KR101446295B1 - Composition for anti-aging containing sargassum yezoense extract - Google Patents

Abstract

 The present invention relates to an anti-aging or moisturizing composition comprising a natural extract.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an anti-aging composition comprising an extract of Saga isozyme,

The present invention relates to an anti-aging or moisturizing composition comprising a natural extract.

Over time, the secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are decreased, and the biosynthesis of immune proteins and biologic proteins necessary for the living body is reduced , Externally, the content of ultraviolet ray reaching the surface of the sun ray is increased due to destruction of the ozone layer, and as the environmental pollution is further intensified, the thickness of the skin is reduced, the wrinkles are increased and the elasticity is decreased, , Freckles and black spots are also increasing. Skin aging processes are generally caused by intrinsic aging and photoaging (Gilchrest, 1989; Ma et al ., 2001; Jenkins, 2002).

Among the constitutive layers of the skin, the epidermis plays an important role in preventing water evaporation inside the human body. The epidermis is divided into the stratum corneum, granular layer, the superficial layer and the basal layer in order from the outside. Cells of the stratum corneum act like bricks, and the intercellular lipids between keratinocytes act like mortar to constitute skin barrier Invest., Dermatol 80 (Suppl.), 44-49, 1983). In addition, a healthy human keratinocyte has a high concentration of natural moisturizing factor (NMF) to help maintain moisture in the skin. For example, substances such as amino acids are water-soluble, (J. Invest. Dermatol. 54, 24-31, 1970).

However, as it is nowadays, there is a tendency to adjust the temperature of the air / heating due to the change of environment or life pattern, the stress caused by various stresses and environmental pollution in the social life, frequent washing according to make- Due to various causes such as aging of the skin, the skin becomes dry, the surface becomes rough, the skin becomes loose, the moisture is lost, and the appearance of the skin does not appear, and thus the need for a skin moisturizer increases . Excessive physical and chemical stimuli from the outside, ultraviolet rays, stress and nutritional deficiencies lower the normal functions of the skin and promote skin aging phenomenon such as loss of elasticity, keratinization and wrinkle formation. Particularly, It is severely damaged.

The inventors have found that Sargassum < RTI ID = 0.0 > yezoense extract of the present invention can prevent skin aging, skin wrinkles or skin moisturization, and loss of elasticity of the skin. Thus, the present invention has been completed.

Gilchrest, 1989; Ma et al., 2001; Jenkins, 2002

It is an object of the present invention to provide a natural antioxidant or moisturizing composition.

To achieve the above object, the present invention provides a composition for anti-aging or moisturizing comprising an extract of Sargassum yezoense as an active ingredient.

The composition according to the present invention contains a natural product obtained from nature as an effective ingredient and has a low adverse effect when applied to a human body, and the possibility of resistance to long-term use is low. Particularly, the composition of the present invention has excellent anti-aging and moisturizing ability and can be usefully used.

Specifically, the composition of the present invention inhibits the biosynthesis of at least one cytokine or enzyme selected from the group consisting of gelatinase, proinflammatory cytokine, matrix metalloprotease, and elastase, And can be used for moisturizing by increasing the biosynthesis or increasing the activity of one or more proteins selected from the group consisting of pillared green and inbolucrine.

FIG. 1 is a graph showing the effect of the above-mentioned Saga isoenzene extract on the expression of a pill green, inbolucrin.
FIG. 2 is a graph showing the results of the above-described assays for the inhibitory effect of the above-mentioned Saga isoenzene extract on gelatinase.
FIG. 3 is a graph showing Western blotting of the inhibition of biosynthesis against MMP-1 by the Sagas island ezoense extract.

In one aspect, the present invention relates to a method of treating sargassum yezoense extract as an active ingredient.

The term " Sargassum " yezoense) belong to feuding cap (Fucales), sargassaceae (Sargasaceae), genus Sargassum (Sargassum). Saga Island Ezoans is distributed on the eastern coast of Korea, the southern coast, and the lower part of the intertidal zone of Jeju Island, and forms a community on rocks affected by waves. In Japan, it is distributed along the coast between Hokkaido and Goto Islands and Hokkaido. It dominates the 0-2 m depth of the coast of the Pacific Tohoku coast. Small and young individuals begin to grow in September and grow very rapidly in February when water temperature starts to rise and grow maximum in June.

