KR102292114B1 - Composition for Improving Skin Conditions Having Moisturizing, Anti-Inflammatory Caused by Fine Dust and Pore Shrinkage Property Comprising Plant Complex Extracts as Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for moisturizing the skin, alleviating fine dust-induced inflammation, and pore contraction comprising a complex extract of Euphrasia officinalis, Angelica archangelica, and Alchemillavulgaris as an active component. The complex extract of three kinds including Euphrasia officinalis, Angelica archangelica, and Alchemillavulgaris according to the present invention has no cell toxicity, exhibits a synergistic effect on an increase of hyaluronic acid synthesis, reduces the production of nitric oxide, which plays an important role in the skin inflammatory response and IL-6, an inflammatory cytokine induced by fine dust, and exhibits a synergistic effect on pore contraction, thereby being usefully used for the purpose of moisturizing the skin, treating and preventing inflammatory diseases, and reducing skin pores in the cosmetic, food, and pharmaceutical fields.

Description

Composition for Improving Skin Conditions Having Moisturizing, Anti-Inflammatory Caused by Fine Dust and Pore Shrinkage Property Comprising Plant Complex Extracts as Active Ingredient comprising a plant complex extract as an active ingredient }

The present invention relates to a composition for moisturizing skin, improving inflammation induced by fine dust, and reducing pores, and more specifically, to skin moisturizing, improving inflammation induced by fine dust, and It relates to a composition for reducing pores.

In general, a moisturizing cosmetic composition used for skin makes it soft and lively by maintaining a certain amount of moisture in human hair or skin, and serves to prevent damage such as cracking and dryness. That is, the cosmetic composition for moisturizing the skin is used for the purpose of beautifying the skin or hair by supplying a certain amount of moisture or maintaining the moisture to the skin or hair, and keeping the skin or hair healthy. The skin is an organ that plays an essential role in maintaining moisture, and it is known that it accounts for about 2/3 of the weight in moisture control. Therefore, in the cosmetic field, functional research related to the supply and maintenance of moisture, that is, moisturizing, is being actively conducted. In addition, in recent years, the development of cosmetics that increase the moisturizing power of the skin by introducing physiologically active substances obtained from natural products into cosmetics to further strengthen the inherent defense function of the skin itself is being actively developed.

In addition, the inflammatory response involves various inflammatory mediators and immune cells in local blood vessels and body fluids when cells or tissues are damaged or infected with external infectious agents such as bacteria, fungi, viruses, and various types of allergens, so that enzyme activation, It exhibits a series of complex physiological reactions such as secretion of inflammatory mediators, infiltration of body fluids, cell migration, and tissue destruction, and external symptoms such as erythema, edema, fever, and pain. The usual inflammatory reaction removes external infectious agents and regenerates damaged tissues to restore the function of living things, but if antigens are not removed or internal substances are the cause and the inflammatory reaction is excessive or continuous, it rather promotes mucosal damage. In some cases, it leads to diseases such as cancer.

It is known that various biochemical phenomena are involved as the cause of inflammation. In particular, nitric oxide synthase (NOS), an enzyme that generates nitric oxide (NO), and enzymes related to the biosynthesis of prostaglandins are It is known to play the most important role in mediating the inflammatory response. Therefore, blocking inflammation by inhibiting the activity of nitric oxide synthase (NOS), an enzyme that produces NO from L-arginine, or cyclooxygenase (COX), an enzyme involved in the synthesis of prostaglandins from arachidonic acid, is an anti-inflammatory treatment. is targeted for development.

In addition, recently, the problem of skin damage due to external environmental factors is increasing. According to the particle size, dust is divided into total suspended particles (TSP) with a particle size of 50 μm or less and particulate matter (PM) with a very small particle size. Then, fine dust is further divided into fine dust (PM10) with a diameter of less than 10 μm and ultrafine dust (PM2.5) with a diameter of less than 2.5 μm. When fine dust enters the body, immune cells remove and discharge the dust, and as a side effect, inflammation can accompany it. Fine dust penetrates into the pores of the skin as well as the respiratory tract, bronchi, and lungs, stimulating the skin, causing troubles and inflammation, further exacerbating the skin condition, and further negatively affecting chronic skin diseases such as atopic dermatitis. can give Therefore, blueberry and black rice extract (Patent Registration No. 10-2106440), Nahansong extract (Patent Publication No. 10-2019-0133634), etc. are presented as alternatives for the treatment and prevention of inflammatory diseases caused by fine dust However, research results of related natural substances for the treatment of inflammatory diseases caused by fine dust are still insignificant.

