KR102403229B1 - Composition for improving skin containing Moringa extract and Opuntia humifusa extract thereof as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and an external preparation for skin for preventing or treating skin diseases, and a cosmetic composition and a food composition for preventing or alleviating skin diseases, all of which contains a Moringa oleifera extract and an Opuntia humifusa extract as active components. The composition can be used as a composition for preventing, alleviating or treating skin diseases through anti-inflammatory effects by promoting skin cell growth and reducing the expression of inflammatory cytokines.

Description

Composition for improving skin containing moringa extract and chrysanthemum extract as active ingredients {Composition for improving skin containing Moringa extract and Opuntia humifusa extract thereof as an active ingredient}

The present invention relates to a composition for improving skin containing a moringa extract and a chrysanthemum extract as active ingredients.

Moringa ( Moringa ) is a tree reaching a height of 5 to 10 m and is cultivated in Asia, Africa, Arabia, etc. It is rich in protein and vitamins, has high nutritional value, and has various pharmacological actions such as hyperglycemia and cancer. It is known to be abundant. Among the various parts of Moringa, the leaves are rich in β-carotene, protein, vitamin C, and calcium, and are used as antioxidants. Moringa seed extract also contains bioactive substances such as flavonoids, isothiocyanates, glucosinolates, and thiocarbonates, and has a liver function protective effect. The superiority of Moringa leaves has been revealed as apoptosis and proliferation inhibitory effects of cancer cells have been reported.

Opuntia humifusa is a cactus that can overwinter inland among the cacti of the genus Opuntia native to Korea, and is distinguished from the cactus baeknyeoncho, which is native to Jeju Island. Since the shape of the stem is similar to that of the palm, it is also called a palm cactus, and it is a fleshy perennial grass, that is, a perennial flowering plant. There are many shapes, such as a shape, a ball shape, and an oval shape, and they are classified according to the shape. It is known that by reducing its moisture by less than half, it can overwinter without freezing to death even at -20°C, so it can maintain its vitality. Cheonnyeoncho contains various nutrients important to the human body, such as phenolic substances, flavonoids, dietary fiber, vitamin C calcium, minerals and amino acids, and complex polysaccharides.

On the other hand, inflammation is an immune response of the human body in response to wounds or diseases, and oxidative stress such as ultraviolet rays, free radicals, and free radicals activates inflammatory factors to cause various diseases and skin aging. Vasoactive polypeptides such as kinin, plasmin, and complement act as vasodilators, contractiles, and chemotaxis, and in addition, lymphocytic such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for the inflammatory response. Arachidonic acid is metabolized to prostaglandin and leukotriene, which are inflammatory mediators, through two pathways, cyclooxygenase or lipooxygenase, and mediates various inflammatory responses.

Anti-inflammatory agents are those that remove inflammatory sources and reduce biological reactions and symptoms in order to eliminate inflammation. Substances that have been used for anti-inflammatory purposes so far include flufenamic acid, ibuprofen, benzydamine, indomethacin, etc. as a nonsteroidal type, and prednisolone as a steroid type. , and dexamethasone. In addition, allantoin, azene, hydrocortisone, etc. are known to be effective in anti-inflammatory, but these substances have limitations in usage due to safety issues for the skin, or have insignificant effects, so that no inflammatory relief effect can be expected. There is this.

In addition, in response to the recent well-being trend of eating well and living well, cosmetics for skin are becoming an essential element, not an option, as an industry with high added value. development is taking place. Recently, as problems due to genetic modification of cosmetic ingredients or environmental hormones have emerged, expectations for functional cosmetics for natural substances that are not harmful to the human body and can be safely used in the long term are increasing, and the demand is also increasing.

Accordingly, the present inventors have made efforts to develop a substance that is non-toxic from natural resources and has an excellent skin disease improvement effect, and as a result, a composition containing a low concentration of Moringa extract and chrysanthemum extract as an active ingredient promotes skin cell growth and By confirming that it exhibits an anti-inflammatory effect, and revealing that the composition can be used as a skin improvement composition, the present invention has been completed.

Republic of Korea Patent Publication No. 10-2020-0106893 (2020.09.15.) Republic of Korea Patent Registration No. 10-1771702 (2017.08.21.) Republic of Korea Patent Publication No. 10-2015-0020466 (2015.02.26)

It is an object of the present invention to provide a composition for alleviating skin wounds, regenerating skin, improving skin, or alleviating inflammatory skin diseases, containing a moringa extract and chrysanthemum extract as active ingredients.

