WO2022102869A1 - Composition for antioxidation, whitening, anti-wrinkle or anti-inflammation containing egg yolk oil and agastache rugose stem cells - Google Patents

Abstract

The present invention relates to a cosmetic composition, a pharmaceutical composition or a food composition for antioxidation, whitening, anti-wrinkle or anti-inflammation containing egg yolk oil and Agastache rugose stem cells as active ingredients. The composition comprising egg yolk oil and Agastache rugose stem cells as active ingredients of the present invention has excellent DPPH free radical scavenging activity and thus has an antioxidant effect, has a tyrosinase inhibitory effect to thus have a whitening effect as well as an effect of enhancing collagen activity to improve wrinkles, and has an anti-inflammatory effect by inhibiting the production of NO and inflammatory cytokines. In addition, the egg yolk oil and Agastache rugose stem cells of the present invention do not have cytotoxicity and side effects on the skin and thus can be safely used in cosmetics, pharmaceuticals, and food compositions.

Description

Antioxidant, whitening, anti-wrinkle or anti-inflammatory composition containing egg yolk oil and baechu-scented stem cells

The present invention relates to a cosmetic composition, a pharmaceutical composition or a food composition for antioxidant, whitening, anti-wrinkle or anti-inflammatory containing egg yolk oil and baecho-hyang stem cells as active ingredients.

Recently, with the improvement of living and income level, there is a lot of interest in improving the quality of life. In particular, interest in the development of bioactive substances for body diseases and metabolism, including antioxidants, has increased as a countermeasure for aging, various inflammatory diseases, and cancer (KB Kim, et al., Journal of Nutrition and Health. 50(5)) ;415-425 (2017)). In order to supply the necessary energy in the living body, biochemical oxidation reactions occur constantly, and reactive oxygen species (ROS), which are generated during this process, is the most stable form of oxygen, triplet oxygen, in the oxidation and reduction process. Singlet oxygen produced by superoxide anion, hydroxyl radical, free radical in an unstable state such as hydrogen peroxide and hydrogen peroxide (H2O2). Because it reacts easily with substances, if it is not removed from the body, oxidative stress is caused. Since this oxidative stress is linked to the inflammatory response, it induces lipid peroxidation in various metabolic processes and damages proteins, cell membranes, and DNA to induce cell aging and transformation, leading to stroke, cancer, arteriosclerosis, Alzheimer's disease, Parkinson's disease, etc. , atherosclerosis, etc. (Valko M, et al., Int J Biochem Cell Biol. 39:44-84 (2007); and Halliwell B, et al., NY, USA. 968 (1999)). Physiological antioxidant enzymes that remove the activated carbon thus generated to protect the living body include superoxide dismutase (SOD), catalase, glutathione-peroxidase (GSHpx) and glutathione S -Transferase (glutathione S-transferase, GST), etc., and α-tocopherol, β-carotene, ascorbic acid and glutathione are known as small molecule antioxidants or free radical scavenging agents (Kim SM, et al. ., Life Sci. 90 (21-22); 874-882 (2012)).

A person's skin color is determined by the concentration and distribution of melanin inside the skin. Melanin is synthesized in melanocytes in the epidermal layer of the skin. In the melanosome, an organelle in melanocytes, an enzyme called tyrosinase acts on tyrosine, a type of amino acid, to produce DOPA and dopaquinone. ) and then undergoes a non-enzymatic oxidation reaction. When such melanin synthesis occurs in the skin excessively, the skin tone is darkened, and spots, freckles, and the like occur. Therefore, by inhibiting the tyrosinase activity in the skin to inhibit the synthesis of melanin, not only can the skin tone be brightened to realize skin whitening, but also hyperpigmentation of the skin such as spots and freckles caused by UV rays, hormones and genetic causes It can improve sedation.

On the other hand, the inflammatory response is a biological defense reaction against external physical and chemical stimuli or bacterial infection, and nitric oxide (NO), which is called an inflammatory mediator, tumor necrosis factor (TNF)-α, interleukin (IL) Produces and secretes various inflammatory cytokines such as -1β, granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-6 (YC Yoo, et al., Journal of Life Science. 26(3);338-345 (2016)). In addition, by activating nuclear factor kappa B (NF-κB), an inflammatory transcription factor, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX- 2) is expressed, prostaglandin E2 (PGE2) is produced, and the secreted inflammatory cytokine activates NF-κB again to amplify the cytokine cascade and expand the inflammatory state. In particular, NO can exhibit antibacterial and antitumor effects, but when it is produced and secreted in excess due to pathological causes, it can lead to risks of septic shock and nerve tissue damage due to excessive vasodilation (S Han, et al., Immunopharmacol. Immunotoxicol. 35;34-42 (2013)). These inflammatory cytokines and inflammatory mediators play an important role in the inflammatory process, and proper regulation of their abnormal production has been proposed as a treatment for inflammatory diseases.

