KR100511114B1 - Composition including antifungal-agents of external application for skin-whitening and external application having the same - Google Patents

Abstract

Disclosed is a skin whitening external preparation comprising an antifungal agent having an action of inhibiting melanin production and tyrosinase activity as an active ingredient, and an external preparation for skin whitening described above. Provided is an external preparation for skin whitening containing 0.001 to 5.00 wt% of an antifungal agent and an external preparation for skin whitening comprising such an active ingredient. The anti-fungal clotrimazole, biponazole, methyl isothiazolinone, sulfonazole, and itraconazole effect on the melanin production and the new use as a whitening agent through the inhibition of tyrosinase have the effect of whitening and melanin production. It is possible to provide an excellent external preparation composition for skin whitening and safety and excellent side effects and external preparations containing the same.

Description

External preparation for skin whitening containing antifungal agent and external preparation for whitening including the same TECHNICAL APPLICATION FOR SKIN-WHITENING AND EXTERNAL APPLICATION HAVING THE SAME}

The present invention relates to an external preparation for skin whitening and an external preparation for skin whitening using the same, and more particularly, to an external preparation for skin whitening containing an antifungal agent as an active ingredient and an external preparation for whitening comprising the same.

Human skin color is genetically determined by the concentration and distribution of melanin in skin cells, but may also be influenced by environmental or physiological conditions such as exposure to sun UV rays, the use of female hormones, pregnancy, cosmetics, fatigue and stress. The melanin pigment is deposited on the skin and blackening of the skin may progress. Due to these causes, pigmented diseases such as blemishes, freckles, etc. occurring on the face have emerged as a major problem of female beauty.

The mechanism of skin blackening is as follows. An internal and external factor activates an enzyme called tyrosinase, which acts on tyrosine, a type of amino acid present in skin tissue, producing dopa and then dopaquinone. The melanin polymer is synthesized from melanocytes in melanocytes, which are skin pigment cells, through the non-enzymatic automatic oxidation process. do.

As the causes and mechanisms of skin blackening have been revealed, melanin is inhibited by incorporating substances having an inhibitory effect of tyrosinase, an enzyme involved in skin blackening, into cosmetics or medicines or inhibiting some reactions during the production of melanin. It is used to reduce the production to improve skin whitening, blemishes, freckles and the like. Representative materials used for this purpose are tocopherol, vitamins, ascorbic acid (ascorbic acid), kojic acid (kojic acid), hydroquinone (hydroquinone) and the like.

However, ascorbic acid is not suitable as a melanogenesis inhibitor due to its low tyrosinase activity inhibitory effect and low stability of the molecule itself. In addition, kojic acid has a strong inhibitory effect on tyrosinase activity, but has a problem in stability such as discoloration and a decrease in titration due to changes over time when combined with pharmaceuticals, and because of high skin irritation, it denatures melanin pigment or melanocytes. There is a limit. In addition, although hydroquinone has been recognized for its effectiveness, its use is generally limited due to safety issues because of its sensitivity to skin.

On the other hand, the antifungal agent has a function of inhibiting or sterilizing the growth of higher organisms having eukaryotic cells such as fungi, and examples thereof include imidazole antifungal agents and triazole antifungal agents.

The imidazole antifungal agents include clotrimazole (Arzneim.-Forsch. 22: 1280, 1972), miconazole (Arzneim.-Forsch. 21: 256, 1971), econazole (Arzneim. -Forsch. 25: 224, 1975), isoconazole (Arzneim.-Forsch. 29: 1344, 1979), triconazole (Antimicrobial Agents Chemotherapy 15: 597-602, 1979), sulconazole (sulconazole) (Eumycetes and Mycosis 23: 314-317, 1982), Arzneim.-Forsch. 32: 17-24, 1982, cloconazole (J. Med. Chem. 26: 768 -770, 1983), bifonazole (Arzneim.-Forsch. 33: 517-524, 1983), butoconazole (J. Med. Chem. 21: 840, 1978), penticonazole ( fenticonazole) (Arzneim.-Forsch. 31: 2127, 1981), zinoconazole (J. Med. Chem. 26: 442-445, 1983), methylisothiazolione (J. Appl. Bacteriol. 69: 569-577, 1990) and ketoconazole (J. Med. Chem. 22: 1003-1005, 1979).