And, for cosmetic improvement of the skin, functional cosmetics for preventing or treating aging of the skin and cosmetics or food materials having an effect of reducing pores of the skin are widely consumed regardless of age or sex. A pore is a small hole through which the oil secreted from the sebaceous gland flows to the surface of the skin. There are ^{100-120 pores per cm 2 in the} skin, so the human face has about 20,000 pores. In general, the diameter of the pores is about 0.02~0.05mm, which is very small and usually inconspicuous, but if sebum secretion is excessive due to age, season, female menstrual cycle, pregnancy, stress, etc. becomes Skin with wide pores can make the face look messy, which has a negative cosmetic image. In addition, if the pores are open, bacteria can easily enter the hair follicles, and it is easy to cause skin troubles. Accordingly, many people became interested in pore reduction, and the management of pores using cosmetics became important.

Causes of enlarged pores include excessive sebum secretion, skin aging, and accumulation of waste products in the pores. In addition, ultraviolet rays promote sebum secretion, and people with oily or acne-prone skin generally secrete a lot of sebum, so the pores gradually increase.

In general, pores become larger from the late 20s, and as the skin ages, the activity of keratinocytes and fibroblasts decreases, collagen synthesis and elastin synthesis ability decrease, and water retention ability decreases. As it decreases, the elasticity of the skin decreases, resulting in enlarged pores. As the elasticity of the skin decreases, the collagen and elastic fibers that supported the pore wall gradually decrease, and as the force to tighten the pores decreases, the pores increase.

Accordingly, many studies have been conducted on compositions for reducing pores using natural extracts such as Wireungchae extract (Patent Registration No. 10-1273066) and ginseng fruit extract (Patent Registration No. 10-1704485), but they are still used in the field. It is necessary to develop a combination of natural materials with sufficient skin pore shrinking effect.

Under this background, the present inventors conducted research on substances having excellent antitussive and expectorant effects, and among the combinations of natural materials, the complex extracts of Eyebright, Angelica, and St. Mary's were excellent in skin moisturizing, anti-inflammatory, and pore-reducing activity. By identifying the effect, the present invention was completed.

Therefore, the main object of the present invention is to provide a cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient.

Another object of the present invention is to provide a food or pharmaceutical composition for skin moisturizing, anti-inflammatory, and pore reduction comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient.

The present inventors conducted a study on a combination of natural plant extracts with excellent skin moisturizing, anti-inflammatory, and pore-reducing effects, and the complex extracts of Eyebright, Angelica, and St. Mary's of the present invention have no cytotoxicity and increase hyaluronic acid synthesis It has a synergistic effect on the skin, directly inhibits the production of nitric oxide, which plays an important role in the skin inflammatory response, and IL-6, an inflammatory mediator induced by fine dust, and has a synergistic effect in reducing skin pores. could check

"Euphrasia officinalis " of the present invention means an annual grass of the genus ginseng family, "Angelica ( Angelica archangelica )" means a dicotyledonous plant Umbilicalaceae Buttercup, " Alchemilla vulgaris" )" means a perennial plant of the Rosaceae family of Rosaceae. The extracts of the eyebright, angelica, and St. Mary's plants of the present invention may be extracted from their natural, hybrid or varietal plants.

As used in the present invention, the term "extract" is an extract obtained by extraction treatment of eyebright, angelica, and St. Mary's plant, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or It includes extracts of all formulations that can be formed using the extract itself and the extract, such as mixtures thereof. The extract of the present invention may be prepared and used in the form of a dry powder after extraction. In the extraction of the eyebright, angelica, and St. Moth, the method of extracting the extract is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, reflux extraction, and the like, and these may be performed alone or in combination of two or more extraction methods.

