KR102475143B1 - Composition for Skin Regeneration Comprising Extract of Vanilla Bean - Google Patents

Abstract

The present invention provides a composition for skin regeneration comprising a vanilla bean extract extracted using at least one extraction solvent selected from the group consisting of water, organic solvents, and mixtures thereof. Since the vanilla bean extract of the present invention promotes cell migration, it can be used for skin regeneration.

Description

Composition for Skin Regeneration Comprising Extract of Vanilla Bean}

The present invention relates to a composition for skin regeneration comprising a vanilla bean extract.

The skin, as the largest organ of the human body, serves as a barrier to protect the body and is the body's first line of defense against diseases. In addition, the skin is composed of three layers: epidermis, dermis, and subcutaneous fat layer, and the epidermis provides a barrier against microbial invasion. Therefore, the primary objective in treating skin wounds is rapid closure and wound healing to prevent infection.

The migration of fibroblasts, which are essential for skin wound healing or tissue regeneration, is accomplished through various signal transduction processes, including concentration gradients of chemotactic substances, growth factors, and extracellular matrix (Ridley AJ et al (2003) Cell migration: integrating signals from front to back. Science 302: 1704-1709)

Integrin, a receptor molecule present on the cell surface, is a transmembrane protein that attaches to the extracellular matrix and transmits signals from other cells to the inside as well as transmits necessary signals from the inside to the outside of the extracellular matrix. It also transmits and interacts with the extracellular matrix to mediate a bidirectional reaction.

The extracellular matrix stores and adequately supplies biochemical factors necessary for cell growth and differentiation, and provides a physical environment for cells to recognize. Major components of the extracellular matrix include collagen, fibronectin, and the like. Collagen forms most of the organic substances of skin, tendon, bone, and teeth, and is particularly high in bone and skin. Collagen is reduced by age and photoaging by ultraviolet irradiation, which is known to be closely related to the formation of wrinkles in the skin.

In this way, since not only cells but also various factors adjacent to the cells are involved in skin regeneration in a complex manner, studies related to the promotion of skin regeneration are continuously being conducted in various fields.

On the other hand, as consumers' interest in skin health continues to increase, the number of smart consumers who meticulously examine various information even when purchasing a product is increasing in the cosmetics industry. Consumers now find products suitable for their individual skin types by looking for and comparing product ingredients one by one, actively sharing information and enjoying consumption. In addition, as consumers' interest in cosmetic compositions using ingredients derived from natural products with little or no irritation rapidly increased, the experience of not using cosmetics was once an issue.

Vanilla (Vanilla Planifolia) is an orchid family native to Mexico, tropical America, and the fruit part "vanilla bean" has been widely used as a spice since ancient times. Vanilla bean is obtained through artificial insemination by humans, and has no taste or smell by itself. To use vanilla as a spice, the vanilla bean must be steamed and dried several times, which usually takes 3 to 6 months. It is only after going through this processing family that vanilla processed beans with a unique aroma of black color are produced.

The present inventors have found that the vanilla bean extract extracted using water or an organic solvent has an excellent effect on skin regeneration and completed the present invention.

Ridley AJ et al (2003) Cell migration: integrating signals from front to back. Science 302: 1704-1709

An object of the present invention is to provide a natural product-derived ingredient capable of promoting skin regeneration.

In order to solve the above problems, the present invention provides a composition for skin regeneration comprising a vanilla bean extract extracted using at least one extraction solvent selected from the group consisting of water, organic solvents, and mixtures thereof.

Since the vanilla bean extract of the present invention promotes cell migration, it can be used for skin regeneration.

1 shows the result of quantifying the degree of cell migration by analyzing images taken according to a wound healing assay.
Figure 2 shows the results of evaluating the cytotoxicity or proliferation efficacy of the vanilla bean extract according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

In the present invention, "skin regeneration" may refer to any action that replenishes a part of the skin when it is lost or promotes the proliferation of skin cells. Specifically, skin regeneration may refer to regeneration of skin tissue at a skin wound site.

In the present invention, "skin wound" may be at least one selected from the group consisting of wounds, burns, abrasions, ultraviolet (UV) light damage, and scars.

In the present invention, "promotion" may mean any action that helps skin regeneration, for example, an action that at least reduces the degree of skin injury.

1. Preparation of vanilla bean extract

Vanilla (Vanilla Planifolia) is an orchid plant native to Mexico, tropical America, and the fruit part is commonly called vanilla bean. In the present invention, vanilla bean is used as a meaning including vanilla processing beans having a unique aroma of black color obtained by steaming and drying vanilla fruits (vanilla beans) several times.

