KR101746639B1 - Composition with skin lightening and moisturizing effects containing bee venom extracts - Google Patents

Abstract

The present invention discloses a composition containing bee venom extract as an active ingredient. The composition has skin whitening and skin moisturizing effects. Also disclosed are cosmetic compositions having skin whitening and moisturizing effects, including the above compositions.

Description

TECHNICAL FIELD The present invention relates to compositions for skin whitening and moisturizing containing bee venom extracts,

The present invention relates to a skin whitening and skin moisturizing composition containing bee venom extract.

Melanin is a dark brown pigment that acts to protect the skin from ultraviolet rays because it has a function to block ultraviolet rays above a certain amount. Thus, when the skin is exposed to ultraviolet light, a large amount of melanin is produced in the body to protect the skin. Skin color is determined by the amount of melanin and the amount of melanin increases as the amount of the skin becomes black. Therefore, a large amount of melanin is caused by spots, dots, freckles, and black spots. Therefore, substances that inhibit melanin production may have a skin whitening effect.

On the other hand, when the amount of skin moisture is reduced, dry keratin often occurs. Also, the skin's luster disappears, fine wrinkles easily occur, and skin aging begins early. That is, in order to have improved skin, it is essential to maintain skin moisture.

The present invention provides a composition having skin whitening or skin moisturizing effect by containing bee venom extract as an active ingredient.

One aspect of the present invention provides a skin whitening composition containing bee venom extract as an active ingredient.

Another aspect of the present invention provides a skin moisturizing composition containing bee venom extract as an active ingredient.

Another aspect of the present invention provides a cosmetic composition comprising the composition.

The composition according to one aspect of the present invention has an excellent skin whitening effect by inhibiting melanin production and improving pigmentation caused by ultraviolet rays by containing bee venom extract as an effective ingredient. It also induces the differentiation of keratinocytes and promotes the expression of filaggrin, thereby having a skin moisturizing effect.

As used herein, the term "extract" refers to a substance obtained by extracting a component contained therein from a natural product, regardless of the method of extraction or the kind of the component. For example, it is a broad concept including both extracts of components dissolved in a solvent from natural products by using water or an organic solvent, and extracts of specific components of natural products, such as oils, and extracts of specific components.

As used herein, the term "skin" refers to a tissue covering the body surface of an animal, and is a concept of the broadest notion including a scalp and hair as well as a tissue covering a body surface such as a face or a body.

Hereinafter, the present invention will be described in detail.

As used herein, the term "bee venom" refers to a bee venom from a honey bee's ovipositor, which is effective for neuralgia, rheumatism, and back pain, and is often used as a folk remedy. And " bee venom extract " collectively refers to a substance or a fraction of a solvent extracted through various extraction methods known in the art for the bee venom.

To penetrate the bee venom into the body and treat the disease, a needle therapy is used. It is known that such a needle therapy is effective for the treatment of arthritis and the like. The types of use of bongjeong therapy can be classified as follows.

The first is the direct method, which is a method of stabbing the lid of a living bee against the site where it is intended to be treated, and then pouring the bee venom for about 30 to 60 seconds and then picking up the saliva. The second method is a needle method, in which tweezers are pulled from a bee with tweezers and then pulled slightly to the part to be treated, and then pulled out to another part. The foot pricking method is a relatively popular method because it can treat 3 ~ 5 places with one bee without strong stimulus. Third, there is a method of spreading a method to treat a wide range of diseases such as herpes zoster. This method is similar to the method of using a triple-needle needle, and it is a treatment that takes advantage of the characteristics of a needle treatment that treats a large part of the lesion with a small amount of needle by repeatedly piercing and pulling the needle quickly. And fourth is injecting the diluted solution into the human acupuncture point directly through the syringe while diluting the bee venom with the saline solution. This method has the advantage that the pain perceived by the patient is much less, and it is also effective to penetrate the bee venom directly to the acupuncture point.

However, the above methods are insufficient to inject bee venom in order to find accurate blood, and it is difficult to precisely control the dose of bee venom for ideal treatment.

In order to solve such a problem, the form of the created treatment is to find the blood of the bee venom in the acupuncture needles and to inject the bee venom into acupuncturists together with acupuncture. The acupuncture method using the one-handed acupuncture method is an excellent method which has a much improved medical effect due to the characteristic of the acupuncture method, but there is also a problem therefrom.

