KR102563733B1 - Composition for wound treatment or skin regeneration containing Aloe vera flower extract as active ingredients - Google Patents

Abstract

The present invention relates to a composition for wound treatment or skin regeneration containing an aloe vera flower extract as an active ingredient, wherein the aloe vera flower extract of the present invention significantly increases migration of dermal fibroblasts, which is an important feature in wound healing Since it does not exhibit cytotoxicity even at high concentrations, it can be usefully used as an active ingredient in a composition for treating skin wounds or regenerating skin.

Description

Composition for wound treatment or skin regeneration containing Aloe vera flower extract as active ingredients {Composition for wound treatment or skin regeneration containing Aloe vera flower extract as active ingredients}

The present invention relates to a composition for wound healing or skin regeneration containing an aloe vera flower extract as an active ingredient.

The skin is a tissue that covers the outermost part of the human body and serves as a primary defense function to protect the body from external chemical and physical shocks by protecting the human body and functioning as a barrier. The skin is composed of three layers: the epidermis, the dermis, and the subcutaneous layer. When the skin is aged by ultraviolet rays, stress, etc., or damaged by wounds, its function is weakened, and at this time, the skin regeneration process begins. The skin regeneration process begins with an acute inflammatory response, then undergoes contraction of connective tissue through proliferation, migration, and proliferation of epidermal cells, generation of new blood vessels, and synthesis of collagen. At this time, migration and proliferation of keratinocytes and dermal fibroblasts, which occupy more than 90% of the epidermis, not only provide wound healing and skin regeneration, but also play an important role in promoting skin elasticity and forming a basement membrane by inducing collagen synthesis.

Aloe is a plant belonging to the genus Aloineae of the family Liliaceae, which lives in the tropics, and is used worldwide for food and medicine. Among them, the most well-known material is Aloe vera, and juice or gel processed from its leaves is one of the representative health functional foods, and is also one of the natural materials widely used as pharmaceutical or cosmetic materials. Aloe vera gel is about 99-99.5% water and about 0.5-1.0% solids, of which polysaccharides are 55%, sugars 17%, minerals 16%, proteins 7%, fats 4%, and phenols 1%. has been The polysaccharide that accounts for a large portion of the solid content of the aloe vera gel is the main physiologically active component of the aloe vera gel, and typically includes acetylated mannan (or acemannan).

On the other hand, as a prior art using aloe vera, Korean Patent Publication No. 10-2020-0140520 describes the respiratory disease improvement effect of aloe vera gel, and Korean Patent Publication No. 10-2016-0086809 discloses the ratio of aloe vera extract The treatment effect of alcoholic chronic fatty liver is disclosed, and Korean Patent Registration No. 10-1520643 only discloses the treatment effect of aloe vera gel on diabetes, but nothing is known about its activity against aloe vera flowers.

Accordingly, the present inventors have made efforts to develop a natural product-derived wound healing agent. As a result, it was confirmed that the aloe vera flower extract significantly increases the migration of dermal fibroblasts, which is an important feature for wound healing, and does not exhibit cytotoxicity even at high concentrations, By revealing that it can be used as an active ingredient of a skin wound treatment or skin regeneration composition, the present invention was completed.

An object of the present invention is to provide a composition for skin wound treatment or skin regeneration containing an Aloe vera flower extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for wound treatment or skin regeneration containing an Aloe vera flower extract as an active ingredient.

In addition, the present invention provides a cosmetic composition for skin regeneration containing an aloe vera flower extract as an active ingredient.

In addition, the present invention provides a food composition for skin regeneration containing an aloe vera flower extract as an active ingredient.

Since the Aloe vera flower extract of the present invention significantly increases the migration of dermal fibroblasts, which is an important feature in wound healing, and does not exhibit cytotoxicity even at high concentrations, it is an active ingredient of a composition for treating skin wounds or skin regeneration. can be useful

1 is a diagram confirming cell viability in NHDF cells treated with water, 50% ethanol (50% EtOH), and 100% ethanol extract (EtOH) of Aloe vera flowers through MTT assay.
Figure 2 is a diagram confirming migration of NHDF cells for 72 hours in order to confirm the wound healing activity of water, 50% ethanol (50% Ethanol), and 100% ethanol extract (Absolute Ethanol) of Aloe vera flowers.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art. Accordingly, the embodiments of the present invention can be modified in many different 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 it may further include other components without excluding other components unless otherwise stated.

