KR101205186B1 - The cosmetic compositions with the improved permeation of hydrophilic active ingredients - Google Patents

Abstract

The present invention relates to a cosmetic composition that promotes skin permeation of a hydrophilic bioactive substance having low skin permeability by using a low phototoxic antiviral extract.

Description

The cosmetic compositions with the improved permeation of hydrophilic active ingredients

The present invention relates to a cosmetic composition for promoting skin penetration of hydrophilic bioactive substances, more specifically 0.1 to 5.0% by weight of lauryl acid derivative, methylpyrrolidone (N-Methyl-2-pyrrolidone) 0.1 to 10.0 It contains the weight%, ethyl alcohol 2.0 ~ 20.0% by weight, and 0.001 ~ 30.0% by weight of the dandelion ball mojaban extract prepared by the method for extracting phototoxic substances, it is excellent in promoting the skin penetration of hydrophilic bioactive substances and phototoxicity by UV And to cosmetic compositions having a very low likelihood of inflammation.

The skin is histologically composed of epidermis, dermis, subcutaneous fat and the like, and its thickness varies according to the site, age and sex. Although various skin conditions exist, these skins each play a role of external barrier function and other physiological functions. The stratum corneum is a layer of keratinized dead cells in which a relatively high hydrophobicity-insoluble protein keratin is concentrated, interposed between an intercellular lipid bilayer, a thin layer of lipids. As a structure, the unique hierarchical structure of the matrix, rich in lipids and insoluble proteins, makes it impermeable to chemical factors, including various compounds from external environments. In particular, the hydrophilic bioactive material is very difficult to permeate due to this hydrophobic multiple structure of the stratum corneum.

Therefore, in general, the transport of substances through keratin, especially the hydrophilic bioactive substances in healthy skin, is not easily achieved due to various protective actions of the skin, and only a very low concentration is transmitted even if permeated. However, in the case of cosmetics, the bioactive substances contained in the cosmetics must pass through the stratum corneum, the outermost layer of the skin in order to show its efficacy, and in the case of hydrophilic bioactive substances, due to the hydrophobic multiple structure of the stratum corneum and its barrier function , Bioactive substances contained in cosmetics have a problem that does not sufficiently exhibit its function. In order to overcome these skin barriers and the hydrophilic bioactive substances contained in cosmetics to function, cosmetics must contain high concentrations of bioactive substances. Due to the high concentration of physiologically active substances, there is a high possibility of causing skin irritation, and a large number of active substances have a characteristic of inhibiting the stability of the cosmetics, and when prescribed at high concentrations, problems appear in productization. Therefore, in order to overcome such a problem, it is essential to develop a skin permeation promoting cosmetic that can be effectively absorbed through the skin by adding a small amount of bioactive substance to the cosmetics stably.

Various studies have reported skin permeation accelerators that promote the permeation of active substances, and their permeation promotion is largely related to ① intercellular lipid bilayer ② desmosome involved in intercellular binding. ) And related proteins, ③ can be divided into mechanisms that act directly on the internal structure of keratinocytes. The action of a delivery enhancer on the intercellular lipid bilayer is primarily a mechanism that loosens the structure of the lipid bilayer, facilitating the delivery of the active substance. A large number of skin penetration promoters facilitate delivery through this function. It is known. Some dermal penetration promoters are known to act on desmosomes and related proteins involved in intercellular binding of keratinocytes, thereby facilitating delivery through mechanisms that separate or loosen keratinocyte intercellular bonds and increase intercellular spacing. Other skin penetration promoters are known to facilitate delivery through mechanisms that penetrate inside keratinocytes to denature keratin or affect internal structure, facilitating delivery through the transcellular route.

In other words, the skin permeation accelerator inevitably alters the structure of the stratum corneum, the outermost skin layer, to promote skin permeation of the bioactive substance. However, due to this, most skin permeation accelerators have high cytotoxicity to skin cells and have a high possibility of causing skin irritation such as causing skin burning and erythema when applied. Therefore, there is a need for measures to alleviate skin irritation such as minimizing the amount of use through effective combination of skin permeation accelerators.