As used herein, Saguar isoenzyme may be contained in the form of an extract, or may be contained as a pulverized product of the herbal medicine itself or as a dried pulverized product of a herbal medicine, but the present invention is not limited thereto. The Sagas island ezoens used in the present invention is not limited to the method of obtaining it, and may be cultivated or used, or may be purchased and used partially or wholly.

The active ingredient of the present invention, Sagas island Ezoens extract, a fraction thereof, or a compound isolated from the fraction can be produced by a separation and purification method comprising the following steps.

1) extracting Saga isoenzene with an extraction solvent;

2) filtering and concentrating the Saga isoenzo extract of step 1);

3) further extracting the concentrate of step 2) with an organic solvent;

4) obtaining an active fraction by fractionating the fraction of step 3) via a praparative chromatography method; And

5) obtaining the active compound from the active fraction of step 4) via HPLC method.

In the above production method, the extraction solvent of step 1) is characterized by being water, alcohol or a mixture thereof. As the alcohol, C ₁ to C ₄ lower alcohol is preferably used, and as the lower alcohol, ethanol or methanol is preferably used. It is preferable to extract the solvent by 5 to 15 times of the amount of the sago isostearase at the time of extraction, and it is more preferable to extract it by 10 times, but not limited thereto. It is preferable to add the solvent and then ultrasonically extract it, but it is not limited thereto. The temperature of the solvent during the extraction is preferably 40 to 100 ° C, more preferably 80 ° C, but is not limited thereto. The extraction time is preferably 2 to 4 hours, more preferably 3 hours, but not always limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 times, but is not limited thereto. Sagase isoenzene was extracted and filtered by the above method, and the filtrate was concentrated and lyophilized to obtain an extract.

In the above production method, the organic solvent in step 3) may be ethyl acetate, hexane, dichloromethane, acetone, or acetonitrile, but is not limited thereto.

In the above preparation method, the praparative chromatography in step 4) can be separated and purified by performing preparative chromatography using a filler selected from the group consisting of YMC pack and the like. The preparative chromatography can be carried out several times by selecting appropriate fillers as necessary.

In the present specification, the Sago isoenzene extract may contain a crude extract of Sago isozyme as described above, and may be included as a soluble fraction of an organic solvent obtained by further extracting the crude extract with an organic solvent having a low polarity. Examples of the organic solvent include, but are not limited to, hexane, methylene chloride, ethyl acetate, n-butanol, and the like. The extract or the soluble fraction of the extract may be used as it is, or may be used as an extract form by concentration after filtration, and may be used as a form of a lyophilizate by concentration and freeze-drying.

As used herein, the term " active ingredient "alone refers to an ingredient that exhibits the desired activity or that can exhibit activity with a carrier that is not itself active.

In the composition for anti-aging or moisturizing, which is one aspect of the present invention, the extract may contain at least one compound selected from the group consisting of the following compounds 1 and 2, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(2)

As used herein, the term " pharmaceutically acceptable "refers to the approval of a government or equivalent regulatory agency that can be used on animals, and more specifically on humans, by avoiding significant toxic effects when used in conventional medicinal dosage Received, approved, or listed in pharmacopoeia or other general pharmacopoeia.

As used herein, "pharmaceutically acceptable salts" are useful as addition salts formed by free acids. Suitable inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid. Examples of the organic acids include citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, There may be mentioned fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid or aspartic acid Etc. may be used. Further, the Saga isoenzo extract, the fractions thereof, or the compounds separated from the fractions of the present invention can be prepared not only by pharmaceutically acceptable salts, but also by conventional methods All salts, hydrates and solvates may be included.

In the composition for anti-aging or moisturizing, which is one aspect of the present invention, the Sagami isoenzene extract may be contained in an amount of 0.001 to 10% by weight based on the total weight of the composition. When the extract of Saga isoenzoin is contained in an amount of less than 0.001% by weight based on the total weight of the composition, the anti-aging effect such as skin elasticity improvement and wrinkle relaxation is insufficient and the skin is dried and pulled out. , Thereby limiting the content of other constituents on the drug. In view of the above, the extract of Sagami isoenzene is preferably used in an amount of 0.0011 to 9 wt%, 0.0012 to 8 wt%, 0.0013 to 7 wt%, 0.0014 to 6 wt%, 0.0015 to 5 wt%, 0.0016 0.0017 to 3 wt.%, 0.0018 to 2 wt.%, Or 0.0019 to 1 wt.%, And specifically 0.002 wt.%.