The triazole antifungal agents include terconazole (J. Med. Chem. 26: 611-613, 1983), itraconazole (Antimicrobial Agents and Chemotherapy 26: 5-9, 1984), vibunazole (Arzneim.-Forsch. 33: 546, 1983), Fluconazole (Antimicrobial Agents and Chemotherapy 27: 815-818, 1985), and in the form of free or salt, or metal tea (UK Patent No. 221683). do.

Among the antifungal agents listed above, clotrimazole, biponazole, myconazole, ketoconazole, itraconazole, econazol, oxyconazole, fluconazole, sulfonazole, penticonazole, haloprozin, nystatin, tolnaftate, glyceroplin , A technique for using amphotericin B in medicinal soap is disclosed in Korean Patent No. 1996-0015160, and Korean Patent Application No. 1999-70074460 discloses clotrimazole, itraconazole, myconazole, ketoconazole, and sulfona Disclosed is the use of sol and the like to improve the health of hair and scalp.

However, the melanin production inhibitory effect of the above-mentioned antifungal agent, especially the application to a whitening agent, is not known at all.

Therefore, a first object of the present invention is to provide an external preparation for skin whitening containing an antifungal agent having melanin production inhibitory activity and tyrosinase activity inhibitory activity as an active ingredient.

A second object of the present invention is to provide an external preparation for skin whitening comprising the external preparation for external preparation for skin whitening described above.

In order to achieve the first object of the present invention, the present invention provides an external composition for skin whitening, characterized in that it contains 0.001 to 5.00% by weight antifungal agent.

Herein, clotrimazole, biponazole, methylisothiazolinone, sulfonazole and itraconazole may be used alone or as a mixture of two or more antifungal agents.

In order to achieve the second object of the present invention, the present invention provides an external preparation for skin whitening, comprising an active ingredient for an external preparation for skin whitening containing 0.001 to 5.00% by weight antifungal agent.

The external preparation for skin whitening preferably includes 0.01 to 5.00 wt% of antifungal agents, and antifungal agents that can be used for whitening purposes include clotrimazole, biponazole, methylisothiazolinone, sulfonazole, and Itraconazole can be illustrated.

The external preparation for skin whitening includes, in addition to the external preparation for skin whitening, a whitening agent, a moisturizing agent, an antioxidant, an oily ingredient, an ultraviolet absorber, a surfactant, a thickener, an alcohol, a powder ingredient, a colorant, a perfume, an aqueous ingredient, water, and various skin nutrients. One or more materials may be included. In addition, the external preparation for skin whitening is prepared in the form of external skin creams, ointments, solutions, lotions and packs as necessary.

According to the present invention, by using a novel use of clotrimazole, biponazole, methyl isothiazolinone, sulfonazole, itraconazole as a whitening agent, the skin whitening effect is excellent, the safety and the melanin production inhibitory effect is excellent, and there are no side effects. The external preparation composition for external use and the external preparation containing the same can be provided.

Hereinafter, the present invention will be described in detail.

The external preparation composition for skin whitening according to the present invention contains 0.001 to 5.00 parts by weight of an antifungal agent. If the amount of the antifungal agent is less than 0.001% by weight, the whitening effect is not sufficiently exhibited. If the amount of the antifungal agent is more than 5.00%, it is not preferable because there is a problem in skin safety. More preferably, the external preparation for skin whitening contains 0.01 to 2.0% by weight of an antifungal agent.

Preferred antifungal agents used in the present invention are clotrimazole, biponazole, methylisothiazolinone, sulfonazole, itraconazole and include one or two or more selected from them.

Examination of antifungal melanoma cell line inhibitory effect on tyrosinase activity and melanin production inhibitory effect can be confirmed its whitening effect and melanin production inhibitory effect.

Specifically, the whitening efficacy test of the antifungal agent is performed in the following order. First, the antifungal agent powder is dissolved in dimethyl sulfoxide (DMSO) and diluted while controlling the concentration to prepare an antifungal sample. After that, a B16 melanoma cell line derived from a mouse is selected and cultured as an experimental cell line. At the cellular level, the effects of antifungal agents that inhibit tyrosinase enzyme activity and melanin production are examined, and the effects of hydroquinone and antifungal agents known as whitening agents and cell nontoxic tyrosinase activity inhibition are compared.

In the present invention, since the antifungal agent is used as a topical coating agent at a low concentration, there is no side effect of the antifungal agent, and the whitening effect of the antifungal agent of the present invention appears in a short time.