In order to achieve the object of the present invention, the present invention has found that a composition containing a Moringa extract and an extract of chrysanthemum as an active ingredient has an anti-inflammatory effect by promoting the growth of skin cells and reducing the expression of inflammatory cytokines, It provides a pharmaceutical composition, quasi-drug, external preparation for skin, cosmetic composition and food composition for skin wound treatment, skin regeneration, or prevention, improvement or treatment of inflammatory skin disease containing Moringa and natural herb extract as active ingredients.

In addition, the present invention provides a quasi-drug for skin improvement, an external preparation for skin, a cosmetic composition and a food composition containing the moringa and natural herb extract as active ingredients.

The composition containing the moringa extract and chrysanthemum extract of the present invention as an active ingredient has a skin cell growth promoting effect and anti-inflammatory effect, so it can relieve skin wounds, regenerate skin, and prevent, improve or treat skin diseases including inflammatory skin diseases. It can be usefully used as a target drug, quasi-drug, cosmetic, food, etc.

1 is a diagram confirming the cytotoxicity of 1% and 5% moringa extract and cheonnyeoncho extract.
Figure 2 is a diagram confirming the skin cell growth promoting effect of 0.1%, 0.5% and 1% of Moringa extract and cheonnyeoncho extract.
3 is a diagram confirming the reduction effect of inflammatory cytokines in human skin keratinocytes of 0.5% and 1% of Moringa extract alone, Cheonnyeoncho extract alone, or Moringa extract and Cheonnyeoncho extract.
Figure 4 is a diagram confirming the reduction effect of inflammatory cytokines in macrophages of 1% Moringa extract and cheonnyeoncho extract.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art to which the present invention pertains can easily carry out the present invention. The embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art. Accordingly, the embodiment of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below.

Throughout the specification of the present invention, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

The present invention provides a pharmaceutical composition for treating skin wounds, regenerating the skin, or preventing or treating inflammatory skin diseases, containing an extract of Moringa and Opuntia humifusa as active ingredients.

As used herein, the term "prevention" refers to any action that suppresses the onset or delays the onset by administration of the composition.

As used herein, the term “treatment” refers to any action in which the symptoms of the disease are improved or beneficially changed by administration of the composition.

As used herein, the term “improvement” refers to any action that at least reduces a parameter related to the condition being treated, for example, the severity of symptoms.

As used herein, the term "administration" means providing a predetermined substance to a patient by any suitable method, and the route of administration of the composition of the present invention is oral through all common routes as long as it can reach the target tissue. or parenterally. In addition, the composition may be administered in a form mounted on any device capable of transporting the active substance to a target cell.

As used in the present invention, the term "extract" refers to an extract obtained by the extraction treatment of Moringa and Chrysanthemum, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, and a prepared or purified product of the extract. , or a mixture thereof, including extracts of all formulations that can be formed using the extract itself and the extract. The extract may be extracted from a natural, hybrid, or mutated plant of Moringa or Chrysanthemum, and may also be extracted from a plant tissue culture.

In the present invention, the moringa may be cultivated or commercially available without limitation. In addition, the Moringa may use one or more parts selected from the group consisting of flowers, leaves, branches, stems, roots, fruits and seeds of Moringa. Moringa may be specifically Moringa oleifera ( Moringa oleifera ).

In the present invention, the cultivated or commercially available cheonnyeoncho may be used without limitation. In addition, the Cheonnyeoncho may use one or more parts selected from the group consisting of flowers, leaves, stems, roots, fruits and seeds of Cheonnyeoncho.

In the present invention, the extract may use water, an organic solvent, or a mixed solvent thereof, and the organic solvent is a C ₁ to C ₅ polar solvent such as lower alcohol, ethyl acetate and acetone, and non-polar such as hexane and dichloromethane. A solvent or a mixed solvent thereof may be used, and preferably, it may be an extract extracted with water, C ₁ to C ₄ lower alcohol, or a mixed solvent thereof. The alcohol may be methanol, ethanol, propanol, butanol and isopropanol, preferably ethanol.

In addition, the mixed solvent may be a mixed solvent of water and C ₁ to C ₄ lower alcohol, and the concentration of C ₁ to C ₄ lower alcohol in the mixed solvent is 50 to 99%, more specifically 60 to 90%, 62 to 87%, 65 to 80%, 65 to 75%. Preferably, it may be a mixed solvent of water and ethanol, and the concentration of ethanol in the mixed solvent may be 50 to 99%, more specifically 60 to 90%, 62 to 87%, 65 to 80%, 65 to 75%. have.