Recently, in order to reduce skin irritation caused by various chemicals, a lot of attention has been focused on functional cosmetics having functions such as wrinkle relaxation and whitening obtained from natural extracts. Natural materials have less side effects on the skin, and as the consumer's response to external skin preparations using natural materials increases, the development value as a raw material of the cosmetic composition is increasing. For example, Japanese Patent Application Laid-Open No. 2-245087 describes the antioxidant effect of Sargassum genus extract, and Japanese Patent Application Laid-open No. Hei 3-294384 describes an antioxidant composition extracted from Hijikia genus. have. Japanese Patent Laid-Open No. 7-10769 describes a extract of Phaeophyceae having an astringent effect. U.S. Patent No. 5,508,033 describes a cosmetic having anti-radical activity, including mother and child, and Japanese Laid-Open Patent Publication No. 2008-056643 discloses a sargassum fulvellum having functions such as antioxidant activity, cell metabolism activity, and skin aging prevention. ) Cosmetics containing extracts are disclosed, and Japanese Patent Laid-Open Publication No. 1998-330280 discloses a mother-child extract containing cosmetics that inhibits collagen production promotion, wrinkles or reduced elasticity.

It is an object of the present invention to provide a cosmetic composition that increases the skin permeability of the hydrophilic bioactive substance and minimizes the possibility of inflammation by using the mother leaf extract with reduced phototoxicity.

In order to develop such a skin permeation promoting cosmetic composition, the present inventors screened materials having excellent skin absorption ability through data research and lauryl acid derivatives after a long study of performing skin absorption capacity tests on hydrophilic bioactive substances. Methylpyrrolidone, ethanol and phototoxic substances Excluded skin penetration promoting cosmetic composition containing the extract produced by the method of extract extraction method has a low skin permeability significantly lower the functionality of the hydrophilic bioactive substances in the skin effectively through the skin Permeation can maximize the functionality of physiologically active substances and minimize the possibility of phototoxicity and inflammation caused by skin due to the inclusion of lauryl acid derivatives, methylpyrrolidone, and ethyl alcohol due to the addition of mothban extract. I found it.

In the present invention, the term "hat hat" refers to various types of hat bands belonging to the genus Sargassum Sp. Maternity hat, large berry hat, zangari hat, slipper, Selman hat (Miyabe hat), black hat, twin hat, double leaf hat, beetle hat, saw blade hat, pretzel hat, thin leaf hat , Worms, torsional hats and warp ball hats.

In the present invention, "hydrophilic bioactive material" refers to a material that is hydrophilic and has an excellent skin improvement effect, and is not particularly limited as long as it is known to those skilled in the art to which the present invention belongs, for example, arbutin, Adenosine, vitamin C and its hydrophilic derivatives, other hydrophilic vitamins and derivatives thereof including vitamin B3, vitamin B5 and vitamin H, centella asiatica extract including acetylglucosamine, madecassoside, selenium aspartate, various plant extracts and Refers to various peptide components or mixtures of two or more components. The content of these hydrophilic bioactive substances is determined according to the activity and solubility of each bioactive substance.

The present invention relates to a cosmetic composition for promoting skin permeation of a hydrophilic bioactive material using a low phototoxic mothban extract prepared by an extract manufacturing process capable of effectively excluding only phototoxic substances from the mothball.

The present invention is a hydrophilic physiological activity, characterized in that it contains at least one of lauryl acid and lauryl acid derivatives, methylpyrrolidone, ethanol, low phototoxic low alban extract and hydrophilic bioactive substances prepared by the above method. Provided is a cosmetic composition for promoting skin penetration of a substance.

In the present invention, the lauryl acid derivative is characterized in that one or more selected from the group consisting of lauryl acid alcohol, lauryl acid amine, lauryl acid PEG-12, lauryl acid sucrose and glycerol lauryl acid.

In addition, the lauryl acid or lauryl acid derivative in the present invention is characterized in that it contains 0.1 to 5.0% by weight relative to the total composition. When the content of the lauryl acid or lauryl acid derivative is less than 0.1% by weight, there is little effect of promoting skin absorption on the hydrophilic bioactive substance, and when it exceeds 5.0% by weight, it is precipitated in the cosmetics due to low solubility and thus cosmetics. There is a possibility of destroying the formulation of the composition or causing skin sensitization and skin irritation.

In addition, the methylpyrrolidone in the present invention is characterized in that it contains 0.1 to 10.0% by weight relative to the total composition. When the content of methylpyrrolidone is less than 0.1% by weight, there is little effect of promoting skin absorption on hydrophilic bioactive substances, and when it exceeds 10.0% by weight, skin sensitization and skin irritation may be caused.

