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WO 2017/094916 Al 8 June 2017 (08.06.2017) P O P C T

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WO 2017/094916 Al 8 June 2017 (08.06.2017) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/094916 Al 8 June 2017 (08.06.2017) P O P C T (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every CUP 1/04 (2006.01) A61K 35/744 (2015.01) kind of national protection available): AE, AG, AL, AM, A61K 8/97 (2017.01) A61P 17/16 (2006.01) AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A61K 8/99 (2017.01) A61Q 19/00 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (21) International Application Number: HN, HR, HU, ID, IL, IN, IR, IS, KE, KG, KN, KP, KR, PCT/JP20 16/086094 KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (22) International Filing Date: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 5 December 2016 (05.12.2016) OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, (25) Filing Language: English TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, (26) Publication Language: English ZW. (30) Priority Data: (84) Designated States (unless otherwise indicated, for every 2015-237848 4 December 2015 (04. 12.2015) JP kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, (71) Applicants: ICHIMARU PHARCOS CO., LTD. [JP/JP]; TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, 318-1, Asagi, Motosu-shi, Gifu, 5010475 (JP). COMBI TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, CORPORATION [JP/JP]; 2-6-7, Moto-asakusa, Taito-ku, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, Tokyo, 1110041 (JP). LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, (72) Inventors: TAKAYAMA Kazue; c/o ICHIMARU PHAR GW, KM, ML, MR, NE, SN, TD, TG). COS CO., LTD., 318-1, Asagi, Motosu-shi, Gifu, 5010475 (JP). MAKIOKA Yuko; c/o Combi Corporation, 5-2-39, Published: Nishibori, Sakura-ku, Saitama-shi, Saitama, 3380832 (JP). — with international search report (Art. 21(3)) (74) Agents: NAGAI Hiroshi et al; Kyowa Patent & Law Of — with (an) indication^) in relation to deposited biological fice, Nippon Life Marunouchi Building, Marunouchi 1-6- material furnished under Rule 13bis separately from the 6, Chiyoda-ku, Tokyo, 1000005 (JP). description (Rules 13bis.4(d)(i) and 48.2(a)(viii)) (54) Title: GLYCEROL PRODUCTION PROMOTER DERIVED FROM Staphylococcus Epidermidis, ANTIMICROBIAL PEPTIDE PRODUC TION PROMOTER DERIVED FROM SKIN EPIDERMAL KERATINOCYTES, AND UTILIZATION THEREOF IN EXTERNAL PREPARA TION FOR SKIN PROTECTION (57) Abstract: Provided are: a skin-protecting agent and [Fig. an external preparation for skin protection, which not only synergistically work in combination with a plant polysac Type strain (ATCCi222i charide but also has an effect of promoting the production of glycerol derived from Staphylococcus epidermidis, 200 which is a skin resident bacterium useful on the skin, and the production of antimicrobial peptides derived from skin epidermal keratinocytes and exhibits excellent effect of 1000 improving the touch feeling when applied to the skin. More specifically, the present invention provides a skin 300 protective agent and an external preparation for skin pro tection, which comprise a lactic acid bacterium belonging to the genus Enterococcus. 3 6 o — 40 u 200 Sample 1 Sample 2 Sample 3 Sample 4 Description Title of Invention: GLYCEROL PRODUCTION PROMOTER DERIVED FROM Staphylococcus Epidermidis, ANTIMICROBIAL PEPTIDE PRODUCTION PROMOTER DERIVED FROM SKIN EPIDERMAL KERATINOCYTES, AND UTILIZATION THEREOF IN EXTERNAL PREPARATION FOR SKIN PROTECTION Technical Field [0001] Field of the Invention The present invention relates to drug agents and an external preparation for skin protection, which protect the skin from external stresses such as drying and harmful bacteria through a function of promoting one or a series of the production of glycerol derived from skin resident bacteria, namely Staphylococcus epidermidis that is a useful bacterium, and the production ofantimicrobial peptides derived from skin epidermal keratinocytes. Background Art [0002] On the human skin, commensal microorganisms form a group as a skin resident bacterial flora. As skin resident bacteria, Staphylococcus epidermidis, Staphylococcus aureus, corynebacteria and the like inhabit the skin, and the protective function of the skin against the outside world is maintained by the balance of these bacteria. However, it is known that the balance of the flora of these bacteria is impaired and such protective function is thus compromised by an environmental factor such as a rapid temperature change, dryness or a strong detergent or chemical agent, or by a physical/mental factor such as injury, emotional stress or poor systemic condition due to a disease (Non-patent Document 1). [0003] For example, Staphylococcus epidermidis known as a useful bacterium of the skin breaks down serum to produce glycerol and thereby plays a role in physical protective functions, such as inhibition of dry skin and suppression of propagation of saprophytic