Composition for the treatment of tinea capitis

Abstract

A novel composition for treating tinea capitis is provided. The composition contains at least one strain selected from Staphylococcus aureus, Staphylococcus hemolyticus, and Staphylococcus wartii.

Description

Technical Field

[0001] This invention relates to compositions for the treatment of tinea capitis, etc. Background Technology

[0002] Tinea capitis is caused by Trichophyton spp. ( Trichophyton Tinea capitis is a skin infection caused by fungi (also known as Trichophyton). Based on the location of the infection, tinea capitis is classified into tinea pedis (athlete's foot), onychomycosis (nail fungus), tinea manuum (hand fungus), tinea corporis (body fungus), tinea cruris (groin fungus), and tinea capitis (scalp fungus), among others.

[0003] Tinea capitis is caused by infection with Trichophyton mentagrophytes present in patients with tinea capitis. For example, in frequently occurring diseases such as tinea pedis (athlete's foot) or onychomycosis (nail fungus), Trichophyton mentagrophytes, which are known to be present in swimming pools, bathhouses, restaurants, slippers, etc., are the main routes of infection.

[0004] Trichophyton floccosum is a transient fungus that lives in the stratum corneum or subcutaneous tissue of the skin. If Trichophyton floccosum proliferates and invades the interior of the stratum corneum, it can cause inflammation or blisters, resulting in symptoms such as itching. It should be noted that Trichophyton floccosum uses keratin, a major component of the stratum corneum, as its nutrient source.

[0005] Previously, antifungal drugs such as itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, iriconazole, fluconazole, fluxaconazole, terbinafine, and butenafine were used as treatments for tinea vulgaris.

[0006] For example, Non-Patent Literature 1 describes the following: Tinea corporis (ringworm) and tinea capitis (ringworm of the scalp) are more common in children before puberty. On the other hand, tinea corporis (ringworm), tinea cruris (ringworm of the groin), and tinea pedis (tinea pedis) are more common in young adults and adults. Since there are diseases that show the same clinical symptoms, it is sometimes preferred to make a diagnosis by potassium hydroxide preparation or culture. The appropriate treatment drugs can be creams or oral preparations.

[0007] Furthermore, Patent Document 1 discloses an invention relating to a topical preparation for the treatment of onychomycosis containing a) an antifungal active substance, b) a volatile component, c) medium-chain triglycerides, and d) ethyl lactate. According to the topical preparation described in Patent Document 1, by using medium-chain triglycerides as a non-volatile component and ethyl lactate as a permeability enhancer, it possesses excellent properties from the viewpoints of nail permeability, efficacy, and formulation properties.

[0008] It should be noted that patent document 1 describes a topical formulation for the treatment of onychomycosis that is superior to oral formulations in terms of fewer systemic side effects and fewer drug interactions.

[0009] Existing technical documents Patent documents Patent Document 1: International Publication No. 2019 / 088005.

[0010] Non-patent literature Non-patent literature 1: American Family Physician, Volume 90, Number 10, 702-711. Summary of the Invention

[0011] The problem that the invention aims to solve In this context, further development of treatments for tinea versicolor is underway.

[0012] Methods for solving problems The present invention provides, for example, a composition for treating tinea capitis.

[0013] [1] A composition for treating tinea capitis, containing a strain of Staphylococcus aureus (Staphylococcus faecium). Staphylococcus hominis ) strains, hemolytic staphylococci ( Staphylococcus haemolyticus ) strains and Staphylococcus aureus ( Staphylococcus warneri At least one strain.

[0014] [2] The tinea pedis treatment composition according to [1] above contains a human staphylococcal strain.

[0015] [3] The tinea pedis treatment composition according to [2] above, wherein the human staphylococcal strain has 16S rDNA containing the base sequence of sequence number 1 or 16S rDNA containing the base sequence having more than 95% identity with sequence number 1, and has anti-tinea pedis activity.

[0016] [4] The tinea pedis treatment composition according to [2] or [3] above, wherein the Staphylococcus aureus strain contains at least one selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus aureus ATCC700236 and Staphylococcus aureus A9 (ATCC commission number: PTA-125203).

[0017] [5] The tinea pedis treatment composition according to any one of [1] to [4] above contains a hemolytic Staphylococcus aureus strain.

[0018] [6] The tinea treatment composition according to [5] above, wherein the hemolytic staphylococcal strain has a 16S rDNA containing the base sequence of sequence number 10 or a 16S rDNA containing a base sequence having more than 95% identity with sequence number 10, and has anti-tinea activity.

[0019] [7] The tinea treatment composition according to [5] or [6] above, wherein the hemolytic staphylococcal strain contains at least one selected from hemolytic staphylococcal ATCC29970, hemolytic staphylococcal ATCC700564 and hemolytic staphylococcal ATCC29969.

[0020] [8] The tinea pedis treatment composition according to any one of [1] to [7] above contains Staphylococcus aureus strain.

