KR20200060822A - Composition comprising sea mustard extract and collagen peptide - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening or wrinkle alleviation and a pharmaceutical composition for alleviating or treating inflammatory diseases, wherein the compositions contain Undaria pinnatifida extracts or a mixture of the Undaria pinnatifida extracts and a collagen peptide extracted from starfish as an active component. An embodiment of the present invention provides the cosmetic composition for skin whitening or wrinkle alleviation which contains at least one selected from a group consisting of the Undaria pinnatifida extract and the collagen peptide as the active component.

Description

Composition comprising sea mustard extract and collagen peptide

The present invention relates to a composition comprising at least one selected from the group consisting of seaweed extract and collagen peptide, the composition relates to a composition having whitening, wrinkle improvement, and anti-inflammatory activity.

Seaweeds inhabit the world and are used for food in some Asian regions, including Korea and Japan. About 800 seaweeds have been identified so far in Korea, of which 50 are used for food and are widely used in various functional food ingredients. In particular, Korea is a three-sided sea and has abundant tidal flats, so it has abundant marine resources, and the aquaculture technology of marine resources, including seaweed, is steadily researched and developed as a green growth high value-added industry.

Seaweed has low protein (6-42%) and fat content (1-3%), high carbohydrate content (6-74%), and various inorganic salts such as potassium, iodine, and calcium, and vitamin-A It contains high amounts of (β-carotene), vitamin C, and niacin. In particular, it is consumed as an essential food for children and mothers during the growing season because of its high iodine content and easy toxin release. In particular, fucoidan among seaweed components is a polymer polysaccharide with a molecular weight of about 200,000 Da or higher that is separated from seaweed, which causes cancer cell-specific apoptosis and regulates the expression of HGF (hepatocyte growth factor) to restore damaged cells and tissues. It is said to have the ability to. It is known that lowering the molecular weight of fucoidan increases the solubility of fucoidan in the body and increases the absorption capacity, thereby increasing pharmacological efficacy.

When the age of 25 to 30, when aging begins in earnest, the ability to produce collagen in the body decreases rapidly, and the skin becomes dry, and the makeup is excited and the blemishes and blemishes increase. Therefore, from this point on, it is helpful to supply the skin with nutrients and collagen capable of restoring cells and tissues to maintain moist and elastic skin. Cosmetics currently on the market contain a large amount of chemical components that irritate the skin, so when used on sensitive skin, rather often cause problems such as skin troubles or skin damage. Accordingly, there is a need to develop cosmetics that contain active ingredients derived from natural resources that are safe for the human body, have little irritation to the skin, and have excellent skin whitening or wrinkle improvement effects. The whitening, wrinkle improvement and anti-inflammatory effects of cosmetic compositions containing collagen peptides extracted from seaweed extract and starfish in seaweed have not been proven, and especially from seaweed extract and starfish than using seaweed extract alone. The synergistic effect of the above-mentioned efficacy when using the extracted collagen peptide is not known until now.

An example of the present invention provides a cosmetic composition for improving whitening or wrinkles comprising at least one selected from the group consisting of seaweed extract and collagen peptide as an active ingredient.

An additional example of the present invention provides a pharmaceutical composition for improving or treating an inflammatory disease comprising at least one selected from the group consisting of seaweed extract and collagen peptide as an active ingredient.

Another example of the present invention provides a method for preparing a composition comprising as an active ingredient one or more selected from the group consisting of seaweed extract and collagen peptide, which has an effect of improving whitening or wrinkles and having an effect of improving inflammation.

The present invention is a composition comprising at least one selected from the group consisting of seaweed extract and collagen peptides as an active ingredient, for example, in a cosmetic composition or anti-inflammatory pharmaceutical composition having anti-inflammatory activity, skin whitening, and wrinkle improvement activity It is about.

 The present invention can provide a cosmetic composition for improving whitening or wrinkles comprising at least one selected from the group consisting of seaweed extract and starfish collagen peptide as an active ingredient. Preferably, a seaweed extract or a mixture of seaweed extract and a starfish collagen peptide as an active ingredient may provide a cosmetic composition for whitening or wrinkle improvement. Preferably, the cosmetic composition for improving wrinkles may include a mixture of seaweed extract and starfish collagen peptide as an active ingredient.

The composition of the present invention may include a seaweed extract as an active ingredient, and the seaweed extract may include a solvent extract prepared by extracting seaweed as an extraction solvent, a solvent fraction thereof, a purified or dried product of the solvent extract or solvent fraction. Can be.

The seaweed extract, based on the solid content based on 100 parts by weight of the total composition, 0.01 to 99.99 parts by weight, 0.1 to 99.0 parts by weight, 0.1 to 90.0 parts by weight, 0.1 to 80.0 parts by weight, 0.1 to 70.0 parts by weight, 0.1 to 60.0 parts by weight, 0.1 to 50.0 parts by weight, 0.1 to 40.0 parts by weight, 0.1 to 30.0 parts by weight, 0.1 to 20.0 parts by weight, 0.1 to 10.0 parts by weight, 0.1 to 5.0 parts by weight, 0.1 to 3.0 parts by weight, 0.1 to 2.0 parts by weight Parts, 0.1 to 1.0 parts by weight, 0.1 to 0.5 parts by weight, 0.1 to 0.3 parts by weight, 0.2 to 3.0 parts by weight, 0.2 to 2.0 parts by weight, or 0.2 to 1.0 parts by weight may be included. The seaweed extract may be in the form of a dried product, a concentrated product, or a concentrated dried product.

The content of seaweed extract contained as the active ingredient in the composition can be appropriately adjusted according to the type and purpose of use, the patient's condition, the type and severity of symptoms, and the like.

