cucumber at 80 to 90 占 폚 to obtain a sea cucumber juice-
Filtering the sea cucumber juice solution under reduced pressure, and
A step of separating a fraction containing a glycoprotein having a molecular weight of 50 kDa or more
Wherein the cosmetic composition is prepared by a process comprising
A cosmetic comprising the composition of any one of claims 1 to 3.
The cosmetic composition according to claim 4, wherein the cosmetic composition is at least one selected from the group consisting of a lotion, a milky lotion, a cream, an essence, a cosmetic lotion, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation, a powder, .
Dipping the sea cucumber at 80 to 90 占 폚 to obtain a sea cucumber juice-
Filtering the sea cucumber juice solution under reduced pressure, and
Comprising fractionating a fraction comprising a glycoprotein having a molecular weight of at least 50 kDa.
A method for producing the cosmetic composition of claim 1.

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