"Extract" referred to in the present invention means a material extracted from raw materials by any method, and is used in the sense of including all of the extracted extract, the concentrate obtained therefrom, and the dried product and powder of the concentrate without limitation.

The extract may be obtained by extracting from a raw material or a dried product thereof, and the raw material of the extract may be used without limitation, such as cultivated or commercially available.

When the extract is obtained by extraction from a raw material, all known conventional extraction methods such as solvent extraction, ultrasonic extraction, filtration and reflux extraction may be used as extraction methods, preferably prepared by using solvent extraction or reflux extraction. can do. The extraction process may be repeated several times, and then additional steps such as concentration or lyophilization may be performed. Specifically, the obtained extract may be concentrated under reduced pressure to obtain a concentrate, and the concentrate may be lyophilized and then a high-concentration extract powder may be prepared using a grinder. The extract also includes fractions obtained by further fractionating the extract.

In the present invention, the extract may be extracted using at least one selected from the group consisting of water, organic solvents, and mixtures thereof as an extraction solvent. The organic solvent is an alcohol, preferably a C1-C4 lower alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene and It may be any one selected from the group consisting of these mixed solvents, preferably ethanol or dichloromethane, but is not limited thereto. In addition, when a mixture of water and organic solvent is used as the extraction solvent, the mixture of water and organic solvent may preferably be a mixture of water and a C1-C4 lower alcohol, more preferably a mixture of water and ethanol. can

The mixture of water and ethanol is 1% (v / v) to 60% (v / v) ethanol aqueous solution, or 1% (v / v) to 40% (v / v) ethanol aqueous solution, or 5% (v / v) v) to 50% (v/v) aqueous ethanol solution, or 10% (v/v) to 50% (v/v) aqueous ethanol solution, or 10% (v/v) to 30% (v/v) aqueous ethanol solution , or 25% (v/v) to 35% (v/v) ethanol aqueous solution. When the concentration of the aqueous ethanol solution is less than the lower limit, the stability of the formulation particles decreases and the skin regeneration effect decreases when included in the composition. In addition, when the aqueous ethanol solution has a concentration exceeding the upper limit, the extract contains a high concentration of ethanol, and when added to the composition without a separate ethanol removal process, skin irritation or toxicity may be caused, and stability of physical properties may be reduced. have. In order to solve this problem, when ethanol is removed through a separate process, the skin regeneration efficacy of the extract may be reduced, and the raw material production cost is increased by adding an ethanol removal process.

In one embodiment, the extraction solvent is at least one selected from the group consisting of water, 1% (v/v) to 60% (v/v) ethanol aqueous solution, and dichloromethane; one or more selected from the group consisting of water, 1% (v/v) to 40% (v/v) ethanol aqueous solution, and dichloromethane; 10% (v/v) to 30% (v/v) at least one selected from the group consisting of aqueous ethanol and dichloromethane; It may be at least one selected from the group consisting of 25% (v/v) to 35% (v/v) ethanol aqueous solution and dichloromethane.

When the mixture of water and ethanol or dichloromethane is used as the extraction solvent, the skin regeneration composition of the present invention may or may not further include a water extract of vanilla bean. When the water extract of vanilla bean is not included, it may be advantageous in terms of skin regeneration effect, and when the water extract is included separately, it may be advantageous in terms of cost or miscibility with other ingredients in the composition.

Extraction may be carried out at 0 ° C to 100 ° C, preferably at 10 ° C to 50 ° C, more preferably at 10 ° C to 35 ° C, but is not limited thereto. Extraction may be carried out for 12 hours to 36 hours, preferably for 15 hours to 30 hours, and more preferably for 20 hours to 25 hours, but is not limited thereto. Extraction may be performed 1 to 8 times, preferably 1 to 6 times, and more preferably 1 to 5 times, but is not limited thereto, and the extract is obtained from each extraction. It may be a single extract or a mixed extract of extracts obtained from each extraction. Among the extraction conditions, when the extraction temperature and time are less than the lower limit or the number of times of extraction exceeds the upper limit, the skin regeneration effect may be reduced.