In the composition for antiaging or moisturizing, which is one aspect of the present invention, the anti-aging composition is a gelatinase, a proinflammatory cytokine, a matrix metalloproteinase (MMP), and an elastase and elastase, or may reduce the activity of the enzyme. Specifically, in an anti-aging or moisturizing composition which is one aspect of the present invention, the proinflammatory cytokine may be a tumor necrosis factor, and more specifically, TNF- alpha). The anti-aging composition of the present invention can inhibit or reduce the production of the cytokine or enzyme, and even if it is produced, the activity can be reduced to inhibit or reduce its function, so that it is useful for preventing aging.

In the composition for anti-aging or moisturizing, which is one aspect of the present invention, the extract may be a Saga isoenzene-C ₁ -C ₆ alcohol extract, and specifically, a Saga isoenzyme- methanol extract or a Saga isoenzyme- Lt; / RTI >

In an anti-aging or moisturizing composition according to one aspect of the present invention, the composition may be one which increases the biosynthesis of at least one protein selected from the group consisting of filaggrin and involucrin, . The moisturizing composition of the present invention can inhibit or reduce the production of the protein, and even if it is produced, its activity can be reduced and its function can be inhibited or lowered, so that it is useful for moisturizing the skin.

In one aspect of the invention, the composition may be a health food composition.

The composition may be processed into a drink, fermented milk, cheese, yogurt, juice, probiotic agent, and health supplement food or the like, and may be used in various other food additives.

In one embodiment, the composition may contain other ingredients or the like that can give synergistic effects to the main effect within a range that does not impair the intended main effect of the present invention. For example, additives such as perfume, coloring agent, bactericide, antioxidant, preservative, moisturizing agent, thickening agent, inorganic salt, emulsifier and synthetic polymer substance may be further added for improvement of physical properties. In addition, it may further contain auxiliary components such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides and seaweed extract. The above components may be mixed and selected without difficulty by those skilled in the art depending on the purpose of formulation or use, and the amount thereof may be selected within a range that does not impair the objects and effects of the present invention.

The composition according to the present invention may be in various forms such as a solution, an emulsion, a viscous mixture, a tablet, a powder, etc., and may be administered by various methods such as simple drinking, injection administration, spraying or squeezing.

In one aspect of the invention, the composition may be a pharmaceutical composition.

In one embodiment, the pharmaceutical composition may be a pharmaceutical composition effective for skin moisturization or anti-aging.

When the composition according to the present invention is applied to medicines, it may be formulated into an oral or parenteral dosage form in the form of solid, semi-solid or liquid by adding an inorganic or organic carrier to the composition as an active ingredient.

Examples of the agent for oral administration include tablets, pills, granules, soft capsules, powders, fine granules, powders, emulsions, syrups and pellets. Examples of the parenteral administration agent include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like. In order to formulate the active ingredient of the present invention, it can be easily formulated according to the conventional method. Surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffering agents, suspending agents and other adjuvants can be suitably used.

The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like.

In addition, the dosage of the active ingredient will vary depending on the age, sex, and weight of the subject to be treated, the specific disease or condition to be treated, the severity of the disease or condition, the route of administration, and the judgment of the prescriber. Dosage determinations based on these factors are within the level of those skilled in the art. Typical dosages are from 0.001 mg / kg / day to 2000 mg / kg / day, more specifically from 0.5 mg / kg / day to 1500 mg / kg / day.

In one aspect of the invention, the composition may be a cosmetic composition.

When the composition according to the present invention is formulated in the form of a cosmetic, it may be formulated in the form of a softening agent, a convergent lotion, a nutritional lotion, an eye cream, a nutritional cream, a massage cream, a cleansing cream, a cleansing foam, a cleansing water, a powder, And the formulations thereof are not particularly limited. In addition, the composition according to the present invention can also be used as a lubricant, an organic solvent, a solubilizer, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, The active ingredient may be combined with any other ingredients commonly used in cosmetics, such as ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, And may contain adjuvants commonly used in the same cosmetic or dermatological fields. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields. In addition, the composition of the present invention may contain a skin absorption promoting substance to increase the skin improving effect.