As the extraction method, ultrasonic extraction, shaking extraction, Soxhelt extraction, or reflux extraction method may be used, but is not limited thereto. It is preferable to wash the extraction solvent and extract by adding 1 to 20 times the amount of well-dried moringa and chrysanthemum, and more preferably adding 5 to 20 times to extract, but is not limited thereto. In addition, the extraction time is preferably 1 to 72 hours, more preferably 2 to 48 hours, but is not limited thereto. In addition, the number of extractions is preferably 1 to 5 times, but is not limited thereto.

In the present invention, the moringa extract and chrysanthemum extract may be formulated in a ratio of 1 to 3 weight ratio of moringa extract 1: cheonnyeoncho extract. In more detail, the weight ratio of the moringa extract and the chrysanthemum extract may be 1: 1 to 2.8, 1: 1.2 to 2.6, 1: 1.5 to 2.5.

In the present invention, the moringa extract and chrysanthemum extract may have cytotoxicity at 5% by weight based on 100% by weight of the total pharmaceutical composition, so less than 5% by weight of the composition may be used, specifically, 100% by weight of the total pharmaceutical composition 4 wt% or less, preferably 0.000001 to 3 wt% or less, preferably 0.00001 to 1 wt% or less, more preferably 0.0001 to 1.5 wt%, most preferably 0.0001 to 1 wt% .

In the present invention, the composition can promote the growth of skin cells safely without toxicity to the human body, and has an anti-inflammatory effect due to a decrease in the expression of inflammatory cytokines TNF-α, IL-1β and IL-6. It may have the effect of treating wounds, regenerating the skin, or preventing or treating inflammatory skin diseases.

The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, chronic lichen simplex, intertidal dermatitis, exfoliative dermatitis, papular urticaria, psoriasis, solar dermatitis, allergic skin disease , systemic lupus erythematosus, acne, and acne may be any one or more skin diseases selected from the group consisting of.

In a specific embodiment of the present invention, the present inventors found that the composition comprising the Moringa extract and the chrysanthemum extract as an active ingredient has no cytotoxicity (see Fig. 1) and can promote skin cell growth (see Fig. 2), It was confirmed that there is an anti-inflammatory effect by suppressing the expression of inflammatory cytokines (see FIGS. 3 to 4). Therefore, the composition comprising the moringa extract and chrysanthemum extract as an active ingredient can be usefully used as a pharmaceutical composition for treating skin wounds, regenerating skin, or preventing or treating inflammatory skin diseases.

In addition, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, and drug of the patient. activity, sensitivity to drugs, administration time, administration route and excretion rate, duration of treatment, factors including concurrent drugs, and other factors well known in the medical field. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent, or may be used in combination with surgery, hormone therapy, drug therapy, and a biological response modifier, and may be administered simultaneously, separately, or sequentially with the agent, single or Multiple doses may be administered. Taking all of the above factors into consideration, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, weight, absorption of the active ingredient in the body, inactivation rate, excretion rate, disease type, and drugs used in combination, generally 0.1 mg to 100 mg per 1 kg of body weight, preferably 0.3 mg to 80 mg, may be administered daily or every other day, or divided into 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of obesity, sex, weight, age, etc., the dosage is not intended to limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intramuscularly, intravenously, intraperitoneally, subcutaneously, intradermally, or locally applied) according to a desired method, and the dosage may be determined by the patient's Although it varies depending on the state and weight, the degree of disease, the drug form, the route of administration and time, it may be appropriately selected by those skilled in the art.

In addition, the composition of the present invention can be delivered using a pharmaceutically acceptable carrier such as colloidal suspension, powder, saline, lipid, liposome, microspheres, or nanospherical particles. They may form a complex with a vehicle, linked to each other or contained in a carrier, with lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation reagents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances or fatty acids. The same can be delivered in/in vivo using delivery systems known in the art. In addition, pharmaceutically acceptable carriers include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, which are commonly used in the formulation. polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. In addition, a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. may be additionally included in addition to the above components.