For example, in the process of penetrating the epidermis and penetrating into the inside of the acupuncture when the bee venom is embedded in the tip of the acupuncture needle, if the bee venom spreads to the epidermis and penetrates into the inside of the acupuncture point, it may be difficult to expect a sufficient therapeutic effect. Specifically, in the treatment of oriental medicine, the needle is used to accurately place the needle in the hemisphere. However, in order to reduce the pain of the patient, when the needle is punctured to pierce the skin at a rapid rate, the coated bee venom solution is absorbed into the epidermal layer before it reaches the acupuncture point, so that the original efficacy can not be expected.

On the basis of this, the present inventors have discovered a new Yonex effect of bee venom while studying bee venom.

One aspect of the present invention provides a skin whitening composition containing bee venom extract as an active ingredient. The composition can inhibit melanin production and improve skin pigmentation caused by ultraviolet light to exhibit a skin whitening effect.

Another aspect of the present invention provides a skin moisturizing composition containing bee venom extract as an active ingredient. The composition can induce the differentiation of keratinocytes and promote the expression of filaggrin, a protein which is an important source for the production of natural moisturizing factors possessed by keratinocytes, thereby exhibiting skin moisturizing effects. On the other hand, a composition containing bee venom itself may exhibit the same or similar effect as a composition containing bee venom extract.

In one aspect of the present invention, bee venom can undergo various pretreatments routinely recognized in the art. For example, purified beeswax can be used by pouring distilled water into the bee venom obtained through electric shock method to suspend bees or debris in bee venom, and to filter out suspended bees or foreign matter.

In one aspect of the present invention, bee venom extract can be obtained by extracting bee venom in a conventional manner in the art. In another aspect of the present invention, bee venom extract can be obtained by extracting bee venom into water, an organic solvent or a mixed solvent of water and an organic solvent, and purifying and separating the solvent. The organic solvent is not particularly limited and may be a C ₁ -C ₅ lower alcohol, ether, ethyl acetate or chloroform. The C ₁ -C ₅ lower alcohol may be any one or a mixture of two or more selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol and isobutanol . In another aspect of the present invention, the volume of the organic solvent in the mixed solvent of water and the organic solvent may be 10 to 90% (v / v). Specifically, the bee venom extract is mixed with a mixed solvent containing 10 to 90% (v / v) ethanol, more specifically 70% (v / v) ethanol, based on the total volume of the mixed solvent May be an extracted extract.

One aspect of the present invention provides a cosmetic composition comprising a bee venom extract as an active ingredient. The cosmetic composition contains a bee venom extract as an active ingredient, thereby exhibiting skin whitening and skin moisturizing effects.

The cosmetic composition may be, for example, a cosmetic composition, and the cosmetic composition includes a cosmetically or dermatologically acceptable medium or base. It may be any formulation suitable for topical application, for example, a solution, a gel, a solid, an anhydrous product of a paste, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing a water phase in an oil phase, a multi-emulsion, Capsules, microgranules, ionic (liposomes) and non-ionic follicular dispersants, foams or compressed propellants. These compositions may be prepared according to conventional methods in the art.

The cosmetic compositions may also be in the form of tablets or capsules prepared by dissolving or dispersing the active ingredient in a diluent such as a lipid, an organic solvent, a solubilizer, a thickener, a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, , A chelating agent, a preservative, a vitamin, a barrier, a wetting agent, an essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, a lipid vesicle or an adjuvant conventionally used in the field of cosmetic or dermatological science have. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

The formulation of the cosmetic composition is not particularly limited, and the formulation can be appropriately selected according to the purpose. For example, softeners, convergent lotions, nourishing lotions, nutritional creams, massage creams, essences, eye creams, eye essences, cleansing creams, cleansing lotions, cleansing foams, cleansing waters, packs, powders, cone sticks, body lotions, Hair lotion, hair conditioner, hair conditioner, hair shampoo, hair lotion, toothpaste, mouthwash, regular detergent, body lotion, body lotion, body lotion But are not limited to, one or more formulations selected from the group consisting of, for example, tablets, capsules, tablets, dragees, lotions, ointments, gels, creams, patches and sprays.