The composition of the present invention may be a composition for external application for skin, more specifically, for external application for anti-inflammatory and / or moisturizing purposes.

The external preparation for skin may contain, in addition to the above-mentioned essential ingredients, a component commonly used for external skin preparations such as cosmetics or medicines such as an aqueous component, an oily component, a powder component, an alcohol, a moisturizer, a thickener, an ultraviolet absorber, a whitening agent, an antiseptic, , Perfumes, coloring agents, various skin nutrients, and the like can be appropriately compounded as needed.

In addition, there are other metal sequestrants such as sodium edetate, sodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, Vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, trehalose and the like, and the like, Sugars and the like can also be appropriately compounded.

The external preparation for skin may be cream, gel, ointment, skin emulsion, skin suspension, transdermal patch, drug-containing bandage.

The external preparation for skin may be formulated by using pharmaceutically acceptable carriers, excipients, binders, disintegrants, solubilizing agents, suspending agents, preservatives or extenders at the time of clinical administration. In the above external preparation for skin, the effective dose of the composition of the present invention is 0.01 to 300 mg / cm ² , preferably 10 to 50 mg / cm ² , and can be administered 1 to 4 times a day. However, the dosage level for a particular patient may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease,

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Saga Island Ezoens  ( Sargassum yezoense ) Preparation of extract

Sagas island ezoens used in the present invention were those native to the East Coast of Gangwon Province, and the outposts were dried and used.

[ Example  1-1] Saga Island Ezoens - Preparation of methanol extract

200 g of dried Saga isoenzyme was immersed in 1 L of 100% methanol and refluxed for 3 hours. This extraction was repeated three times. The extract was dried under reduced pressure and concentrated to obtain 24.6 g of a concentrate.

[ Example  1-2] Saga Island Ezoens - Preparation of water extract

200 g of dried Saga isoenzyme was immersed in 1 L of water and refluxed for 3 hours. The extract obtained by repeating this three times was dried under reduced pressure and concentrated to obtain 18.4 g of a concentrate.

[ Example  1-3] Saga Island Ezoens - Preparation of ethanol extract

200 g of dried Saga isoenzyme was immersed in 1 L of 95% ethanol and refluxed for 3 hours. The extract obtained by repeating this three times was dried under reduced pressure and concentrated to obtain 22.8 g of a concentrate.

Saga Island Ezoens  From the extract Fraction  Produce

2 g of the extract of Example <1-1> was suspended in distilled water and fractionated three times with ethylacetate. The fraction was dried under reduced pressure to obtain 1.26 g of a fraction.

Saga Island Ezoens From the fraction  activation Fraction  Produce

1 g of the fraction of Example 2 was subjected to preparative ODS column chromatography (YMC-Pack ODS-AQ, 250 X 10 mm, YMC Co., Japan). The solvent used was 80% methanol containing 0.05% trifluoroacetic acid. When the flow rate was 3 ml / min, the activity of the fraction obtained after 30 to 35 minutes elapsed was the best. The fractions were collected, dried under reduced pressure, and concentrated to obtain fractions (201 mg).

Separation of the compound of formula (1)

The active fractions obtained in Example 3 above were analyzed by high performance liquid chromatography, i.e., HPLC (TSK gel silica-60 column, 4.6 X 250 mm, Tosoh, Japan). A solvent mixture of chloroform: hexame: acetone: methanol in a ratio of 72: 18: 9: 1 was prepared and flowed at a mean flow rate of 1 ml. Lt; / RTI &gt; of the title compound were obtained. The structure of the compound was confirmed by NMR ( ¹ H and ¹³ C-NMR, Varian 500 MHz spectrometer, Varian USA). As a result, it was confirmed that the compound separated in the 5th minute had the structure of formula (1).

Separation of the compound of formula (2)

The active fractions obtained in Example 3 above were analyzed by high performance liquid chromatography, namely HPLC (TSK gel silica-60 column, 4.6 X 250 mm, Tosoh, Japan). A solvent mixture of chloroform: hexame: acetone: methanol in a ratio of 72: 18: 9: 1 was prepared and flowed at a mean flow rate of 1 ml per minute. Lt; / RTI &gt; of the title compound was obtained. The structure of the compound was confirmed by NMR ( ¹ H and ¹³ C-NMR, Varian 500 MHz spectrometer, Varian USA). As a result, it was confirmed that the compound separated in 10 minutes had the structure of formula (2).