In the present invention, the type of extraction solvent used to extract the eyebright, angelica, and St. Mary's is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water; C1-C4 lower alcohols, such as methanol, ethanol, propyl alcohol, and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Alternatively, a mixture thereof may be used, and preferably, water, lower alcohol, 1,3-butylene glycol, and ethyl acetate may be used alone or in combination of two or more.

In the present invention, the extract extracted with hot water or chilled extract is filtered to remove floating solid particles, for example, using nylon or the like to filter the particles, or filtered using freeze filtration, etc. It can be used by drying it using spray drying or the like.

As used herein, the term "fraction" refers to a result obtained by performing fractionation in order to separate a specific component or a specific component group from a mixture including various components.

Non-limiting examples of the fractionation method may include a method of obtaining a fraction from the extract by treating an extract obtained by extracting eyebright, angelica, and St. Mary's herb with a predetermined solvent. The kind of solvent used to obtain the fraction in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohol; and non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. In the case of using alcohol in the fractionation solvent, specifically C1 to C4 alcohol may be used.

The cosmetic composition of the present invention may be used in combination with the Eyebright, Angelica, and St. Mary's extracts. In the case of combining each of the components, the combination ratio is not particularly limited thereto, but the mixing ratio of the Eyebright, Angelica, and St. Mary's herbs is not particularly limited. Silver can be used in combination in a weight ratio of 1: 0.5 - 2: 0.5 - 2.

On the other hand, the cosmetic composition of the present invention is characterized in that it has a skin moisturizing activity by promoting the synthesis of hyaluronic acid. The term "moisturizing the skin" means to increase the feeling of moisture in the skin and to maintain a moist state. In a preferred embodiment of the present invention, it has been confirmed that the three types of complex extracts have a synergistic effect on the inhibition of nitric oxide production compared to the single extracts of Eyebright, Angelica, and St. Mary's (see FIG. 2).

In addition, the cosmetic composition of the present invention is characterized in that it has anti-inflammatory activity through inhibition of nitric oxide (NO) production. In a preferred embodiment of the present invention, it can be confirmed that the complex extract of Eyebright, Angelica, and St. Moth can effectively act in the treatment and prevention of inflammation-related diseases by directly inhibiting the production of nitric oxide, which plays an important role in the inflammatory response. There was (see Figure 3).

In addition, the cosmetic composition of the present invention is characterized in that it has anti-inflammatory activity by inhibiting the production of the inflammatory cytokine IL-6 caused by fine dust. In a preferred embodiment of the present invention, the complex extract of Eyebright, Angelica, and St. Mary's Cord shows a significantly improved IL-6 protein production inhibitory effect compared to their single extract treatment group, and It was confirmed that the combination had a synergistic effect in inhibiting the generation of IL-6, an inflammation-mediated cytokine induced by fine dust (see FIG. 4 ).

In addition, the cosmetic composition of the present invention is characterized in that it has skin moisturizing and anti-inflammatory activity as well as pore shrinking activity. In a preferred embodiment of the present invention, in order to confirm the pore-constricting effect of the complex extract of Eyebright, Angelica, and St. Mary's, the pore-constricting effect was tested using hemoglobin as a protein to replace the skin, and compared to each single extract. It was confirmed that these complex extracts showed an excellent synergistic effect in reducing pores.

In the cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction of the present invention, the composition is a mask pack, solution, ointment, cream, foam, nourishing lotion, softening lotion, softening water, emulsion, makeup base, essence, soap, liquid Containing detergents, bath products, sunscreen creams, sun oils, suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing agents, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays It may be prepared in a formulation selected from the group, but is not limited thereto.

In addition, the cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction of the present invention may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and as a common ingredient, for example, oil, Water, surfactant, humectant, lower alcohol, thickener, chelating agent, color, preservative, fragrance, and the like may be appropriately mixed, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction of the present invention varies depending on the formulation.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures thereof may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or a mixture thereof may be used as a carrier component. In particular, in the case of a spray, additional chlorophyll propellants such as fluorohydrocarbons, propane/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, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty ester, fatty acid ester of polyethylene glycol or sorbitan may be used. have.