bacteria by adjusting the skin surface pH. Accordingly, when the ratio of Staphylococcus epidermidis decreases, these protective functions may no longer be demonstrated and this may cause a skin disease. In order to inhibit such a skin disease, moisturizing agents, antibacterial agents, disinfectant components and the like are widely used in skin external preparations; however, depending on the drug components, there is a concern that the protective functions provided by Staphy lococcus epidermidis, which is an aerobic bacterium, may also be inhibited. [0004] Moreover, Staphylococcus epidermidis also plays a role in enhancing the b i ological protective functions of living body by stimulating skin epidermal ker- atinocytes and promoting the production of antimicrobial peptides therefrom. As the antimicrobial peptides produced by skin epidermal keratinocytes, human (hBD2) and human β-defensin-S (hBD3) are known, and it has been reported that these antimicrobial peptides show an antibacterial action against Staphylococcus aureus, which is a harmful bacterium, and serve as a biological barrier to protect the skin from such harmful bacteria (Non-patent Document 2). [0005] Conventionally, for the purpose of promoting the proliferation of Staphy lococcus epidermidis, saccharides such as glucooligosaccharides have been used as a s similable sources (Patent Document 1); however, since not only useful Staphylococcus epidermidis but also other harmful bacteria such as Staphylococcus aureus assimilate such saccharides, a skin protective function is not attained in some cases. In addition, many saccharides give a sticky feel when applied to the skin, and this presents another problem of the difficulty in formulation in terms of the feel. [0006] Staphylococcus aureus is known to exist in a large number on the skin of people having dry skin or rough skin and atopic dermatitis patients. It is also known that toxins produced by Staphylococcus aureus cause a further deterioration of the skin condition. For keeping the skin surface condition healthy, it is thus believed to be important to effectively kill Staphylococcus aureus and to maintain Staphylococcus epidermidis having a skin protective function. [0007] The roles of Staphylococcus epidermidis on the skin are similar to those of lactic acid bacteria and bifidobacteria that are useful intestinal bacteria. These useful in testinal bacteria are known to adjust the intestinal pH, inhibit the proliferation of harmful bacteria such as Clostridium perfringens and Escherichia coli, and have a protective function in the intestinal tract. [0008] For improvement of the intestinal environment, useful bacteria such as lactic acid bacteria and bifidobacteria are taken into the intestinal tract; however, generally, in order to allow these useful bacteria to reach the intestine alive and colonize there, it is required to select acid-resistant strains and to encapsulate them into an enteric coated-type capsule. In addition, in order to increase the number of useful bacteria in the intestine, a method of ingesting a food or drink containing dietary fibers and various oligosaccharides that are selectively assimilated by the useful bacteria has also been employed. Yet, depending on the individual constitution, these methods sometimes cause excessive intestinal fermentation and diarrhea. As a method, which solves such a problem, the use of heat-killed cells of a lactococcus bacterium has been proposed (Patent Document 2). [0009] Ingestion of heat-killed cells of Enterococcus faecalis in the form of a food or drink enables to facilitate the proliferation of bifidobacteria in the intestinal tract and to thereby suppress the proliferation of harmful bacteria and improve the intestinal protective function. Furthermore, since the dead bacterial cells are easily processed as a food or drink, unnecessary fermentation does not proceed, so that the storage stability of the food or drink is hardly affected. [0010] Preferred examples of heat-killed cells of a lactic acid bacterium include those of a microbe classified as Enterococcus faecalis, and it is particularly preferred to use En- terococcus faecalis EC- 12 strain (accession No. FERM BP- 10284), whose 16S rDNA has been registered as "AB 15482" at the National Institute of Genetics. PRIOR ART REFERENCES PATENT DOCUMENTS [001 1] [Patent Document 1] Japanese Laid-open Patent Application (Kokai) No. 2005-002087 [Patent Document 2] Japanese Laid-open Patent Application (Kokai) No. 2004-051530 NON-PATENT DOCUMENTS [0012] [Non-patent Document 1] Gallo, R.L. et al., Journal of Investigative Der matology 131:1974-1980 (2011) [Non-patent Document 2] Kisich, K.O. et al., Journal of Investigative Der matology 127:2368-2380 (2007) SUMMARY OF THE INVENTION Problems to be Solved by the Invention [0013] For maintenance of a healthy skin surface condition, a means and a method for protecting the skin from external stresses such as drying and harmful bacteria are indis pensable.
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