Therefore, in the cosmetic field, research and development of functional cosmetics having antioxidant, whitening, anti-wrinkle or anti-inflammatory effects are being actively conducted, and in particular, research using natural substances is continued in order not to cause toxicity or irritation to the skin. .

For example, Korean Patent Registration No. 10-1952943 reports the antioxidant effect of avocado seed extract, and Korean Patent Registration No. 10-1950384 discloses skin whitening containing extracts of horseradish extract, yellow lily extract, and rhododendron extract as active ingredients. A composition having an effect and anti-wrinkle effect has been described, and Korean Patent Registration No. 10-1949430 discloses the anti-inflammatory and anti-wrinkle effect of the ginseng extract.

On the other hand, egg yolk oil is an oily extract extracted from yolk (yolk) of bird eggs, and is called egg yolk oil, egg oil, egg oil, and the like. Egg yolk oil is known to contain lipids, vitamin A, vitamin E, linoleic acid, and the like, and lipids are known to consist of neutral lipids, phospholipids, sterols, and the like.

In addition, "Baechohyang (排草香, Agastacherugosa)" means a perennial plant belonging to the Lamiaceae family. It is an indigenous food resource that is wild or partially cultivated in the southern regions. In the present invention, the baecho scent by itself has little or no insecticidal effect, but when used in combination with pyrethrum, it serves as a kind of effect enhancer that maximizes the insecticidal effect possessed by the pyrethrum.

Accordingly, the present inventors continued their research to develop a new natural substance that has antioxidant, whitening, anti-wrinkle or anti-inflammatory effects while having fewer side effects on the human body. The present invention was completed by confirming the fact that it exhibits an inflammatory and anti-wrinkle effect.

The technical problem to be solved in the present invention is to provide a composition having excellent human stability while having antioxidant, whitening, anti-wrinkle or anti-inflammatory effects, including.

In order to solve the above technical problem, the present invention provides an antioxidant, whitening, anti-wrinkle or anti-inflammatory composition comprising egg yolk oil and baecho-hyang stem cells as active ingredients.

Preferably, in the present invention, the composition comprising egg yolk oil and baecho-hyang stem cells may be a cosmetic composition, a pharmaceutical composition, or a food composition.

In the present invention, the egg yolk contains isoflavone as a plant estrogen and lecithin, a component of human cell membrane, and the oil extracted from the egg yolk supplies nutrients to the skin and forms a protective film on the skin. Thus, it exhibits excellent moisturizing power and improves skin health.

The egg yolk oil is an oily substance extracted from algae eggs, and the egg yolk oil contains lecithin, phospholipids, vitamin A, vitamin E, and linoleic acid. The bird may be a chicken. Since ancient times, egg yolk oil has been known to be effective in stroke, heart disease, high blood pressure, dysentery, asthma, athlete's foot, prolapse, hemorrhoids, and nourishment and has been used as a folk remedy.

The egg yolk oil may be extracted by a socho method in which egg yolk oil is extracted by burning the yolk of a boiled egg, a solvent extraction method in which egg yolk oil is extracted using a separate solvent, an extraction method by a milking method, a supercritical extraction method, and the like. For example, it may be manufactured using the method described in Japanese Patent Application Laid-Open No. Hei 1-5474, Korean Patent Application Laid-Open No. 87-6843, Korean Patent Application Laid-Open No. 92-11388, and Korean Patent Publication No. 1995-14959. .

As used herein, the term 'stem cells' refers to master cells that can reproduce without limitation to form specialized cells of tissues and organs. Stem cells are pluripotent or pluripotent cells capable of developing. Stem cells can divide into two daughter stem cells, or one daughter stem cell and one derived ('transit') cell, which then proliferates into mature, complete cells of the tissue.