The cosmetic composition according to one aspect of the present invention may contain 0.0001 to 30% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. The cosmetic composition according to another aspect of the present invention may contain 0.001 to 15% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. The cosmetic composition according to another aspect of the present invention may contain 0.01 to 5% by weight of a composition containing bee venom extract as an active ingredient based on the total weight of the composition. When used in the above range, it is not only suitable for exhibiting the intended effect of the present invention but also can satisfy both the stability and safety of the composition and may be suitably used within the above range in terms of cost effectiveness. More specifically, when the content of the composition containing the bee venom extract as an active ingredient is lower than the above range, skin whitening or skin moisturizing effect due to the bee venom ingredient can not be sufficiently obtained, and when it exceeds the above range, The increase in effectiveness may not be significant.

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to examples, comparative examples and test examples. However, these examples, comparative examples and test examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

[Example 1] Production of bee venom and preparation of bee venom extract

Bee venom was obtained by electric shock method. The bee venom obtained from the electric shock method was poured with distilled water to float bees or debris in bee venom. Filtered suspended bees or debris to obtain purified beacon.

The obtained bee venom was extracted with 70% ethanol. Specifically, 70% ethanol corresponding to 3 times the volume of the bee venom obtained was added, and the mixture was extracted and degreased with stirring at room temperature five times. 70% ethanol was added to the degreased bees poison four times, and the mixture was refluxed three times and then immersed at 15 ° C for 2 days. The precipitate was filtered and centrifuged to separate the residue and filtrate, and the separated filtrate was concentrated under reduced pressure to obtain an extract. After the resulting extract was suspended in water, the pigment was removed using ether, and the aqueous layer was extracted with butanol five times. The obtained organic solvent layer was concentrated under reduced pressure to give an extract. The resulting extract was dissolved in methanol, ethyl acetate was added, and the resulting precipitate was dried to obtain a bee venom extract to give Example 1.

[Test Example 1] Evaluation of skin whitening effect

1) Inhibitory effect of melanin on melanin production in rat

To determine the inhibitory effect of bee venom extract on melanogenesis, rat pigment cells were used. First, in a medium supplemented with 10% fetal bovine serum, 100 nM 2-O-tetradecanoyphorbol-13-acetate and 1 nM cholera toxin in DMEM (Dulbeccos modified Eagles media), C57BL / 6 Mouse-derived mouse pigment cells (Dooley, TP et al, Skin pharmacol, 7, pp 188-200) were cultured at 37 ° C and 5% CO ₂ . The cultured Mel-Ab cells were removed with 0.25% trypsin-EDTA, and the cells were cultured at a concentration of 10 ⁵ cells / well (cells / well) in a 24-well plate. 1ppm and 10ppm of the extract were added and cultured. At this time, hydroquinone was used as a positive control. After the culture medium was removed, the cells were washed with PBS, and the cells were dissolved with 1 N sodium hydroxide. The absorbance at 400 nm was measured, and the inhibition rate of melanin production was calculated according to the following formula (1) Dooley's method).

Inhibitory effect of bee venom extract and hydroquinone on the production of melanin Test substance Melanin production inhibition rate (%) Example 1 (1 ppm) 13.5 Example 1 (10 ppm) 35.2 Hydroquinone (control group) 43.6

As shown in Table 1, the bee venom extract of Example 1 according to the present invention showed a degree of inhibition of melanin formation similar to that of hydroquinone. Thus, it can be confirmed that bee venom extract has excellent whitening effect.

2) Whitening effect on human skin

The following experiment was conducted to examine the whitening effect of bee venom extract on human skin. First, 12 healthy boys were exposed to a 1.5 cm diameter opaque tape at the upper part of the subject's body, and then they were exposed to ultraviolet rays (UVB) of about 1.5 to 2 times the minimum erythema dose of each subject. To induce blackening of the skin. The bee venom extract of Example 1, which is the test substance after the ultraviolet ray irradiation, and hydroquinone were each applied, and the state change was observed for 10 weeks without applying anything to one place. The color of skin was measured with a colorimeter CR2002 (Minolta Co., Japan) every 1 week. Then, the skin color difference (DELTA L *) at the time of initiation of application and completion of application according to each test substance was calculated according to the following equation (2), and the results are shown in Table 2 below. On the other hand, the whitening effect is judged by comparison of DELTA L * between the sample application region and the control region. When the DELTA L * value is 2, the whitening of the deposited pigment is noticeable. .