The present invention provides a pharmaceutical composition for wound treatment or skin regeneration containing an Aloe vera flower extract as an active ingredient.

As used herein, the term "extract" refers to an extract obtained by the extraction treatment of the aloe vera flower, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or mixtures thereof, and extracts of all formulations that can be formed using the extract itself and the extract. The extract may be extracted from natural, hybrid, or mutant plants of Aloe vera flowers, and may also be extracted from plant tissue culture.

In the present invention, cultivated or commercially available aloe vera flowers may be used without limitation.

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 lower alcohol, a polar solvent such as ethyl acetate and acetone, and a non-polar solvent such as hexane and dichloromethane. A solvent or a mixed solvent thereof may be used, and preferably 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. 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 perform extraction by adding 1 to 20 times the amount of washed and well-dried aloe vera flowers with the extraction solvent, and more preferably to perform extraction by adding 5 to 20 times, but is not limited thereto. In addition, the extraction time is preferably 1 to 72 hours, more preferably 1 to 48 hours, but is not limited thereto. In addition, the number of times of extraction is preferably 1 to 5 times, but is not limited thereto.

In addition, the composition of the present invention is any one selected from the group consisting of solutions, suspensions, granules, tablets, capsules, pills, extracts, emulsions, syrups, gels, pastes, ointment frosts, powders, emulsions and aerosol forms. It is preferable that it is a formulation, but it is not limited thereto.

In a specific embodiment of the present invention, the present inventors confirmed the effects of water, 50% ethanol and 100% ethanol extract of aloe vera flowers on NHDF cell migration, and as a result, 50% ethanol extract and 100% ethanol extract of aloe vera flowers were significant. It was confirmed that the wound closure rate was increased. In addition, it was confirmed that it does not exhibit cytotoxicity at high concentrations (see FIGS. 1 and 2).

Therefore, the aloe vera flower extract of the present invention significantly increases the migration of dermal fibroblasts, which is an important feature in wound healing, and does not exhibit cytotoxicity even at high concentrations, so it is useful as an active ingredient in a composition for treating skin wounds or skin regeneration. can be used

The pharmaceutical composition according to the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient. The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention is one commonly used in formulation, and carbohydrate compounds (eg, lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, cellulose, etc.) ), gum acacia, calcium phosphate, alginates, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, salt solutions, alcohol, gum arabic, vegetable oils (e.g. corn oil, cotton) seed oil, soy milk, olive oil, coconut oil), polyethylene glycol, methyl cellulose, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate and mineral oil, and the like, but are not limited thereto.

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 can be administered by various routes such as oral or parenteral, and all modes of administration can be expected, for example, it can be administered by oral, rectal or intravenous, intramuscular, or subcutaneous injection. there is. At this time, as a parenteral route, transdermal administration is preferred, and among them, topical application by application is most preferred.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and response sensitivity. It can be. The oral dosage of the pharmaceutical composition of the present invention is within the range of 0.001-100 mg/kg (body weight) for adults. In addition, in the case of an external preparation, it can be applied 1 to 5 times a day in an amount of 1.0 to 3.0 ml based on adults and continued for 1 month or longer. However, the dosage is not limited to the scope of the present invention.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by those skilled in the art. or it may be prepared by incorporating into a multi-dose container. At this time, 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 additionally contain a dispersing agent or stabilizer.

In addition, the composition of the invention may be an external agent. External skin preparations may be formulated as a dosage form selected from the group consisting of external powders, external tablets, external solutions, ointments, creams, gels, warning agents, dressings, patches, sprays, and suppositories.

In addition, the external skin preparation may be a parenteral preparation formulated in a solid, semi-solid or liquid form 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, gum acacia, 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 cosmetic composition for skin regeneration containing an Aloe vera flower extract as an active ingredient.

Since the aloe vera flower extract of the present invention significantly increases the migration of dermal fibroblasts, which is an important feature in wound healing, and does not exhibit cytotoxicity even at high concentrations, it can be usefully used as an active ingredient in a cosmetic composition for skin regeneration.

As used herein, the term "cosmetic composition" is a composition that is topically applied 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 cosmetic formulation, for example, basic product cosmetics (toilet, cream, essence, cleansing foam, cleansing water, pack, soap), body product cosmetics (body lotion, body oil, Body gel, soap), color product cosmetics (foundation, lipstick, mascara, makeup base), hair product cosmetics (shampoo, conditioner, hair conditioner, hair gel), etc. In addition, transdermal dosage forms such as ointments, liquids, dressings, patches, or sprays may be prepared, but are not limited thereto.

