JP2012206973A - Biofilm formation inhibitor of acne bacterium - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a preparation for external use that inhibits biofilm formation of an acne bacterium as a precipitating cause of pimple, effectively prevents or treats pimple, and has high safety and stability.SOLUTION: Culture solution of malassezia genus fungus inhibits the biofilm formation of the acne bacterium. By using the culture solution of malassezia genus fungus together with an antimicrobial agent, the antifungal to the acne bacterium is improved comparing with the case of the antimicrobial agent only. In addition, the preparation for external use containing the culture solution of malassezia genus fungus and the preparation for external use containing the culture solution of malassezia genus fungus and the antimicrobial agent exhibit an excellent pimple prevention and symptom moderating effect, and has high safety and stability. Thus, the acne bacterium biofilm formation inhibitor is effective for pimple prevention and the symptom moderation.

Description

The present invention relates to an external preparation that effectively inhibits acne by inhibiting biofilm formation of acne bacteria, which is a cause of acne exacerbation.

Acne (acne vulgaris), one of the skin diseases, develops when acne bacteria grow in sebaceous hair follicles with closed pores and sebum. Acne bacteria form a biofilm in the hair follicle, and the development of this biofilm is considered to be deeply involved in the development of acne (Non-patent Document 1).

A biofilm is an aggregate composed of extracellular polysaccharides produced by microorganisms, bacterial cells, and the like, and microorganisms in the environment survive in the biofilm. If a biofilm is formed, the antibacterial agent does not penetrate into the inside even if an antibacterial agent is administered, and thus the expected effect cannot be obtained. In other words, it is important not only to use an appropriate antibacterial agent but also to inhibit biofilm formation for the control of microorganisms in the environment.

As biofilm formation inhibitors, methods using chemical substances such as amide compounds and polyhydric alcohols have been reported (Patent Documents 1 and 2). However, natural products, particularly microorganism-derived biofilm formation inhibitors have not been known.

Int J Dermatol. Vol. 42 (12), PP. 925-927 (2003) JP2010-43003 JP 2005-225830 A

There has been a demand for an external preparation that inhibits biofilm formation of acne bacteria, which is one of the causes of acne, and is excellent in safety and stability.

As a result of intensive studies aimed at solving the above problems, the present invention has found that a culture solution of the genus Malassezia inhibits biofilm formation of Acne bacteria. Furthermore, the external preparation containing the Malassezia fungus culture solution, and the external preparation containing the Malassezia fungus culture and the antibacterial agent are excellent in safety and stability, and the present invention has been completed.

The culture solution of the genus Malassezia used in the present invention was produced by culturing Malassezia furfur, Malassezia sympodialis, Malassezia globosa, Malassezia restricta, or Malassezia dermatis.

In the external preparation for skin of the present invention, oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, preservatives, fragrances, moisturizing agents are within the range not impairing the effect. Components such as agents, powders, ultraviolet absorbers, thickeners, dyes, antioxidants, whitening agents, chelating agents, and the like can also be blended.

The external preparation for skin of the present invention can be used for any of cosmetics, quasi-drugs and pharmaceuticals. Examples of the dosage form include skin lotions, creams, emulsions, gels, aerosols, essences, packs, Examples include detergents, bath preparations, foundations, powders, lipsticks, ointments, poultices, mouthwashes, liquid toothpastes, and toothpastes.

The Malassezia fungal culture inhibited the biofilm formation of Acne. Moreover, the antibacterial effect of the external preparation containing the main culture solution and the antibacterial agent was enhanced as compared with the external preparation containing only the antibacterial agent.

Next, in order to describe the present invention in detail, examples and experimental examples are given, but the present invention is not limited thereto.

Production Example 1 Furfur culture solution Malassezia furfur NBRC0656 in Leeming & Notman medium (total amount 1000 mL, peptone 10 g, bovine bile extract 4 g, yeast extract 0.1 g, glucose 5 g, glycerol 1 g, glycerol monostearate 0.5 g, Tween 60 0.5 g, 32.5. % Fat milk (10 mL) (Non-patent Document 2), the mixture was cultured at 32 ° C. for 1 week, filtered through a 0.45 μm filter, A furfur culture medium was prepared.