The extract may be obtained by performing a conventional fractionation process in order to increase the skin regeneration effect. For example, the fraction obtained by passing the extract obtained according to the extraction through an ultrafiltration membrane having a certain molecular weight cut-off value, various chromatography (made for separation according to size, charge, hydrophobicity or affinity) Active fractions obtained through various additional purification methods such as separation are also included in the extract of the present invention. A specific active fraction with a higher skin regeneration effect can be prepared by separating an extract in which various components are mixed according to the nature of the active component through concentration gradient column chromatography or the like.

The column chromatography can separate and purify the active fraction using a filler selected from the group consisting of silica gel, Sephadex, LH-20, ODS gel, RP-18, polyamide, Toyopearl and XAD resin. . Column chromatography may be performed several times by selecting an appropriate filler as necessary, but is not limited thereto. In using the chromatography, the elution solvent, elution rate, and elution time may apply solvents, rates, or times generally used in the art.

2. Cosmetic composition for skin regeneration containing vanilla bean extract

One embodiment of the present invention provides a cosmetic composition for skin regeneration comprising a vanilla bean extract extracted using at least one extraction solvent selected from the group consisting of water, organic solvents, and mixtures thereof.

The preparation method of the vanilla bean extract is the same as that mentioned in " 1. Preparation of the vanilla bean extract ", so the description is omitted.

The cosmetic composition of the present invention contains the vanilla bean extract in an amount of 0.0001 to 10% by weight, or 0.001 to 5% by weight, or 0.01 to 3% by weight, or 0.1 to 2% by weight, or 0.5 to 1.5% by weight based on the total weight of the cosmetic composition. can do.

The cosmetic composition of the present invention may be prepared in liquid or solid form using bases, adjuvants and additives commonly used in the field of cosmetics. Cosmetics in liquid or solid form may include, but are not limited to, cosmetic products, creams, lotions, and bath products.

Bases, adjuvants and additives commonly used in the field of cosmetics are not particularly limited, and examples thereof include water, alcohol, propylene glycol, stearic acid, glycerol, cetyl alcohol, and liquid paraffin.

When the composition of the present invention is prepared as a cosmetic composition, the composition of the present invention may include not only vanilla bean extract, but also components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, and pigments. and conventional adjuvants such as perfumes, and carriers.

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

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

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

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan.

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

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl Amidobetaine, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

The cosmetic composition of the present invention may be used alone or overlappingly applied, or may be used overlappingly with other cosmetic compositions other than the present invention. In addition, the cosmetic composition with excellent skin regeneration effect according to the present invention can be used according to a conventional method of use, and the number of times of use can be varied according to the user's skin condition or taste.

When the cosmetic composition of the present invention is a soap, surfactant-containing cleansing or surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, removed, or washed with water. As specific examples, the soap is liquid soap, powder soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel, and a cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream. , cleansing lotion, cleansing water and cleansing gel, but are not limited thereto.

3. Pharmaceutical composition for skin regeneration containing vanilla bean extract

Another embodiment of the present invention provides a pharmaceutical composition for skin regeneration comprising a vanilla bean extract extracted using at least one extraction solvent selected from the group consisting of water, organic solvents, and mixtures thereof.

The preparation method of the vanilla bean extract is the same as that mentioned in " 1. Preparation of the vanilla bean extract ", so the description is omitted.

Specifically, the pharmaceutical composition for skin regeneration of the present invention may be a pharmaceutical composition for preventing or treating skin diseases caused by skin wounds. The skin wound may be at least one selected from the group consisting of wounds, burns, abrasions, ultraviolet (UV) light damage, and scars.

The term "prevention" includes inhibiting the occurrence of skin disorders. The term "treatment" includes the inhibition, alleviation, or elimination of the development of a skin disorder.

The pharmaceutical composition of the present invention, in addition to the vanilla bean extract, preferably further contains other ingredients that can give a synergistic effect to the effect of the vanilla bean extract within a range that does not impair the effect of the present invention. can do. conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, or carriers.

The route of administration of the pharmaceutical composition includes oral, intravenous, intramuscular, intraarterial, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal, for example topical application by application method can be applied. The parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intralesional injection or infusion techniques.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

In the present invention, "pharmaceutically effective amount" means an amount sufficient to promote the regeneration of damaged skin or treat skin diseases caused by skin wounds, as a reasonable benefit / risk ratio applicable, and the effective dose level depends on the type of subject and severity, age, sex, type of disease, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including concomitantly used drugs, and other factors well known in the medical field. have. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be single or multiple administration. Considering all of the above factors, it is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects, and can be easily determined by a person skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an object for the prevention or treatment of skin diseases caused by skin wounds, and any may be applied. For example, the pharmaceutical composition of the present invention can be used not only for humans but also for non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds and fish. do.

