WO2019009663A1 - Composition for preventing and ameliorating athlete's foot - Google Patents

Abstract

The present invention relates to a composition for preventing and ameliorating athlete's foot. More specifically, the present invention relates to a cosmetic or pharmaceutical composition for preventing and ameliorating athlete's foot, which has skin protection, antioxidant and antimicrobial activity simultaneously by comprising a natural oil as a main ingredient. Since the composition for preventing or ameliorating athlete's foot according to the present invention comprises a natural oil as a main ingredient, and thus has few side effects such as toxicity and skin irritation and exhibits skin protection, antioxidant and antibacterial activity at the same time, said composition can be preferably used as a cosmetic or pharmaceutical composition for prevention and improvement of athlete's foot.

Description

Composition for prevention and improvement of athlete's foot

The present invention relates to a composition for prevention and improvement of athlete's foot. More particularly, the present invention relates to a cosmetic or an external-use composition for prevention and improvement of athlete's foot having skin protection, antioxidant and antimicrobial action simultaneously with natural oil as a main ingredient.

The study and efficacy of physiological mechanism through dermatological science has been studied from extracting and purifying the active ingredient from natural materials as a new high material, from the study of merely staying in the dimension of skin protection, There is a surge in demand for.

The hormone abnormality due to excessive stress in modern society and sudden changes in the external environment may cause the abnormal keratinization of the stratum corneum to fall off due to atmospheric pollutants or ultraviolet rays, The skin tone becomes unstable and the skin becomes rough and dull.

Skin aging is caused by oxidative damage of intracellular substances by free radicals. That is, highly reactive free radicals, such as active radicals such as hydroxy radicals and superoxide radicals, oxidize the lipid components constituting the cell membrane to destroy the cell membrane, thereby oxidizing the protein and stopping the function of the nucleic acid, It destroys its components and causes cell death. Such skin aging can be caused by excessive exposure to ultraviolet rays, nutritional deficiency of skin cells, and side effects of cosmetics.

By inhibiting oxidative damage in vivo, aging of the skin can be prevented or reduced. Antioxidant cosmetics, on the other hand, inhibit the oxidative damage of skin, including the oxidation of fat components, to prevent or delay skin aging.

In particular, the production amount and the market growth rate of a multi-functional cosmetic having two or more functionalities (for example, including a wrinkle-improving function and a whitening function at the same time) in one product among functional cosmetics are much higher than those of a single functional cosmetic product.

Therefore, according to the environment-friendly and nature-oriented trend, effective substances used in cosmetics are based on the usefulness of not only chemical substances but also natural substances derived from plants, and natural materials having few side effects on the skin are compounded in various forms in cosmetics .

Korean Patent Registration No. 10-1117564 (Feb. 10, 2012) discloses that excessive salt and / or sugar; Water phase component; Anionic, amphoteric and nonionic surfactants; And a polyol, wherein the salt is supersaturated and dissolved in the cosmetic composition, and the supersaturated and dissolved salt is present in the form of recrystallized particles, the water component is purified water, and the anionic surfactant is ammonium laureth sulfate , The amphoteric surfactant is cocamidopropyl betaine, the nonionic surfactant is cocamide di-is, and the polyol is glycerin.

The patent is based on the finding that the patent is capable of obtaining exfoliation, skin amelioration, blood circulation promoting effect and formulation safety by recrystallized salt and / or sugar particles and of remaining in a saturated solution of salt and / Salt and / or sugar exhibits a moisturizing function. It contains anionic, amphoteric and nonionic surfactant, and is excellent in skin cleansing effect and rinsability. Also, it is excellent in ease of use in the form of a gel. However, There is a problem that the function of the normal cells due to the sterilization action peculiar to the salt is deteriorated and the collagen is destroyed thereby causing excessive aging of the skin.

Therefore, it is required to overcome the limitations of the prior art, to have fewer side effects, and to develop a product of a natural material capable of improving the condition of the skin, which is excellent in antimicrobial, antioxidant and anti-inflammatory effects, which are essential for skin health.

The inventors of the present invention have made intensive studies to develop a cosmetic or composition for prevention and improvement of athlete's foot which has skin protection, antioxidant and antimicrobial action simultaneously with natural products, and as a result, Of course, it has been confirmed that it has an antioxidative and antimicrobial action and completed the present invention.