In addition, the external preparation for skin whitening according to the present invention includes the external preparation for skin whitening containing the antifungal agent described above as an active ingredient.

The external preparation for skin whitening includes other ingredients commonly used for preparing cosmetics or medicines in addition to the external preparation for skin whitening including the antifungal agent, for example, other whitening agents, humectants, antioxidants, oily ingredients, ultraviolet rays. Absorbents, surfactants, thickeners, alcohols, powder components, colorants, fragrances, aqueous components, water, various skin nutrients and the like can be used in combination in appropriate amounts as necessary. In addition, all formulations used in conventional skin external preparations, such as creams, ointments, solutions, lotions, and packs, are applicable so that they can be conveniently used according to the use of external preparations.

Hereinafter, the present invention will be described in detail through preferred embodiments.

Example 1

Antifungal agents  Sample Preparation

Antifungal agents clotrimazole, biponazole, methylisothiazolinone, sulconazole, (manufactured by Sigma Co., Ltd.), itraconazole (manufactured by Janssen Korea) to be used for the experiment were prepared, and each antifungal agent was dissolved in dimethyl sulfoxide. After diluting the solution in the medium to adjust the concentration, the following experiment was performed.

Antifungal  At the cellular level Tyrosinase  Active inhibitory effect

The antifungal agent was added to the culture solution of melanoma cells (B-16 mouse melanoma cells) and observed to inhibit the tyrosinase activity at the cellular level. B16 mouse melanoma cells derived from the mouse were used by the Korea Cell Line Bank. Cultivation was performed using a DMEM medium containing 10% fetal bovine serum (Hassa) in a carbon dioxide incubator (37 ° C., 5%) and incubating 10 ⁵ cells per culture vessel for one day.

After 24 hours of culture, 1 μg / ml and 10 μg / ml of the above antifungal sample solutions were added, and the culture was continued for 2 days, and the effect of inhibiting tyrosinase activity was measured by the following method. First, harvested melanocytes to make cell infiltrates, add 100 μl of cell lysate (1% Triton X-100, 10 mM sodium phosphate, pH 7.0, 0.1 mM PMSF) and destroy the cells by shaking occasionally for 30 minutes at 4 ° C. After centrifugation, the supernatant was taken and used as a Tyrosinase activity measurement solution. 100 μl of 100 mM sodium phosphate (pH 7.0) was added to 50 μl of the supernatant, and kept in a water bath at 30 ° C. for 5 minutes. Then, 50 μl of 100 mM L-dopa was added and the absorbance was measured at 37 ° C. and 405 nm with a temperature control device. The change was observed for 1 hour. Tyrosinase activity inhibition rate was calculated according to the following formula (1), the results are shown in Table 1.

                                                   ---Formula (1)

Inhibitory activity of tyrosinase activity at the cellular level of antifungal agents sample Inhibitory activity of tyrosinase activity 1 μg / ml 10 μg / ml Comparative Example 1 - - Clotrimazole 43.8 74.5 Biponazole 37.2 81.1 Methylisothiazolinone 23.6 90.9 Sulconazole 11.5 77.5 Itraconazole 87.1 90.4 Hydroquinone 46.3 92.3

As can be seen in Table 1, it can be seen that the antifungal agent has a tyrosinase activity inhibitory activity against melanoma cells of the cultured mice. Hydroquinone has a strong tyrosinase activity inhibitory ability, whereas cytotoxicity is severe, antifungal agents can be seen to be a safe substance having a tyrosinase activity inhibitory effect without showing cytotoxicity.

Antifungal  Inhibitory effect of melanin production at the cellular level

The antifungal agent prepared above was added to the culture of melanoma cells to test the whitening effect at the cellular level (Lotan R., Lotan D., Cancer Res., 40: 3345-3350, 1980). Antifungal agent and hydroquinone were added to the culture medium of B-16 melanoma cells at each concentration and cultured for 3 days, and then the cells were trypsinized to remove the melanin after centrifugation. 300 μl of 1N sodium hydroxide to which 10% DMSO was added was added, and the melanin was dissolved by treatment at 80 ° C. for 1 hour, and the absorbance was measured at 475 nm. The relative melanin production relative to Comparative Example 2 was shown as inhibition rate (%). Calculations and results are shown in Table 2 below. Melanin production inhibition was computed according to following formula (2).