In addition, the ethyl alcohol in the present invention is characterized in that it contains 2.0 to 20.0% by weight relative to the total composition. When the content of the ethyl alcohol is less than 2.0% by weight, there is almost no skin absorption promoting effect on the hydrophilic bioactive substance, and due to the low solubility of the lauryl acid derivative, the lauryl acid derivative is precipitated in the cosmetic to form a formulation of the cosmetic composition. It may be destroyed, and if it exceeds 20.0% by weight, it may cause skin sensitization and skin irritation.

In addition, in the present invention, the low phototoxic molar algae extract is characterized in that it contains 0.001 to 30% by weight based on the freeze-drying weight of the total composition. When the content of the extract is less than 0.001% by weight, there is little effect of inhibiting phototoxicity and inflammation-induced effect by UV rays and skin permeability, and when the content of the extract is more than 30.0% by weight, the effect of increasing the content is not economical.

In addition, the hydrophilic bioactive material in the present invention is characterized in that it contains 0.001 to 30% by weight relative to the total composition. When the hydrophilic bioactive substance content is less than 0.001% by weight, the bioactive function is weak, and when more than 30.0% by weight, the effect of increasing the effect of the content is not economical.

The cosmetic composition for promoting skin permeation of the hydrophilic bioactive substance may be applied to various functional cosmetic compositions such as aging functional cosmetics and whitening functional cosmetics in addition to general cosmetic compositions such as lotions, lotions and creams. In addition, even if the hydrophilic bioactive material has an effect of promoting hair growth, the composition of the present invention may be applied to promote skin absorption thereof.

In addition, the cosmetic composition may be prepared in a variety of formulations, there is no particular limitation on the formulation, for example, it can be prepared in a cream, lotion, emulsion, essence and the like.

In addition, the present invention is characterized in that, in the preparation of the low light phototoxic extract, at least one of the reduced pressure concentration process and the freeze-drying process after the extract preparation process.

In addition, the present invention is characterized in that at least one of the low pressure concentration step and the freeze-drying step after the solvent fractionation step in the preparation of low phototoxic low moth extract.

In addition, the present invention, in the preparation of the low phototoxic low moss extract, the solvent fractionation process by adding a high polar solvent to the prepared moss extract first and then mixed with a low polar solvent to mix, disperse, settle the low Removing the polar solvent layer.

In addition, the present invention is characterized in that the extraction solvent is at least one solvent selected from purified water, methanol, ethanol, butanol, ethyl acetate, glycerin, butylene glycol, propylene glycol in the preparation of low phototoxicity.

In addition, the present invention is characterized in that the high polar solvent is composed of one or more solvents selected from purified water, methanol, ethanol, butanol.

In addition, the present invention is characterized in that the low polar solvent is composed of one or more solvents selected from hexane, dichloromethane, chloroform.

In addition, the present invention is characterized in that the high polar solvent: low polar solvent in the ratio of 1: 5 to 3: 1 by volume ratio during mixing and dispersion of the high polar solvent and the low polar solvent.

In addition, the present invention is characterized in that the hat platter is a dandelion ball hat.

More specifically, in the present invention, the low phototoxic molar algae extract is prepared by using one or more solvents selected from purified water, methanol, ethanol, butanol, ethyl acetate, glycerin, butylene glycol, and propylene glycol as extraction solvents. Extract obtained by cold condensation or heat filtration at room temperature using a solvent or additionally concentrated under reduced pressure or lyophilization of the solvent, step 2: extract prepared by using at least one solvent selected from purified water, methanol, ethanol, butanol After uniformly dispersing in the configured high polar solvent, the low polar solvent composed of one or more solvents selected from hexane, dichloromethane and chloroform and mixed and dispersed in a ratio of 1: 5 to 3: 1 by volume ratio, and then allowed to stand, Solvent fractionation step to remove the layer, step 3: The solvent was further concentrated under reduced pressure It is prepared by freeze-drying.

In addition, the high polar solvent used in the preparation method of the phototoxic agent-excluded mother-ban extract of the present invention is characterized in that the content of purified water is in the range of 5.0-90.0% by weight, when the content of purified water is less than 5.0% by weight It is impossible to maintain the separated phase, so that it is not possible to effectively separate phototoxic substances, and if the content of purified water exceeds 90.0% by weight, the active ingredient contained in the Maban extract cannot be sufficiently dissolved in the high polar solvent, and thus the low polar solvent. Since the active ingredients are distributed in layers, the physiological activity of the prepared phototoxic inducer-excluded mother extract is low and the production yield is not economical.