Accordingly, it is an object of the present invention to provide a method for producing a flavor enhancer which comprises 50 to 75% by weight of tea tree oil, 10 to 30% by weight of avocado oil, 3 to 15% Athlete's foot (50%), vitamin E acetate (0.5 to 2 wt%), polysorbate (20) 0.5 to 2 wt%, benzyl alcohol 0.01 to 0.05 wt% and thymol 0.01 to 0.05 wt% Or an external composition for improvement.

The composition for prevention or improvement of athlete's foot according to the present invention is a cosmetic or pharmaceutical composition for prevention and improvement of athlete's foot since the composition for preventing or improving athlete's foot is natural oil as a main ingredient and exhibits skin protection, antioxidant and antibacterial action simultaneously as well as less side effects such as toxicity and skin irritation Can be preferably used.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing DPPH free radical scavenging performance of the example compositions.

2 is a graph showing the hydroxy radical scavenging activity of the example composition.

3 is a graph showing the results of Fe ²⁺ chelating assay analysis of the composition of the examples.

4 is a graph (pH 1.2) showing the nitrite scavenging activity of the composition of Example.

In one aspect, the present invention provides a method for producing an extract, comprising, in one aspect, 50 to 75 wt% of tea tree oil, 10 to 30 wt% of avocado oil, 3 to 15 wt% of windshield extract or cottonwood extract, 3 to 15 wt% A thickening agent for athlete's foot comprising 0.5 to 2% by weight of vitamin E acetate, 0.5 to 2% by weight of vitamin E acetate, 0.5 to 2% by weight of polysorbate 20, 0.01 to 0.05% by weight of benzyl alcohol and 0.01 to 0.05% Prevention or amelioration.

Hereinafter, the composition for prevention and improvement of athlete's foot according to the present invention will be described in more detail.

Tea Tree Oil, one of the main components of the composition of the present invention, is a plant native to Australia and belongs to the genus Myrtaceae. It contains particulate matter in the leaves. When the leaves are crushed, Lt; / RTI > is obtained. The tea trioyl contained 2.5% of alpha-pinene, 9.1% of alpha-terpinene, 3.9% of para-cymene, 4.3% of 1.8-cineol, 24.6% of gamma-terpinene, 2.3% of alpha-terpineol, 42.1%, terpinolene 4.1%, and trace amounts of residual components. The effect of antibacterial as well as antiinflammatory, analgesic and wound treatments is effective. Its action mechanism is not clear as its effect is similar to that of ginseng. In the present invention, tea tree oil is preferably compressed or rectified using leaves.

Avocado (Persea americana Mill.) Is a fruit originating in Mexico and Central America, and is widely consumed worldwide. Avocado oil is a vegetable oil made by squeezing flesh and skin. Avocado is rich in vitamin E called tocopherol, which is one kind of fat-soluble vitamin. It removes active oxygen from our body and suppresses aging. And is known to have the effect of preventing heart disease and cancer. In the present invention, the avocado oil is preferably used by pressing or rectifying the flesh of the flesh.

The windshield (Saposhnikoviae divaricata Schiskin) is a perennial herb that belongs to the mountain-type Umbelliferae (Umbelliferae) and is a perennial herb called jinhwa wind, mountain wind wind, screen wind herb, mountain windbreak tree, It is widely distributed in Korea, China, Vietnam and Mongolia. It is known that the main effects of windblown by oriental medicine are sweating, fever, analgesic, diuretic, paralysis, limb paralysis, shinhae, geumdam, toothache, neuralgia, rheumatism and so on.

Chamaecyparis obtusa (Chamaecyparis obtusa) is an evergreen arborescent tree, and it is also called ashiko, hinoki and cypress. It is 30 ~ 40m in height and 1 ~ 2m in width. The bark is reddish brown, and small needle-shaped leaves are densely grown on branches. It is distributed in Japan and southern part of Korea. It contains γ-cadinene, α-terpineol and borneol, and leaves and wood contain 1% essential oil. . It is used as a natural insecticide for the prevention of house dust mites causing allergic diseases such as atopy, asthma and rhinitis. It is recommended to use the leaves, stems, and branches extracted from the leek. The windshield and the leathery extract may be used singly or in the form of a mixture.