---Formula (2)

[Control Example 1]

In the same manner as in Example 1, but using a buffer solution instead of the antifungal agent was used as Comparative Example 1.

Inhibition of melanin production at the cellular level of antifungal agents sample Melanogenesis inhibitory activity (%) 1 μg / ml 10 μg / ml Control - - Clotrimazole 33.2 90.7 Biponazole 20.5 85.8 Methylisothiazolinone 34.8 97.7 Sulconazole 14.0 95.4 Itraconazole 65.3 73.9 Hydroquinone 63.6 97.7

As shown in Table 2, it can be seen that the antifungal agent has melanin production inhibitory ability against melanoma cells of the cultured rats. While hydroquinone has potent melanin production inhibitory ability but severe cytotoxicity, antifungal agents have high melanin production inhibitory effect without showing cytotoxicity.

Example 2

Preparation of cream

After preparing a whitening medicine containing an antifungal agent confirmed that the tyrosinase inhibitory effect and the melanin production inhibitory effect was excellent by the above examples, the stability test and skin hyperpigmentation inhibition ability was tested.

Specifically, it was prepared by including the antifungal clotrimazole and other ingredients as shown in Table 3 as a prescription for cream in medicine.

Medicine  Stability Test of the Composition

In order to determine the stability of the prepared pharmaceutical composition, the prepared medicine was put in a transparent glass container, and then, the degree of discoloration was observed at 1, 2, 4, and 6 weeks while standing at room temperature. The results are shown in Table 4 below.

Skin Pigmentation  Inhibitory effect

Twenty healthy men and women were selected and aluminium foil with two holes of 6 diameters of 7 mm in the lower part of both arms was attached, and 60m / cm using the Oriel solar simulator 1000W at a distance of 10 cm from the arm. The light quantity of ² was investigated. At this time, the irradiation site was washed with 70% ethanol aqueous solution before irradiation. Prepared according to Example 2, twice a day from 3 days before irradiation to 3 weeks after irradiation, prepared according to Comparative Example 2 with a sample containing the antifungal clotrimazole containing ascorbic acid and containing the antifungal clotri Bases containing no masol were applied in pairs to the same row. In each case, the degree of pigmentation was visually determined when using the creams according to Example 2 and Comparative Example 2, and Example 2 was compared to Comparative Example 2 in three stages, in which the degree of pigmentation inhibition was remarkable, slight, and no effect. Evaluation was made, and the results are shown in Table 5.

[Comparative Examples 1 and 2]

As shown in Table 3 below, in Comparative Example 1, the cream was prepared by the formulation containing antifungal clotrimazole and sodium hydrogen sulfite as in Example 1, and in Comparative Example 2, the antifungal clotrimazole from Example 1 Except sodium bisulfite and a cream containing ascorbic acid was prepared, the safety experiment was carried out in the same manner as in Example 2, and the results are shown in Table 4.

Cream composition (unit: weight) Compounding ingredient Example 3 Comparative Example 1 Comparative Example 2 Clotrimazole 1.0 1.0 - Ascorbic acid - - 1.0 Sodium bisulfite - 0.1 - ethanol 10.0 10.0 10.0 Stearic acid 2.0 2.0 2.0 Methylparaben 0.25 0.25 0.25 Mineral oil 6.0 6.0 6.0 Propylene glycol 5.0 5.0 5.0 antiseptic a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount Purified water Remaining weight Remaining weight Remaining weight

Stability Test of Pharmaceutical Compositions Test substance 1 day 2 weeks 4 Weeks 6 Weeks Example 3 Light white Light white Light white Light white Comparative Example 1 Light white Light white Light white Light white Comparative Example 2 Light white Light white Brown mum

As shown in the results of Table 4, it can be seen that the test substance containing the antifungal agent does not discolor much after oxidation for 6 weeks at room temperature.

Skin pigmentation inhibition effect (unit: person) Experimental substance A noticeable effect Slight effect no effect Example 3 8 8 4 Comparative Example 2 0 4 16

As shown in the results of Table 5, the drug containing the antifungal drug methyl isothiazolinone prepared according to Example 3 shows a whitening effect on 16 out of 20 subjects, ascorbic acid instead of the antifungal agent Compared with the pharmaceutical composition containing excellent whitening effect, and no side effects in the skin can be seen that the antifungal agent is a safe and effective blemish or freckles or skin whitening agent.

Example 3

Preparation of Ointment

A skin external ointment containing an antifungal agent was prepared as follows. The antifungal agent used clotrimazole.