In addition, the volume ratio of the high polar solvent and the low polar solvent used in the method for preparing phototoxic inducing substance banban extract of the present invention is 1: 5 to 3: 1 ratio, the volume ratio of the high polar solvent is less than 1: 5 In this case, the phase separated from the low polar solvent cannot be maintained, or a sufficient amount of maban extract cannot be used in a limited manufacturing container condition, so that the yield of the extract is not effective, and the volume ratio of the high polar solvent exceeds 3: 1. In this case, the phototoxic inducers contained in the Mabanban extract cannot be sufficiently dissolved in a low polar solvent, and thus the phototoxicity of the prepared Maban extract excludes the Mabanban extract, which is not effective.

The present invention provides a skin permeation promoting cosmetic composition containing a lauryl acid derivative, methylpyrrolidone, ethyl alcohol and low phototoxic extract, and as a skin external preparation or cosmetic that exhibits a predetermined functional effect while having less skin irritation It is available.

In addition, the present invention can be used as a skin permeation cosmetic composition that can significantly maximize the functionality of the bioactive material by effectively penetrating through the skin of the hydrophilic bioactive material having a low functional skin penetration is low.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples. In addition, in the present embodiment, the extract was prepared by selecting the hard-end bead hat jar among the hat bands, but it is clear that the scope of the present invention is not limited to the hard-end bead hat disc. In addition, in the present embodiment, the cosmetic composition was prepared by selecting arbutin as a hydrophilic bioactive substance model drug having a low skin permeability, but it is clear from the present invention that the range of the hydrophilic bioactive substance is not limited to arbutin.

Example 1: Mabanban extract preparation

The dried hard ball bead hatch was refluxed three times for 5 hours in an 80% (V / V) ethanol aqueous solution, cooled, and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C. or lower to prepare Hatban extract.

Example 2: Preparation of low half phototoxic extract

After dissolving and dissolving 10 g of the lyophilized lyophilized extract obtained in Example 1 in purified water, methanol and ethanol in a volume ratio of 3: 4: 3 evenly, 1 l of a highly polar mixed solvent was dissolved hexane, dichloromethane, chloroform The mixture was stirred with 1 L of the low polar mixed solvent mixed at a volume ratio of 7: 2: 1. After stirring the mixed liquid for 8 hours in a sealed state at room temperature, the low polar mixed solvent layer and the high polar mixed solvent layer were carefully separated. The recovered high polar mixed solvent layer was filtered through Whatman # 5 filter paper, and concentrated under reduced pressure and freeze-dried at 50 ° C. or lower to prepare a mothban extract that excludes phototoxic substances.

Example 3: Skin permeation promoting effect confirmation experiment

This example was carried out to determine the effect of each sample on the skin permeability of the model drug. The skin used in the experiment was to remove the dorsal skin of 8-10 weeks old hairless mouse, remove subcutaneous fat and tissues carefully to avoid damaging the skin with saline solution, and store it at -70 ℃ until the skin is flattened. It was thawed and used before the experiment. The frozen hairless mouse skin was slowly thawed at room temperature prior to the experiment and placed between the receptor compartment with the stratum corneum facing the donor compartment of the Franz diffusion cell and then placed on each surface of the sample and on the surface above the skin. Arbutin selected as a hydrophilic bioactive substance model was administered. In the receiver compartment, pH 7.4 phosphate buffered saline was added and stirring was continued at 300 rpm while maintaining the temperature at 37 ° C. The area of skin in contact with the recipient compartment was 2.0 cm 2, and the capacity of the recipient compartment was 5 ml. After administering 1.0 ml of each sample to the skin surface, the samples were collected using a fraction collector.

All samples taken were filtered before scanning the column with a 13 mm disk filter (pore size 0.45 μm) and analyzed with a Waters 2690 alliance separation module (Waters, USA) equipped with a photodiode array detector system 996 (Waters, USA). . The Xterra RP18 (5 μm, 4.6 x 205 mm, Waters, USA) column was used as the stationary phase, and the arbutin mobile phase was mixed with methanol, water and acetic acid buffer with a solvent of 10:80:10, and the flow rate was 0.8 ml / min. Was detected at a wavelength of 280 nm.