The type of seaweed for the production of the seaweed extract is not particularly limited, but can be used seaweed (UNDARIA PINNATIFIDA). Dried sea bream contains 12.7% of protein, 52% of sugar, 1.1% of fat, 1300mg of Ca, vitamin A　140IU, B1 0.11mg%, B2 0.14mg%, nicotinic acid 10mg%, vitamin C 15mg%, calcium content This is high, and iodine also contains about 100 mg%. Other alginic acid and carotenoid pigments include fucoxanthine, violaxanthine, lutein, and beta-carotene.

The type of solvent for the seaweed extract is not particularly limited, and a commonly usable solvent may be used. For example, water, alcohols of C1 to C6, or mixtures thereof can be used, preferably water. The seaweed extract may mean a solvent extract, a fraction of an extract, a concentrate of an extract, or a dry matter of an extract.

In order to obtain the seaweed extract, any seaweed extract preparation method known in the art at the time can be used, and for example, the number of times of extraction of seaweed extract can be extracted repeatedly 2-5 times. As an extraction method, hot water extraction, ultrasonic extraction, or the like can be used under pressure, and hot water extraction is appropriate.

Specifically, the seaweed extract is a step of adding an extraction solvent to dry seaweed and primary extraction at 55 to 75°C for 2 to 6 hours; After the first extraction, the seaweed is removed and a solvent is added for a second extraction at 55 to 75° C. for 2 to 6 hours; And mixing the primary extract and the secondary extract in a ratio of 1:1 to 5:1.

The extraction solvent may be extracted by adding 5 to 15 times the total weight of seaweed, preferably 7 to 12 times, or 8 to 11 times to extract. In addition, the extraction temperature may be 55 ℃ to 75 ℃, preferably 60 ℃ to 70 ℃, or 65 to 70 ℃. By maintaining the temperature range, it is possible to effectively prevent decay, discoloration and odor of seaweed. The primary extraction time is preferably performed for 2 to 6 hours, preferably 3 to 5 hours, or 3 to 4 hours, and the secondary extraction time is also 2 to 6 hours, preferably 3 to 5 hours, or It is preferred to perform for 3 to 4 hours. When the extraction time exceeds the above range, the risk of seaweed excessively retreating and the destruction of nutrients extracted from seaweed rises rapidly, and when the extraction time is less than the above range, the active ingredient of seaweed is not sufficiently extracted, so within the above range It is preferred to extract.

The seaweed extract of the present invention exerts an anti-inflammatory effect by reducing the NO ₂ concentration of cells, and based on 100% of the tyrosinase activity inhibition rate of the untreated control group, the inhibition rate of the sample treatment group is 15% or more, for example, 15 to 15 It exhibits 40% and has a high inhibition rate of tyrosinase activity, so it has an excellent skin whitening effect. In addition, based on the high elastase activity inhibition rate of the untreated control group, the elastase activity inhibition rate of the treatment treatment group of the present invention is 10% or more, for example, a high elastase activity inhibition rate of 10 to 30%. It has an excellent wrinkle improvement effect.

The composition according to an embodiment of the present invention may include a seaweed extract and a collagen peptide mixture.

The method for producing the collagen peptide is not particularly limited, but may be an aminopeptide decomposition product of starfish or a hot water extract thereof. In one example, the collagen peptide may be provided as an extract extracted by treating the starfish pulverized product with an aqueous alkali solution, processing aminopeptidase by adjusting the pH to a pH range of 5.0 to 7.0, and hot water extraction with hot water at 50°C to 90°C. have. In a specific embodiment, the starfish collagen peptide, removing the salt of the starfish pulverized; Alkali treatment by dipping the salt-removed starfish pulverized in an aqueous alkali solution such as NaOH; Neutralizing by adding water to the alkali-treated starfish pulverized product and adding citric acid to a final pH of the mixture from 5.0 to 7.0; Adding 1 to 2 parts by weight, preferably 1.6 to 1.8 parts by weight, and hydrolyzing the aminopeptidase to the neutralized mixture based on 100 parts by weight of the total mixture, and adding distilled water to the hydrolyzate and temperature of 60°C to 80°C It may be a starfish collagen peptide extract prepared through the step of extracting with hot water.

In the step of hydrolyzing by treating the aminopeptidase, it is preferable to further treat the membrane-modified nonionic surfactant, and 0.01 to 5 parts by weight of the membrane-modified nonionic surfactant may be treated with respect to 100 parts by weight of the starfish. The membrane-modified nonionic surfactant may include, for example, any one or two or more selected from aliphatic sugar-based nonionic surfactants and polyethylene oxide-based aliphatic ethers. The aliphatic saccharide-based nonionic surfactant may be dodecylmaltoside, dodecylglucoside, heptylglucoside, or the like. In addition, the polyethylene oxide-based aliphatic ether may be polyoxyethylene octylphenyl ether, octaethylene glycol n-dodecyl monoether (octa(ethyleneglycol) ndodecylmonoether), or the like. The starfish may be an amour starfish, a starfish, a spider starfish, or the like. When the above extraction method is used, collagen peptides and fat-soluble peptides effective for preventing skin aging can be effectively extracted.

The collagen peptide of the present invention may mean a starfish collagen peptide extract, a concentrate of an extract, a powder of an extract or a concentrate, or a pure starfish collagen peptide contained in the extract. Preferably it may be a starfish collagen peptide extract.