In addition, when formulating the composition of the present invention, it may be formulated using usually used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, and these solid preparations include at least one excipient (starch, calcium carbonate, water Cross (sucrose) or lactose (lactose) or gelatin) and lubricant (magnesium styrate talc), etc. can be mixed and prepared. Liquid formulations for oral administration include suspensions, internal solutions, emulsions or syrups, and these liquid formulations are prepared by mixing at least one or more diluents (water, liquid paraffin), excipients, wetting agents, sweeteners, fragrances, preservatives, etc. can do. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, etc., and the non-aqueous solvents and suspensions include vegetable oils such as propylene glycol, polyethylene glycol, and olive oil; Injectable esters such as ethyl oleate may be used, and the suppository may include witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, and the like.

In addition, the present invention provides a quasi-drug for improving the skin or preventing or treating inflammatory skin disease containing a moringa extract and a chrysanthemum extract as active ingredients.

As used in the present invention, the term "quasi-drug" refers to items with a milder action than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals, for example, the Pharmaceutical Affairs Act. According to the article, quasi-drugs exclude products used for medicinal purposes, and may be products used for the treatment or prevention of diseases in humans or animals, or products with minor or no direct action on the human body.

The skin improvement may be any one or more skin improvement effects selected from the group consisting of skin wound alleviation, skin regeneration, skin wrinkle improvement, skin aging improvement, skin elasticity increase, or anti-inflammatory.

The quasi-drug composition is a body cleanser, disinfectant cleaner, cleaner, kitchen cleaner, cleaning agent, toothpaste, mouthwash, wet tissue, detergent, soap, hand wash, hair cleaner, hair softener, humidifier filler, mask, ointment, and filter filler from the group consisting of It can be prepared in the formulation of choice.

In addition, the present invention provides a skin external preparation for improving the skin or preventing or treating inflammatory skin diseases, which contains a moringa extract and a chrysanthemum extract as active ingredients.

As used herein, the term “external preparation” refers to a preparation provided for external use.

In the present invention, the external preparation for skin may be formulated in a dosage form selected from the group consisting of an external acid, an external tablet, an external solution, an ointment, a cream, a gel, a warning agent, a dressing, a patch, a spray, and a suppository.

In addition, the external preparation for skin may be a parenteral administration preparation formulated in the form of a solid, semi-solid, or liquid by adding a commercially available inorganic or organic carrier, excipient, and diluent. The preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions and emulsions, but is not limited thereto. Carriers, excipients and diluents that may be included in the external preparation include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the present invention provides a moringa extract and Provided is a cosmetic composition for improving skin or preventing or improving inflammatory skin disease containing a cheonnyeoncho extract as an active ingredient.

As used herein, the term "cosmetic composition" is a composition that is applied topically to human skin to improve the appearance and health of the skin without affecting the structure or function of the body.

In the present invention, the cosmetic composition is a skin-adhesive type cosmetic formulation, for example, basic product cosmetics (lotion, cream, essence, cleansing foam, cleansing water, pack, soap), body product cosmetics (body lotion, body oil, body gel, soap), cosmetics for color products (foundation, lipstick, mascara, makeup base), cosmetics for hair products (shampoo, conditioner, hair conditioner, hair gel), and the like. In addition, it may be prepared as a transdermal dosage form, for example, an ointment, a liquid, a dressing, a patch, or a spray, but is not limited thereto.

The cosmetic composition may include an acceptable carrier in a cosmetic formulation, and the carrier is a compound or composition that is already known and used that can be included in a cosmetic formulation, or a compound or composition to be developed in the future, which the human body adapts upon contact with skin. It means that there is no toxicity, instability or irritation beyond possible. The carrier may be included in an amount of about 1% to about 99.99% by weight based on the total weight of the cosmetic composition, preferably about 90% by weight to about 99.99% by weight of the weight of the cosmetic composition. However, since the ratio varies depending on the formulation as described above in which the composition for external application for skin of the present invention is prepared, its specific application site (face, neck, etc.) or its desired application amount, the ratio is viewed from any aspect. It should not be construed as limiting the scope of the invention.

On the other hand, as the carrier, alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscosity modifier, emulsion, stabilizer, sunscreen, color developer, fragrance, and the like may be exemplified. Compounds/compositions that can be used as alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizing agents, viscosity modifiers, emulsifiers, stabilizers, sunscreens, coloring agents, fragrances, etc. are already known in the art, so those skilled in the art An appropriate substance/composition may be selected and used.

In addition, the present invention provides a moringa extract and Provided is a food composition for improving skin or preventing or improving inflammatory skin disease containing a chrysanthemum extract as an active ingredient.