The pharmaceutical composition may contain at least one active ingredient exhibiting the same or similar function in addition to the vanilla bean extract. The composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively.

Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills and the like. Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included. have.

Formulations for parenteral administration include powders, granules, tablets, capsules, sterilized aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc., and sterile injection preparations according to conventional methods, respectively. It can be formulated and used in the form of a cream, gel, patch, spray, ointment, warning, lotion, liniment, pasta, or cataplasma. , but is not limited thereto. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogeratin and the like may be used.

The pharmaceutical composition may further contain adjuvants such as preservatives, stabilizers, hydration agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, which are conventional methods such as mixing and granulation. It can be formulated according to the coating or coating method.

The dose of the pharmaceutical composition may vary depending on various factors including age, body weight, general health, sex, administration time, route of administration, excretion rate, drug combination, and severity of a specific disease. For example, when the pharmaceutical composition is for external use, it is recommended to apply the pharmaceutical composition 1 to 5 times a day in an amount of 1.0 to 3.0 ml for adults and continue for 1 month or more, but the dosage is not intended to limit the scope of the present invention. .

The pharmaceutical composition may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

4. Food composition for skin regeneration containing vanilla bean extract

Another embodiment of the present invention provides a food composition for skin regeneration comprising a vanilla bean extract extracted using at least one extraction solvent selected from the group consisting of water, organic solvents, and mixtures thereof.

The preparation method of the vanilla bean extract is the same as that mentioned in " 1. Preparation of the vanilla bean extract ", so the description is omitted.

The vanilla bean extract is preferably contained in an amount of 0.0001 to 20.0% by weight based on the total weight of the food composition, more preferably 0.001 to 10.0% by weight, or 0.01 to 5% by weight, but limited thereto. It doesn't work. If the effective content of the vanilla bean extract is less than 0.0001% by weight, it is difficult to implement a skin regeneration effect, and if it exceeds 20% by weight, the effect of skin regeneration due to the increase in content is poor, which is uneconomical.

The food composition may further contain, in addition to the vanilla bean extract of the present invention, other ingredients that may give a synergistic effect to the effect of the vanilla bean extract, preferably within a range that does not impair the desired effect of the present invention. can Conventional adjuvants such as, for example, stabilizers, solubilizers, vitamins, pigments and flavors, or carriers may be included.

In addition, it may include ingredients commonly added during food preparation, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of such carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides such as conventional sugars such as dextrins and cyclodextrins and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, natural flavoring agent thaumatin, stevia extract, and synthetic flavoring agent may be used. For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, natural extract and the like may be further included in addition to the vanilla bean extract of the present invention.

There is no particular limitation on the type of food using the food composition. Examples of foods to which the food composition containing the vanilla bean extract of the present invention can be added include meat, sausages, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, There are various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, and the like, and includes all foods in a conventional sense.

Hereinafter, the present invention will be described in detail with reference to preparation examples and examples.

However, the following Preparation Examples and Examples are only for exemplifying the present invention, and the contents of the present invention are not limited by the following Preparation Examples and Examples.

Preparation Example 1. Preparation of vanilla bean extract

After drying/pulverizing the vanilla bean, 10 times the solvent by the weight of the vanilla bean was purified water, 10% ethanol, 30% ethanol, 50% ethanol, 70% ethanol, 90% ethanol, 100% ethanol, ethyl acetate (EA) and dichloromethane (DCM) were added differently and stirred at room temperature.

Thereafter, each was filtered and then subjected to forced concentration to obtain a concentrate, and the concentrate was freeze-dried to obtain a high-concentration extract powder. However, when 90% ethanol, 100% ethanol, ethyl acetate and dichloromethane were used as extraction solvents, oily samples were obtained.

Experimental Example 1. Wound healing measurement method

1. Experiment method

After dispensing human dermal fibroblasts into the scar block and removing the block, 100 μg/ml of human Epidermal Growth Factor (hEGF) (Sigma), 125 μg/ml of the water extract obtained in Preparation Example 1, 10% (v/v ) Ethanol aqueous solution extract 125 μg/ml, 30% (v/v) ethanol aqueous solution extract 125 μg/ml, 50% (v/v) ethanol aqueous solution extract 125 μg/ml, 70% (v/v) ethanol aqueous solution extract 125 ㎍/ml, 90% (v/v) ethanol aqueous solution extract 0.00125%, 100% (v/v) ethanol aqueous solution extract 0.00125%, ethyl acetate extract 0.00125%, and dichloromethane extract 0.00125% were respectively treated and left to stand for 24 hours. cultured.