The collagen peptide is 0.1 to 99.9 parts by weight, 0.5 to 99.5 parts by weight, 1 to 99 parts by weight, 1 to 90 parts by weight, 1 to 80 parts by weight, 1 to 70 parts by weight, 1 to 100 parts by weight of the total cosmetic composition To 60 parts by weight, 1 to 50 parts by weight, 1 to 40 parts by weight, 1 to 30 parts by weight, 1 to 20 parts by weight, 1 to 10 parts by weight, 2 to 98 parts by weight, 2 to 90 parts by weight, 2 to 80 parts by weight Parts by weight, 2 to 70 parts by weight, 2 to 60 parts by weight, 2 to 50 parts by weight, 2 to 40 parts by weight, 2 to 30 parts by weight, 2 to 20 parts by weight, 2 to 10 parts by weight, 3 to 97 parts by weight , 3 to 90 parts by weight, 3 to 80 parts by weight, 3 to 70 parts by weight, 3 to 60 parts by weight, 3 to 50 parts by weight, 3 to 40 parts by weight, 3 to 30 parts by weight, 3 to 20 parts by weight, or It may be included in 3 to 10 parts by weight, preferably 1 to 8 parts by weight, 1 to 6 parts by weight, 2 to 8 parts by weight, 2 to 6 parts by weight, parts by weight, 3 to 8 parts by weight, or 3 to 6 parts by weight It can be included as a wealth.

The content may be appropriately adjusted according to the use, application form, and purpose of use of the cosmetic composition. However, if it is less than the above range, a substantial anti-inflammatory or anti-wrinkle effect cannot be obtained, and the effect is remarkably deteriorated, and if it exceeds the above range, the stability of the cosmetic formulation is lowered, so it is preferable that it is the above range.

In a preferred embodiment according to the present invention, the mixing ratio of the seaweed extract and collagen peptide mixture is 1:99 to 99:1, 10:90 to 90:10, 20:80 to 80:20, 30:70 based on the solid content. It may be from 70:30 to 40:60 to 60:40. Preferably 1:0.3 to 1:99, 1:1.5 to 1:80, 1:2 to 1:60, 1:3 to 1:40, 1:3.3 to 1:30, 1:4 to 1:25 , 1:5 to 1:20, 1:6 to 1:25, 1:6 to 1:20, or 1:6 to 1:15 may be mixed in a weight ratio (above seaweed extract is solids weight, starfish Collagen peptides can be solids weight or liquid extract weights.).

The extract containing the starfish collagen peptide contains components such as butylene glycol in addition to the collagen peptide, and based on 100 parts by weight of the extract, the pure starfish collagen peptide is 1 to 70 parts by weight, preferably 2 to 65 parts by weight, 3 to 60 parts by weight, 4 to 60 parts by weight, 5 to 60 parts by weight, 6 to 60 parts by weight may be included.

The starfish collagen peptide may be a peptide having a gelation temperature of 30°C to 40°C and a weight average molecular weight of 200 to 2,500 Daltons (Da).

The seaweed extract or the mixture of seaweed extract and starfish collagen peptide of the present invention exhibits excellent anti-inflammatory effect by reducing the NO ₂ concentration of cells. Specifically, the LPS concentration can be reduced by 20 to 80%, preferably 20 to 70%, 30 to 80%, or 30 to 70% compared to the control treated with LPS only.

In addition, the mixture of the present invention has a high inhibition rate of tyrosinase activity, and thus has an excellent skin whitening effect. Specifically, it may have a high tyrosinase activity inhibition rate of 10 to 40%, preferably 15 to 40%, 20 to 40%, or 25 to 40% compared to the untreated control group. More preferably, the mixture may have a tyrosinase activity inhibition rate of 20 to 35% compared to an untreated control group.

The mixture of the present invention has an excellent effect of improving wrinkles by having an elastase activity inhibition rate. Specifically, it has an inhibition rate of elastase activity of 5 to 40%, preferably 5 to 30%, 10 to 30%, or 15 to 30%, compared to the untreated control, preferably the mixture is 15 to 15 to the untreated control It may have an elastase activity inhibition rate of 25%, or 20-30%.

In addition, the seaweed extract of the present invention or a mixture of seaweed extract and starfish collagen peptide does not irritate the skin and is safe for the human body.

The cosmetic composition of the present invention has an effect of inhibiting intracellular anti-inflammatory, tyrosinase activity and elastase activity, and also improves whitening and wrinkles without irritating the skin through skin clinical experiments It was confirmed that the efficacy is excellent. Therefore, the composition of the present invention can be used as a cosmetic composition or pharmaceutical composition for anti-inflammatory, anti-wrinkle and/or whitening.

In the present specification, whitening refers to a function that helps to whiten the skin by preventing the deposition of excessive melanin pigment on the skin or by suppressing the formation of famine and freckles by thinning the color of the existing melanin pigment. Melanin pigment in human skin is a major mechanism to protect against UV damage, but it causes undesirable problems such as the formation of abnormal pigments such as melasma, freckles, senile lentigines or excessive pigmentation. , Tyrosinase causes melanin biosynthesis in human skin. Therefore, inhibitors of tyrosinase are known as important materials used in cosmetics for the whitening effect. The seaweed extract of the present invention, or a cosmetic composition comprising a mixture of seaweed extract and a collagen peptide as an active ingredient, has been shown to have an inhibitory effect of tyrosinase and has a skin whitening effect. In particular, the mixture of seaweed extract and collagen peptide exerts a synergy effect in inhibiting tyrosinase by mixing the seaweed extract and collagen peptide mixture, and thus has better skin whitening efficacy than cosmetic compositions containing seaweed extract alone.