[ Experimental Example  1] Skin moisturizing and hydration related Pillar Green  ( filaggrin ) And Inboluklein  Expression of involucrin

Human keratinocytes (Lonza, switzerland) isolated directly from the epidermis of newborn infants were cultured in 100-fold dishes at 10 ⁶ concentrations and replaced with medium containing 10 μM of each test substance and 10 μg / ml of extract Respectively. After 24 hours of culture, the cells were harvested to obtain proteins, and the amount of protein was quantified using a BCA kit (Pierce Biotechnology, USA). The amounts of filaggrin and Involucrin produced using the Western Blot method were quantified. The western blot was prepared by loading 30 μg of protein per lane on an SDS-PAGE gel, electrophoresing, and transferring the protein to a PVDF membrane (Amersham (USA)) using a protein transfer kit (Invitrogen, USA) , USA). After transfer, the antibodies against PPARα, pillar and inbolucrin (Santacruz Biotechnology, USA) were stuck at 37 ℃ for 1 hour, washed three times with PBS, and the secondary antibody with HRP was incubated at 37 ℃ For about 1 hour. After washing three times with PBS, the degree of luminescence was exposed to the film using an ECL kit, and the intensity of the band was quantified using a densitometer. The value was 100 for the UV control group.

Saga Island Ezoens  Extract, objection Fraction , Or In the fraction  Biosynthesis effect of moisturizing factors on isolated compounds sample density Pillar Green Inboluklein Untreated group 100 100 WY14643 10 uM 120.1 101.4 Example <1-1> 10 [mu] g / ml 128.3 107.6 Example 2 10 [mu] g / ml 138.8 106.7 Example 3 10 [mu] g / ml 141.3 109.4 Example 4 10 uM 128.4 108.2 Example 5 10 uM 130.5 110.5

As a result, as shown in Table 1 and Fig. 1, the extracts of Saga isoenzoides, fractions thereof, or the compounds of the formula (1) or (2), which were isolated from the fractions, It was found that the skin moisturization can be improved by increasing the biosynthesis amount of the bolucrin.

[ Experimental Example  2] UV-induced Gelatinase A ( MMP -9) and Gelatinase B ( MMP -2) to inhibit the biosynthesis and to measure the inhibitory effect

Human keratinocytes (Lonza, Switzerland) were cultured in a 24-well plate incubator at a concentration of 1 × 10 ⁴ , and irradiated with ultraviolet B at 30 mJ / cm ² after 24 hours. Each test substance was then treated with 10 μM of extracts and fractions Was replaced with a medium containing 10 ug / ml. After 48 hours of culture, the supernatant was harvested, harvested, and subjected to zymography using gelatin gel. The amount of MMP-2 and MMP-9 produced was measured by densitometry.

In order to measure the effect on the activity, the supernatant irradiated with ultraviolet ray B at 30 mJ / cm ² was dipped in gelatin gel, renaturation was carried out in buffer A for 30 minutes, After development, the degree of inhibition of activity was quantified by measuring the vest size of MMP-2 and MMP-9 using a densitometer. The values are 100 for the ultraviolet control group, and the comparative values are shown in Table 2.

Saga Island Ezoens  Extract, objection Fraction , Or In the fraction  MMP-2 &lt; RTI ID = 0.0 &gt; and MMP -9 expression and activity inhibition sample density Degree of expression inhibition Degree of activity inhibition MMP -2 (%) MMP -9 (%) MMP -2 (%) MMP -9 (%) Control group 100 100 100 100 Example <1-1> 10 [mu] g / ml 51.2 55.4 56.3 54.8 Example 2 10 [mu] g / ml 52.7 56.8 49.8 47.7 Example 3 10 [mu] g / ml 53.2 57.8 50.7 48.6 Example 4 10 uM 45.2 47.3 53.1 52.8 Example 5 10 uM 58.9 57.8 60.4 64.1

As a result, as shown in Table 2 and FIG. 2, the Sagas island ezoens extract, fractions thereof, or the compounds of formula (1) or (2) isolated from the fractions inhibited the biosynthesis or activity of MMP-2 and MMP-9 It is possible to prevent skin wrinkles, enhance elasticity and enhance skin adhesion by protecting or preventing the epidermal dermal boundary.