Production Example 2 After cultivating the globosa culture solution Malassezia globosa by the method of Production Example 1, it was filtered in the same manner. A globosa culture solution was prepared.

Production Example 3 After culturing malastezia restricta by the method of Production Example 1, filtration was performed in the same manner. A restrictic medium was prepared.

Production Example 4 After culturing Symposialis culture solution Malassezia symposialis by the method of Production Example 1, the same was filtered, A symposialis culture solution was prepared.

Production Example 5 After cultivating the dermatis culture solution Malassezia dermatis by the method of Production Example 1, it was filtered in the same manner. A dermatis broth was prepared.

Formulation Example 1 Lotion Formulation Amount (parts)
1. M.M. furfur culture medium (Production Example 1) 5.0
2. Potassium aluminum sulfate 0.3
3. 1,3-butylene glycol 8.0
4). Glycerin 2.0
5. Xanthan gum 0.02
6). Citric acid 0.01
7). Sodium citrate 0.1
8). Ethanol 5.0
9. Methyl paraoxybenzoate 0.1
10. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
11. Fragrance 0.1
12 Make the total volume 100 with purified water
[Production method] Components 1 to 7 and 12 and components 8 to 11 are uniformly dissolved, and both are mixed and filtered to obtain a product.

Comparative Example A conventional lotion was obtained by replacing component 1 with 5.0 parts of purified water in conventional lotion formulation example 1.

Formulation Example 2 Cream Formulation Amount (parts)
1. M.M. Globosa culture solution (Production Example 2) 3.0
2. Salicylic acid 0.1
3. 1,3-butylene glycol 8.5
4). Ethyl paraoxybenzoate 0.05
5. Methyl paraoxybenzoate 0.2
6). Sodium hydroxide 0.17
7). Potassium dihydrogen phosphate 0.68
8). Squalane 6.0
9. Olive oil 3.0
10. Stearic acid 2.0
11. Beeswax 2.0
12 Octyldodecyl myristate 3.5
13. Polyoxyethylene cetyl ether (20E.O.) 3.0
14 Behenyl alcohol 1.5
15. Glycerol monostearate2.5
16. Fragrance 0.1
17. [Manufacturing method] Components 8 to 15 are heated and dissolved and mixed with purified water to 100, and then kept at 70 ° C to obtain an oil phase. Ingredients 2-7 and 17 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. Components 1 and 16 are added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Formulation Example 3 Latex Formulation Amount (parts)
1. M.M. restricta culture solution (Production Example 3) 3.0
2. Triclosan 0.05
3. Squalane 5.0
4). Olive oil 5.0
5. Jojoba oil 5.0
6). Cetanol 1.5
7). Glycerol monostearate 2.0
8). Polyoxyethylene cetyl ether (20E.O.) 3.0
9. Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
10. Fragrance 0.1
11. Propylene glycol 1.0
12 Glycerin 2.0
13. Methyl paraoxybenzoate 0.2
14 Sodium borate decahydrate 1.0
15. Hydrochloric acid 0.17
16. [Manufacturing method] Components 2 to 9 are heated and dissolved and mixed with purified water to a total amount of 100, and kept at 70 ° C to obtain an oil phase. Components 11 to 16 are dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. Components 1 and 10 are added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Formulation Example 4 Gel formulation Formulation amount (parts)
1. M.M. Symposialis culture solution (Production Example 4) 4.0
2. Pionine 0.001
3. 1,3-butylene glycol 5.0
4). Glycerin 5.0
5. Xanthan gum 0.1
6). Carboxyvinyl polymer 0.2
7). Potassium hydroxide 0.2
8). Tris (hydroxymethyl) aminomethane 0.80
9. Hydrochloric acid 0.12
10. Ethanol 5.0
11. Methyl paraoxybenzoate 0.1
12 Polyoxyethylene hydrogenated castor oil (60 EO) 0.1
13. Fragrance 0.1
14 [Manufacturing method] Components 1 to 9 and 14 and components 10 to 13 are uniformly dissolved in purified water to a total amount of 100, and both are mixed to obtain a product.