As used herein, wrinkle improvement includes maintaining or enhancing the ability of skin to relate to wrinkles and elasticity. In this regard, since elastase activity is known to increase skin aging and reduce skin elasticity, the activity inhibitor of elastin degrading enzyme can be used to suppress skin aging and improve wrinkles. The seaweed extract of the present invention, or a cosmetic composition comprising a mixture of seaweed extract and collagen peptide as an active ingredient, has an excellent effect of inhibiting elastin activity, and thus has a wrinkle improvement effect. In particular, the mixture of seaweed extract and collagen peptide is a seaweed extract. In particular, the mixture of seaweed extract and collagen peptide exerts a synergy effect on the elastase inhibitory efficiency by mixing the seaweed extract and collagen peptide mixture to produce seaweed extract. It has a better wrinkle improvement effect than the cosmetic composition containing alone.

The cosmetic composition of the present application is a functional cosmetic composition for improving whitening or wrinkles, and depending on the route of administration, it is possible to apply for external application to the skin, transdermal or subcutaneous, preferably for external application to the skin or transdermally, more preferably for external application to the skin. Can be The cosmetic composition can be applied percutaneously to the skin, scalp or hair, such as basic cosmetics, makeup cosmetics, body products (body cleanser, body lotion, etc.), shaving products, hair products (shampoo, conditioner, hair essence, etc.) It means a composition that can be used in the manufacture of all cosmetic products, and may be formulated in the form of warnings, sprays, suspensions, emulsions, creams, gels, foams, etc., but there is no particular limitation on the form. For example, it may have a formulation such as lotion, emulsion, cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, foundation, powder, makeup remover and detergent.

The cosmetic composition, in addition to the active ingredient, all kinds of ingredients that can be used for normal productization or formulation, such as fragrances, pigments, fungicides, antioxidants, preservatives, moisturizers, thickeners, excipients, diluents, inorganic salts and synthetic polymer materials, etc. It may further include, the type and content can be appropriately adjusted according to the purpose and purpose of use of the final product.

According to an example of the present invention, it is possible to provide a pharmaceutical composition for improving or treating an inflammatory disease comprising seaweed extract or seaweed extract and a starfish collagen peptide as an active ingredient. Preferably, it is possible to provide a pharmaceutical composition for improving or treating an inflammatory disease comprising seaweed extract and starfish collagen peptide as an active ingredient.

Description of the seaweed extract and starfish collagen peptide and the content range included in the composition are the same as described above. When treating the seaweed extract or the mixture of seaweed extract and starfish collagen peptide of the present invention with cells, it was experimentally proved to have an excellent NO ₂ inhibitory effect, especially when treated with seaweed extract alone, seaweed extract and starfish collagen peptide When the mixture of was treated, it was confirmed that the NO ₂ inhibitory effect was increased nearly twice.

The inflammatory diseases include rhinitis, gastritis, enteritis, nephritis, hepatitis, burns, wounds caused by surgical or dental surgery, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iris, scleritis, Uveitis, dermatitis, atopic dermatitis, gingivitis, and eczema, and may preferably be dermatitis, or gum inflammation.

The present invention provides a cosmetic composition for improving skin whitening or wrinkles, and a pharmaceutical composition for improving or treating an inflammatory disease, comprising a seaweed extract or a mixture of a seaweed extract and a starfish collagen peptide as an active ingredient.

Figure 1 shows the experimental results confirming the effect and anti-inflammatory effect on the NO ₂ content in the cells of the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and starfish collagen peptide mixture.
Figure 2 shows the experimental results confirming the cytotoxic effect of the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and starfish collagen peptide mixture.
Figure 3 shows the experimental results of measuring the inhibitory effect of tyrosinase of the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and starfish collagen peptide mixture.
Figure 4 shows the experimental results of measuring the elastase inhibitory efficacy of the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and starfish collagen peptide mixture.
5 is a result of measuring the overall size wrinkle improvement rate after using the cosmetic composition containing the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and starfish collagen peptide mixture as an active ingredient, the result of evaluating the degree of wrinkle improvement around the eyes Indicates.
6 is a result of evaluating the degree of improvement in wrinkles around the eyes by measuring the improvement rate of wrinkles after using the cosmetic composition containing the seaweed extract according to Example 1 and the seaweed extract according to Example 2 and the starfish collagen peptide mixture as an active ingredient. Indicates.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only to illustrate the present invention, the present invention is not limited by the following examples.

Example 1. Preparation of seaweed extract

Seaweed extract was prepared using natural seaweed supplied from Gopo-ri fishing village in Yeolam-ri, Uljin, Gyeongbuk.

After processing the dried seaweed, the falling phage (by-product) seaweed was washed several times and freeze-dried. To the freeze-dried seaweed, water of 10 times the weight of seaweed was added and the mixture was first extracted for 4 hours at a temperature of 60 to 65°C.

Next, the seaweed after the first extraction was removed, and water 3 times the weight of seaweed was added to the seaweed, followed by secondary extraction at a temperature of 60 to 65°C for 4 hours. A final seaweed extract was prepared by mixing the primary extract and the secondary extract. It is appropriate to reduce the amount of water added during the second extraction rather than the first extraction because the seaweed that has completed the first extraction contains a large amount of water. When performing the second extraction, the production yield can be increased, and the active ingredient of seaweed that is not dissolved in the first extraction step is dissolved during the second extraction to extract water extract of seaweed containing a high content of nutrients of seaweed such as fucoidan. Can be obtained. Next, after freezing the water extract of seaweed at -20°C, the ice crystals were sublimated with a high vacuum of 1 mmHg and dried and powdered.