[ Experimental Example  3] Tumor necrosis factor alpha (UV) TNF -α) to measure biosynthesis inhibitory effect

Human keratinocytes (Lonza, switzerland) isolated directly from the newborn foreskin were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , irradiated with ultraviolet B at 30 mJ / cm ² , 10 μM of each test substance, In the case of the fraction, the medium was replaced with a medium containing 10 ug / ml. Cells were harvested 6-24 hours after incubation and the amount of tumor necrosis factor alpha (TNF-a) produced was quantitated using ELISA (Pharmingen 555212) method. The values are 100 for the ultraviolet control group, and the comparative values are shown in Table 3.

Saga Island Ezoens  By ultraviolet rays TNF -α inhibitory effect on biosynthesis sample density TNF -a biosynthesis (%) Untreated group 100 Wy-14,643 10 uM 40 Example <1-1> 10 [mu] g / ml 35 Example 2 10 [mu] g / ml 32 Example 3 10 [mu] g / ml 31 Example 4 10 uM 42 Example 5 10 uM 28

As a result, as shown in Table 3, it can be seen that the extract of Sagas island ezoens, fractions thereof, or the compounds of formula (1) or (2) isolated from the fractions reduced the biosynthesis of tumor necrosis factor alpha by UV.

[ Experimental Example  4] Type I by UV Collagenase Metalloftaez -1 (Metalloelastase-1, MMP -1) to inhibit biosynthesis and inhibition of collagen degradation

Human fibroblasts (Lonza, Switzerland), a primary cell directly isolated from the neonatal foreskin, were cultured in a 48 well plate incubator at a concentration of 10 ⁴ , and irradiated with ultraviolet A at 15 J / cm ² after 24 hours , 10 μM of each test substance or 10 μg / ml of each test substance. On the second day of culture, the supernatant was harvested and the amount of type I collagenase produced using the ELISA (AP biotech RPN2610) method was quantitated. The same supernatant was also used to quantify the remaining undissociated amount of procollagen using the ELISA (Takara MK101) method. The values were compared with those of ultraviolet irradiation group 100.

Saga Island Ezoens  The ultraviolet rays of the extracts and components MMP -1 for inhibiting biosynthesis sample
Type I collagenase biosynthesis (%) Amount of remaining procollagen (%) UV A group 100 100 Example < 1-1 > (10 ug / ml) 30 200 Example 2 (10 [mu] g / ml) 25 205 Example 3 (10 [mu] g / ml) 22 208 Example 4 (10 uM) 35 180 Example 5 (10 uM) 20 220

As a result, as shown in Table 4 and FIG. 3, the Sagas island ezoense extract, its fractions, or the compounds isolated from the fractions decreased the biosynthesis of type I collagenase by ultraviolet light, and the inflammation mediator (TNF- α) due to decreased biosynthesis. In addition, inhibition of the degradation of procollagen was inhibited by inhibiting the biosynthesis of MMP-1.

[ Experimental Example  5] affect skin elasticity Elastase  ( Elastase ) Effect on activity

Human leukocyte elastase (HNE, Sigma, E8140 (1 U)) was prepared by adding 1 ml of 0.2 M Tris-Cl (pH 8.0) to 1 vial to make 1 U / ml and incubating the substrate (N- (Methoxysuccinyl) Ala-Pro-Val 4-nitroanilide (M4765) MW 590.6) was diluted to 120 mM in DMSO and diluted 1:20 with 0.2 M Tris-Cl (pH 8.0) mM. The 96-well plate was incubated with 0.2 M Tris-Cl, test substance and enzyme for 10 min. After incubation at room temperature for 30 min, the amount of p-nitroanilide decomposed was measured at 405 nm.

A pancreatic pancreatic elastase (PPE, Sigma (E1250 (19 mg / ml)) is made up to 3X in a 0.2 M Tris-Cl (pH 8.0) After adding 96 μl of 0.2 M Tris-Cl, test solution and enzyme to 96 well plate, add 10 μl of 10 mM Tris-HCl (pH 7.4) After incubation, the substrate was placed and reacted at room temperature for 30 minutes. The amount of p-nitroanilide decomposed was measured at 405 nm.

The degree of inhibition of each enzyme was determined by the following formula.

Blank; Only the enzyme in the total reaction solution is excluded.

