Bactericidal composition for propionibacterium acnes in sebum plugs

Abstract

To provide a composition that exhibits superior bactericidal activity against Propionibacterium acnes present in sebum plugs.SOLUTION: The present invention provides a bactericidal composition for Propionibacterium acnes in sebum plugs, comprising ethanol and isopropyl methylphenol.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] It relates to a bactericidal composition for Propionibacterium acnes in comedones.

Background Art

[0002] Acne vulgaris is one of the inflammatory diseases that occur on the face and the like. Acne is mainly known to be caused by Cutibacterium acnes (hereinafter sometimes referred to as "acne bacteria"). Conventionally, various preparations for treating acne have been reported. For example, Patent Document 1 describes an anti-acne bacterial composition containing cis-6-hexadecenoic acid or its salt and isopropylmethylphenol or trichlorocarbanilide as active ingredients, and it has been reported that the active ingredients have an excellent bactericidal effect against acne bacteria.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] Conventionally, the bactericidal effect of a preparation is often evaluated for acne bacteria suspended in a liquid, for example. However, acne bacteria are present in large numbers in comedones in pores on the skin. Comedones are known as solids formed by a mixture of sebum, old keratin, and the like in pores. The present inventor has noticed that even when using the same preparation, the bactericidal effect against acne bacteria present in comedones is inferior compared to the bactericidal effect against acne bacteria suspended in a liquid in a test tube or the like. And the inventor has noticed that it is important to provide a preparation for treating acne that is useful against acne bacteria present in comedones.

[0005] Accordingly, an object of the present invention is to provide a composition having an excellent bactericidal action against acne bacteria present in comedones.

Means for Solving the Problems

[0006] As a result of intensive studies, the present inventors have found that the combined use of ethanol and isopropylmethylphenol can effectively reduce acne bacteria present in comedones. The present invention has been completed through further studies based on this finding, and the present disclosure includes inventions represented, for example, by the following. Item 1. A bactericidal composition for acne bacteria in comedones, containing ethanol and isopropylmethylphenol. Item 2. The bactericidal composition for acne bacteria in comedones according to Item 1, wherein the content of ethanol in the composition is 8% by mass or more, and the content of isopropylmethylphenol is 0.03% by mass or more. Item 3. The bactericidal composition for acne bacteria in comedones according to Item 1, wherein the content of ethanol in the composition is 11% by mass or more, and the content of isopropylmethylphenol is 0.01% by mass or more. Item 4. The bactericidal composition for acne bacteria in comedones according to any one of Items 1 to 3, further containing propylene glycol. Item 5. A method for reducing acne bacteria in comedones, including applying an effective amount of a composition containing ethanol and isopropylmethylphenol to the skin where comedones are present in a subject who needs to reduce acne bacteria in comedones. Item 6. Use of ethanol and isopropylmethylphenol for the production of a bactericidal composition for acne bacteria in comedones. Item 7. Ethanol and isopropylmethylphenol for use in reducing acne bacteria in comedones.

Effects of the Invention

[0007] It can exhibit an excellent bactericidal action against acne bacteria present in comedones.

Brief Description of the Drawings

[0008]

Figure 1

Figure 2

Mode for Carrying Out the Invention

[0009] Hereinafter, the embodiments included in the present disclosure will be described in more detail. In the present disclosure, "containing" also includes the meanings of "substantially consisting of" and "consisting of".

[0010] The bactericidal composition for acne bacteria in comedones of the present disclosure contains ethanol and isopropyl methylphenol. Hereinafter, the bactericidal composition for acne bacteria in comedones of the present disclosure may be described as "the composition of the present disclosure".

[0011] In the composition of the present disclosure, the content of ethanol is not limited, but 3% by mass or more is preferably exemplified, 8% by mass or more is more preferably exemplified, and 11% by mass or more is even more preferably exemplified. Also, in the composition of the present disclosure, the content of ethanol is preferably exemplified as 3% by mass or more and 15% by mass or less. In the composition, the content of ethanol is more preferably 5% by mass or more and 15% by mass or less, even more preferably 8% by mass or more and 15% by mass or less, and particularly even more preferably 8% by mass or more and 12% by mass or less, etc.