Rhizoma Polygoni Cuspidati is a plant belonging to the family Polygonaceae and grows in mountainous areas and is distributed in Korea, Japan, Taiwan and China. In one room, the roots are called hojanggun, and they are used as relaxation, diuretic, and tongue economy, and as a sedative in the private sector. When it is young, the stem looks like a hoof, and it has the name Ho Chang Geun. Root can be obtained by extracting roots and rootstocks.

Sasae is a plant of various ages of paddy field and grows frequently in the southern part of Korea and the mountains of Jeju Island. The stem is straight up to 1 ~ 2m in height, 3 ~ 6mm in diameter, and the bract covers the stem for 2 ~ 3 years. The nodes are covered with inverted hair and white powder, but the leaves of the sheath-shaped leaves disappear after 4 years. Leaves are long, oval-shaped, 10 ~ 25cm long, pointed like a tail or long as the tail. There are no hairs on both sides of the leaf, there are hairs on the base of the back side, there are saw teeth like thorn on the edge, and the sheath has hairs. It is known to reduce fever and poison, remove sputum, urinate well, and smooth metabolism. Sasa can be obtained by extracting leaves, stems and roots.

The vegetable oil containing the tea tree oil and the avocado oil may be obtained by squeezing a leaf, a fruit or a fruit into plants or by adding an extraction solvent to the raw material at a weight ratio of 1: 2 to 1:20, and adding the extract to a reflux condenser After cooling by heating distillation for 2 to 3 hours, the extracted extract can be separately filtered for 5-7 hours. Alternatively, the crude oil may be purchased commercially and used.

Leaves, stems, branches and the like can be extracted by hot water extraction, cold-watering or warm-up extraction. In this case, the extraction solvent is mixed with the extraction solvent at a weight ratio of 2 to 20 times, and the mixture is extracted at 70 to 125 ° C for 2 to 24 hours, filtered and concentrated to obtain an extract

The bamboo shoots, bamboo shoots, and windshield roots are thoroughly cleaned, dried, and then added with an extraction solvent 5 to 10 times the weight of the raw materials. The resulting mixture is placed in a autoclave autoclave and heated at 65 to 125 ° C for 2 to 6 hours, Extraction by heating for 5 hours, filtration and concentration may be obtained

The extraction solvent may be at least one selected from water, lower alcohols having 1 to 4 carbon atoms, polyhydric alcohols, and mixtures thereof. As the lower alcohol having 1 to 4 carbon atoms, methanol, ethanol and the like can be used. As the polyhydric alcohol, butylene glycol, propylene glycol, pentylene glycol and the like can be used. Mixtures of water and lower alcohols, mixtures of water and polyhydric alcohols, mixtures of lower alcohols and polyhydric alcohols, or mixtures of water and lower alcohols and polyhydric alcohols can be used as the mixture.

The present invention is mainly characterized in finding ingredients having multifunctional properties such as skin protection, antioxidant activity and antibacterial property from natural plants and an optimal combination ratio thereof. The tea tree oil, avocado oil, Extracts and vitamin E acetates are the main components of skin protection, antioxidant, and antibacterial activity. Sasa extract acts as a preservative. Polysorbate acts as a dispersant. Benzyl alcohol and thymol act as preservatives and stabilizers. do.

The composition ratio of the composition according to the present invention is set considering the effective dose and side effects of each component, and if the ratio is out of the range, the effect may be deteriorated.

The desired effects of the present invention can be achieved by mixing the above-mentioned natural plant extracts at an optimum ratio, thereby preventing growth of various microorganisms and maximizing cell protection and antioxidative effects.

When the amount of the tea trioyl is less than 50 wt%, the antibacterial effect is insufficient. When the amount of the tea trioyl is more than 75 wt%, the effect of increasing the amount of the tea trioyl is insufficient. In addition, the antioxidative and preservative effect of avocado oil is less than 10% by weight, and when it is more than 30% by weight, the synergistic effect by combination with each ingredient may not be achieved. The skin protection, antioxidant and germicidal effect are less than 3% by weight of the roots extract or the cotton bud extract and the silkworm extract, and less than 15% by weight are uneconomical because the composition is not balanced with the whole composition. The content of the sage extract is 1 to 10% by weight, the content of vitamin E acetate is 0.5 to 2% by weight, the content of polysorbate 20 is 0.5 to 2% by weight, the content of benzyl alcohol is 0.01 to 0.05% by weight and the content of thymol is 0.01 to 0.05% %, It is possible to optimally exhibit stable coordination and various functions of the entire composition.