2. Experimental results

Cell migration was observed with a phase contrast microscope, and after 24 hours of stationary culture, the cell migration results were photographed, and the area of empty space reduced by cell migration was measured with cell image software (ImageJ v.1.50e), and the cell migration ratio was quantified and Table 1 and shown in Figure 1.

untreated group positive control
(hEGF) water extract group 10%
EtOH group 30%
EtOH group 50%
EtOH group 70%
EtOH group 90%
EtOH group 100%
EtOH group EA group DCM group Cell Migration Rate (%) 0 83.9 17.4 26.2 34.2 15.0 -6.7 -9.0 -13.2 3.6 31.6

In the results of Table 1 and Figure 1, the vanilla bean water extract group, 10% (v / v) ethanol aqueous solution extract group, 30% (v / v) ethanol aqueous solution extract group, 50% (v / v) ethanol aqueous solution In the case of the extract group and the DCM extract group, the cell migration rates were 17.4%, 26.2%, 34.2%, 15.0%, and 31.6%, respectively, showing superior effects compared to the untreated group. In particular, the 30% (v / v) ethanol aqueous solution extract group and the DCM extract group showed excellent wound healing efficacy.

Experimental Example 2. Cytotoxicity or proliferative efficacy evaluation

To confirm the cytotoxicity or proliferative effect, MTT reduction method (3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction method) was performed. Specifically, human fibroblasts (ATCC 2076) were seeded in a 24-well plate at a density of 1×10 ⁵ cells using DMEM medium (Gibco, USA) containing 10% bovine serum. After 1 day, while changing to a new medium (DMEM, bovine serum 0%), the water extract obtained in Preparation Example 1, 10% (v / v) ethanol aqueous solution extract, 30% (v / v) ethanol aqueous solution extract, 50% (v/v) ethanol aqueous solution extract and 70% (v/v) ethanol aqueous solution extract at concentrations of 7.8 μg/ml, 15.6 μg/ml, 31.3 μg/ml, 62.5 μg/ml and 125 μg/ml, respectively, and 90% (v/v) ethanol aqueous solution extract, 100% (v/v) ethanol aqueous solution extract, ethyl acetate extract and dichloromethane extract at concentrations of 0.000078125%, 0.00015625%, 0.0003125%, 0.000625% and 0.00125%, respectively. After treatment, it was incubated at 37°C for 24 hours. At this time, the positive control group used a medium containing 10% FBS, and the untreated and experimental groups used a medium containing no FBS. Thereafter, the cells were washed with PBS, and 1 ml of a 10-fold diluted MTT (2.5 mg/ml) in DMEM medium was added, followed by incubation at 37° C. for 4 hours. After the incubation was completed, the medium was removed, 0.5 ml of DMSO was added, completely dissolved at room temperature for 10 minutes, and absorbance was measured at 570 nm, and the results are shown in FIG. 2 .

In FIG. 2, it was confirmed that all extracts did not exhibit cytotoxicity or proliferation efficacy at concentrations of 125 μg/ml or 0.00125% or less.

Preparation Example 2. Preparation of a cosmetic composition containing vanilla bean extract

2-1. Preparation of Essence

Using the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1, an essence was prepared according to the contents (parts by weight) shown in Table 2 below.

Furtherance Content (parts by weight) triethanolamine 0.25 carboxyvinyl polymer 0.22 glycerin 4 Butylene Glycol 2 Vanilla Bean Extract 1.5 beeswax 0.5 Cetostearyl Alcohol One Glyceryl Monostearate One squalene 4 Purified water balance

2-2. Manufacture of softening lotion

Softening lotion containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient was prepared as shown in Table 3 below.

Raw material Content (parts by weight) 1,3-butylene glycol 1.00 Disodium EDT 0.05 allantoin 0.10 dipotassium glycyrrhizate 0.05 Citric Acid 0.01 sodium citrate 0.02 glycereth-26 1.00 Arbutin 2.00 PEG-40 Hydrogenated Castor Oil 1.00 ethanol 30.00 Vanilla Bean Extract 0.5 coloring agent a very small amount flavoring agent a very small amount Purified water balance

2-3. Manufacture of nourishing cream

A nourishing cream containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient was prepared as shown in Table 4 below.