The amount of drug permeated per unit area of skin was plotted against time and then the drug flux at equilibrium was calculated from the slope of the site showing linearity. The delay time (Lag time, T _L ) was obtained from the time when the straight line passed the x axis. The effects of the base and skin permeation accelerators were compared and calculated by calculating the transdermal penetration ratio (ER), drug flow rate and permeability at equilibrium using the following equations (1) and (2).

J _s is the drug flow at equilibrium, T _L is the delay time, S is the area where permeation occurs, (dQ / dt) _ss is the slope obtained by plotting the amount of drug passing through the skin at equilibrium over time, C _d is the saturation concentration in the base of the drug, and K _p is the transmission coefficient. Indicates.

Example 4: Confirmation of skin permeation promoting effect of saturated fatty acid

This example was conducted to determine the effect of saturated fatty acid on the skin permeability of arbutin, a model drug. Dermal by the method of Example 3 using 1.0 ml each of the model drug 2% arbutin, 0.5% of each saturated fatty acid sample, 50% of ethyl alcohol and 47.5% of purified water (control group added 0.5% of purified water instead of saturated fatty acid) The amount of permeated arbutin was measured to confirm the skin permeation promoting effect of the model drug arbutin by saturated fatty acids (Table 1).

sample Latency (hr) Drug flow rate (ug / cm ² / hr) Percutaneous penetration rate (%) Percutaneous penetration rate Control group 5.9 2.33 1.12 One Octanoic acid 5.1 37.6 18.05 16.16 Decanoic acid 4.5 82.99 39.84 35.77 Lauryl acid (dodecanoic acid) 3.1 85.27 40.93 36.65 Myristic acid 2.6 83.88 40.26 36.06

As can be seen from Table 1, all carbonic acid saturated fatty acids, octanoic acid, decarbonic acid with 10 carbon atoms, lauryl acid with 12 carbon atoms and myristic acid with 14 carbon atoms, all promote transdermal permeation compared to the control group for arbutin, a hydrophilic active ingredient. The rate was confirmed to be 16 or more. In particular, the rate of transdermal penetration of lauryl acid was the best at 36.65, and the delay time was 3.1 hours. In addition, transdermal permeability also showed the best result with 40.93%.

Example 5 Skin Permeation Promoting Effect of Lauryl Acid Derivative

In this embodiment, the lauryl acid derivatives of the present invention are lauryl alcoho, lauryl amine, lauryl acid PEG-12, sucrose laurate and glycerol laurate. To determine the effect of arbutin, a drug model of glyceryl laurate, on skin permeability. 1.0 ml each of the sample drug 2% arbutin, 0.5% of each lauryl acid derivative, 50% of ethanol and 47.5% of purified water (control group added 0.5% of purified water instead of lauryl acid derivative), Example 3 By measuring the amount of arbutin permeated percutaneously by the method of to determine the skin permeation promoting effect of the arbutin model drug by the lauryl acid derivative (Table 2).

sample Delay time
(hr) Drug flow rate
(ug / cm ² / hr) Transdermal Permeability
(%) Percutaneous penetration rate
Control group 7.6 10.37 2.49 One Lauryl acid 1.8 215.43 51.71 20.77 Lauric acid alcohol 1.2 293.58 70.46 28.31 Lauric acid amine 1.1 441.12 89.87 42.52 Lauryl Acid PEG-12 5.3 53.03 12.73 5.11 Lauryl acid sucrose 5.1 91.23 21.91 8.79 Glycerin Lauric Acid 4.4 203.95 48.95 19.66

As can be seen in Table 2, among the lauric acid derivatives, arbutin promoted the skin absorption of lauryl acid amine. The rate of transdermal penetration was 42.52, and the delay time was decreased by about 7 times compared to the control group. Percutaneous permeation rates of uryl acid alcohol, laurylcin PEG-12, lauryl acid sucrose, and glycerol glycerol were 20.77, 28.31, 5.11, 8.79, and 19.66, respectively, which significantly increased skin absorption compared to the control group.