Since the composition according to the present invention contains an extract of a natural substance, which is not a chemical synthesis agent, it can be seen from the results of Examples and Test Examples described later that there is no irritation or toxicity to the skin and thus it is excellent in stability. But also can be used as a cosmetic or therapeutic agent requiring such functionality since it has antimicrobial activity against bacteria including aceotrophic bacteria, healing of atopic dermatitis, therapeutic treatment of infant's dirt, protection against skin and improvement of skin texture have.

Accordingly, the present invention provides, in a further aspect, a composition for improving athlete's foot comprising the composition and cosmetics or a pharmaceutically acceptable carrier as an active ingredient.

The formulations of the cosmetics may be prepared by conventional methods well known in the art of cosmetics.

The excipients of these cosmetics may vary depending on the purpose of use, but they generally contain a preservative such as glycerin, butylene glycol or propylene glycol or a vegetable preservative, a purified water, a moisturizer, a bulking agent, a cream agent, a surfactant, a pH adjuster, It is preferable to mix them according to the form of the desired formulation.

The composition of the present invention can be suitably formulated in the form of an external preparation such as a suspension, emulsion or aerosol according to a conventional method.

When the composition of the present invention is prepared from a pharmaceutical composition, it may be preferable to further include a conventional pharmaceutically acceptable carrier.

The dosage of the pharmaceutical composition of the present invention is suitably set in accordance with the form of the preparation, the method of administration, the purpose of use, and the age, body weight, and symptoms of the patient to which the active ingredient is administered. For example, 10 mu g to 200 mg / kg per day for an adult. However, those skilled in the art will appreciate that the pharmacokinetic properties of a particular reagent and its mode and route of administration; Age, health, weight; It will be understood that the dosage varies depending on known factors such as the nature and extent of the symptoms, the type of concurrent treatment, the frequency of treatment and the desired effect.

<Examples>

Hereinafter, the present invention will be described in more detail with reference to examples. However, these embodiments are described for the purpose of helping understanding of the present invention, and do not limit the content and scope of the invention.

Example 1: Preparation of plant essential oil

Add 1 Kg of finely ground tea tree leaves to a reflux condenser by adding 5 times of extraction solvent (purified water and 30% alcohol) with a mixer, heat distill for 2 ~ 3 hours, cool, The mixture was filtered (Toyo No. 2) for 6 hours to obtain an extract. The extraction yield of tea tree was 18.6%, and the brix measured by refractometer was 2.32%. The filtrate was then concentrated under a reduced pressure of -0.08 MPA at a temperature of 60 DEG C for 3 to 8 hours. The brix of solids was measured at 35.6 (purified water) and 37.8 (alcohol), respectively. Similarly, avocado oil was obtained by treating avocado pulp. The brix of solids was measured as 53.6 (purified water) and 57.4 (alcohol), respectively.

Example 2: Preparation of fern extract

Five milliwatts of extraction solvent (purified water and 30% alcohol) was added to 1 kg of soft white leaf, followed by extraction at 70 to 125 ° C for 2 to 12 hours to obtain a cottonwood extract. The yield of the whiteness was 30.5% and the brix was 4.1%. Subsequently, the extract was filtered with a paper filter paper to filter off the foreign substances, and the filtrate was concentrated at a temperature of 60 ° C under a reduced pressure of -0.08 MPA for 8 hours. The brix of solids was measured as 40.1 (purified water) and 42.3 (alcohol), respectively.

Example 3: Preparation of Hojangun, Root Root and Sasa extract

After washing 1 Kg of silkworm roots and bamboo shoots thoroughly, it was dried, and then, purified water of 5 times of the raw material weight and 30% of the raw material were added to the autoclave and heated in a high pressure sterilizing pot at a temperature of 65 to 125 ° C for 2 to 6 hours Extracted, filtered and concentrated to obtain an extract. The brix of solids content was measured as 43.6 (purified water) and 45.8 (alcohol), respectively, and the brix of the extract of windshield was 55.2 (purified water) and 56.8 (Purified water) and 56.8 (alcohol).