As used herein, the term "plant stem cell" is a cambium-derived plant stem cell. In particular, it includes pure plant stem cells (CMC Cambial Meristematic Cell) in a state that is not physically damaged at all in the cambium sandwiched on the boundary line between the xylem and the phloem of a plant.

As used herein, the term “callus” (or callus, or callus, callus, wound tissue, callus) is an undifferentiated amorphous cell mass, representative of which is a tumor tissue formed by meristems formed around the wound when a plant is wounded. am. Plants are largely composed of meristems that undergo cell division and permanent tissues that do not. When cells of meristems are first put in a nutrient medium and grown, callus is formed. After that, an embryonic embryo is formed and it differentiates into a plant body. These callus are often referred to as "plant stem cells". There is no limitation on the type of callus used in the present invention.

As used herein, the term “differentiation” refers to a phenomenon in which cells divide and proliferate and specialize in structure or function, that is, cells, tissues, etc. of living organisms change form or function to perform a given task. say changing In general, it is a phenomenon in which a relatively simple system is divided into two or more qualitatively different subsystems. For example, in ontogenesis, a distinction such as a head or a trunk arises between egg parts that are initially homogeneous, or a distinction arises between cells such as myocytes and nerve cells. Differentiation occurs, or as a result, a state of being divided into subdivisions or subsystems that can be distinguished qualitatively is called differentiation.

The present invention includes culturing plant stem cells or all types of induced pluripotent plant stem cells induced by various methods and preparing an extract therefrom, which can be performed according to a general method known in the art. For example, by culturing plant stem cells or all types of induced pluripotent plant stem cells induced by various methods, collecting the suspension after proteolytic enzyme treatment, or filtering the plant stem cells by reacting them at 65° C. for 2 hours, each Proteins derived from cells can be extracted.

According to one embodiment of the present invention, callus powder was used as baechohyang stem cells. There is no limitation on the callus powder that can be used in the present invention.

According to one embodiment of the present invention, it is preferable that the egg yolk oil and baechohyang stem cells are used in a weight ratio of 1:1 to 2:1.

The present invention provides an antioxidant, whitening, anti-wrinkle or anti-inflammatory composition comprising egg yolk oil and baecho-hyang stem cells as active ingredients.

In the present invention, the "active ingredient" refers to a component that can exhibit the desired activity alone or can exhibit activity together with a carrier that does not have the activity itself.

In the composition of the present invention, egg yolk oil and pear herb stem cells may be included in an amount of 0.005 to 50% by weight, more preferably 0.01 to 30% by weight, and most preferably 0.1 to 10% by weight based on the total weight of the composition. At this time, if the content of egg yolk oil and baecho-hyang stem cells is less than 0.005% by weight, the desired effect of the present invention cannot be obtained, and if it exceeds 50% by weight, the effect is not proportional as the content increases, so it may be inefficient and There is a problem in that the stability of the

The composition containing egg yolk oil and baecho-hyang stem cells of the present invention exhibits an antioxidant effect, a whitening effect, an anti-wrinkle effect and an anti-inflammatory effect, and has almost no cytotoxicity as a natural substance.

According to one embodiment of the present invention, there is provided a cosmetic composition comprising egg yolk oil and baecho-hyang stem cells as active ingredients.

In the cosmetic composition of the present invention, in addition to egg yolk oil and pear extract stem cells as active ingredients, components commonly added to cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances, such as conventional adjuvants, and carriers are additionally added. can be added.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , oil, powder foundation, emulsion foundation, wax foundation, spray, etc., but is not limited thereto. More specifically, it may be prepared in the form of a nourishing cream, an astringent lotion, a softening lotion, a lotion, an essence, a nutrition gel or a massage cream.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, gum tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. are used as carrier components. can be

When the formulation of the present invention is a powder or a spray, toss, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar or gum tracanth may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.

The cosmetic composition of the present invention further comprises a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, a film forming agent, water, Ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any conventionally used in cosmetics It may contain adjuvants commonly used in the field of cosmetics or dermatology, such as other ingredients of And, the above ingredients may be introduced in an amount generally used in the field of dermatology.

According to one embodiment of the present invention, there is provided a pharmaceutical composition comprising egg yolk oil and baecho-hyang stem cells as active ingredients.