As used herein, the term "food composition" refers to any kind of composition that can be eaten and/or drunk without causing toxic symptoms in eating or drinking each composition.

As used in the present invention, the term "functional food" is the same term as food for special health use (FOSHU), and in addition to nutritional supply, medicine processed to efficiently exhibit bioregulatory functions, It is a food with high medical effect.

In the present invention, the food composition is beverage, meat, sausage, bread, biscuit, rice cake, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, soup, beverage, alcoholic beverage and vitamin complexes, dairy products and dairy products, and processed foods and health functional foods in a conventional sense, but is not limited thereto. The beverage composition is not particularly limited in other ingredients, and as in conventional beverages, various flavoring agents (natural flavoring agents, for example, thaumatin, stevia extract, rebaudioside A, glycyrrhizin, etc.; and synthetic flavoring agents, such as saccharin, aspartame, etc.) or natural carbohydrates (monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin; Ordinary sugars such as cyclodextrin and sugar alcohols such as xylitol, sorbitol, and erythritol) may be contained as additional ingredients. The proportion of the natural carbohydrate is generally from about 1 g to 20 g, preferably from about 5 g to 10 g per 100 g of the composition of the present invention.

In addition, in addition to the above, the food of the present invention is nutritional, vitamin, mineral (electrolyte), synthetic flavor, natural flavor, colorant, thickener (cheese, chocolate, etc.), pectic acid, facic acid salt, alginic acid, alginate, organic acid , protective colloidal thickener, pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent and pulp, etc. These components may be used independently or in combination, and the proportion of these additives may also be appropriately determined by those skilled in the art. can be selected. The proportion of the additive may be generally from about 0.1 g to 20 g, preferably from about 1 g to 20 g per 100 g of the composition of the present invention.

In addition, the health functional food may be used as it is, or may be used with other food or food ingredients by adding an active ingredient to food, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of its use (for prevention or improvement). In general, in the production of food or beverage, the health functional food of the present invention may be added in an amount of preferably 15% by weight or less, preferably 10% by weight or less, based on the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than or equal to the above range.

In the present invention, the content, content ratio, and the type of inflammatory skin disease of the moringa extract and chrysanthemum extract of the present invention in quasi-drugs, external preparations for skin, cosmetic compositions and food compositions for skin improvement or prevention, improvement or treatment of inflammatory skin diseases are The same as the description of the pharmaceutical composition for preventing or treating skin diseases, the specific description is incorporated herein by reference.

Hereinafter, the present invention will be described in more detail through Examples, Experimental Examples and Preparation Examples. However, the following Examples, Experimental Examples, and Preparation Examples are provided to aid the understanding of the present invention, and are not intended to limit the scope of the present invention.

<Example 1> Preparation of moringa extract and chrysanthemum extract

<1-1> Moringa ( Moringa oleifera ) extract preparation

Using 70% ethanol in 16 times the amount of Moringa leaves, extraction was performed at 60° C. for 10 hours, extracted twice, filtered under reduced pressure (Whatman filter paper #2), concentrated, and then freeze-dried to powder.

<1-2> Cheonnyeoncho ( Opuntia humifusa ) extract preparation

16 times the amount of 70% ethanol of the chrysanthemum sample was extracted at 55°C for 12 hours, extracted twice, filtered under reduced pressure (Whatman filter paper #2), concentrated, and then freeze-dried to powder.

<Experimental Example 1> Confirmation of cytotoxicity of moringa extract and chrysanthemum extract

In order to confirm the cytotoxicity of the Moringa extract and the chrysanthemum extract, the cell viability was confirmed after the human dermal cells were treated with the moringa extract and the chrysanthemum extract.

The experimental procedure was as follows:

1) Human dermal cells, HS68 cells, were cultured in RPMI-1640 medium (Gibco) containing 10% FBS (fetal bovine serum) at 37° C. and 5% CO ₂ conditions.

2) The HS68 cells cultured in step 1) were aliquoted in a 96-well plate at a concentration of 1×10 ⁴ cells/well, and cultured at 37° C. and 5% CO ₂ conditions for 18 hours.