Example 2. Preparation of a mixture of seaweed extract and collagen peptide

2-1. Preparation of collagen peptides from starfish

The starfish collagen peptide was purchased from Amur Collagen Co., Ltd., or the starfish collagen peptide was prepared by the following manufacturing method.

Specifically, the intestines of the North Pacific seastar were removed, washed several times, crushed to a volume size of about 10 cm ³ , immersed in distilled water for 12 hours, and dried for 2 days to remove salt. After 30 minutes of salt-removed Amur Bulgari crushed product is immersed in 270 g of 2.5% by weight sodium hydroxide (NaOH) aqueous solution and alkali-treated for 12 hours at room temperature, the supernatant containing non-active and other components is removed and the remaining Amur Bulgari pulverized product Was washed with distilled water. Subsequently, alkali-treated Amurbulasari crushed product and 500 g of distilled water were added to the reactor and mixed, and citric acid was added so that the final pH of the mixture was 6.0. Subsequently, 1.7 g of aminopeptidase (Recombinant Aeromonas Aminopeptidase, PEPROTECH) based on the total weight of the aqueous solution and 5 g of polyethylene oxide-based aliphatic ether (Triton x-100, SIGMAALDRICH) were added as a membrane-modified nonionic surfactant, and 4 hours at 47°C. The mixture was stirred for hydrolysis and reacted at 80° C. for 20 minutes to deactivate the enzyme and terminate the hydrolysis reaction.

 Subsequently, 1,000 g of distilled water was added to the hydrolyzed Amurbulasari crushed product, and hot water extraction was performed at 70°C for 7 hours. Filter the filtered mixture of hot water extract (6 ㎛) to finally obtain a starfish collagen peptide-containing extract.

2-2. Preparation of seaweed extract and starfish collagen peptide mixture

1 g of the seaweed extract powder prepared in Example 1 and 10 mL of the starfish collagen peptide-containing extract prepared in Example 2-1 were mixed to finally prepare a mixture of seaweed extract and starfish collagen peptide.

Example 3. Confirmation of anti-inflammatory efficacy

In order to confirm the anti-inflammatory efficacy of the mixture of the seaweed extract prepared in Example 1 and the seaweed extract prepared in Example 2 and the starfish collagen peptide extract, after processing the mixture sample in a RAW 264.7 cell line, it is one of inflammatory cytokines. The concentration of NO ₂ (nitro oxide) was measured. Treatment of LPS, known as a bacterial toxin, in the RAW 264.7 cell line produces mediators of inflammatory reactions such as the inflammatory cytokines NO ₂ (nitro oxide) and PG (cytokine prostaglandin), resulting in inflammation-related pathological reactions.

As a cell for measuring anti-inflammatory efficacy, a RAW 264.7 cell line was dispensed into a 24-well plate containing DMEM medium and cultured for 24 hours in a 37°C incubator. After the cultivation was completed, the existing medium of each well was replaced with fresh DMEM medium, and the seaweed extract prepared in Example 1 and the mixed sample prepared in Example 2 were diluted by concentrations of 1.0 mg/ml and 5.0 mg/ml, respectively. And treated 10 μl each. Subsequently, after 1 hour, 0.2 ug/ml of Lipopolysaccharide (LPS) was treated and 37° C. Incubation was carried out for 24 hours in an incubator. The RAW 264.7 cell line (Con) and LPS grown in a medium not treated with any sample as a control group were treated alone, and the samples of Examples 1 and 2 were treated with RAW 264.7 cell line.

Next, 100 μl of the supernatant of the obtained culture solution and 100 μl of Griess reagent were mixed and dispensed in a 96-well plate, and allowed to stand at room temperature for about 10 minutes, followed by irradiation with 540 nm light to measure absorbance. The concentration of nitrate was calculated using NaNO ₂ as a standard. The concentration (μM) of Nitrate (NO ₂ ) according to the treatment of LPS, Example 1 or Example 2 is shown in Table 1 and FIG. 1 below.

division sample Sample concentration (mg/ml) NO ₂ concentration (μM) LPS LPS 0 18.02 Example 3-1 Example 1 1.0 11.79 Example 3-2 Example 1 5.0 9.12 Example 3-3 Example 2 1.0 8.10 Example 3-4 Example 2 5.0 7.09

As shown in Table 1 and Figure 1, when processing the seaweed extract of Example 1 or the mixture sample of Example 2, it was confirmed that the concentration-dependent NO ₂ production inhibitory effect appears. Particularly, when the sample at a concentration of 1.0 mg/ml was treated, the mixture of Example 2 exhibited an excellent NO ₂ inhibitory ability twice as much as when the seaweed extract of Example 1 was treated alone. That is, when the mixture of seaweed extract and starfish collagen peptide was treated, rather than when the seaweed extract was treated alone, it was confirmed that it has a relatively excellent NO ₂ inhibitory ability and excellent anti-inflammatory efficacy.

Example 4. Stability evaluation

In order to confirm the stability in the body of the seaweed extract of Example 1 and the seaweed extract of Example 2 and the starfish collagen peptide mixture, cell viability (%) was measured.

To prepare melanocytes for the test, 100 µl of a suspension (5000cells/well) of B16-F10 melanocytes (manufacturer ATCC LOT. 60508145) was dispensed into a 96-well plate and pretreated for 24 hours at 37° C. and 5% CO ₂ conditions. Cells were attached to the well plate by -incubation. In a blank well, 100 µl of DMEM (Dulbecco's Modified Eagle Medium, ATCC) medium containing no B16-F10 melanocytes was dispensed.