Control group; Includes elastase and substrate only (without sample to be tested).

Inhibition (%) = (1-B / A) X 100

(A: activity of the control group, B: activity of the sample containing the extract)

Saga Island Ezoens  Of extracts and ingredients Elastase  Active inhibitory effect sample
Human leukocyte elastase Pig pancreatic elastase Control group 100 100 Example < 1-1 > (10 ug / ml) 80 30 Example 2 (10 [mu] g / ml) 81 38 Example 3 (10 [mu] g / ml) 83 40 Example 4 (10 uM) 78 25 Example 5 (10 uM) 90 45

As a result, as shown in Table 5, the extracts of Saga isoenzoides, fractions thereof, or the compounds of formula (1) or (2) isolated from the fractions inhibited the elastase activity and prevented the collapse of the elastic fibers, Can be prevented.

[ Experimental Example  6] Saga Island Ezoens  Extract, objection Fraction  And In the fraction  Acute Toxicity Test of Orally Administered Compounds

The present inventors conducted the following experiments in order to examine the acute toxicity of the extract of Sagami isoenzene, the fractions thereof and the compounds isolated from the fractions. Acute toxicity tests were carried out using 6-week-old SPF SD rats. The extracts of Saga isozymes, their fractions and the compounds isolated from the fractions were each suspended in a 0.5% methylcellulose solution and administered once orally at a dose of 5 g / kg / 10 ml. Hematologic tests and blood biochemical tests were performed by observing the mortality, clinical symptoms, and weight changes of the animals after the administration of the test materials. The autopsy was performed to visually examine the abdominal organs and thoracic organs.

As a result of the experiment, there were no clinically symptomatic or dead animals in all animals treated with the test substance, and no toxic change was observed in weight change, blood test, blood biochemical test, and autopsy findings.

As a result, no toxic change was observed in rats up to 5 g / kg, and the oral dose of LD ₅₀ was found to be a safe substance of 5 g / kg or more.

The composition containing Sagitta mayoezone extract according to the present invention may be applied to various formulations as described below, but is not limited thereto.

[Formulation Example 1] Tablets

The granules were prepared by mixing 150 mg of Saga isoenzole extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate and 40 mg of ethanol was added thereto, followed by drying at 60 ° C. And tableted by tableting.

[Formulation Example 2]

Granules were formed by mixing 150 mg of Saga isoenzo extract, 100 mg of glucose, 50 mg of red ginseng extract and 600 mg of starch, and 100 mg of 30% ethanol was added thereto, followed by drying at 60 ° C to form granules, Respectively. The final weight of the contents was 1 g.

[Formulation Example 1] Lotion

Compounding ingredient weight% Saga Island Ezo Ense extract 3.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Water soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.10 Citric acid 0.05 1,3-butylene glycol 3.00 Purified water to 100

[Formulation Example 2] Pack

Compounding ingredient weight% Saga Island Ezo Ense extract 5.00 Polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Lauroylhydroxyproline 1.00 Water soluble collagen (1% aqueous solution) 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water to 100

[Formulation Example 3]

Compounding ingredient weight% Saga Island Ezo Ense extract 2.00 Hydroxyethylene cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 Concentrated glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 5.00 Purified water to 100

[Formulation Example 4] ointment

Compounding ingredient Content (% by weight) Saga Island Ezo Ense extract 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Squalane 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.0 Wax 4.0 Preservative, coloring, fragrance Suitable amount Purified water Balance

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (9)

A cosmetic composition for moisturizing comprising an extract of Sargassum yezoense as an active ingredient. A health food composition for moisturizing comprising an extract of Sargassum yezoense as an active ingredient. 3. The method according to claim 1 or 2,
Wherein the extract comprises at least one compound selected from the group consisting of the following compounds 1 and 2 or a salt thereof.
[Chemical Formula 1]

(2)

3. The method according to claim 1 or 2,
Wherein said composition comprises said Saggassum yezoense extract in an amount of 0.001 to 10% by weight based on the total weight of the composition. 3. The method according to claim 1 or 2,
Wherein the extract is a Saga isoenzene-C ₁ -C ₆ alcohol extract. 3. The method according to claim 1 or 2,
Wherein the composition increases or increases the biosynthesis of at least one protein selected from the group consisting of filaggrin and involucrin.
delete delete delete

Images