Skin Ointment Compounding ingredient Content (% by weight) Clotrimazole 1.0 Prepayment 5.0 Diethylsebagate 8.0 Squalane 5.0 Propylene glycol 5.0 Vaseline 75.5 Sodium benzoate 0.5 Total 100

Of the present invention skin external ointment against blemishes Whitening  effect

Thirty people with blemishes regardless of age and gender were applied to the affected area twice daily for 60 days, and then applied to the affected area at the composition ratio shown in Example 3, and then every 10 days. The state was observed. The results are shown in Table 7.

Decontamination effect of the external preparation for skin whitening of the present invention on blemishes (unit: persons) case Very useful utility Slightly useful Availability (%) 30 8 13 6 90

As shown in Table 7 above, the usefulness is the result of a combination of very useful, useful, and slightly useful. Therefore, the external preparation for skin whitening of the present invention is very excellent in the removal of blemishes, that is, the whitening effect.

Example 4

The external preparation for skin whitening of the present invention can be used in various formulations such as liquids, milk lotions, and packs in addition to the creams and ointments according to Examples 2 and 3, and the liquids, milk lotions, and packs were prepared by the following methods. .

Liquid  Produce

Liquids containing antifungal agents were prepared in the proportions shown in Table 8. The antifungal agent used clotrimazole.

Liquid Ingredient Content (% by weight) Clotrimazole 1.0 ethanol 75 Purified water 24

Preparation of Milk Croissants

Milk crocs containing antifungal agents were prepared in the proportions shown in Table 9. The antifungal agent used clotrimazole.

Milk Croissant Ingredient Content (% by weight) Clotrimazole 0.5 Propylene glycol 5.4 glycerin 4.0 Triethanolamine 1.2 Tocopheryl Acetate 3.0 Liquid paraffin 5.0 Squalane 3.0 Macadamia Nut Oil 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.0 Carboxy Vinyl Polymer 1.0 Incense, preservative a very small amount Purified water Remaining weight

Pack  Produce

Packs containing antifungal agents were prepared as follows. The antifungal agent used clotrimazole.

Pack Compounding ingredient Content (% by weight) Clotrimazole 0.5 Polyvinyl alcohol 15.0 glycerin 5.0 Propylene glycol 4.0 ethanol 8.0 Polyoxyethylene oleylethyl 1.0 Methyl paraoxybenzoate 0.2 Pigment Quantity incense Quantity Purified water Remaining weight

According to the present invention, the anti-fungal agent clotrimazole, biponazole, methyl isothiazolinone, sulfonazole, itraconazole and the whitening effect using the novel use as a whitening agent through the inhibitory action of melanin production and tyrosinase inhibition It is possible to provide an external preparation comprising an external preparation for skin whitening having an excellent melanin production inhibitory effect, excellent safety, and no side effects.

Although described above with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and changed within the scope of the invention without departing from the spirit and scope of the invention described in the claims below I can understand that you can.

Claims (8)

For skin whitening, characterized in that it contains at least one antifungal agent selected from the group consisting of 0.001 to 5.00% by weight of clotrimazole, biponazole, methylisothiazolinone, sulfonazole, and itraconazole, based on the total weight of the composition. External preparation composition. According to claim 1, wherein the content of the antifungal agent is an external composition for skin whitening, characterized in that 0.01 to 2.00% by weight based on the total weight of the composition. delete 0.001 to 5.00% by weight of an external composition for skin whitening containing at least one antifungal agent selected from the group consisting of clotrimazole, biponazole, methylisothiazolinone, sulfonazole, and itraconazole as an active ingredient The external preparation for skin whitening characterized by the above-mentioned. The external preparation for skin whitening according to claim 4, wherein the content of the antifungal agent is 0.01 to 2.0 wt% of the total weight of the external preparation composition for skin whitening. delete The method of claim 4, wherein the external preparation for skin whitening further includes a whitening agent, a moisturizing agent, an antioxidant, an oily ingredient, an ultraviolet absorber, a surfactant, a thickener, an alcohol, a powder ingredient, a colorant, a perfume, an aqueous ingredient, water, and a skin nutrient. External preparation for skin whitening, characterized in that. The external preparation for skin whitening according to claim 4, wherein the external preparation for skin whitening has any one of a skin external cream, an ointment, a liquid, a lotion, and a pack.