Example 6: Confirmation of skin permeation promoting effect of the combination of amine lauryl acid and methylpyrrolidone

This example was performed to determine the effect of the optimized transdermal penetration composition on the skin permeability of arbutin, a model drug, through the combination of lauryl acid amine and methylpyrrolidone, the lauryl acid derivative of the present invention. Percutaneous permeation was carried out by the method of Example 3 using 1.0 ml of each of the sample drug, which was mixed with a model drug, 2% arbutin, 1.0% lauryl acid amine, 5.0% methylpyrrolidone 5.0% ethanol and purified water. The amount of arbutin was measured to confirm the skin permeation promoting effect of the model drug arbutin.

sample Delay time
(hr) Drug flow rate
(ug / cm ² / hr) Transdermal Permeability
(%) Percutaneous penetration rate Control group 4.98 1.88 0.26 One Lauric acid amine 1.0% 1.76 218.49 30.59 116.39 Methylpyrrolidone 5.0% 4.62 2.89 0.4 1.54 Lauric acid amine 1.0%
+ Methylpyrrolidone 5.0% 1.09 231.19 32.37 123.16

As can be seen in Table 3, the methylpyrrolidone 5.0% treatment group was found to have a weak transdermal penetration promoting effect of the model drug arbutin. In contrast, when a combination of 1.0% lauryl acid amine and 5.0% methylpyrrolidone was treated, it was confirmed that the percutaneous permeation promoting effect was increased by at least about 7% compared to the lauryl acid amine 1.0% alone treatment group.

Examples 7 to 8 and Comparative Example 1

This embodiment is to prepare a cosmetic containing a reciprocal lauryl amine derivative and lauryl acid amine and methylpyrrolidone to prepare a transdermal permeation composition and a extract of a transdermal bead capsulban extract prepared by a method for extracting phototoxic stimulant exclusion extract Through the experimental method of Example 3, the effect on the skin permeability of the arbutin as a model drug was evaluated by performing a comparative experiment with Comparative Example 1.

Cosmetics used in the comparative experiments are in the form of a gel, the composition of which is shown in Table 4. First, the a) phase recorded in Table 4 is heated to dissolve and mix. A powder of b) was slowly added thereto, uniformly dispersed in a homomixer, and then cooled slowly to prepare a gel (Examples 7 to 8 and Comparative Example 1).

division Raw material name Example 7 Example 8 Comparative Example 1 end Laurylamine 0.15% 0.15% Methylpyrrolidone 5.0% 5.0% Citric acid 0.06% 0.06% Ethyl Alcohol 20% 20% 20% Purified water 61.38% 56.38% 66.59% Arbutin 2.0% 2.0% 2.0% Application of phototoxic exclusion process 5.0% - Fiji-26-Bootes-26 / PG-40 Hydrogenated Castor Oil 0.4% 0.4% 0.4% Glycereth-26 10% 10% 10% Triethanolamine 0.01% 0.01% 0.01% I Polyquaternium-37 1.0% 1.0% 1.0%

Note) Unit: wt%

sample Delay time
(hr) Drug flow rate
(ug / cm ² / hr) Transdermal Permeability
(%) Percutaneous penetration rate Control group 3.95 3.38 0.88 One Comparative Example 1 1.98 6.19 1.59 1.8 Example 7 1.11 113.22 29.44 33.45 Example 8 0.98 123.68 32.16 36.55

As can be seen in Table 5, the gel formulation without the addition of laurylamine and methylpyrrolidone showed a transdermal permeability similar to that of the control group, indicating a weak transdermal permeation effect of the model drug arbutin. On the contrary, in Examples 7 to 8, in which 0.15% of lauryl acid amine, 5.0% of methylpyrrolidone, and 20.0% of ethyl alcohol were added, transdermal permeation rate was found to be 33 or more, which has a very good transdermal permeation promoting effect. It was confirmed.

Example 9 Cytotoxicity Confirmation Experiment of Skin Permeation Promoting Formulation

This example was carried out to evaluate the cytotoxicity of skin cells by treatment of the transdermal penetration gel formulations obtained in Examples 7-8 and the blank test gel formulation of Comparative Example 1.

Fibroblasts (fibroblasts) were placed in a 24-well test plate 1 x 10 ⁵ and attached for 24 hours. Each well was washed once with PBS (Phosphate buffered saline), and each well was treated with 1 ml of cell culture medium (10% FBS added to DMEM) and a sample diluted to an appropriate concentration, followed by incubation for 24 hours. . After 24 hours, the medium was removed, and 500 μl of the cell culture medium and 2.5 μl / ml of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) And then cultured in a 37 ° C CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added. After shaking for 5 minutes, the cells were lysed and 100 쨉 l of the supernatant was transferred to a 96-well test plate, and the absorbance at 565 nm was measured in a microplate reader. Cell survival rate (%) was measured by Equation 3, and the concentration of the sample that did not affect the survival of the cells was determined.