3) Cell viability was confirmed by measuring the absorbance of the cultured HS68 cells in step 2). Specifically, in step 3), a 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho), 1:3 mixture of the moringa extract and chrysanthemum extract prepared in <Example 1> in step 3) (Moringa:Cheonnyeoncho) was treated at a concentration of 1% or 5%, respectively, and after culturing for an additional 72 hours, a cell viability assay kit (EX-cytox; DaeiLab Service, Korea) solution was added according to the manufacturer's procedure and ELISA Cell viability was confirmed by measuring the absorbance at 450 nm three times using a reader. The survival rates confirmed in Tables 1 to 2 are shown below (FIG. 1).

5% control Moringa: Millennial herb = 1:1 Moringa: Millennial herb = 1:2 Moringa: Millennial herb = 1:3 1 time 0.4155 0.078 0.0805 0.0715 100% 19% 19% 17% Episode 2 0.4205 0.0775 0.077 0.0715 100% 18% 18% 17% 3rd time 0.4055 0.0705 0.0645 0.0735 100% 17% 16% 18%

One% control Moringa: Millennial herb = 1:1 Moringa: Millennial herb = 1:2 Moringa: Millennial herb = 1:3 1 time 0.4935 0.377 0.3945 0.389 100% 76% 80% 79% Episode 2 0.3995 0.3315 0.3795 0.368 100% 83% 95% 92% 3rd time 0.3905 0.384 0.3785 0.3675 100% 98% 97% 94%

As a result, as shown in FIGS. 1 to 1, it was confirmed that 1% of the moringa extract and the chrysanthemum extract had no cytotoxicity.

<Experimental Example 2> Moringa extract And confirmation of skin cell growth promotion of extracts of chrysanthemum

In order to confirm the skin cell growth promoting effect of the Moringa extract and the chrysanthemum extract, the human skin keratinocytes were treated with the moringa extract and the chrysanthemum extract, and then the cell growth rate was measured by performing the MTT assay.

The experimental procedure was as follows:

1) After preparing in <Example 1>, freeze-dried to powder Moringa and ethanol extract 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho), 1:3 mixture (Moringa:Cheonnyeoncho) was weighed and dissolved in DMSO to prepare a sample.

2) Human skin keratinocytes, HaCaT keratinocytes, were seeded in a 96-well plate to 1×10 ⁴ cells/well and then cultured for 24 hours.

3) Remove the culture medium of HaCaT keratinocytes in step 2), and add a 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa) of the Moringa extract of step 1) and ch. :Cheonnyeoncho), 1:3 mixture (Moringa:Cheonnyeoncho) was mixed so as to be 1%, 0.5%, and 0.1%, respectively, and the cells were treated.

4) After 3 days of culture, the skin cell growth promoting effect was confirmed through MTT assay. Specifically, the medium was removed from the cells cultured for 3 days in step 4), 100 μl of MTT reagent was added and incubated for 30 minutes, and 150 μl of DMSO was added per well after removing the MTT reagent. It was incubated at ℃ for 15 minutes. Thereafter, the absorbance was measured at 540 nm using a microplate reader equipment, and the cell growth rate was calculated from the measured absorbance value and obtained.

As a result, as shown in FIG. 2, compared to the control group (control, ctrl) cultured only with the cell culture medium, the Moringa extract and the chrysanthemum extract (Moringa extract: cheonnyeoncho extract = 1:1, 1:2 and 1:3 ratio) In all three types, a cell growth promoting effect was observed at a low concentration of 0.1%. In addition, the 0.1% concentration showed a growth promoting effect of 15 to 22% compared to the control (see FIG. 2).

<Experimental Example 3> Anti-inflammation of moringa extract and chrysanthemum extract check the effect

qPCR was performed to confirm the anti-inflammatory effect of the moringa extract and chrysanthemum extract.

<3-1> Confirmation of anti-inflammatory in human keratinocytes (HaCaT keratinocytes)

Specifically, the amount of cytokine production in response to inflammatory stimulation induced by TNF-α and IFN-γ in human keratinocytes was measured by qPCR method.

The experimental procedure was as follows:

1) After preparing in <Example 1>, freeze-dried to powder A 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho), 1:3 mixture (Moringa:Cheonnyeoncho) of ethanol extract of Moringa and chrysanthemum was weighed and dissolved in DMSO to prepare a sample . In addition, in order to use as a comparative group, it was prepared in <Example 1> and then lyophilized and powdered. The samples were prepared by dissolving each of the ethanol extracts of moringa and chrysanthemum in DMSO.

2) HaCaT keratinocytes, which are human skin keratinocytes, were seeded in a 96-well plate to 1×10 ⁵ cells/well and then cultured for 24 hours.