Raw material Content (parts by weight) 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Magnesium aluminum silicate 0.3 PEG-40 Stearate 1.2 Stearic Acid 2.0 polysorbate 60 1.5 Lipophilic Glyceryl Stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 mineral oil 4.0 squalene 3.8 Vanilla Bean Extract 1.5 vegetable oil 1.8 dimethicone 0.4 dipotassium glycyrrhizate a very small amount allantoin a very small amount sodium hyaluronate a very small amount Tocopheryl Acetate Appropriate amount triethanolamine Appropriate amount flavoring agent Appropriate amount Purified water balance

2-4. manufacture of lotion

A lotion containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient was prepared according to the composition shown in Table 5 below.

Raw material Content (parts by weight) Cetostearyl Alcohol 1.6 stearic acid 1.4 Glycerin lipophilic monostearate 1.8 PEG-100 Stearate 2.6 Sorbital Sesquioleate 0.6 squalene 4.8 macadamia oil 2 jojoba oil 2 Tocopherol Acetate 0.4 Methylpolysiloxane 0.2 Tocopherol Acetate 0.4 1,3-butylene glycol 4 xanthan gum 0.1 glycerin 4 d-pandenol 0.15 Vanilla Bean Extract 1.0 allantoin 0.1 Carbomer (2% aq. Sol) 4 triethanolamine 0.15 ethanol 3 Purified water balance

Preparation Example 3. Preparation of pharmaceutical compositions and oral compositions containing vanilla bean extract

3-1. Manufacture of injectables

Injections were prepared according to the contents (parts by weight) shown in Table 6 below using the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1.

Furtherance Content (parts by weight) Vanilla Bean Extract 100 sodium metabisulfite 3.0 Methylparaben 0.8 Propylparaben 0.1 Sterile distilled water for injection Appropriate amount

The above components were mixed and prepared to a final volume of 2 ml by a conventional method, filled into an ampoule and sterilized to prepare an injection.

3-2. manufacture of tablets

Tablets containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient were prepared according to the composition shown in Table 7 below.

Furtherance Content (parts by weight) Vanilla Bean Extract 200 potato starch 100 lactose 100 colloidal silicic acid 16 magnesium stearate Appropriate amount

According to a conventional tablet manufacturing method, the above ingredients were mixed and tableted to prepare tablets.

3-3. Manufacture of capsules

Capsules containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient were prepared according to the composition shown in Table 8 below.

Furtherance Content (parts by weight) Vanilla Bean Extract 100 lactose 50 starch 50 talc 2 magnesium stearate Appropriate amount

Capsules were prepared by mixing the ingredients and filling them into gelatin capsules according to a conventional capsule manufacturing method.

3-4. manufacture of pills

Pills containing the vanilla bean extract extracted with 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient were prepared according to the composition shown in Table 9 below.

Furtherance Content (parts by weight) Vanilla Bean Extract 120 corn starch 100 sterile distilled water Appropriate amount

A pill was prepared by mixing the above components and forming a sphere having an appropriate size according to a conventional pill preparation method.

3-5. Preparation of composition for oral use

3-5-1. manufacture of toothpaste

An oral composition containing the vanilla bean extract extracted with a 30% (v / v) ethanol aqueous solution of Example 1 as an active ingredient was prepared as shown in Table 10 below.

Furtherance Content (parts by weight) Vanilla Bean Extract 20 glycerin 6 sodium saccharin 0.1 silica 7 precipitated calcium carbonate 39 sodium lauryl sulfate 2 menthol 0.4 peppermint oil 0.1 Purified water Appropriate amount

The above ingredients were mixed, and toothpaste was prepared according to a conventional method for preparing toothpaste.

Claims (5)

A cosmetic composition for skin regeneration comprising a vanilla bean extract obtained by extraction with dichloromethane and further promoting the migration of fibroblasts than a vanilla bean water extract. delete The method of claim 1,
The skin regeneration is the regeneration of the skin tissue of the skin wound, a cosmetic composition for skin regeneration. The method of claim 3,
The skin wound is at least one selected from the group consisting of wounds, burns, abrasions, ultraviolet (UV) light damage and scars, a cosmetic composition for skin regeneration. A pharmaceutical composition for wound healing comprising a vanilla bean extract obtained by extraction with dichloromethane and further promoting the migration of fibroblasts than a vanilla bean water extract.

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