Examples 4-8: Preparation of a composition comprising a natural extract

Each of the components obtained in Examples 1 to 3 was formulated in a formulation ratio shown in Table 1 below to prepare a composition for preventing athlete's foot. Each plant oil and extract was mixed with purified water extract and alcohol extract at a weight ratio of 1: 1.

Component Content (g)	Example 4	Example 5	Example 6	Example 7	Example 8	Comparative Example
Tea trio day	50	55	53	65	70	70
Avocado oil	10	30	10	10	20	30
Windshield extract	15	3.0	10	3.0	-	-
Leek extract	-	3.0	15	-	4.0	-
Hojangun extract	12.0	13	8.0	15	3.0	-
Sasa extract	10	5.5	2.5	6.0	1.0	-
Vitamin E-acetate	0.94	0.94	0.50	2.0	1.0	-
Polysorbate (20)	2.0	0.50	0.90	0.9	0.9	-
Benzyl alcohol	0.01	0.05	0.05	0.05	0.05	-
Thymol	0.05	0.01	0.05	0.05	0.05	-
Sum	100	100	100	100	100	100

Test Example 1: Antioxidative effect analysis of natural extract composition

1.1) Analysis of DPPH radical scavenging ability

The in vitro antioxidant activity was measured by reacting the free radical DPPH having the maximum absorbance at 517 nm with the compositions of Examples 4 to 8 to determine the antioxidative effect using the absorbance value lowering in yellow. DPPH free radical scavenging activity was determined using a reaction mixture comprising the composition of the example dissolved in methanol in a 96-well microplate and 0.1 mM DPPH dissolved in ethanol. The extracts were mixed with 0.6 mL of DPPH solution and incubated at room temperature for 20 minutes. The absorbance of the reaction mixture was then measured at 517 nm. The percent inhibition of radical scavenging was determined in comparison to the ethanol treated group or the ascorbic acid (100 [mu] g) treated group. The percentage of DPPH free radical scavenging activity was calculated by the following equation.

(%) = (Abs _{control without sample -} Abs _sample ) / Abs _{control without sample} Х 100

The results are shown in Fig. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing DPPH free radical scavenging performance of the example compositions. Con, vehicle treated group as a negative control group; AA, treated with ascorbic acid (1 mg / ml) as a positive control group; Sample of 100 시 sample (hereinafter, the same as in the test example).

Positive control ascorbic acid (AA) inhibited free radicals by 98.36 ± 1.32% at 1 mg / ml concentration. The results are shown in Table 1. The sample treatment groups were 91.25 ± 0.12 (Example 4), 92.45 ± 0.65 (Example 5), 93.66 ± 0.23 (Example 6), 96.21 ± 0.15 (Example 7), 92.57 ± 0.24 47.35 +/- 0.27 (comparative) inhibition (Figure 1).

1-2) Analysis of hydroxyl radical scavenging ability

Hydroxyl radical scavenging activity was measured by reacting hydroxyl radicals generated by Fenton ' s reagent with the composition of the examples to lower the absorbance value.

Embodiment the composition 100 μM EDTA, it was added to the reaction mixture containing oxy-ribose 1 mM H ₂ O _2, and 2.8 mM to. The volume was supplemented with 20 mM pH 7.4 phosphate buffer to make 1 mL and then incubated at 37 ° C for 1 hour.

The mixture was then heated in the bath to 95 캜 for 15 minutes, and each of the 2.8% TCA and 1% 0.5% TBA was added. After cooling, the mixture was centrifuged at 4800 rpm for 15 minutes. The absorbance of the supernatant was measured at 532 nm. For comparative analysis, negative control group was treated with vehicle (phosphate buffer) and positive control group was treated with ascorbic acid to analyze hydroxyl radical scavenging ability.

2 is a graph showing the hydroxy radical scavenging activity of the example composition. Positive control ascorbic acid (AA) inhibited free radicals by 51.69 ± 1.13% at 1 mg / ml concentration. The results are shown in Table 1. The sample treatment groups were 41.25 ± 0.67 (Example 4), 42.75 ± 0.76 (Example 5), 47.87 ± 0.83 (Example 6), 45.21 ± 0.96 (Example 7), 39.01 ± 0.89 23.50 ± 1.23 (comparative) (FIG. 2).