[0012] Isopropylmethylphenol (IPMP) contained in the composition of the present disclosure is also referred to as 4-isopropyl-3-methylphenol. In the composition of the present disclosure, the content of IPMP is not limited, but preferably exemplified as 0.01% by mass or more, more preferably exemplified as 0.03% by mass or more. Further, in the composition of the present disclosure, the content of IPMP is preferably exemplified as 0.01% by mass or more and 0.5% by mass or less. In the composition of the present disclosure, the content of IPMP is more preferably 0.01% by mass or more and 0.3% by mass or less, still more preferably 0.01% by mass or more and 0.1% by mass or less, etc.

[0013] Although not limiting the present disclosure, as one embodiment of the composition of the present disclosure, a composition in which ethanol is 8% by mass or more and IPMP is 0.03% by mass or more in the composition is preferably exemplified. Further, although not limiting the present disclosure, as one embodiment of the composition of the present disclosure, a composition in which ethanol is 11% by mass or more and IPMP is 0.01% by mass or more in the composition is preferably exemplified.

[0014] In the composition of the present disclosure, the content of ethanol relative to IPMP is not limited either, but preferably exemplified as 6 parts by mass or more and 1500 parts by mass or less of ethanol per 1 part by mass of IPMP. As the content of ethanol per 1 part by mass of IPMP, more preferably exemplified is 10 parts by mass or more and 1500 parts by mass or less of ethanol. As the content of ethanol per 1 part by mass of IPMP, still more preferably is 10 parts by mass or more and 1200 parts by mass or less, particularly preferably 50 parts by mass or more and 1200 parts by mass or less, particularly more preferably 80 parts by mass or more and 1200 parts by mass or less, etc.

[0015] In the composition of the present disclosure, in addition to ethanol and IPMP, any components such as pharmaceutically acceptable components and components applicable to the skin may be contained as necessary within a range that does not interfere with the effects of the present disclosure.

[0016] As an example of the optional component, propylene glycol is exemplified. When containing propylene glycol, the content of propylene glycol in the composition of the present disclosure is not limited, but preferably 1% by mass or more and 15% by mass or less is exemplified. As the content of propylene glycol, more preferably 2% by mass or more and 12% by mass or less, still more preferably 2% by mass or more and 10% by mass or less, etc. are exemplified.

[0017] As an example of the optional component, 1,3-butylene glycol is exemplified. When containing 1,3-butylene glycol, the content of 1,3-butylene glycol in the composition of the present disclosure is not limited, but preferably 1% by mass or more and 15% by mass or less is exemplified. As the content of 1,3-butylene glycol, more preferably 2% by mass or more and 12% by mass or less, still more preferably 2% by mass or more and 10% by mass or less, etc. are exemplified.

[0018] As an example of the optional component, water is exemplified. When containing water, the content of water in the composition of the present disclosure is not limited, but preferably 92% by mass or less is exemplified. As the content of water, more preferably 60% by mass or more and 92% by mass or less, still more preferably 60% by mass or more and 90% by mass or less are exemplified. As the content of water, more preferably 70% by mass or more and 90% by mass or less, still more preferably 75% by mass or more and 85% by mass or less, etc. are exemplified.

[0019] As an example of the optional component, in addition to propylene glycol, 1,3-butylene glycol and water, solvents (lower alcohols having 1 to 5 carbon atoms such as methanol, isopropanol, glycerin, etc., alcohols such as polyhydric alcohols (regardless of anhydrous or hydrous)), fragrances, coloring agents, emulsifiers, buffers, penetration enhancers, stabilizers, preservatives, thickeners, solubilizers, pH adjusters, surfactants, antioxidants, cooling agents, astringents, gelling agents, skin softeners, ultraviolet absorbers, moisturizers, whitening agents, anti-inflammatory agents, various pharmacologically active ingredients such as vitamins, etc. are exemplified. The optional component may be used alone or in combination of two or more. The content of the optional component may also be determined appropriately. Ethanol and IPMP are not included in the optional component.