Bo: Absorbance at 565 nm of wells that had undergone color development of the cell culture medium

Bt: absorbance at 565 nm of the well that underwent chromogenic reaction without treatment of the sample

St: absorbance at 565 nm of the well treated with the sample and color-

As can be seen in Table 6, Example 7, which is a gel formulation containing lauryl acid amine and methylpyrrolidone, showed a significant increase in cytotoxicity compared to Comparative Example 1, which contained 100% cell viability and 0.05%. It became. In contrast, the formulation of Example 8 containing the phototoxic agent exclusion light bulb bead capillary extract of Example 2 in addition to lauryl acid amine and methylpyrrolidone was significantly improved in safety with 100% cell survival concentration of 0.2%. It was confirmed. Therefore, it was confirmed that the cosmetics containing the skin permeation accelerator exhibited a safety improvement effect due to the improvement of skin cytotoxicity, because the extract containing the phototoxic moxibustion ball extract mothball extract of Example 2 when the skin was actually applied.

Name of sample 100% cell survival concentration Comparative Example 1 0.2% Example 7 0.05% Example 8 0.2%

Example 10: Experiment for confirming phototoxicity and cell damage protection by ultraviolet irradiation

This Example was carried out to evaluate the phototoxicity by the treatment of the sample after UV irradiation by the treatment of the transdermal penetration gel formulations obtained in Examples 7 to 8 and the blank test gel formulation of Comparative Example 1 and cell damage protection by UV rays. It was.

By comparing the cell death rate of cells treated with only UV light and those treated with each sample at the same time after UV treatment, the cell death rate was used to determine the phototoxicity and protection against UV damage of cells against cell damage caused by UV light. You can check it. Keratinocyte was added to the 24-well test plate at 1 × 10 ⁵ cells for 24 hours. Each well was washed once with PBS (phosphate buffered saline), and 500 μl of PBS (Phosphate buffered saline) was added to each well. The keratinocytes were irradiated with 10 mJ / cm 2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki Co., Japan), and then PBS (phosphate buffered saline) 10% FBS added) was added. Each sample was treated with 0.2% and incubated for 24 hours. After 24 hours, the medium was removed, and 500 μl of the cell culture medium and 2.5 μl / ml of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) And then cultured in a 37 ° C CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added. After shaking for 5 minutes, the cells were lysed and 100 쨉 l of the supernatant was transferred to a 96-well test plate, and the absorbance at 565 nm was measured in a microplate reader. Cell survival rate (%) was measured by Equation 3, and the concentration of the sample that did not affect the survival of the cells was determined.

Name of sample Cell survival rate (%) UV alone treatment group 76.2% Comparative Example 1 54.3% Example 7 22.6% Example 8 68.3%

As a result, as shown in Table 7, it was confirmed that about 24% of cells were killed by UV treatment alone. In addition, when the gel formulation which does not contain a skin permeation accelerator such as ultraviolet rays and lauryl acid amine and UV irradiation was added in parallel, about 46% of the cells were killed to confirm the phototoxic effect by the gel formulation and ultraviolet irradiation. In particular, in Example 7, containing lauryl amine, methylpyrrolidone, etc., about 77% of the cells were killed, and the phototoxicity due to ultraviolet irradiation was greatly increased by the addition of lauryl amine, methylpyrrolidone, etc. I could confirm that. On the other hand, the formulation of Example 8 containing the phototoxic trigger material exclusion light bulb bead cap extract of Example 2 significantly improved the cell viability to 68.3%, which was higher cell death rate compared to the UV alone treatment group, but Comparative Example 1 Compared to the formulation of, the phototoxicity caused by ultraviolet light was canceled. It was confirmed that the cell damage caused by ultraviolet rays was greatly reduced due to the inclusion of the phototoxic molecule-excluded light bulb bead cap extract.

Example 11: Skin irritation potential test

This example was carried out to evaluate the possibility of irritation in the use of the actual skin application of the transdermal penetration gel formulations obtained in Examples 7 to 8 and the blank test gel formulation of Comparative Example 1.