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 esters and polyoxyethylene sorbitan esters; Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

When the formulation of the present invention is a soap, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolysates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. may be used as carrier components. can

In the cosmetic composition for skin moisturizing, anti-inflammatory, and pore-reducing of the present invention, the complex extract of eyebright, angelica, and St. Mary's is specifically included in dry weight, 0.0001% to 50% by weight of the total weight of the cosmetic composition. and, more specifically, may be included in an amount of 0.0005 wt% to 10 wt%. Within the above range, there is an advantage of exhibiting excellent efficacy for skin moisturizing, anti-inflammatory, and pore reduction, and there is an advantage in that the formulation of the composition is stabilized.

According to another aspect of the present invention, the present invention provides a food composition for skin moisturizing, anti-inflammatory, and pore reduction comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient. The complex extract of Eyebright, Angelica, and St. Mary's herb of the present invention has excellent skin moisturizing activity and at the same time exhibits excellent anti-inflammatory and pore-reducing activity. It can be usefully used as a composition. The complex extract of Eyebright, Angelica, and St. Mary's herb of the present invention and the effects on moisturizing, anti-inflammatory and pore reduction are the same as described above. The food composition may be used in the form of health functional food, but is not limited thereto.

The food composition of the present invention may be included in the form of a fraction of the complex extract of eyebright, angelica, and St. Mary's or a processed product thereof. In addition, the composition may include a food pharmaceutically acceptable food supplement additive in addition to the active ingredient.

In the present invention, "food supplementary additive" means a component that can be added to food as an auxiliary, added to the manufacture of health functional food of each formulation can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.

The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, the term “functionality” refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological actions with respect to the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.

In addition, the dosage form of the health functional food may also be manufactured without limitation as long as it is a dosage form recognized as a health functional food. The food composition of the present invention can be prepared in various types of dosage forms, and unlike general drugs, edible herbs are used as raw materials, so there are no side effects that may occur during long-term administration of the drug, and it is excellent in portability and can be applied to the skin. It can be taken as a supplement to increase water content, reduce the occurrence of inflammation, and enhance the effect of shrinking pores.

There is no limitation in the form that the health functional food of the present invention can take, and it may include any food in a conventional sense, and may be used in combination with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in the food according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes, and the like, and it can be prepared by adding to juice, tea, jelly, juice, etc. prepared by using the complex extract of eyebright, angelica, and St. Mary's herb according to the present invention as a main component. Also included are foods used as feed for animals.

According to another aspect of the present invention, the present invention provides an anti-inflammatory pharmaceutical composition comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient.

The complex extract of Eyebright, Angelica, and St. Mary's herb of the present invention can be usefully used as a pharmaceutical composition for the treatment or prevention of inflammation because it exhibits an excellent effect on inflammation. The complex extract and anti-inflammatory effect of eyebright, angelica, and motherwort of the present invention are the same as described above.

In the present invention, "treatment" means any action that improves or benefits an inflammatory disease by administration of the composition, and "prevention" means any action that inhibits or delays the onset of an inflammatory disease by administration of the composition.

In addition, in the present invention, "inflammatory disease" is a generic term for diseases with inflammation as the main lesion, and in the present invention, the inflammatory disease is a nitric oxide (NO)-mediated inflammatory disease. Preferably, the nitric oxide (NO)-mediated inflammatory disease is atopic dermatitis, psoriasis, sinusitis, rhinitis, conjunctivitis, asthma, dermatitis, inflammatory collagen vascular disease, glomerulonephritis, encephalitis, inflammatory enteritis, chronic obstructive pulmonary disease, sepsis, plaque Hematological shock, pulmonary fibrosis, undifferentiated spondyloarthrosis, undifferentiated arthropathy, arthritis, inflammatory osteolysis, chronic inflammatory disease caused by viral or bacterial infection, colitis, ulcerative colitis, inflammatory bowel disease, type 1 diabetes, type 2 diabetes, arthritis , rheumatoid arthritis, reactive arthritis, osteoarthritis, scleroderma, osteoporosis, atherosclerosis, myocarditis, endocarditis, pericarditis, cystic fibrosis, Hashimoto's thyroiditis, Graves' disease, leprosy, syphilis, Lyme disease, Borreliosis, Neurogenic-borreliosis, tuberculosis, sarcoidosis, lupus, classmate lupus, tuberculous lupus, lupus nephritis, systemic lupus erythematosus, macular degeneration, uveitis, irritable bowel syndrome, Crohn's disease, Sjogren's syndrome, fibromyalgia, chronic fatigue syndrome, chronic fatigue immunodeficiency syndrome, myalgia encephalomyelitis, amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, multiple sclerosis, autism spectrum disorder, attention deficit disorder, attention deficit hyperactivity disorder, etc., but are not limited thereto. .

