JP5840437B2 - Anti-inflammatory agent - Google Patents

Description

The present invention is an external preparation for skin, characterized by containing urea and having an effect of inhibiting inflammation due to innate immune reaction of epidermis keratinocytes against Acne bacteria or Malassezia.

Urea is a natural moisturizing factor (NMF component) on the skin, and it is said that it enhances the water retention ability of the stratum corneum and has a keratolytic and exfoliating action. Widely used for pharmaceuticals and the like (Patent Document 1). Although it may be used for an anti-acne preparation (patent document 2), the improvement effect with respect to the inflammation by acne which is a causative agent of acne and malassezia which is a causative agent of Marassezia folliculitis has not been known until now.

Acne is medically called acne vulgaris, and acne bacteria are involved in the inflammation. Further, as an inflammatory skin disease similar to acne, malassezia folliculitis due to malassezia can be mentioned. It is said that the lipase produced by Acne or Malassezia degrades sebum, and the fatty acids generated thereby cause inflammation (Non-patent Documents 1 and 2). In recent years, it has been considered that human innate immune responses against acne or malassezia are deeply involved in inflammation of acne and malassezia folliculitis.

Antibiotics and vitamins are used for the treatment of acne, which is a bacterial acne, and antifungal agents are used for internal and external use in the treatment of malassezia folliculitis, which is targeted for the fungus, malassezia ( Non-patent documents 3, 4). However, inflammation due to innate immune reactions is also caused by bacterial components of Acne and Malassezia killed by antibacterial and antifungal agents, effectively reducing the inflammation of acne and Malassezia folliculitis For this, combined use of anti-inflammatory agents is desired.

JP-A-2005-263666 Special table 2005-517649

Eur J Dermatol, Vol. 14, PP. 4-12 (2004) J Med Vet Mycol, Vol. 31, PP. 77-85 (1993) Sayama Yoshikatsu, Fragrance Journal, 1985, No. 74, p. 21-29 J Clin Microbiol, Vol. 43, PP.2824-2829 (2005)

Therefore, it has been desired to develop an external preparation for skin that controls inflammation induced by acne causing acne or malassezia causing marathetia folliculitis.

As a result of intensive studies aimed at solving the above-mentioned problems, the present inventors have found that urea has an effect of suppressing the innate immune response of human epidermal keratinocytes against Acne bacteria or Malassezia. Furthermore, the skin external preparation containing urea was found to have an effect of improving inflammation in acne and malassezia folliculitis, and the present invention was completed.

Urea used in the present invention is not particularly limited. Urea obtained by chemical synthesis, urine blended in cosmetics such as hand creams and lotions, quasi-drugs, pharmaceutical external preparations for skin, bathing agents, etc. Any of urea and the like concentrated and purified from can be suitably used.

The blending amount of urea in the external preparation for skin in the present invention is preferably 0.1 to 20% by weight. If it is less than 0.1% by weight, it is difficult to obtain a sufficient effect, and if it exceeds 20% by weight, the enhancement of effect is small and uneconomical.

In the external preparation for skin of the present invention, fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, which are components used in ordinary antibacterial agents, within a range that does not impair the effect thereof In addition, components such as metal soaps, preservatives, fragrances, moisturizers, powders, ultraviolet absorbers, thickeners, pigments, antioxidants, whitening agents, chelating agents, and the like can be blended.

In the present invention, urea can be used in any of cosmetics, quasi drugs, and pharmaceuticals. Examples of the dosage form include skin lotion, cream, emulsion, gel, aerosol, essence, pack, and washing. Agents, bath preparations, foundations, dusting, ointments and the like.

Urea suppressed the innate immune response of human epidermal keratinocytes to Acne or Malassezia. Moreover, the skin external preparation containing urea showed the outstanding improvement effect with respect to the inflammation in acne and Malassezia folliculitis.

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

Formulation Example 1 Toner lotion amount (parts)
1. Urea 1.0
2. 1,3-butylene glycol 8.0
3. Glycerin 2.0
4). Xanthan gum 0.02
5. Sodium dihydrogen phosphate 0.064
6). Disodium hydrogen phosphate 1.35
7). Ethanol 5.0
8). Methyl paraoxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
10. Fragrance 0.1
11. Make the total volume 100 with purified water
[Production method] Components 1 to 6 and 11 and components 7 to 10 are uniformly dissolved, and both are mixed and filtered to obtain a product.

Comparative Example 1 A conventional lotion was obtained by replacing component 1 with 1.0 part of purified water in the conventional lotion formulation example 1.

Formulation Example 2 Cream component Blending amount (parts)
1. Urea 5.0
2. Squalane 6.0
3. Olive oil 3.0
4). Stearic acid 2.0
5. Beeswax 2.0
6). Octyldodecyl myristate 3.5
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Behenyl alcohol 1.5
9. Glycerol monostearate2.5
10. Fragrance 0.1
11.1,3-butylene glycol 8.5
12 Ethyl paraoxybenzoate 0.05
13. Methyl paraoxybenzoate 0.2
14 Sodium hydroxide 0.17
15. Potassium dihydrogen phosphate 0.68
16. [Manufacturing method] The components 2 to 9 are heated and dissolved and mixed with purified water, and the oil phase is maintained at 70 ° C. Ingredients 1 and 11 to 16 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. The component 10 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Comparative Example 2 A conventional cream was prepared by replacing Component 1 with 5.0 parts of purified water in Conventional Cream Formulation Example 2.