Formulation Example 5 Ointment Formulation Formulation 1. M.M. dermatis broth (Production Example 5) 5.0
2. Salicylic acid 0.05
3. Triclosan 0.05
4). Polyoxyethylene cetyl ether (30E.O.) 2.0
5. Glycerol monostearate 10.0
6). Liquid paraffin 5.0
7). Cetanol 6.0
8). Methyl paraoxybenzoate 0.1
9. Propylene glycol 10.0
10. [Production method] Components 3 to 7 are heated and dissolved and mixed with purified water to a total amount of 100 and kept at 70 ° C. to obtain an oil phase. Ingredients 2 and 8 to 10 are dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring, and then component 1 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Experimental Example 1 Biofilm formation inhibitory effect of Acne fungus in culture solution of Malassezia fungus clostridia medium (Oxoid) 80 μL, Acne fungus dispersion (10 ⁹ / mL), Malassezia fungus prepared in Production Examples 1-5 The culture solution or 10 μL of Leeming & Notman medium (negative control) was added to a 96-well microplate and anaerobically cultured at 35 ° C. One week later, the supernatant was removed, washed with a phosphate buffer, and 0.5% crystal violet was added to stain the biofilm. After further washing with a phosphate buffer, ethanol was added to elute the dye, and the absorbance at 950 nm was measured to determine the amount of biofilm formed.

J Clin Microbiol. Vol. 25, PP. 2017-2019 (1987)

The results are shown in Table 1.

From Table 1 above, it is clear that when a Malassezia fungus culture solution is included, biofilm formation of Acne is suppressed.

In other Malassezia species, M. Similar to the case of using furfur, the biofilm inhibitory effect of acne bacteria was observed.

Experimental Example 2 Combined Effects of Antibacterial Agent and Malassezia Fungus Culture Solution After culturing Acne bacteria in the presence of Malassezia culture solution or Leeming & Notman medium according to the method of Example 2, 2% (W / V) potassium aluminum sulfate The viable count of acne bacteria after 6 hours was measured by a culture method.

The results are shown in Table 2.

When acne bacteria were cultured with the addition of the Malassezia fungus culture solution, the viable count of Acne bacteria decreased compared to when the Malassezia fungus culture solution was not included. It is clear that the antibacterial effect on the bacteria was increased.

In other Malassezia species, M. Similarly to the case of using furfur, the antibacterial effect of potassium aluminum sulfate against acne bacteria was enhanced.

Even when nadifloxacin, clindamycin phosphate, salicylic acid, triclosan or pionine was used as the antibacterial agent, the culture solution of Malassezia fungus enhanced the antibacterial properties of these antibacterial agents against acne.

Experimental Example 3 Usage Test Using the lotions of Formulation Example 1 and Comparative Example, a 20 month adult usage test (10 males and 10 females) with acne on the face was conducted for 1 month. After use, it was judged about the improvement of acne by questionnaire survey.

The results are shown in Table 3.

The lotion containing the Malassezia fungus culture solution showed an excellent effect of improving acne symptoms. During the test period, there was no skin problem and there was no problem with safety.

As a result of using tests on the cosmetics and pharmaceuticals obtained in Formulation Examples 2 to 5 in the same manner, they all showed an acne prevention and improvement effect.

The external preparation containing the culture solution of the genus Malassezia of the present invention can be used for cosmetics, quasi-drugs, and pharmaceuticals for the purpose of preventing and treating acne.

Claims (4)

A biofilm formation inhibitor of acne that contains a yeast-like fungal culture as an active ingredient. A biofilm formation inhibitor of Acne fungus, comprising a culture solution of Malassezia fungus as an active ingredient. A skin external preparation for the prevention or treatment of acne comprising a culture solution of Malassezia fungus as an active ingredient. An antibacterial composition against an acne bacterium comprising a culture solution of Malassezia fungus and an antibacterial agent.