1-3) Analysis of total polyphenol and total flavonoid content

The compositions of the examples were mixed with 100 μL of Folin-Denis reagent and incubated for 3 minutes at room temperature (RT, 22-24 ° C.) The reaction mixture was neutralized with 100 μl of 10% sodium carbonate and incubated for 1 hour. Absorbance was measured at 760 nm with a plate reader. Polyphenol content was calculated using gallic acid as standard.

The results are shown in Table 2. The polyphenol content and flavonoid content of each composition were significantly higher than those of the comparative examples.

ingredient	Example 4	Example 5	Example 6	Example 7	Example 8	Comparative Example
Polyphenol (mg / g)	56.72 + 1.51	57.32 + - 0.98	52.77 + - 0.54	56.45 + 0.71	52.34 + - 0.35	25.43 + - 0.23
Flavonoid (mg / g)	7.41 ± 0.82	8.55 0.35	8.01 + - 0.78	8.25 + - 0.66	7.15 ± 0.78	2.16 ± 0.35

1-4) Reducing power efficacy analysis

Reducing power efficacy analysis was based on the comparison of the effect of ascorbic acid with the lowering of the absorbance value by reacting the composition of each Example with the induction of oxidative reaction by FeCl ₃ .

Each of the compositions of the examples was mixed with 0.5 mL of 200 mM sodium phosphate buffer (pH 6.6), and 0.5 mL of 1% potassium ferricyanide, then the mixture was incubated at 50 DEG C for 20 minutes, followed by 0.5 mL of 10% TCA . It was centrifuged at 650 g for 10 minutes and 1 mL of supernatant was mixed with equal volume of distilled water and 0.2 mL of 0.1% ferric chloride. The same treatment was performed on standard ascorbic acid solution and the absorbance was measured at 700 nm. The reducing power was calculated and expressed in terms of ascorbic acid.

The results are shown in Table 3. The Reducing power efficacy of each composition was significantly higher than that of the comparative example.

	Example 4	Example 5	Example 6	Example 7	Example 8	Comparative Example
The ascorbic acid equivalent (g)	28.12 ± 1.51	27.23 + - 0.31	22.67 ± 0.51	26.35 ± 0.78	22.36 + - 0.92	13.34 ± 0.53

1-5) Fe ²⁺  chelating activity assay

The activity of Fe ²⁺ chelating activity was measured by reacting the composition of the present invention with the induction of the oxidative reaction by the Fe ²⁺ -ferrozine complex to lower the absorbance value.

150 μl of the prepared 500 μM FeSO ₄ was added to the reaction mixture in which 170 μL of 0.1 M Tris-HCl (pH 7.4), 220 μL of Tris-HCl buffer and the composition of each example were mixed. The reaction mixture was incubated at room temperature for 5 minutes and 10 μL of 0.25% 1,10-phenanthroline (w / v) was added. Absorbance was measured at 510 nm. The Fe ²⁺ chelating ability was calculated by the following equation for the control group.

Fe ²⁺ chelating ability (%) = (Abs _{control without sample -} Abs _sample ) / Abs _{control without sample} Х 100.

For comparative analysis, the negative control group was treated with vehicle (Tris-HCl buffer) and the positive control group was treated with ascorbic acid to analyze Fe ²⁺ chelating activity.

3 is a graph showing the results of Fe ²⁺ chelating assay analysis of the composition of the examples. Positive control ascorbic acid (AA) inhibited the oxidative response of Fe ²⁺ -ferrozine complex by 39.08 ± 3.92% at 1 mg / ml concentration. The compositions of each example inhibited the oxidative response of the Fe ²⁺ -ferrozine complex by 46.21 ± 1.02% to 53.25 ± 2.02% at 100 μl, while suppressing the comparative example by 18.51 ± 1.85%.

1-6) Nitrite scavenging ability analysis

The nitrite scavenging activity was measured by comparing the absorbance of the nitrite with the composition of the examples to determine the antioxidative effect. For comparative analysis, vehicle (PBS) was treated with negative control group and nitrite scavenging ability was treated with ascorbic acid in positive control group.