Test substance The degree of skin color brightness (DELTA L *) Example 1 1.81 ± 0.15 Hydroquinone (positive control) 1.89 ± 0.15 Vehicle (negative control) 0.51 + - 0.13

As shown in Table 2, the bee venom extract of Example 1 showed a degree of skin color brightness similar to that of hydroquinone. Thus, it can be confirmed that bee venom extract has an effect of improving pigmentation caused by ultraviolet light to brighten skin color.

[Test Example 2] Test for inducing differentiation of human keratinocytes

In order to confirm skin barrier function and skin moisturizing ability of bee venom extract, the test using absorbance was carried out. First, primary keratinocytes were placed in a culture flask and adhered to the bottom. The cells were then added to the culture solution prepared as shown in Table 3 below, and the cells were cultured for 5 days until the cells reached 70-80% of the bottom area . The cells were harvested and washed with PBS (phosphate buffered saline). The cells were washed with 10 mM Tris-HCl buffer (Tris-HCl buffer, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiothreitol) , pH 7.4), sonication was performed for 3 minutes, and the mixture was boiled for 10 minutes. The mixture was centrifuged at 1200 rpm for 30 minutes, and the separated precipitate was suspended in 1 ml of PBS and the absorbance at 340 nm was measured. Separately, a portion of the solution after the sonication was taken to measure the protein content and used as a criterion when evaluating the degree of cell differentiation. The test results are shown in Table 3 below, in which the low calcium (0.03 mM) treatment group and the high calcium (1.2 mM) treated group were used as negative / positive control groups and the test substance was added to low calcium concentration.

Concentration used The ability to differentiate in keratinocytes (%) Control group Low Ca < ^{2 +} & gt ^; (0.03 mM) 100 And Ca ^{2 +} (1.2 mM) 206 Example 1 0.1 ppm 120 Example 1 0.05 ppm 105

As shown in Table 3, the amount of CE (cornified envelope) produced upon keratinocyte differentiation was measured to compare the promoting effect of cell differentiation It was confirmed that the bee venom extract of Example 1 promotes differentiation in keratinocytes. Therefore, bee venom extract can enhance skin barrier function and protect skin moisture.

[Test Example 3] Expression of filaggrin by RT-PCR analysis

In order to test the skin moisturizing ability of bee venom extract, the degree of fila green expression was measured. The cell lines used in the experiments were the Dr. As the human kerationocyte HaCaT cell line from Fusenig, the cells were divided into 1 × 10 ⁵ cells in a 60 mm dish and cultured for 1 day. Then, the test materials were treated as shown in Table 4, and then cultured for 24 hours . Total RNA was extracted from the cells cultured and tested by the above method using a triazole (Trizol, Gibco Laboratories, USA), and the total RNA was extracted from the cells by the method provided in One Step RNA PCR Kit (AMV) (Takara Bio Inc., Japan) RT-PCR (reverse transcriptional polymerase chain reaction) was performed. Primers for filaggrin (Bioneer, Korea) were used as primers and beta-actin was used as an internal control for comparison. . After the image analysis, the control group was set at 100, and the comparative values are shown in Table 4 below.

Concentration used Pillared green production (%) Untreated control group - 100 Example 1 0.1 ppm 180 Example 1 0.05 ppm 112

As shown in Table 4, it was confirmed that the bee venom extract of Example 1 promotes the expression of pilgar green in keratinocytes. Therefore, bee venom extract promotes the production of natural moisturizing factors of the skin and enhances the skin moisturizing ability.

[Example 2 and Comparative Example 1]

Example 2 and Comparative Example 1, which are external preparations of nutritional cream formulations, were prepared according to the composition ratios shown in Table 5 below. The unit of the content ratio of the following ingredients is% by weight.