3) After removing the culture medium of the cultured HaCaT keratinocytes in step 2), TNF-α and IFN-γ were added to a new culture medium at a concentration of 10 ng/ml, respectively, and the cells were treated and incubated for 1 hour.

4) 1 hour later, a 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho) of the Moringa extract and cheonnyeoncho extract of step 1) in cells stimulated with TNF-α and IFN-γ , 1:3 mixture (Moringa: cheonnyeoncho), moringa extract, and cheonnyeoncho extract were mixed so as to be 1% and 0.5%, respectively, and additionally treated with cells.

5) After culturing for 24 hours, mRNA was obtained from the cells, and the production amount of IL-1β, an inflammatory cytokine, was confirmed through qPCR. Specifically, in step 5), the medium was removed from the inflammatory cells for 24 hours, and after culturing the cells using a TRIZOL solution, 200 μl of chloroform was added and 15 at 12,000 x g. After centrifugation for minutes, the supernatant containing RNA was obtained and transferred to a new tube. 500 μl of isopropanol was added to the transferred supernatant and mixed, followed by precipitation of RNA at 80° C. for 16 hours. The precipitated RNA was obtained by centrifugation, and then washed with 70% EtOH/DEPC solution, the pellet was dissolved in 30 μl of DEPC water, and the RNA concentration was measured using a nanodrop device. The cDNA was synthesized using reverse transcriptase at 37°C, and the IL-1β expression level was measured by qPCR using the SYBR Green method using the synthesized cDNA as a template.

As a result, as shown in FIG. 3 , it was confirmed that the expression of IL-1β was increased upon stimulation of TNF-α and IFN-γ compared to the control group, so that an inflammatory response occurred by stimulation of TNF-α and IFN-γ. On the other hand, the Moringa and chrysanthemum mixture (Moringa extract: chrysanthemum extract = 1:1, 1:2 and 1:3 ratio) treatment group decreased the expression of IL-1β compared to the control group, It was confirmed that the mixed extract had an excellent effect of inhibiting IL-1β expression. In particular, it was confirmed that the effect was the most excellent in the group treated with Moringa extract: chrysanthemum extract = 1: 2 mixture. In addition, the moringa and chrysanthemum mixture (Moringa extract: chrysanthemum extract = 1:1, 1:2 and 1:3 ratio) treatment group had superior IL-1β expression inhibitory effect compared to the moringa extract and chrysanthemum extract alone treatment group As it appears to be, it was confirmed that the synergistic effect of the moringa and chrysanthemum extract of the present invention (FIG. 3).

<3-2> Confirmation of anti-inflammatory in macrophages

Specifically, the amount of cytokine production in response to the inflammatory stimulus induced by LPS in macrophages was measured by qPCR method.

The experimental procedure was as follows:

1) 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho), 1: 3 The mixture (Moringa: Cheonnyeoncho) was weighed and dissolved in DMSO to prepare a sample.

2) Macrophage RAW 264.7 cells After seeding in a 96-well plate to 1×10 ⁵ cells/well, the cells were cultured for 24 hours.

3) After removing the culture solution of RAW 264.7 cultured in step 2), 100 ng/ml of LPS (Lipopolysaccharide) was added to the fresh culture medium, respectively, to treat the cells, and incubate for 1 hour.

4) After 1 hour, 1:1 mixture (Moringa:Cheonnyeoncho), 1:2 mixture (Moringa:Cheonnyeoncho), 1:3 mixture (Moringa:Cheonnyeoncho), 1:3 mixture ( Moringa:Cheonnyeoncho) was mixed so as to be 1% each, and additionally treated with cells.

5) After culturing for 24 hours, mRNA was obtained from the cells, and the production amount of inflammatory cytokines TNF-α, IL-1β and IL-6 was confirmed through qPCR. Specifically, in step 5), the medium was removed from the inflammatory cells for 24 hours, and after culturing the cells using a TRIZOL solution, 200 μl of chloroform was added and 15 at 12,000 x g. After centrifugation for minutes, the supernatant containing RNA was obtained and transferred to a new tube. 500 μl of isopropanol was added to the transferred supernatant and mixed, followed by precipitation of RNA at 80° C. for 16 hours. The precipitated RNA was obtained by centrifugation, and then washed with 70% EtOH/DEPC solution, the pellet was dissolved in 30 μl of DEPC water, and the RNA concentration was measured using a nanodrop device. cDNA was synthesized using reverse transcriptase at 37° C., and the expression levels of TNF-α, IL-1β and IL-6 were measured by qPCR using the SYBR Green method using the synthesized cDNA as a template.