Next, the seaweed extract of Example 1 was diluted in water to prepare a sample having a concentration of 0.1 mg/ml, 0.2 mg/ml, 5 mg/ml, and then treated with 10 µl for each concentration in the well. Along with this, a negative control containing 10 μl of medium and a positive control in which 10 μl of ethanol (EtOH) was added to the medium were incubated at 37° C. and 5% CO ₂ for 24 hours. Next, 10 µl of the CCK-8 solution was added to each well, followed by incubation for 1 hour, and absorbance was measured by irradiating 450 nm light. Blank was measured by adding a sample containing no melanocytes.

Similarly, after diluting the seaweed extract and the starfish collagen peptide mixture of Example 2 in water to prepare a sample having a concentration of 0.1mg/ml, 0.2mg/ml, 5mg/ml, 10 µl for each concentration in the well Each was treated, and the negative control containing 10 µl of the medium and the positive control with ethanol added to the medium were incubated at 37° C. and 5% CO ₂ for 24 hours, and then 10 µl of the CCK-8 solution was added to each well. After incubation for 1 hour, absorbance was measured by irradiating light of 450 nm. Blank was measured by adding a sample containing no melanocytes. Cell viability was measured using Equation 1 below, and the measurement results are shown in Table 2 and FIG. 2 below.

[Equation 1]

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 by adding distilled water in the presence of melanocytes)

1 experiment sample Sample concentration (μg/ml) Cell viability (%) CON - - 100 EtOH EtOH - 23.07 LPS LPS - 101.99 Example 4-1 Example 1 0.1 98.74 Example 4-2 Example 1 0.2 98.74 Example 4-3 Example 1 5.0 102.85 2 experiments sample Sample concentration (μg/ml) Cell viability (%) CON - - 100 EtOH EtOH - 19.563 LPS LPS - 91.48 Example 4-4 Example 2 0.1 98.10 Example 4-5 Example 2 0.2 98.10 Example 4-6 Example 2 5.0 95.68

As shown in Table 2 and FIG. 2, when melanin cells were treated with the mixture of Example 1 or Example 2, the cell viability was higher than 90% regardless of whether LPS was treated. On the other hand, when 100% of EtOH was treated, the cell viability was 10% or less. Therefore, the stability of the body of the mixture of seaweed extract of Example 1 of the present invention and Example 2 was confirmed.

Example 5. Skin whitening efficacy verification

As a test sample, in order to confirm the skin whitening efficacy of the seaweed extract of Example 1 or the mixture of Example 2, after processing various concentrations of samples in B16-F10 melanocytes, tyrosinase inhibitory activity was measured. Did.

To prepare melanocytes for the test, 500 μl of supernatant (50,000 cells/well) of B16-F10 melanocytes (manufacturer ATCC LOT. 60508145) was dispensed into a 24-well plate for 24 hours at 37° C. and 5% CO ₂ conditions. Cells were attached to the well plate by pre-incubation.

Next, dilute each of vitamin C (Vit-C) with water as a seaweed extract of Example 1, a mixture of Example 2, and a positive control to sample 0.2 mg/ml, 1.0 mg/ml, and 5.0 mg/ml concentrations. After preparation, 50 μl was dispensed into each well and incubated at 37° C. and 5% CO ₂ for 24 hours. Subsequently, 10 μl of 1 mg/ml trypsin was treated and reacted for 10 minutes, followed by centrifugation at 1000 rpm for about 2 minutes to obtain pellets. The pellet obtained therefrom was dissolved in a 0.5 ml solution of 1% triton X-100 PBS, 0.5 ml of 0.2% L-DOPA 0.1M sodium phosphate buffer was mixed and incubated at 37° C. for 2 hours and 490 nm. The absorbance was measured by irradiating the light. The tyrosinase inhibition rate was calculated using Equation 2 below, and the measurement results of the tyrosinase inhibition rate are shown in Table 3 and FIG. 3 below. As an additive-free control group, melanocytes were prepared in the same manner as in Example, but the absorbance was measured by conducting the same experiment without processing the sample.

In Table 3 and Figure 3, the positive control vitamin C (Vit-C), as a result of treating the samples according to Examples 1 and 2 for each concentration of 0.2, 1.0 and 5.0 mg/ml, tyrosina of melanocytes The rate of inhibition of azease activity was indicated. The activity inhibition rate of the untreated control was 100%.

[Equation 2]

Inhibition rate of tyrosinase activity (%)={(absorbance of the control group-absorbance of the sample-added group)/absorbance of the control group} X 100

division sample Sample concentration
(mg/ml) Inhibition rate of tyrosinase activity (%) Example 5-1 Sample of Example 1 0.2 16.26 Example 5-2 Sample of Example 1 1.0 28.50 Example 5-3 Sample of Example 1 5.0 30.52 Example 5-4 Sample of Example 2 0.2 18.07 Example 5-5 Sample of Example 2 1.0 34.69 Example 5-6 Sample of Example 2 5.0 36.31 Vit-C Vit-C 0.2 27.59 Vit-C Vit-C 1.0 56.88 Vit-C Vit-C 5.0 70.95

As can be seen from Table 3 and FIG. 3, when the seaweed extract of Example 1 or the mixture of Example 2 was treated, it was confirmed that tyrosinase activity was inhibited. When the seaweed extract of Example 1 was treated alone, the inhibitory rate of tyrosinase activity was increased depending on the concentration, and the inhibitory rate of tyrosinase activity was approximately 15 to 35%. When the mixture of Example 3 was treated, the tyrosinase activity inhibition rate was also increased depending on the concentration, and showed a tyrosinase activity inhibition rate of about 15 to 40%. Therefore, when a mixture of seaweed extract and starfish collagen peptide was used rather than sole treatment of seaweed extract, it was confirmed that synergistic effect occurred, showing excellent tyrosinase activity inhibitory effect, and excellent whitening effect.