In addition, the treatment and prevention of the inflammatory disease may be to reduce the expression of the inflammatory cytokine IL-6, the treatment and prevention of the inflammatory disease by reducing the expression of the inflammatory cytokine IL-6 caused by fine dust It may be to prevent and treat inflammatory diseases, but is not limited thereto.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. 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, and the like. 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, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like.

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 daily dose of the pharmaceutical composition of the present invention is, for example, 0.001-100 mg/kg.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. Alternatively, it may be prepared by being introduced into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may further include a dispersant or stabilizer.

According to another aspect of the present invention, the present invention provides an anti-inflammatory pharmaceutical composition containing the complex extract of eyebright, angelica, and St. Mary's as an active ingredient. Prevention of inflammatory diseases comprising administering to an individual other than humans or a method of treatment. As used herein, the term "administration" refers to the introduction of a given substance into a subject in an appropriate manner. As used herein, the term "individual" refers to all animals, including humans, rats, mice, livestock, etc., which have or can develop an inflammatory disease. As a specific example, it may be a mammal including a human.

The features and advantages of the present invention are summarized as follows:

(1) The present invention provides a skin moisturizing, anti-inflammatory, and pore-reducing composition comprising a complex extract of eyebright, angelica, and St. Mary's herb as an active ingredient.

(2) The three types of complex extracts composed of Eyebright, Angelica, and St. Mary's grass of the present invention have no cytotoxicity, have a synergistic effect on the increase in hyaluronic acid synthesis, and play an important role in the skin inflammatory response. Because it reduces the expression of the inflammatory cytokine IL-6 caused by dust and has a synergistic effect in reducing skin pores, the purpose of skin moisturizing, treatment and prevention of inflammatory diseases, and skin pores reduction in cosmetics, food, and pharmaceutical fields can be usefully used as

(3) In addition, the composition of the present invention is not only very safe for the human body, but also has excellent stability.

1 is a graph showing eyebright, angelica, and single or complex extracts of St. Mary's chrysanthemum treated by concentration in HaCaT cells, and cell viability was measured.
2 is a graph showing eyebright, angelica, and single or complex extracts of St. Mary's choi, treated by concentration in HaCaT cells, and measuring the amount of hyaluronic acid (Hyaluronan) produced.
Figure 3 is a graph showing eyebright, angelica, and single or complex extracts of St. Mary's Cordyceps treated by concentration in a macrophage cell line (Raw264.7 cells), and measuring changes in nitric oxide (NO) production.
4 is a graph showing the change in the production amount of IL-6 by treating the single or complex extracts of eyebright, angelica, and St. Mary's chords by concentration in HaCaT cells.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and therefore, the scope of the present invention should not be construed as being limited by these examples.

Example 1. Preparation of Eyebright, Angelica, and Virgin Mary extract

1-1. Preparation of Eyebright Extract

After pulverizing 100 g of dried eyebright seedlings, using 3 L of purified water as a solvent, heating and refluxing extraction for 12 hours, cooling, and filtering with filter paper having a 1.2 μm permeation size to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry eyebright powder.

1-2. Preparation of Angelica Extract

After grinding 100 g of dried angelica whole plant, 3 L of purified water was used as a solvent, followed by heating and reflux extraction for 12 hours, followed by cooling, followed by filtration with filter paper having a permeation size of 1.2 μm to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare an angelica dry powder.

1-3. Preparation of St. Mary's extract

After pulverizing 100 g of dried sagebrush, 3 L of purified water was used as a solvent, followed by heating and reflux extraction for 12 hours, followed by cooling, and filtration with filter paper having a 1.2 μm permeation size to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry powder of St. Moss.

1-4. Preparation of complex extracts of Eyebright, Angelica, and St. Mary's

100 g of mixed material mixed with dried Eyebright, Angelica, and St. Mary's outpost in the same amount was pulverized, then heated and refluxed for 12 hours using 3 L of purified water as a solvent, cooled and cooled, and filtered with a filter paper having a 1.2 μm permeation size Filtration gave a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry powder of three kinds of plant materials.