The pharmaceutical composition according to the present invention includes a pharmaceutically acceptable carrier in addition to egg yolk oil and baechuhyang stem cells. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil; it's not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, preferably parenterally, and more preferably applied by topical application.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. can be The dosage of the pharmaceutical composition of the present invention is within the range of 0.001-100 mg/kg for adults. In addition, in the case of an external preparation, it is recommended to apply it once to 5 times a day in an amount of 1.0 to 3.0 ml based on an adult and continue it for at least 1 month. However, the dosage does not limit the scope of the present invention.

The pharmaceutical composition of the present invention is prepared in unit dose form or multi-dose by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person of ordinary skill in the art. It can be prepared by introducing into a container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersing agent or stabilizer.

According to one embodiment of the present invention, there is provided a food composition comprising egg yolk oil and baecho-hyang stem cells as active ingredients.

The food composition according to the present invention may further include egg yolk oil and pear-derived stem cells as active ingredients, as well as ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. there is.

Examples of the carbohydrate include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

For example, when the food composition of the present invention is prepared as a drink, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, cephalopod extract, jujube extract, licorice extract, etc. can be included

On the other hand, the egg yolk oil and baecho flavored stem cells of the present invention are natural substances, harmless to the human body, and have almost no toxicity and side effects, so they can be safely used even when used for a long period of time. can

As described above, the composition comprising egg yolk oil and baecho-hyang stem cells of the present invention as an active ingredient has excellent DPPH free radical scavenging activity, which has an antioxidant effect, and has a tyrosinase inhibitory effect to have a whitening effect as well as increase collagen activity. It has a wrinkle-improving effect, and has an anti-inflammatory effect by inhibiting the production of NO and inflammatory cytokines. In addition, the egg yolk oil and baecho-hyang stem cells of the present invention do not have cytotoxicity and skin side effects, so they can be safely used in cosmetics, pharmaceuticals and food compositions.

Hereinafter, examples and the like will be described in detail to help the understanding of the present invention. However, the embodiments according to the present invention may be modified in various other forms, and the scope of the present invention should not be construed as being limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Preparation of a composition comprising egg yolk oil and baechohyang stem cells

After boiling the eggs in boiling water at 100°C for 15 minutes, the shell and egg white of the eggs were removed and the yolk was prepared. After putting 1 kg of yolk in a stainless steel container, mashing it, and heating it at a temperature of 100 to 300 ° C for 5 to 10 minutes, the yolk swells, burns black, and becomes ashes. Carbonized components such as ash were removed, and oily egg yolk oil was collected in a separate container. 50 g of egg yolk oil was obtained based on 1 kg of egg yolk.

In this experiment, for baechohyang stem cells, commercially available callus powder was used.

A composition comprising the egg yolk oil and baechohyang stem cells in a weight ratio of 2: 1 was prepared.

<Test Example 1> Measurement of antioxidant effect

(1) DPPH (1-1-diphenyl-2-picryl-hydrazyl) radical electron donating ability measurement

100 μl of DPPH solution (5x10-4 M) was added to 100 μl of the composition obtained in Example 1 for each concentration and reacted for 30 minutes in an incubator at 25° C., and then absorbance was measured at 540 nm using a microplate counter. As controls, L-ascorbic acid and butylated hydroxyanisole were used. Electron donating ability was expressed as the absorbance decrease rate of the sample solution and the non-additive group.

As a result, the composition according to the present invention exhibited an antioxidant effect of 85.2% at a concentration of 500 μg/ml of egg yolk oil and baechohyang stem cells.

(2) Measurement of ABTS cation radical scavenging activity

7mM 2,2-azino-bis(3-ethyl-benthiazoline-6-sulfonic acid) and 2.4mM potassium persulfate were mixed and left at room temperature for 24 hours to form ABTS+ㅇ, diluted with ethanol and 100 μl of ABTS+ㅇ 100 μl of the sample was added to the flask, left for 1 minute, and absorbance was measured at 732 nm, and L-ascorbic acid was used as an experimental control. The radical scavenging ability was expressed as the value of the concentration of the sample required for reduction to 1/2.

As a result of ABTS+ㅇ measurement, the composition according to the present invention exhibited an antioxidant effect of 81.2% at a concentration of 50 μg/ml of egg yolk oil and baechohyang stem cells.