0.2 mg of each product was applied to the upper arm of 20 healthy men and women aged 20-50 using FINN CHAMBER (FINLAND) and subjected to a closed patch test for 48 hours to remove the degree of erythema formation and test subjects. The degree of stimulation was confirmed through questions. Determination of skin irritation was made according to the skin irritation index criteria of Table 8, which was made by using sodium lauryl sulfate, and no irritation of skin irritation indexes 0 to 1, no stimulation of 2 to 3 irritation, 4 or more The stimulus was judged to be a strong stimulus. The experimental results are shown in Table 9.

division Stimulus index Criteria stimulus
none 0 No erythema has formed and the investigator does not complain of irritation or itching. One Erythema did not form, but the investigator complained of minor itching. light
stimulus 2 There is a slight skin change or the investigator complains of minor irritation. 3 Weak erythema is formed and the investigator complains of irritation strong
stimulus 4 A clear form of erythema was formed and the investigator complained of irritation. 5 Vivid red erythema formed, and test subject complains of strong irritation

division Causes of irritation Irritation Rate Average Stimulus Index No irritation Slight irritation Strong stimulation Comparative Example 1 4 16 0 80% 2.05 Example 7 0 14 6 100% 2.95 Example 8 3 16 One 85% 2.3

As can be seen from the results of Table 9, the gel formulation of Example 8 added with lauryl acid amine, methylpyrrolidone, etc., Comparative Example 1 containing no skin permeation accelerator with a stimulation incidence of 100% at an average irritation index of 2.95 Compared to the mean stimulation index of 2.05 and 80%, the stimulus incidence rate was significantly higher. In contrast, the formulation of Example 8 containing the phototoxic stimulant exclusion light bulb bead capillary extract of Example 2, in addition to lauryl acid amine, methylpyrrolidone, and the like, had a stimulation incidence of 85% and a similar stimulation rate as that of Comparative Example 1. The results showed that the likelihood of stimulation was significantly reduced.

Claims (15)

a) at least one member selected from the group consisting of lauryl acid, lauryl acid alcohol, lauryl acid amine, lauryl acid PEG-12, lauryl acid sucrose and glycerol lauryl acid;
b) methylpyrrolidone;
c) ethanol;
d) a hydrophilic skin improving substance selected from arbutin, adenosine, hydrophilic vitamins and derivatives thereof that penetrates the percutaneous and has a skin improving effect; And
e) a) preparing a mabanban extract using at least one extraction solvent selected from purified water, methanol, ethanol, butanol, ethyl acetate, glycerin, butylene glycol, and propylene glycol;
B) Add one or more high polar solvents selected from purified water, methanol, ethanol and butanol and one or more low polar solvents selected from hexane, dichloromethane and chloroform to the extract prepared in the above process, and then mix, disperse, and leave the mixture. And a solvent fractionation step of removing one low polar solvent layer. A cosmetic composition for promoting skin permeation of a hydrophilic skin improvement material, comprising: a low phototoxic low half-cover extract prepared by the method.
delete The method according to claim 1,
At least one selected from the group consisting of lauryl acid, lauryl acid alcohol, lauryl acid amine, lauryl acid PEG-12, lauryl acid sucrose, and glycerol lauryl acid contains 0.1 to 5.0 weight of the total composition. Composition.
The method according to claim 1,
The methylpyrrolidone composition is characterized in that it contains 0.1 to 10.0% by weight relative to the total composition.
The method according to claim 1,
The ethyl alcohol is a composition, characterized in that containing 2.0 to 20.0% by weight relative to the total composition.
The method according to claim 1,
The low toxicity photo moth extract is a composition characterized in that it contains 0.001 to 30% by weight relative to the total composition.
The method according to claim 1,
The hydrophilic skin improvement material is a composition, characterized in that it contains 0.001 to 30% by weight relative to the total composition.
The method according to claim 1,
The composition of claim 1, wherein at least one of the reduced pressure concentration step and the freeze-drying step is added after the extract preparation step.
The method according to claim 1,
Wherein b) after the solvent fractionation step, the composition characterized in that the addition of one or more of the reduced pressure concentration process and lyophilization process.
The method according to claim 1,
The b) solvent fractionation process is characterized in that to remove the low-polar solvent layer after mixing and dispersing, standing still by adding a high polar solvent to the prepared Maban extract and then mixed with a low polar solvent.
delete delete delete The method according to claim 1,
The composition of the high polar solvent and the low polar solvent when mixing and dispersing the high-polar solvent: low polar solvent is a volume ratio of 1: 5 to 3: 1.
The method according to claim 1,
The hat band is a composition characterized in that the ball head bead hat.