Example 2. Cell viability test according to the addition of plant extracts

The effect of single or complex extracts of Eyebright, Angelica, and St. Mary's Root on the growth of human immortalized keratinocytes (HaCaT) was confirmed by the following method.

^{First, human keratinocytes were inoculated in a 24-well cell culture dish at a concentration of 1×10 5} in DMEM (Dulbeccos modified Eagles medium) medium, which is a dedicated medium containing 10% FBS (fetal bovine serum, Cambrex), for 24 hours. , 37° C., 5% CO ₂ Incubated under humid conditions. Thereafter, the medium was removed and the single or complex extracts of Examples 1-1 to 1-4 diluted with serum-free DMEM medium were treated by concentration and cultured for 24 hours. 24 hours later, 1 mg/mL of MTT was treated, and 2 hours later, formazan generated by MTT treatment in cells was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, when human keratinocytes were treated with single or complex extracts of Eyebright, Angelica, and St. Mary's, it was confirmed that they did not significantly affect the cell viability of human keratinocytes, and of Eyebright, Angelica, and St. Mary's In the experimental group treated with the complex extract, the number of human keratinocytes was increased in a concentration-dependent manner.

Through the above results, it could be confirmed that the single or complex extracts of Eyebright, Angelica, and St. Mary's herb did not show toxicity to the human body (see FIG. 1 ).

Example 3. Skin moisturizing activity of eyebright, angelica, and virgin motherwort complex extract

In order to verify the skin moisturizing effect of the complex extracts of Eyebright, Angelica, and St. Mary's herb of the present invention, their effect on hyaluronic acid synthesis promotion was tested.

Hyaluronic acid (HA, Hyaluronan, Hyaluronic acid) is mainly synthesized by epidermal keratinocytes and dermal fibroblasts, and is a substance that plays an important role in creating an environment in which cells can function normally by combining with water. A decrease in hyaluronic acid in the skin is one of the direct causes of a decrease in skin elasticity and a decrease in moisture content. Therefore, in this experiment, the amount of hyaluronic acid produced in human fibroblasts was confirmed.

Human fibroblasts (NHDF) were inoculated with 1 ml of a culture solution in a 24-well cell culture dish at a concentration of about 1×10 ⁵ , and then cultured at 37° C., 5% CO ₂ environment for 24 hours. Thereafter, the single extracts of Eyebright, Angelica, and St. Mary's and their complex extracts prepared according to Example 1 were replaced with a medium containing 1, 10, and 100 μg/ml, respectively, and cultured for 24 hours. Thereafter, the culture medium was harvested and centrifuged for 5 minutes at 4°C and 7,500 rpm, and the expression level of Hyaluronan (Hyaluronan kit, R&D Systems, Inc., Minneapolis, MN, USA) was performed using an ELISA method. Change was confirmed.

As a result, referring to FIG. 2, compared with the single extract treatment group of Eyebright, Angelica, and St. Mary's, the complex extract treatment group improved hyaluronic acid by 105%, 103%, and 103%, respectively, at 1 μg/ml dose. Synthesis increased effect was exhibited, and hyaluronic acid synthesis increased by 123%, 127%, and 122% improved at the dose of 10 μg/ml, respectively, and 129%, 135%, and 127% at the dose of 100 μg/ml, respectively It exhibited an improved hyaluronic acid synthesis increasing effect (see FIG. 2).

Through the above results, it was confirmed that the synergistic effect on the increase of hyaluronic acid synthesis occurred according to the three types of mixture of Eyebright, Angelica, and St. Mary's grass of the present invention.

Example 4. Anti-inflammatory activity of eyebright, angelica, and virgin motherwort complex extract

4-1. Inhibition of nitric oxide production

In order to confirm the anti-inflammatory effect of the complex extract of Eyebright, Angelica, and St. Mary's herb of the present invention, the inhibition rate of the production of nitric oxide (NO), which is a representative cytotoxic substance involved in inducing inflammation, was measured.