Example 6. Skin wrinkle improvement effect verification

As a test sample, in order to verify the effect of improving the skin wrinkles of the seaweed extract of Example 1 or the mixture of Example 2, after treating various concentrations of samples in B16-F10 melanocytes, the inhibition rate of Elastase activity was inhibited. It was measured.

To prepare melanocytes for the test, 500 μl of supernatant (50,000 cells/well) of B16-F10 melanocytes (manufacturer ATCC LOT. 60508145) was dispensed into a 24-well plate at 37° C. for 5 hours at 5% CO ₂ Cells were attached to the well plate by pre-incubation.

Next, dilute each of vitamin C (Vit-C) with water as a seaweed extract of Example 1, a mixture of Example 2, and a positive control to sample 0.2 mg/ml, 1.0 mg/ml, and 5.0 mg/ml concentrations. After preparing, 10 μl was dispensed into each well, and 50 μl of porcine pancreas elastase (10 μg/mL) solution dissolved in 50 mM tris-HCl buffer (pH 8.6) was added in 50 μl and 50 mM tris-HCl buffer (pH) as a substrate. After adding 100 μl of N-succinyl-(L-Ala)3-p-nitroanilide (0.5mg/mL) dissolved in 8.6), react for 20 minutes, and then measure the inhibition rate of elastase activity of B16-F10 melanocytes. Did. The elastase activity inhibition rate was measured by the absorbance reduction rate of the sample solution added and non-added. The measured elastase activity inhibition rate is shown in Table 4 and FIG. 4 below. As an additive-free control group, melanocytes were prepared in the same manner as in Example, but the absorbance was measured by conducting the same experiment without processing the sample. The elastase activity inhibition rate was calculated using Equation 3 below.

[Equation 3]

division sample Sample concentration
(mg/ml) Elastase activity inhibition rate (%) Example 6-1 Sample of Example 1 0.2 12.45 Example 6-2 Sample of Example 1 1.0 12.62 Example 6-3 Sample of Example 1 5.0 20.78 Example 6-4 Sample of Example 2 0.2 12.19 Example 6-5 Sample of Example 2 1.0 20.51 Example 6-6 Sample of Example 2 5.0 25.40 Vit-C Vit-C 0.2 36.78 Vit-C Vit-C 1.0 75.91 Vit-C Vit-C 5.0 92.29

As shown in Table 4 and Figure 4, when treating the mixture of the seaweed extract of Example 1 of the present invention and Example 2, it was confirmed that the elastase inhibitory ability was shown in a concentration-dependent manner. When the mixture of Example 2 was treated, it showed an inhibition rate of elastase activity of approximately 10 to 30%, and when the extract of Example 1 was treated, an inhibition rate of elastase activity of 10 to 25% was shown. Therefore, the seaweed extract of the present invention or the seaweed extract and starfish collagen peptide mixture has an excellent elastase inhibitory effect, thereby improving the wrinkles of the skin. This rise, it was confirmed that it shows excellent skin wrinkle improvement effect.

Example 7. Preparation of cosmetic composition comprising a mixture of seaweed extract and starfish collagen peptide

A cosmetic of an essence formulation containing 0.2% by weight of seaweed extract of Example 1 and 5% by weight of the mixture of Example 2 (about 3.3% by weight of a pure starfish collagen peptide) based on 100% by weight of total cosmetics was prepared. The composition of the cosmetics of the prepared essence formulation is shown in Table 5 below. If necessary, only a portion of the constituent components may be selected and mixed in an appropriate composition ratio.

Korean name content Purified water To 100% by weight Seaweed extract of Example 1 0.2 Seaweed extract & starfish collagen peptide mixture of Example 2 5.0 Adenosine 0.04 Gold (CI 77480) 0.05 1,2-hexanediol 2.0 Niacinamide 2.0 Methylglucose-10 0.1 Pentylene glycol 0.1 Carrageenan extract 0.1 Sugar cane extract 0.1 Cacao seed extract 0.1 Sodium hyaluronate 2.0 Dipropylene glycol 0.1 Glyceres-26 0.1 Betaine 0.1 PG-60 Hydrogenated Castor Oil 0.1 Dimethicone 0.1 Ammonium acryloyl dimethyl taurate / VP copolymer 0.1 Tromethamine 0.1 Caprylyl glycol 0.1 Octyldodeseth-16 0.1 Glyceryl polyacrylate 1.0 dextrin 0.1 Ethylhexylglycerin 1.0 Butylene Glycol 0.1 lecithin 0.1 Carbomer 0.1 Disodium LED 0.1 Spices 0.1 glycerin 3.0

Example  8. Cosmetic  Composition skin irritation test

For the skin irritation test of the cosmetic composition prepared in Example 7, a patch test was performed for 24 hours. After sufficiently soaking the cosmetic composition prepared in Example 7 on the patch, it was applied to the skin of a total of 20 subjects (20 randomly selected adult females, average age 43.5 years), and then left for 24 hours. After 24 hours, the patch was removed, and 30 minutes, 24 hours, and 48 hours later, a dermatologist judged whether the subject had primary skin irritation. Skin irritation determination was performed according to the International Contact Dermatology Association (ICDRG) standard and the American Cosmetics Association (PCPC) guidelines, and the irritation index was calculated using the skin reaction score of each subject. Table 7 shows the results of the 24-hour patch test in accordance with the criterion for the patch test in Table 6.