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

Meanwhile, as the carrier, alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscosity modifier, emulsion, stabilizer, sunscreen, coloring agent, fragrance, and the like may be exemplified. Compounds/compositions that can be used as alcohols, oils, surfactants, fatty acids, silicone oils, humectants, humectants, viscosity modifiers, emulsions, stabilizers, sunscreens, coloring agents, fragrances, etc. are already known in the art, so those skilled in the art An appropriate corresponding material/composition can be selected and used.

In addition, the present invention provides a food composition for skin regeneration containing an Aloe vera flower extract as an active ingredient.

Since the aloe vera flower extract of the present invention significantly increases the migration of dermal fibroblasts, which is an important feature in wound healing, and does not exhibit cytotoxicity even at high concentrations, it can be usefully used as an active ingredient in a food composition for skin regeneration.

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

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

In the present invention, the food composition is a beverage, meat, sausage, bread, biscuits, rice cake, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, soup, beverages, alcoholic beverages and vitamin complexes, dairy products and milk-processed products, processed foods in a conventional sense, and health functional foods, but are not limited thereto. The beverage composition is not particularly limited in other ingredients, and as in conventional beverages, various flavors (natural flavors such as thaumatin, stevia extract, rebaudioside A, glycyrrhizin, etc.; and synthetic flavors, such as For example, saccharin, aspartame, etc.) or natural carbohydrates (monosaccharides, such as glucose, fructose, etc.; disaccharides, such as maltose, sucrose, etc.; and polysaccharides, such as dextrin, Conventional sugars such as cyclodextrin and sugar alcohols such as xylitol, sorbitol, and erythritol) may be contained as additional components. The proportion of the natural carbohydrate is generally about 1 g to 20 g, preferably 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 contains nutrients, vitamins, minerals (electrolytes), synthetic flavors, natural flavors, colorants, enhancers (cheese, chocolate, etc.), pectic acid, pectic acid salts, alginic acid, alginates, organic acids. , protective colloidal thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonating agent and fruit flesh, etc. These components may be used independently or in combination, and the ratio of these additives may also be determined appropriately by those skilled in the art. can be chosen The proportion of the additives may be generally about 0.1 g to 20 g, preferably about 1 g to 20 g per 100 g of the composition of the present invention.

In addition, the health functional food may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined depending on the purpose of its use (for prevention or improvement). In general, when preparing 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 the purpose of health and hygiene or health control, the amount may be less than the above range.

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

chemicals and reagents

Dried Aloe vera (A. vera ) flowers (UV-AVF1001) were provided by Univera Co., Ltd. (Seoul, Korea). DMEM (Dulbecco's modified Eagle's medium) medium, fetal bovine serum (FBS), penicillin and streptomycin were purchased from Gibco-BRL (Grand Island, NY, USA).

cell culture

Normal human dermal fibroblasts (NHDF) were purchased from the Korean Cell Line Bank (Seoul National University, Korea). All of the cells were cultured in high-glucose DMEM (Thermo Scientific, Waltham, MA) containing 10% FBS and 1% penicillin-streptomycin (PS), and the cultured cells were humidified at 37°C and 5% CO _{2 .} kept in \. In addition, NHDF cells used in the present invention were used between passage numbers 5 and 10.

statistical analysis

Results are expressed as mean ± standard error from three independent experiments, and statistical analysis was performed using GraphPad Prism 5 (GrahPad Software Inc., La Jolla, CA, USA). In addition, One-way analysis of variance was followed by Newman-Keuls multiple comparison test, and p < 0.05 was considered statistically significant.

<Example 1> Preparation of aloe vera flower extract

1 g of dried and powdered aloe vera flowers were put into 10 mL of 100% ethanol, 50% ethanol, and water, respectively, and extracted with ultrasonic waves at 40° C. for 60 minutes, followed by centrifugation and filtration. Thereafter, each extract was concentrated under reduced pressure using a rotary vacuum concentrator and then lyophilized. Each extract prepared was stored at -4°C until use.