As a result, as shown in FIG. 4 , the expression of TNF-α, IL-1β, and IL-6 increased upon LPS stimulation compared to the control group, so it was confirmed that an inflammatory response occurred by LPS stimulation. And, all three samples significantly inhibited the expression of TNF-α, IL-1β and IL-6, and it was confirmed that there was an anti-inflammatory effect through this (FIG. 4).

Claims (13)

As a pharmaceutical composition for preventing or treating inflammatory skin diseases, comprising Moringa leaf extract and Opuntia humifusa extract as active ingredients,
The composition contains Moringa leaf extract: Cheonnyeoncho extract in a weight ratio of 1:1 to 3,
The moringa leaf extract and chrysanthemum extract are contained in an amount of 0.5 to 1% by weight based on 100% by weight of the total pharmaceutical composition,
The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, intertidal dermatitis, exfoliative dermatitis, urticaria papule, psoriasis, solar dermatitis, allergic skin disease, acne and acne. Any one or more selected from the group consisting of, the composition.
The method of claim 1,
The extract is an extract obtained by extracting C ₁ to C ₅ lower alcohol or a mixture thereof with a solvent, composition.
delete delete The method of claim 1,
The composition will have an anti-inflammatory effect, the composition.
6. The method of claim 5,
The anti-inflammatory effect is by reducing inflammatory cytokines, the anti-inflammatory effect appears, composition.
7. The method of claim 6,
The inflammatory cytokine is TNF-α, IL-1β and IL-6, the composition.
delete As a quasi-drug for the prevention or treatment of inflammatory skin diseases, containing moringa leaf extract and chrysanthemum extract as active ingredients,
The quasi-drug contains Moringa leaf extract: Cheonnyeoncho extract in a weight ratio of 1:1 to 3,
The moringa leaf extract and chrysanthemum extract are contained in an amount of 0.5 to 1% by weight based on 100% by weight of the total pharmaceutical composition,
The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, intertidal dermatitis, exfoliative dermatitis, urticaria papule, psoriasis, solar dermatitis, allergic skin disease, acne and acne. Any one or more selected from the group consisting of, quasi-drugs.
delete As an external preparation for the prevention or treatment of inflammatory skin diseases, containing moringa leaf extract and chrysanthemum extract as active ingredients,
The external preparation for skin contains Moringa leaf extract: Cheonnyeoncho extract in a weight ratio of 1:1 to 3,
The moringa leaf extract and chrysanthemum extract are contained in an amount of 0.5 to 1% by weight based on 100% by weight of the total pharmaceutical composition,
The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, intertidal dermatitis, exfoliative dermatitis, urticaria papule, psoriasis, solar dermatitis, allergic skin disease, acne and acne. Any one or more selected from the group consisting of, external preparation for skin.
As a cosmetic composition for improving skin or preventing or improving inflammatory skin disease, comprising a Moringa leaf extract and an extract of chrysanthemum as an active ingredient,
The composition contains Moringa leaf extract: Cheonnyeoncho extract in a weight ratio of 1:1 to 3,
The moringa leaf extract and chrysanthemum extract are contained in an amount of 0.5 to 1% by weight based on 100% by weight of the total pharmaceutical composition,
the skin improvement is anti-inflammatory,
The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, intertidal dermatitis, exfoliative dermatitis, urticaria papule, psoriasis, solar dermatitis, allergic skin disease, acne and acne. Any one or more selected from the group consisting of, the composition.
As a food composition for skin improvement or prevention or improvement of inflammatory skin disease, comprising a Moringa leaf extract and an extract of chrysanthemum as an active ingredient,
The composition contains Moringa leaf extract: Cheonnyeoncho extract in a weight ratio of 1:1 to 3,
The moringa leaf extract and chrysanthemum extract are contained in an amount of 0.5 to 1% by weight based on 100% by weight of the total pharmaceutical composition,
the skin improvement is anti-inflammatory,
The inflammatory skin diseases include inflammatory dermatitis, skin inflammation, acute/chronic eczema, atopic dermatitis, contact dermatitis, seborrheic dermatitis, intertidal dermatitis, exfoliative dermatitis, urticaria papule, psoriasis, solar dermatitis, allergic skin disease, acne and acne. Any one or more selected from the group consisting of, the composition.

Images