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(++) Moderate erythema with or without partial edema or papules 3 (+++) Moderate erythema with diffuse edema 4 (++++) Strong erythema with blisters and diffuse edema
Intense erythema with diffuse edema with vesicles

division Evaluation after 2 weeks of product use Evaluation after 3 weeks of product use Erythema - - Edema - - Scaling - - Itching - - Stinging - - Burning sensation Burning - - Stiffness - - Prickling - -

As shown in Table 7, erythema, swelling, swelling, itchiness, pain, burning, stiffness, and tingling irritation did not occur on the skin even after 2 and 3 weeks after using the cosmetic composition of Example 7, and the skin irritation index 0 points. Therefore, it was confirmed that the cosmetic composition of Example 7 does not irritate the skin and is a stable material.

Example 9. Cosmetic composition wrinkle improvement efficacy test

In order to test the wrinkle improvement efficacy of the cosmetic composition prepared in Example 7, a clinical trial was conducted to evaluate the efficacy of improving the wrinkles around the eyes for 4 weeks after using the cosmetic composition in a total of 20 subjects, and 20 subjects without dropouts. All completed the test, and the basic information of the subjects is shown in Table 8 below.

Total number of subjects 22 people gender Male: 0 Female: 20 Average age 43.5 years 30s 4 people 40s 8 people 50 cars 10 people

Wrinkle improvement efficacy is ANTERA 3D ^TM (Miravex, Ireland). ANTERA 3D ^TM Is a device that can measure the condition of the epidermis and dermis of the skin using a multi-wavelength LED light source. It can reconstruct 2D and 3D of images using the built-in program and analyze by wavelength.

The cosmetic composition prepared in Example 7 was divided into two groups before, two weeks, and four weeks after use to measure the skin condition of the subject. In order to accurately measure the skin condition of the subject, the use of basic cosmetics was prohibited 12 hours before the start of the first test, and the subject was washed with a standard cleanser provided by the tester, and then lightly patted with a paper towel to remove moisture. Then, after 30 minutes of stability under constant temperature and humidity conditions (20 to 24°C, 40 to 60% RH), images of wrinkles on both sides of the eyes were measured using ANTERA 3DTM. An ANTERA 3D imaging area was set up with a 6cm X 6cm forward area including the entire orbital and both eyes.

Next, using the cosmetic composition of the present invention, after the second and fourth weeks of testing, device evaluation was performed in the same site in the same manner as the first visit. Subsequently, statistical significance between eye wrinkle measurements measured before, 2 weeks after, and 4 weeks after using the cosmetic composition of the present invention was analyzed and verified using Wilcoxon code rank test. Measurement results, Overall size wrinkle improvement rate is shown in Figure 5, Depth wrinkle improvement rate is shown in Figure 6.

As shown in Figures 5 and 6, compared to before using the cosmetic composition of the present invention, after 2 weeks of use, 4 weeks after use, it was confirmed that a statistically significant level of improvement in wrinkles around the eyes appeared (p <0.05). . In addition, during the 4-week test period, no special skin adverse reaction was observed in all subjects, and it was confirmed that a statistically significant level of improvement in eye wrinkles appeared after the second week of use of the cosmetic composition of the present invention.

Claims (12)

Seaweed extract, or a mixture of seaweed extract and starfish collagen peptide containing an active ingredient, whitening or wrinkle improvement cosmetic composition. The cosmetic composition according to claim 1, wherein the seaweed extract is contained in an amount of 0.01 to 99.99 parts by weight based on 100 parts by weight of the cosmetic composition based on the solid content. The cosmetic composition of claim 1, wherein the cosmetic composition includes a mixture of seaweed extract and starfish collagen peptide as an active ingredient, and a mixed weight ratio of seaweed extract and starfish collagen peptide is 1:99 to 99:1. The cosmetic composition according to claim 1, wherein the starfish collagen peptide has a gelation temperature of 30°C to 40°C and a weight average molecular weight of 200 to 2,500 Daltons (Da). The method of claim 1, wherein the starfish collagen peptide
The starch pulverized product is treated with an aqueous alkali solution, adjusted to a pH range of 5.0 to 7.0, and treated with aminopeptidase, and provided as an extract extracted with hot water at a temperature of 50°C to 90°C. The cosmetic composition of claim 1, wherein the seaweed extract is a hot water extract of seaweed. The method of claim 6, wherein the seaweed extract
Adding 5 to 15 times the volume ratio of the solvent to the dried seaweed and primary extraction at 55 to 75°C; A second extraction at 55 to 75° C. by adding a solvent of 3 to 5 times the weight of seaweed to the seaweed obtained after the first extraction; And mixing the first extract and the second extract in a ratio of 1:1 to 5:1. The cosmetic composition according to claim 1, wherein the cosmetic composition exhibits an inhibition rate of tyrosinase activity of 15 to 40% compared to an untreated control group and has a whitening property. The cosmetic composition of claim 1, wherein the cosmetic composition exhibits 10 to 30% of elastase inhibitory ability and has wrinkle improvement properties compared to an untreated control group. Seaweed extract, or a pharmaceutical composition for the improvement or treatment of inflammatory diseases comprising a mixture of seaweed extract and starfish collagen peptide as an active ingredient. The method of claim 11, wherein the inflammatory disease is rhinitis, gastritis, enteritis, nephritis, hepatitis, burns, wounds caused by surgical or dental surgery, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, irisitis , Scleritis, uveitis, dermatitis, atopic dermatitis, gingivitis, and eczema, pharmaceutical composition. The pharmaceutical composition of claim 10, wherein the pharmaceutical composition includes a mixture of seaweed extract and starfish collagen peptide as an active ingredient, and a mixed weight ratio of seaweed extract and starfish collagen peptide is 1:99 to 99:1.

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