<Example 2> Sample preparation and cell treatment

Each extract prepared in <Example 1> was dissolved in PBS, and filtered to make a stock solution at a concentration of 10 mg/mL. In addition, stock solutions were diluted to concentrations of 25 μg/mL and 100 μg/mL in serum-free media and used.

On the other hand, in the cell treatment, NHDF cells were washed twice with PBS at 80% confluency, treated at the desired concentration, and then cultured for 24 hours and 48 hours, respectively. In addition, 10 μg/mL madecassoside was used as a positive control (PC).

<Example 3> Confirmation of cell viability and cell proliferation effect

MTT assay was performed to confirm cell viability. Specifically, NHDF (3 × 10 ³ cells/well) cells were seeded in a 96-well plate. Then, after processing 25 μg/mL and 100 μg/mL of the samples prepared in <Example 2>, they were incubated for 24 hours at 37° C. in 5% CO ₂ . After removing the culture medium and culturing for 24 hours and 48 hours, 0.5 mg/mL of MTT solution was added. After 1 hour, after removing the MTT solution, 100 μL of DMSO was added to each well. Then, absorbance was measured at a wavelength of 570 nm using a microplate reader (Molecular Devices E09090; San Francisco, CA, USA).

<Example 4> Scratch wound-healing assay

A scratch wound was applied to a monolayer of cultured NHDF cells with WoundMaker ^™ (Essen Bioscience, Michigan, USA).

Then, the cells were cultured for 72 hours in a serum-free medium containing 25 μg/mL of the sample prepared in <Example 2>.

The wound healing effect was confirmed by capturing images of cells every 2 hours using IncuCyte ZOOM (Essen Bioscience, MI, USA).

<Experimental Example 1> Confirmation of the effect of aloe vera flower extract on NHDF cell migration

First, through cell viability confirmation, all NHDF cells treated with aloe vera flower 100% ethanol extract (EtOH), aloe vera flower 50% ethanol extract (50% EtOH), and aloe vera flower extract (water), as shown in FIG. It was confirmed that it did not exhibit cytotoxicity as well (FIG. 1).

In addition, an in vitro wound healing assay was performed to determine the wound healing activity using the dose confirmed through the MTT assay.

On the other hand, cell migration is an important feature of skin wound healing, and the NHDF cell migration rate was determined based on the percentage of wound closure relative to the wound gap induced through scratch analysis.

As shown in Figure 2, the aloe vera flower 100% ethanol extract (Absolute Ethanol) and the aloe vera flower 50% ethanol extract (50% Ethanol) significantly increased the wound closure rate compared to the untreated control group (Control). confirmed (FIG. 2).

Claims (12)

A pharmaceutical composition for treating wounds containing an Aloe vera flower extract as an active ingredient,
The pharmaceutical composition for treating wounds, wherein the aloe vera flower extract is extracted with ethanol or a mixed solvent of ethanol and water.
delete delete The pharmaceutical composition for treating wounds according to claim 1, wherein the aloe vera flower extract increases the migration of dermal fibroblasts.
The method of claim 1, wherein the composition is any one selected from the group consisting of solutions, suspensions, granules, tablets, capsules, pills, extracts, emulsions, syrups, gels, pastes, ointment frosts, powders, emulsions and aerosols. A pharmaceutical composition for treating wounds, characterized in that the preparation of.
A pharmaceutical composition for skin regeneration containing an aloe vera flower extract as an active ingredient,
The aloe vera flower extract is a pharmaceutical composition for skin regeneration extracted with ethanol or a mixed solvent of ethanol and water.
delete The pharmaceutical composition for skin regeneration according to claim 6, wherein the aloe vera flower extract increases the migration of dermal fibroblasts.
A cosmetic composition for skin regeneration containing an aloe vera flower extract as an active ingredient,
The aloe vera flower extract is a cosmetic composition for skin regeneration extracted with ethanol or a mixed solvent of ethanol and water.
The cosmetic composition for skin regeneration according to claim 9, wherein the aloe vera flower extract increases the migration of dermal fibroblasts.
A food composition for skin regeneration containing an aloe vera flower extract as an active ingredient,
The aloe vera flower extract is a food composition for skin regeneration extracted with ethanol or a mixed solvent of ethanol and water.
A health functional food composition for skin regeneration containing an aloe vera flower extract as an active ingredient,
The health functional food composition for skin regeneration, wherein the aloe vera flower extract is extracted with ethanol or a mixed solvent of ethanol and water.