First, RAW 264.7 cells, a macrophage cell line, were purchased from the Korea Cell Line Bank (KTCC, Seoul, Korea) and used, 10% FBS, 100 μg in DMEM (Dulbecco's modified Eagle's medium) containing 10% FBS (Fetal bovine serum). The cells were inoculated in a medium supplemented with /ml penicillin and 100㎍/ml streptomycin, and cultured at _{37°C, 5% CO 2 conditions.}

Specifically, the RAW 264.7 cells ^{were inoculated into a 96-well plate at a concentration (in DMEM) of 1×10 6} cells/mL and cultured for 24 hours, with a single extract of Eyebright, Angelica, and St. , A sample diluted to a concentration of 1, 10, and 100 μg/ml of these complex extracts was added, and a fresh medium containing LPS (1 μg/ml), known as an endotoxin, was treated and cultured for 24 hours. After that, 100 μl of the cell culture supernatant and 100 μl of Griess reagent [1% (w/v) sulfanilamide, 0.1% (w/v) naphtylethylenediamine in 2.5% (v/v) phosphoric acid] were mixed in a 96-well plate for 10 After reacting for a minute, the amount of nitric oxide produced was measured by measuring the absorbance at 540 nm using an ELISA reader. The concentration of the produced nitrite (nitrite) was calculated using a standard curve in which sodium nitrite (sodium nitrite) was dissolved in DMEM medium. Based on the difference in the amount of nitrite produced in the control group treated with LPS and the control group not treated with LPS, the nitric oxide production inhibitory activity of each sample was confirmed.

As a result of the experiment, it was confirmed that the three types of complex extracts had a better nitric oxide production inhibitory effect than the single extracts of Eyebright, Angelica, and St. Mary's.

Specifically, the complex extracts of Eyebright, Angelica, and St. Mary's were reduced by 40% (1㎍/㎖), 53% (10㎍/㎖) and 62% (100㎍/㎖), respectively, compared to the LPS-treated control group. By inhibiting the production of nitric oxide, it was observed that the extract showed a superior synergistic effect in inhibiting the production of nitric oxide compared to their single extract (see FIG. 3).

4-2. Inhibition of production of IL-6, an inflammatory mediator caused by fine dust

In order to confirm the effect of inhibiting the secretion of inflammatory cytokines induced by fine dust stimulation of the complex extract of Eyebright, Angelica, and St. Mary's seed according to the present invention, an experiment was performed on human immortalized keratinocytes (HaCaT). HaCaT cells were inoculated in a 96-well plate at a ^{concentration of 1.0×10 5} , and then cultured at 37° C., 5% CO ₂ humid conditions for 24 hours. After washing with PBS solution, 25 μg/ml of ultrafine dust (PM 2.5, Particulate Matter 2.5) with a diameter of less than 2.5 μm was added to each well individually or with samples (10, and 100 μg/ml) of various concentrations. treated together and incubated for 24 hours. The normal control group was treated with only the medium. After incubation, the supernatant was taken and the amount of IL-6 protein produced was analyzed using an enzyme-linked immunosorbent assay (ELISA, Enzyme-Linked Immunosorbent Assay) kit.

Referring to FIG. 4 , it was found that the generation of IL-6 protein induced by ultrafine dust was inhibited in a concentration-dependent manner in the group to which the three types of complex extracts of eyebright, angelica, and St. Mary's grass of the present invention were added. At the /mL concentration, it was confirmed that IL-6 protein production was inhibited by up to 62% compared to the sample untreated group. On the other hand, in the group treated with a single extract of Eyebright, Angelica, and St. Mary's herb, the IL-6 protein production inhibitory activity was found to be rather insignificant. In comparison, the three complex extract treatment groups of the present invention exhibited 58%, 57%, and 58% improved IL-6 protein production inhibitory activity, respectively.

Through the above results, the three types of complex extracts of Eyebright, Angelica, and St. Mary's seed of the present invention show a significantly improved IL-6 protein production inhibitory effect compared to their single extract treatment group, Eyebright, Angelica, And it was confirmed that there was a synergistic effect in inhibiting the production of IL-6, an inflammation-mediated cytokine induced by fine dust, due to the combination of St. Moss.