Each sample was treated with 0.1 N HCl solution (pH 1.2), pH 3.0 and pH 5.0 solution, and 1 mM NaNO ₂ solution was added. The mixture was incubated at 37 [deg.] C for 1 hour. 2% acetic acid and Griess reagent were added and incubated at room temperature for 15 minutes. The absorbance of the mixture was measured at 520 nm.

4 is a graph (pH 1.2) showing the nitrite scavenging activity of the composition of Example. Nitrite inhibition (%) = [1 - (Abs _sample / Abs _{control without sample} )] x 100. Control, group treated with PBS as a negative control group; AA, a group treated with ascorbic acid (1 mg / ml) as a positive control; 100 ㎍, 100 쨉 g of a composition as a sample group.

Positive control ascorbic acid (AA) inhibited nitrite at a concentration of 1 mg / ml by 91.0 ± 3.12% at pH 1.2. The composition of each example inhibited nitrite at 56.73 ± 1.12% (Example 4) to 61.32 ± 1.09% (Example 7) at pH 1.2 and 20.51 ± 0.76% inhibition of the comparative example.

Test Example 2: Antimicrobial activity test

The antimicrobial and antimicrobial properties of the compositions according to Examples 4 to 8 were tested by disc paper test.

The test strain was dispensed into a petri dish, spread over the entire medium using a glass rod, and then placed on a paper disc having a diameter of 5 mm. Then, 30 쨉 l of the composition of each example was placed on a paper disc. After overnight incubation at 30 ° C, the inhibition zone was checked for antimicrobial activity. The results are shown in Table 4 below.

Test strain	Inhibition zone (d): cm
	Ex4	Ex5	Ex6	Ex7	Ex8	Comp. Ex.
Staphylococcus aureus ATCC 10537	2.5	2.4	2.6	2.8	2.5	1.0
Staphylococcus aureus ATCC 256E	2.6	2.5	2.5	3.0	2.5	1.5
Salmonella typhimurium KCTC 1925	2.8	2.9	2.6	3.1	2.7	1.8
Candida albicans ATCC 10231	2.4	2.8	2.8	3.2	2.5	2.0

As can be seen from the above test results, the composition according to the present invention has excellent antimicrobial activity against bacteria such as fungi such as aphthous fungus, though it has a difference in degree. Therefore, it can be used as a cosmetic product or a treatment for athlete's foot.

Test Example 3: Measurement of growth inhibitory activity of fungi

( Eg , Trichophyton rubrum , KCTC 6345), Micosporum audouinii (KCTC 6346), Trichophyton ferrugineum (KCTC6351), Epidermophyton floccosum , KCTC 6586), and Trichophyton mentagrophytes (KCTC 6077) were tested for growth inhibitory activity. Fungal growth inhibitory activity was assayed by the paper disc diffusion method. Each strain was inoculated into a Sabouraud's agar medium and treated with each composition (Examples 4 to 8, treatment concentration 100)) directly on a paper disc. The leaves were placed on the culture medium inoculated with each bacterium and cultured for 5 days at 28 ° C. The size of the clear zone formed around the foliage was measured to check the antimicrobial activity. The results are shown in Table 5 .

Test strain	Inhibition zone (d): cm
	Ex4	Ex5	Ex6	Ex7	Ex8	Comp. Ex.
Trichophyton rubrum	3.0	3.2	3.3	3.8	3.5	0.5
Micosporum audouinii	3.2	3.3	3.1	3.5	2.8	0.5
Trichophyton ferrugineum	2.5	3.0	2.8	3.3	2.5	0.8
Epidermophyton floccosum	2.5	3.5	3.0	3.0	2.5	1.0
Trichophyton mentagrophytes	3.0	3.5	3.0	3.5	3.0	0.8

The size of the growth inhibition ring by the composition of the examples was 2.5 cm or more and showed excellent fungicidal activity. These antimicrobial activities were similar in other fungi.