[0020] The pH of the composition of the present disclosure may be applicable to the skin. For example, when the temperature of the composition is 25°C, the pH is 4 or more and 7 or less, preferably 5 or more and 6.5 or less.

[0021] The form of the composition of the present disclosure is not limited as long as it can be applied to the skin where plugs are present, and is preferably liquid or semi-solid. Therefore, the composition is preferably exemplified by liquids, suspensions, emulsions, sprays, gels, pastes, creams, etc. As the usage modes of the composition, cleansing agents (makeup removers), facial washes, lotions, milky lotions, beauty essences, creams, packs, exfoliating cosmetics, foam cleansing agents, foam facial washes, sunscreen agents, makeup bases, foundations, massage creams, hair tonics, hair styling agents, and other basic cosmetics, and external composition products such as external pharmaceutical products are preferably exemplified. For example, when used as an exfoliating cosmetic or a pack, the composition of the present disclosure can be used by impregnating carriers such as conventionally known cotton pads for makeup, non-woven sheets, woven sheets (such as gauze), makeup puffs, sponges, etc. The composition of the present disclosure may or may not be rinsed with water or the like after application to the skin depending on the usage mode and the like.

[0022] The composition of the present disclosure can be produced according to the conventionally known normal procedures in the above various forms, usage modes, etc. The composition of the present disclosure may be produced by mixing ethanol and IPMP, and also, if necessary, mixing any of the above components.

[0023] The composition of the present disclosure is used by applying it to the skin where plugs are present. The skin site is not limited, and examples include the face, neck, hands, arms, back, scalp, torso, legs, etc. The amount and frequency of applying the composition of the present disclosure to the skin are not particularly limited. For example, depending on the age of the user, the degree of the symptom, the application form, the expected degree, etc., an appropriate amount of the composition of the present disclosure may be applied to the skin once or several times a day.

[0024] In the present disclosure, "inside the comedone" means the surface and the inside of the comedone, and the composition of the present disclosure can sterilize the acne bacteria present on the surface and inside of the comedone. Thus, the composition of the present disclosure has a bactericidal effect on the acne bacteria present in the comedone. Therefore, the composition of the present disclosure can be used for the purpose of preventing or improving acne, or preventing or improving skin diseases caused by acne bacteria infections such as seborrheic dermatitis. In particular, the composition of the present disclosure can be preferably used for preventing or improving acne, or preventing or improving skin diseases caused by acne bacteria infections, based on the bactericidal effect on the acne bacteria present in the comedone.

[0025] Also, from this, it can be said that the present disclosure includes a method for reducing acne bacteria inside the comedone, which includes applying an effective amount of a composition containing ethanol and isopropylmethylphenol to the skin where the comedone exists in a subject who needs to reduce acne bacteria inside the comedone. Also, it can be said that the present disclosure includes the use of ethanol and isopropylmethylphenol for the production of a composition for killing acne bacteria inside the comedone. Also, it can be said that the present disclosure includes ethanol and isopropylmethylphenol for use in reducing acne bacteria inside the comedone. As described above, by using ethanol and isopropylmethylphenol in combination, a bactericidal effect is exerted on the acne bacteria inside the comedone. In the method and the like, each condition such as the composition for killing acne bacteria inside the comedone, ethanol, isopropylmethylphenol, and any component is explained in the same manner as above.

Example

[0026] Hereinafter, the embodiments of the present disclosure will be described more specifically with examples, but the embodiments of the present disclosure are not limited to the following examples.