Compounding ingredient Example 2 Comparative Example 1 Purified water To 100 To 100 Bee venom extract (Example 1) 0.1 - Vegetable hydrogenated oil 1.50 1.50 Stearic acid 0.60 0.60 Glycerol stearate 1.00 1.00 Stearyl alcohol 2.00 2.00 Polyglyceryl-10pentastearate and behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 Caprylic / carboxy triglyceride 11.00 11.00 Cyclomedicone 6.00 6.00 Preservative, incense Suitable amount Suitable amount Triethanolamine 0.1 0.1

[Test Example 4] Measurement of skin moisturizing effect

In order to confirm the skin moisturizing effect of bee venom extract on a human, the following experiment was carried out on Example 2 and Comparative Example 1. 40 healthy women in their thirties were divided into two groups each consisting of 20 persons and the nutritional creams of Example 2 and Comparative Example 1 were applied to the face in the face for 4 weeks twice daily at a temperature of 24-26 DEG C and a humidity of 75% (24 ° C, relative humidity: 40%) after one week, two weeks, four weeks after application, and two weeks after the application was stopped Skin moisture was measured. The increment of the measured value at each measurement is expressed as a percentage based on the coneometer value measured just before the start of the test, and the results are shown in Table 6 below.

Clinical outcome Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Comparative Example 1 30 33 34 15 Example 2 33 41 42 32

As shown in Table 6, it can be seen that Example 2 containing bee venom extract had a larger increase in skin moisture content than that of Comparative Example 1. In addition, the value after 6 weeks after 2 weeks of discontinuation of application of the test substance appeared similar to the value after 1 to 2 weeks after application of the test substance, and even when the test substance was not applied, Of the total amount of water retained for a certain period of time. Therefore, it can be seen that when bee venom extract is applied to humans, the skin moisturizing ability is strengthened.

Formulation examples of compositions according to one aspect of the present invention are described below, but may be applied to various other formulations, which are not intended to be limiting but merely illustrative of the invention.

[Formulation Example 1] Lotion-type preparation

Bee venom extract of Example 1 3.00

L-ascorbic acid-2-phosphate magnesium salt 1.00

Water soluble collagen (1% aqueous solution) 1.00

Sodium citrate 0.10

Citric acid 0.05

Licorice extract 0.20

1,3-butylene glycol 3.00

Purified water balance

(Unit: wt%)

[Formulation Example 2]

Bee venom extract of Example 1 1.00

Polyethylene glycol monostearate 2.00

Self emulsifying monostearate glycerin 5.00

Cetyl alcohol 4.00

Squalane 6.00

Tri-2-ethylhexane glyceryl 6.00

Sphingo glycolipid 1.00

1,3-butylene glycol 7.00

Purified water balance

(Unit: wt%)

[Formulation Example 3] Packed preparation

Bee venom extract of Example 1 5.00

Polyvinyl alcohol 13.00

L-ascorbic acid-2-phosphate magnesium salt 1.00

Lauroylhydroxyproline 1.00

Water soluble collagen (1% aqueous solution) 2.00

1,3-butylene glycol 3.00

Ethanol 5.00

Purified water balance

(Unit: wt%)

[Formulation Example 4] Cosmetic liquid preparations

Bee venom extract of Example 1 2.00

Hydroxyethylene Cellulose (2% aqueous solution) 12.00

Xanthan gum (2% aqueous solution) 2.00

1,3-butylene glycol 6.00

Dark glycerin 4.00

Sodium hyaluronate (1% aqueous solution) 5.00

Purified water balance

Claims (6)

Bee venom is an organic solvent; Or an extract of bee venom extracted with a mixed solvent of water and an organic solvent as an active ingredient,
Wherein the organic solvent is at least one selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol, isobutanol, ether, ethyl acetate and chloroform. A composition for skin moisturizing, comprising bee venom extract as an active ingredient. 3. The method of claim 2,
Bee venom extract is water for bee venom; Organic solvent; Or an extract obtained by extracting with a mixed solvent of water and an organic solvent. The method of claim 3,
Wherein the organic solvent is at least one selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, n-butanol, isobutanol, ether, ethyl acetate and chloroform. 7. A cosmetic composition comprising the composition according to any one of claims 1 to 6. 6. The method of claim 5,
Wherein the cosmetic composition comprises 0.0001 to 30% by weight of the skin whitening composition or the skin moisturizing composition based on the total weight of the cosmetic composition.