Test Example 4: Skin irritation test

Skin irritation experiments were conducted to examine the degree of irritation of each composition according to the present invention to experimental animals. Eighteen New Zealand white rabbits weighing 2.3 to 2.7 kg were used as experimental animals. First, experimental animals were refined in an animal room for one week after their intake, and normal animals were observed during the refinement period and only healthy animals were used for the test. The rabbits were divided into treatment compartments and control compartments 24 hours before application of the test substance, and then 0.5 ml of the test substance per test animal was applied once to the administration site, and sterilized physiological saline was applied to the untreated control compartment in the same amount . After attaching, it was covered with a sheet of solid material, fixed with a tape, and applied for a certain period of time. After the application time, the application portion was gently rinsed with physiological saline. On the seventh day after application, local anesthetics (lidocaine, Gwangmyeong drug) were injected through the ear vein and euthanized, followed by autopsy. And observed with naked eyes.

Skin reactions were evaluated for 24 hours and 72 hours using the " Standards for Toxicological Testing of Drugs and the like ". The degree of irritation to the skin was also calculated according to the method of calculating the Draize's Primary Irritation Index (P.I.I.), which is commonly used. As a result of the test, irritation such as erythema, scar formation and edema of the test substance application site was not recognized, and the primary irritation rate of skin by P. I. Draize was evaluated as "0".

Test Example 5: Acute Toxicity Test

The compositions of the examples were subjected to acute toxicity studies and their toxicity was observed. As experimental animals, male and female SD rats of 105 weight and 105 weight, respectively, were used and tested by using the compositions of the respective examples and distilled water as a negative control. First, the rats were incubated in a laboratory incubation box with a temperature of 22 ° C, a relative humidity of 53% and a fluorescent light (09:00 lit-18: 00 off) contrast cycle and a light intensity of 150-300 Lux for about a week , And then healthy animals were selected and divided into groups so that the average body weight was matched. The animals were orally administered at a dose of 20 ml / kg once a day for 14 days, and then the changes in general condition, poisoning symptoms, , Autonomic nerves, weight change, and presence of dead animals.

According to the experimental results, there was no significant difference in the body weight during the experimental period, but there was no significant difference. The highest concentration of 20 ml per kg, which is the maximum amount of sample to be administered to the test animal, was selected, LD ₅₀ was more than 20 ml / kg BW because of the inability to calculate the lethal dose, and it was found that the composition was not toxic when it was judged according to the general symptoms or the specific lesion upon autopsy.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined in the appended claims. It will be clear to those who have knowledge.

The present invention relates to a cosmetic or pharmaceutical composition for prevention and improvement of athlete's foot since the composition for prevention or improvement of athlete's foot is natural oil as a main component and exhibits skin protection, antioxidant and antimicrobial action simultaneously as well as less side effects such as toxicity and skin irritation It is useful in the production of related products.

Claims (4)

1. Vitamin E acetate 0.5 to 50% by weight, tea tree oil 50 to 75% by weight, avocado oil 10 to 30% by weight, safflower root extract or curd extract 3-15% by weight, Wherein the composition comprises 0.5 to 2% by weight of polysorbate 20, 0.5 to 2% by weight of polysorbate 20, 0.01 to 0.05% by weight of benzyl alcohol and 0.01 to 0.05% by weight of thymol.
2. The method according to claim 1,

   The vegetable oil containing the tea tree oil or the avocado oil is obtained by squeezing a leaf, a fruit or a fruit into plants, or an extraction solvent is added to the raw material in a weight ratio of 1: 2 to 1:20, After cooling, the extract was filtered for 5 to 7 hours to obtain an extract.

   Leaves, stems, branches and the like can be extracted by hot water extraction, cold-watering or warm-up extraction. In this case, the extraction solvent is mixed with the extraction solvent in a weight ratio of 2 to 20 times, and the mixture is extracted at 70 to 125 ° C for 2 to 24 hours, filtered and concentrated to obtain an extract.

   The bamboo shoots, bamboo shoots, and windshield roots are thoroughly cleaned, dried, and then added with an extraction solvent 5 to 10 times the weight of the raw materials. The resulting mixture is placed in a autoclave autoclave and heated at 65 to 125 ° C for 2 to 6 hours, The mixture is heated for 5 hours for extraction, filtered, and concentrated to obtain a composition for external application for preventing or improving athlete's foot.
3. The method according to claim 1,

   Wherein the extraction solvent is at least one selected from the group consisting of purified water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof.
4. The composition for external use for the prevention or improvement of athlete's foot according to claim 1, which has an antioxidative action, an antibacterial action or a pervative protective action.

Images