<Test Example 2> Measurement of whitening effect

Tyrosinase (Tyrosinase) inhibitory activity was measured according to the method of Yagi et al. In the reaction section, 0.2 mL of a substrate solution in which 10 mM L-DOPA was dissolved in 0.5 mL of 1/15 M sodium phosphate buffer (pH 6.8) and 0.2 mL of mushroom tyrosinase (110 U/mL) were added to a mixture of 0.1 mL of sample solution and reacted at 25 ° C for 2 minutes. and DOPA chrome generated in the reaction solution was measured at 475 nm. The tyrosinase inhibitory activity was expressed as the absorbance reduction rate of the group with and without the sample solution.

As a result of measuring tyrosinase inhibitory activity, egg yolk oil and baecho-hyang stem cells showed 76% tyrosinase inhibitory effect compared to arbutin at a concentration of 5,000 μg/ml.

<Test Example 3> Measurement of anti-inflammatory activity

Nitric oxide (NO) was measured as the amount of nitric oxide (NO) in the cell supernatant as nitrite (nitrite) and nitrate (nitrate). Griess reagent, which is a safe form after reduction with nitrate for nitrite, was used, and 2x106 cells in a 6 well plate were washed twice with PBS when the confluence was 80% and then cultured for more than 12 hours using a serum-free medium. Next, 10 μg/ml of lipopolysaccharide (LPS) was added to all wells except for the control group and stimulated. After 2 hours, samples were treated by concentration. The amount of NO production was measured by absorbance at 540 nm after collecting the supernatant after 24 hours and reacting with griess reagent for 10 minutes. The absorbance measured when the sample was added was converted by setting the amount of NO generated in the group to which only LPS was added as 100%. Dexamethasone (Dexa) was used as a positive control.

As a result of measuring nitric oxide (NO) inhibitory activity, egg yolk oil and baechohyang stem cells showed an excellent inhibitory ability of 55% at a concentration of 50 μg/ml.

<Test Example 4> Wrinkle improvement effect measurement

(1) Elastase inhibitory activity measurement

Elastase inhibitory activity was measured as follows. Prepare egg yolk oil and baecho-hyang stem cells to a certain concentration, take 0.5 ml each to test tube, add 0.5 ml of porcine pancreas elastase (2.5 Unit/ml) solution dissolved in 50 mM tris-HCl buffer (pH 8.6), and then 50 mM tris- Substrate N-succinyl-(L-Ala)3-p-nitroanilide (0.5 mg/ml) dissolved in HCl buffer (pH 8.6) was added and reacted for 20 minutes to generate p-nitroanilide from the substrate was measured at 405 nm. The elastase inhibitory activity was expressed as a decrease in absorbance in the group with and without the sample solution.

Elastase is an enzyme that degrades elastin, a protein, and is a non-specific hydrolase that can degrade collagen, another important matrix protein. In the dermal tissue of the skin, collagen and elastin related to the elasticity of the skin form a network structure. Elastase is decomposed by elastase and the bond of the skin's network structure is broken, and elastase is known as the main cause of wrinkles. . In addition, as one of the leukocyte granulation enzymes that decompose elastin in the body, the activity of the enzyme is extremely high in abnormal tissues, which directly causes tissue destruction, causing wrinkles and loss of elasticity of the skin.

For comparison, ursolic acid (UA) was used as a positive control. As shown here, the elastase inhibitory activity increased as the concentration of egg yolk oil and baechohyang stem cells increased, but showed insignificant activity compared to the control Ursolic acid.

(2) Collagenase inhibitory activity measurement

Collagenase inhibitory activity was measured as follows. That is, the reaction group was prepared by adding 4 mM CaCl ₂ to 0.1 M tris-HCl buffer (pH 7.5), 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (0.3 mg/ml) To a mixture of 0.25 ml of the dissolved substrate solution and 0.1 ml of the sample solution, 0.15 ml of collagenase (0.2 mg/ml) was added, left at room temperature for 20 minutes, and then 0.5 ml of 6% citric acid was added to stop the reaction, followed by ethyl acetate 1.5 ml was added and the absorbance was measured at 320 nm. Collagenase inhibitory activity was expressed as a decrease in absorbance between the group with and without the sample solution.