[0027] Test Example 1 The bactericidal effect on the acne bacteria inside the comedone was evaluated according to the following procedure. 1) Experimental Procedure · Preparation of Inoculum Solution Cutibacterium acnes (NBRC107605 strain) was revived from a glycerol stock into modified GAM agar medium (manufactured by Nissui Pharmaceutical Co., Ltd., hereinafter referred to as "agar medium"). It was cultured at 37°C for 3 to 5 days under anaerobic conditions (cultured in an anaerobic box containing AnaeroPack (registered trademark) Kenki (manufactured by Mitsubishi Gas Chemical Company, Inc.)). Colonies that appeared on the agar medium were scraped off with a 10 μL loop for 1 platinum loopful and suspended in physiological saline. The resulting suspension was further diluted 10-fold to prepare a bacterial solution. The bacterial solution was spread on the agar medium and cultured at 37°C for 5 days under anaerobic conditions in the same manner as described above. When the number of colonies that appeared on the agar medium was visually counted and calculated, the number of bacteria in the bacterial solution was approximately 10 8 CFU / mL.

[0028] · Acne Bacteria Inoculation into Pseudocorns 1 g of pseudo-comedo was weighed, 0.1 mL of the bacterial solution was inoculated into the pseudo-comedo, and after mixing the bacterial solution into the pseudo-comedo with a medicine spoon, it was uniformly stirred for 10 minutes with a stirring device (stirring and defoaming device Kakuhunter, manufactured by Shashin Kagaku Co., Ltd.). It has been previously confirmed by preliminary studies using a coloring solution that the bacterial solution is uniformly mixed into the pseudo-comedo by performing this mixing and stirring operation. The resulting stirred product was used as a pseudo-comedo inoculated with bacteria. In this test example, a mixture of proteins and lipids prepared with reference to the literature describing the comedo composition (Koji Mizukoshi et al., Journal of the Society of Cosmetic Scientists of Japan, 41, 262 - 268 (2007); Toshinori Iida et al., Fragrance Journal, 45(2) 22 - 27 (2017); F. Hashimoto, et al., Journal of Society of Cosmetic Chemists of Japan, 23, 2 (1989)) was used as the pseudo-comedo. The composition of the pseudo-comedo is shown in Table 1. In the table, "%" means mass%.

[0029]

Table 1

[0030] · Contact with Test Solution and Bacterial Count Measurement 0.1 g of the inoculated pseudo-comedo was placed in a 1.5 mL tube, and 1 mL of the test solution was added. The mixture was allowed to stand at room temperature (25 °C) for 60 minutes.

[0031] The test solutions are as follows. The test solutions were prepared by mixing the following components with water. For example, Example 1 is an aqueous solution containing IPMP and ethanol. Example 1 Isopropylmethylphenol (IPMP) + Ethanol (EtOH) aqueous solution Example 2 Isopropylmethylphenol (IPMP) + Ethanol (EtOH) + Propylene glycol aqueous solution Comparative Example 1 Dimethyl sulfoxide (DMSO) aqueous solution Comparative Example 2 Propylene glycol (PG) aqueous solution Comparative Example 3 EtOH aqueous solution Comparative Example 4 IPMP + DMSO aqueous solution

[0032] After the standing, the total amount of 0.1 g of the inoculated pseudo-comedo and 1 mL of the test solution was mixed with 1 mL of a neutralizing agent (TSB + TLHC (Inactivator B.), manufactured by Merck KGaA). Then, a vortex mixer was used to finely suspend the pseudo-comedo to obtain a suspension. 0.2 mL of the suspension and 0.8 mL of the neutralizing agent were mixed and further diluted with the neutralizing agent to prepare a 10-fold dilution series, which was spread on an agar medium. Culturing was carried out under anaerobic conditions at 37 °C for 3 to 5 days in the same manner as described above, and the number of colonies was visually counted to determine the number of bacteria per 1 g of the pseudo-comedo (this number of bacteria was defined as the "test value"). The number of bacteria in the inoculated pseudo-comedo immediately before contact with the test solution was counted and calculated in the same manner using an agar medium to obtain the number of bacteria per 1 g of the pseudo-comedo (10 6 CFU / mL), which was defined as the "control value".