In summary, the complex extract of Eyebright, Angelica, and St. Mary's grass of the present invention directly inhibits the production of nitric oxide, which plays an important role in the inflammatory response, and the generation of IL-6, an inflammatory cytokine induced by ultrafine dust. It was confirmed that it can effectively act in the treatment and prevention of inflammation-related diseases.

Example 5. Pore shrinking activity of eyebright, angelica, and virgin motherwort complex extract

In order to confirm the pore-constricting effect of the complex extract of Eyebright, Angelica, and St. Mary's herb of the present invention, the pore-constricting effect was tested using hemoglobin as an experimental protein to replace the skin.

The skin is mostly composed of protein, and the protein components bind with polyphenols, pulling and contracting the pores. As such, the pore-contraction effect was measured by using the chemical astringent action in which sweat glands and pores are temporarily contracted by a protein-coagulating substance. Hemoglobin was used as a protein to replace the skin.

A hemoglobin solution (0.05 g/50 ml) was prepared by adding a 0.9% PBS (Phosphate Buffer Saline, 0.1 mM, pH 7.4) solution to hemoglobin, and to 2 ml of the hemoglobin solution, Eyebright, Angelica prepared in Example 1 , and a single or complex extract of St. Mary's was added at concentrations of 0.5%, 1%, and 2% (v/v), respectively.

Then, after shaking and mixing for 30 seconds, centrifugation was performed at 3,500 rpm for 10 minutes, and 2 ml of purified water was added to 1 ml of the supernatant, and absorbance was measured at 407 nm by UV-Visible spectrum. For the comparative experiment, a test group to which 2 ml of Phosphate Buffer Saline (PBS) was added instead of the sample of Example 1 was used as a negative control group. The amount of hemoglobin precipitation was measured according to [Equation 1] below and summarized in [Table 1], and it was determined that the higher the hemoglobin precipitation (%), the better the pore-contraction effect.

[Equation 1]

Pore contraction effect (%) = (1 - absorbance of test group / absorbance of control group) × 10

[Table 1]

Referring to [Table 1], it was confirmed that the hemoglobin precipitation level was higher in the test group treated with these three complex extracts compared to the single extracts of Eyebright, Angelica, and St. Mary's, and through this, the It was confirmed that the complex extracts of Evelight, Angelica, and St. Mary's herb had a synergistic effect in reducing skin pores.

Specifically, the complex extract of Eyebright, Angelica, and St. Mary's Cord had a hemoglobin precipitation of 22.1% at a 0.5% treatment concentration, a hemoglobin precipitation of 28.5% at a 1% treatment concentration, and a hemoglobin precipitation of 32.9% at a 2% treatment concentration. was observed, and it was observed that they showed a superior synergistic effect in reducing pores compared to their single extract.

Through the above results, the complex extract of Eyebright, Angelica, and St. Moth of the present invention has no cytotoxicity, exhibits a synergistic effect on the increase in hyaluronic acid synthesis, and the production of nitric oxide, which plays an important role in skin inflammatory response, and It was confirmed that it directly inhibited IL-6, an inflammatory mediator induced by fine dust, and furthermore, had a synergistic effect in reducing skin pores.

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

Eyebright, angelica, and a complex extract formulated in a weight ratio of 1:0.5-2:0.5-2,
The complex extract is characterized in that it has a skin moisturizing activity by promoting the synthesis of hyaluronic acid,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of nitric oxide (NO) production,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of the production of inflammatory cytokine IL-6 caused by fine dust,
A cosmetic composition for skin moisturizing, anti-inflammatory, and pore reduction. delete delete delete delete Eyebright, angelica, and a complex extract formulated in a weight ratio of 1:0.5-2:0.5-2,
The complex extract is characterized in that it has a skin moisturizing activity by promoting the synthesis of hyaluronic acid,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of nitric oxide (NO) production,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of the production of inflammatory cytokine IL-6 caused by fine dust,
A food composition for skin moisturizing, anti-inflammatory, and pore reduction. Eyebright, angelica, and a complex extract formulated in a weight ratio of 1:0.5-2:0.5-2,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of nitric oxide (NO) production,
The complex extract is characterized in that it has anti-inflammatory activity through inhibition of the production of inflammatory cytokine IL-6 caused by fine dust,
Pharmaceutical composition for anti-inflammatory.

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