Collagen is mostly present in the dermal layer of the skin, and accounts for about 70-80% of the total dry weight of the skin, and plays a role in supporting the skin while occupying most of the extracellular matrix. However, collagen biosynthesis is reduced by internal factors such as a decrease in cellular activity due to natural aging, and degradation is accelerated by external factors such as an increase in stress caused by various harmful environments and an increase in reactive oxygen species by sunlight. As the skin matrix is destroyed, wrinkles are formed. This collagen decreases with age and photoaging by UV irradiation, which is known to be closely related to skin wrinkle formation, and thus the collagenase inhibitory activity of egg yolk oil and pear-oriented stem cells was measured. For comparison, ursolic acid (UA) was used as a positive control.

For comparison, ursolic acid (UA) was used as a positive control. As a result of measuring the collagenase inhibitory activity of egg yolk oil and baecho-hyang stem cells, the collagenase inhibitory activity was higher than 50% at a concentration of 500 ug/ml.

<Formulation Example 1> Preparation of cosmetic composition

(1) A cream of the following ingredients was prepared.

Ingredient name	content(%)
Purified water	59.45
glycerin	14
Butylene Glycol	4
Sorbitan Stearate	3
cetyl alcohol	2
Hydrogenated Poly (C6-14 Olefin)	2
PEG-32	2
Cetearylethylhexanoate	2
niacinamide	2
1,2-Hexanediol	2
cyclopentasiloxane	2
Phytosteryl isostearyl dimer dilinoleate	One
Macadamia Seed Oil	One
Glyceryl Stearate SE	One
Polyglyceryl-6 distearate	One
shea butter	0.5
tromethamine	0.35
Carbomer	0.3
Spices	0.2
egg yolk	0.1
baecho-hyang stem cells	0.1

(2) A toner of the following components was prepared.

Ingredient name	content(%)
Purified water	85.47550
Butylene Glycol	6.00000
propanediol	3.00000
pentylene glycol	3.00000
1,2-Hexanediol	2.00000
Ethylhexylglycerin	0.05000
Polyglyceryl-10 Laurate	0.18750
Polyglyceryl-4 Laurate	0.13750
Caprylyl/Capryl Glucoside	0.08750
Spices	0.06000
egg yolk oil	0.00100
baecho-hyang stem cells	0.00100

(3) A serum of the following components was prepared.

Ingredient name	content(%)
Purified water	81.718795
propanediol	10
glycerin	5
1,2-Hexanediol	1.2
Glyceres-26	0.7
Ammonium acryloyldimethyl taurate/vpicopolymer	0.2
panthenol	0.1
Butylene Glycol	0.0975
xanthan gum	0.08
Ethylhexylglycerin	0.6
Polyglyceryl-10 Laurate	0.0375
Polyglyceryl-4 Laurate	0.0275
Caprylyl/Capryl Glucoside	0.0175
tocopherol	0.00005
Spices	0.22
egg yolk oil	0.001
baecho-hyang stem cells	0.001

In the composition ratio, suitable ingredients are mixed with a preferred formulation example, but the ingredients or the mixing ratio may be arbitrarily modified according to regional and national preferences such as demanding class, demanding country, and use.

The above bar is only a preferred embodiment of the present invention, it is not limited to the present invention, and various modifications and changes may be possible in the present invention by those of ordinary skill in the art. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention are all included within the protection scope of the present invention.

Claims (6)

1. A cosmetic composition for antioxidant, whitening, anti-wrinkle or anti-inflammatory, comprising egg yolk oil and baecho-hyang stem cells as active ingredients.
2. The method of claim 1,

   Antioxidant, whitening, anti-wrinkle or anti-inflammatory cosmetic composition, characterized in that the egg yolk oil and baecho-hyang stem cells are mixed in a weight ratio of 1:1 to 2:1.
3. A pharmaceutical composition for antioxidant, whitening, anti-wrinkle or anti-inflammatory, characterized in that it contains egg yolk oil and baecho-hyang stem cells as active ingredients.
4. 4. The method of claim 3,

   Antioxidant, whitening, anti-wrinkle or anti-inflammatory pharmaceutical composition, characterized in that the egg yolk oil and baecho-hyang stem cells are mixed in a weight ratio of 1:1 to 2:1.
5. A food composition for antioxidant, whitening, anti-wrinkle or anti-inflammatory, characterized in that it contains egg yolk oil and baecho-hyang stem cells as active ingredients.
6. 6. The method of claim 5,

   Antioxidant, whitening, anti-wrinkle or anti-inflammatory food composition, characterized in that the egg yolk oil and baecho-hyang stem cells are mixed in a weight ratio of 1:1 to 2:1.