[0033] The reduction rate of Propionibacterium acnes in the pseudo-comedo after contact with the test solution was calculated according to the following formula. In this formula, when the test value is smaller than the control value, the calculated value is positive (plus), and when the test value is larger than the control value, the calculated value is negative (minus). When the calculated value is negative, it means that no reduction in Propionibacterium acnes is observed (specifically, in the case of Comparative Examples 1 and 4), so it was shown as a reduction rate of 0% in Figure 1 described later.

[0034]

Number

[0035] 2) Results The results are shown in Fig. 1. The larger the value of the acne bacilli reduction rate in the comedo, the fewer the acne bacilli present in the comedo. As shown in Fig. 1, in Comparative Example 1 using DMSO, no reduction in acne bacilli in the comedo was observed. Also, in Comparative Example 2 using PG, which is conventionally known to have a bactericidal effect, the reduction rate of acne bacilli in the comedo was as low as about 9%. In Comparative Example 3 using EtOH, which is conventionally known to have a bactericidal effect, the reduction rate was higher than that in Comparative Example 2, but it only reached about 78%. Further, in Comparative Example 4 using a combination of IPMP, which is conventionally known to have a bactericidal effect, and DMSO, no reduction in acne bacilli in the comedo was observed. In contrast, in Example 1 using a combination of IPMP and EtOH, the reduction rate was very high at about 94%, and an excellent reduction effect was observed. In Example 2 using a combination of IPMP, EtOH, and PG, the reduction rate was very high at 99.9% or more, and an even more excellent reduction effect was observed. As described above, in this test example, since a pseudo-comedo in which acne bacilli were uniformly mixed therein was used, it can be seen that in Example 1 and Example 2, the test solution acted not only on the acne bacilli present on the surface of the pseudo-comedo but also on the acne bacilli inside the pseudo-comedo. From this, it can be seen that the test solution penetrated into the comedo and exerted a bactericidal effect on the acne bacilli present inside the comedo.

[0036] Furthermore, for the comedones (not pseudo-comedones, using comedones collected from humans, and it was confirmed that Cutibacterium acnes was present inside the collected comedones) contacted with the test solution of Example 2, fluorescence staining was performed using the LIVE / DEAD BacLight Bacterial Viability Kit (manufactured by Molecular Probes). After staining, the viability of Cutibacterium acnes inside the comedones was observed using a confocal laser scanning microscope (CLSM). This kit determines cells stained green by SYTO 9 as viable bacteria and cells stained red by Propidium Iodide as dead bacteria. As a result, most of the Cutibacterium acnes cells inside the comedones were dead bacteria showing red fluorescence. In contrast, when distilled water was contacted with the comedones instead of the test solution of Example 2 and the same staining and observation were performed, most of the Cutibacterium acnes inside the comedones were viable bacteria showing green fluorescence. From this, it can also be seen that the test solution penetrates inside the comedones and exerts a bactericidal effect on Cutibacterium acnes present inside the comedones.

[0037] From the results of this test example, it was found that the combined use of IPMP and EtOH has an excellent bactericidal effect on Cutibacterium acnes inside the comedones.

[0038] Test Example 2 The bactericidal effect on Cutibacterium acnes inside the comedones was evaluated according to the following procedure. 1) Experimental Procedure · Preparation of Inoculum Solution and Acne Bacteria Inoculation into Pseudocorns An inoculum solution was prepared in the same manner as in Test Example 1. Also, in the same manner as in Test Example 1, Cutibacterium acnes was inoculated into the pseudo-comedones to prepare pseudo-comedones inoculated with bacteria.

[0039] · Contact with Test Solution and Bacterial Count Measurement Using the following test compositions (Examples 3 to 10, Reference Examples 1 to 2, and Comparative Examples 5 to 10) as test solutions, the contact time between the inoculated pseudo-comedones and the test solutions was changed to 120 minutes, and the pseudo-comedones were contacted with the test solutions in the same manner as in Test Example 1 to measure the number of bacteria. The following test compositions were prepared by mixing the following components with water. For example, Example 3 is an aqueous solution containing 0.01% by mass of IPMP and 8% by mass of EtOH. The case where sterilized water was contacted instead of the test composition was used as a control.