Formulation Example 3 Emulsion Component Amount (parts)
1. Urea 3.0
2. Squalane 5.0
3. Olive oil 5.0
4). Jojoba oil 5.0
5. Cetanol 1.5
6). Glycerol monostearate 2.0
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12 Methyl paraoxybenzoate 0.2
13. Sodium borate decahydrate 1.0
14 Hydrochloric acid 0.17
15. [Manufacturing method] Components 2 to 8 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 1 and 10 to 15 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 with stirring.

Formulation Example 4 Gel component content (parts)
1. Urea 2.0
2. Ethanol 5.0
3. Methyl paraoxybenzoate 0.1
4). Polyoxyethylene hydrogenated castor oil (60 EO) 0.1
5. Fragrance 0.1
6.1,3-Butylene glycol 5.0
7). Glycerin 5.0
8). Xanthan gum 0.1
9. Carboxyvinyl polymer 0.2
10. Potassium hydroxide 0.2
11. Tris (hydroxymethyl) aminomethane 0.8
12 Hydrochloric acid 0.12
13. [Production method] Ingredients 2 to 5 and ingredients 1 and 6 to 13 are dissolved uniformly in purified water, and the two are mixed to obtain a product.

Formulation Example 5 Pack component Blending amount (parts)
1. Urea 3.0
2. Polyvinyl alcohol 12.0
3. Ethanol 5.0
4.1,3-Butylene glycol 8.0
5. Methyl paraoxybenzoate 0.2
6). Polyoxyethylene hydrogenated castor oil (20 EO) 0.5
7). Citric acid 0.1
8). Sodium citrate 0.3
9. Fragrance 0.1
10. Potassium dihydrogen phosphate 0.09
11. Disodium hydrogen phosphate 0.09
12 [Production method] Components 1 to 12 are uniformly dissolved in purified water to make a total amount of 100 to obtain a product.

Experimental Example 1 Inhibition test of innate immune response of human epidermal keratinocytes with urea Propionibacterium acnes JCM6425 was used as an acne bacterium. After anaerobic culture at 37 ° C. for 2 days using a modified GAM agar medium (Nissui Pharmaceutical), it was dispersed in phosphate buffered saline (PBS) to a turbidity of 1.0 (660 nm), and the test bacteria Liquid. As Malassezia, Malassezia furfur NBRC 0656 was used. After culturing at 32 ° C. for 2 days using Leeming & Notman agar medium (J Clin Microbiol, Vol. 25, PP. 2017-2019, 1987), the mixture was dispersed in PBS to a turbidity of 1.0 (660 nm), and tested. Bacterial fluid was used. Neonatal foreskin-derived human epidermal keratinocytes (Invitrogen) are cultured in 37-well 5% CO ₂ conditions using 700 μL of 154S medium (Invitrogen) with a predetermined amount of HKGS (Invitrogen) added in a 12-well plate. Tested at 60%. After removing the medium used for the pre-culture, 560 μL of medium, 70 μL of medium containing 10% by weight of urea (Wako Pure Chemical Industries) and 70 μL of the test bacterial solution are added, and cultured at 37 ° C. under 5% CO ₂ for 3 hours. did. MRNA extraction using Trizol reagent (Invitrogen), reverse transcription reaction using SYBR Green Two-step qRT-PCR Kit (Invitrogen), and then using Step One Plus Real-Time PCR system (Applied Bios) The expression level of sex cytokine IL-8 was measured (J Dermatol, Vol. 36, PP. 213-223, 2009). The data was obtained as a ratio (%) when the expression level of IL-8 was 100 when urea was not added (when only acne bacteria were added).

The results when acne bacteria are used as test bacteria are shown in Table 1.

From Table 1 above, when urea is added, it is clear that the innate immune response of human epidermal keratinocytes to acne bacteria is suppressed.

Table 2 shows the results when malassezia was used as the test bacterium.

From Table 2 above, it is clear that the innate immune response of human epidermal keratinocytes to Malassezia is suppressed when urea is added.

Experiment 2 Use test 1
Using the conventional lotions of Formulation Example 1 and Comparative Example 1, a use test for one month was conducted on 30 adults (15 men and 15 women) with acne on the face. Half of the subjects used Formulation Example 1, and the other half used Comparative Example 1. After use, a questionnaire survey was conducted to determine improvement in inflammation in acne. As a result, the external preparation for skin characterized by containing urea showed an excellent improvement effect on inflammation in acne (Table 3). During the test period, there was no skin problem and there was no problem with safety.

Experiment 3 Use test 2
Using the conventional creams of Formulation Example 2 and Comparative Example 2, a one month use test was conducted on 30 adults (15 males and 15 females) who had malatian folliculitis in their body parts. Half of the subjects used Prescription Example 2, and the other half used Comparative Example 2. After use, a questionnaire survey was conducted to determine improvement in inflammation in Malassezia folliculitis. As a result, the external preparation for skin characterized by containing urea showed an excellent improvement effect on inflammation in Malassezia folliculitis (Table 4). During the test period, there was no skin problem and there was no problem with safety.

As a result of the same usage test for the cosmetics and pharmaceuticals obtained in Formulation Examples 3 to 5, all of them showed an excellent improvement effect on inflammation in acne and malassezia folliculitis.

The external preparation for skin characterized by containing urea of the present invention can be used for cosmetics, quasi-drugs, and pharmaceuticals for the purpose of improving inflammation in acne and malassezia folliculitis.

Claims (1)

An anti-inflammatory agent for external use against inflammation of Malassezia folliculitis caused by Malassezia , comprising 0.1 to 1% by weight of urea.