[0040] According to the following formula, the reduction rate of acne bacteria in the pseudo-comedones after contact with the test solution or sterilized water was calculated. In this test example, the number of bacteria after contact with the test solution (number of bacteria per 1 g of pseudo-comedones) was defined as the "test value", and the number of bacteria after contact with sterilized water (number of bacteria per 1 g of pseudo-comedones) was defined as the "control value". In this formula, when the test value is smaller than the control value, the calculated value is positive (plus), and when the test value is larger than the control value, the calculated value is negative (minus).

[0041] [Number]

[0042] [Test Composition] Example 3 0.01% IPMP + 8% EtOH Aqueous Solution Example 4 0.03% IPMP + 8% EtOH Aqueous Solution Example 5 0.05% IPMP + 8% EtOH Aqueous Solution Example 6 0.08% IPMP + 8% EtOH Aqueous Solution Example 7 0.01% IPMP + 11% EtOH Aqueous Solution Example 8 0.03% IPMP + 11% EtOH Aqueous Solution Example 9 0.05% IPMP + 11% EtOH Aqueous Solution Example 10 0.08% IPMP + 11% EtOH Aqueous Solution Reference Example 1 8% EtOH Aqueous Solution Reference Example 2 11% EtOH Aqueous Solution Comparative Example 5 0.03% IPMP + 8% DMSO Aqueous Solution Comparative Example 6 0.05% IPMP + 8% DMSO aqueous solution Comparative Example 7 0.08% IPMP + 8% DMSO aqueous solution Comparative Example 8 0.03% IPMP + 11% DMSO aqueous solution Comparative Example 9 0.05% IPMP + 11% DMSO aqueous solution Comparative Example 10 0.08% IPMP + 11% DMSO aqueous solution

[0043] 2) Results The results are shown in Figure 2. The reduction rate of acne bacteria in Example 3 using a combination of 8% EtOH and 0.01% IPMP exceeded the reduction rate of acne bacteria in Reference Example 1 using 8% EtOH. Further, in Example 4 (using a combination of 8% EtOH and 0.03% IPMP) where the blending amount of IPMP was increased to 0.03% in Example 3, the reduction rate of acne bacteria further improved to 93%. Also, in Example 5 using a combination of 8% EtOH and 0.05% IPMP and Example 6 using a combination of 8% EtOH and 0.08% IPMP, the reduction rate of acne bacteria further improved depending on the concentration of IPMP. Further, compared with the reduction rate of acne bacteria in Reference Example 2 using 11% EtOH, the reduction rate in Example 7 using a combination of 11% EtOH and 0.01% IPMP was significantly improved, and the reduction rate in Example 7 was 99%. Also, very high reduction rates of acne bacteria were observed in Example 8 (using a combination of 11% EtOH and 0.03% IPMP), Example 9 (using a combination of 11% EtOH and 0.05% IPMP), and Example 10 (using a combination of 11% EtOH and 0.08% IPMP) where the blending amount of IPMP was further increased in Example 7. In contrast, as described above, although IPMP is conventionally known to have a bactericidal action, in Comparative Examples 5 to 10 using a combination of IPMP and DMSO, the reduction rate of acne bacteria was low, and effective reduction of acne bacteria in the horny plugs was not observed.

[0044] From the results of this test example, it was also found that the combined use of IPMP and EtOH has an excellent bactericidal action against acne bacteria in the horny plugs.

Claims

1. A bactericidal composition for preventing acne bacteria in keratin plugs, comprising ethanol and isopropyl methylphenol.

2. 2. The composition for disinfecting acne bacteria in keratin plugs according to claim 1, wherein the composition contains 8% by mass or more of ethanol and 0.03% by mass or more of isopropyl methylphenol.

3. 2. The composition for disinfecting acne bacteria in keratin plugs according to claim 1, wherein the composition contains 11% by mass or more of ethanol and 0.01% by mass or more of isopropyl methylphenol.

4. The antibacterial composition for acne bacteria in keratin plugs according to any one of claims 1 to 3, further comprising propylene glycol.

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