[0021] [9] The tinea treatment composition according to [8] above, wherein the Staphylococcus aureus strain has a 16S rDNA containing the base sequence of sequence number 17 or a 16S rDNA containing a base sequence having more than 95% identity with sequence number 17, and has anti-tinea activity.

[0022]

[10] The tinea treatment composition according to [8] or [9] above, wherein the Staphylococcus aureus strain contains Staphylococcus aureus ATCC27836.

[0023]

[11] The tinea treatment composition according to any one of [1] to

[10] above, wherein the pathogenic fungus of tinea comprises Trichophyton spp.

[0024]

[12] The tinea corporis therapeutic composition according to

[11] above, wherein the pathogenic fungus of tinea corporis comprises Trichophyton rubrum ( Trichophyton rubrum Fungal strains, Trichophyton mentagrophytes ( Trichophyton mentagrophytes Fungal strains and Trichophyton tonsurans ( Trichophyton tonsurans At least one fungal strain.

[0025]

[13] The tinea pedis treatment composition according to any one of [1] to

[12] above is a topical agent.

[0026]

[14] A method for manufacturing a composition for treating tinea capitis, comprising: The process of collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (1), Step (2) involves isolating and culturing the cultured bacterial flora to obtain candidate bacteria. The steps (3) for evaluating the antifungal activity of the candidate bacteria and identifying bacteria with antifungal activity, and Step (4) of preparing a tinea treatment composition containing bacteria having the anti-tinea fungal activity; The bacteria having the aforementioned anti-Trichophyton floccosum activity contain at least one strain selected from Staphylococcus aureus, Staphylococcus hemolyticus, and Staphylococcus variabilis.

[0027] The effects of the invention According to the present invention, a novel composition for the treatment of tinea capitis is provided. According to a preferred embodiment of the invention, it can be used even when the artificial chemical synthesis and extraction of the active compound is difficult, or when the efficacy is based on a biological response derived from live bacteria, and can have one or more of the following effects: prevention or reduction of side effects, prevention or inhibition of rashes on the skin surrounding the affected area, improvement of skin adhesion, maintenance of relief, prevention of recurrence, reduction of the frequency of application, improvement of compliance, and higher therapeutic efficacy. Attached Figure Description

[0028] [ Figure 1 Typical examples of various evaluations of antifungal activity against Trichophyton mentagrophytes.

[0029] [ Figure 2 [Graph showing the antifungal activity of various species of candidate bacteria (91 strains) obtained from the ankle and sole.]

[0030] [ Figure 3 Staphylococcus spp. Staphylococcus The phylogenetic tree of known and candidate bacteria.

[0031] [ Figure 4 Phylogenetic tree of known and candidate strains of Staphylococcus aureus.

[0032] [ Figure 5 Phylogenetic tree of known and candidate strains of hemolytic Staphylococcus. Detailed Implementation

[0033] The present invention will now be described in detail.

[0034] <Composition for the Treatment of Tinea> The tinea capitis treatment composition of this invention contains at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii. According to the tinea capitis treatment composition of this invention, tinea capitis can be treated. It should be noted that, in this specification, "treatment" means preventing or inhibiting the proliferation of Trichophyton spp. fungi (so-called Trichophyton) through bactericidal or bacteriostatic action.

[0035] It is known that the onset and recurrence of tinea capitis exhibit individual differences. The inventors predicted that these individual differences lay in the resident skin flora and investigated the effects of resident skin flora and known strains of Staphylococcus on tinea capitis collected from the ankles and soles of healthy individuals. The results showed that Staphylococcus aureus (hereinafter also referred to as "Staphylococcus hominis"), a resident skin flora, was significantly affected. S. hominis )”) strain, hemolytic staphylococci (hereinafter also referred to as “hemolytic staphylococci”) S. haemolyticus strains of Staphylococcus aureus and Staphylococcus vortex (hereinafter also referred to as "Staphylococcus vortex").S . warneri The strain is effective in treating tinea capitis. According to the present invention, a new composition for the treatment of tinea capitis can be provided.

[0036] It should be noted that because Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains are common skin flora, they produce few or no side effects. Therefore, for example, they can prevent or inhibit rashes on the skin around the affected area that may have occurred with conventional topical agents.

[0037] Furthermore, since Staphylococcus aureus, Staphylococcus hemolyticus, and Staphylococcus wartii are resident bacteria of the skin, they easily adhere to the skin. Therefore, compared with conventional topical agents, they can have effects such as a longer duration of effect on the affected skin and a reduction in the frequency of application.

[0038] In addition, since Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus variabilis strains are live bacteria that reside on the skin, they can have therapeutic effects even when the artificial chemical synthesis and extraction of the active pharmaceutical compound is difficult or when the efficacy is based on the life response derived from live bacteria.

[0039] Therefore, the tinea pedis treatment composition of the present invention has the advantages of fewer side effects, better compliance, and higher efficacy compared with other topical agents.

[0040] [Staphylococcus aureus strain] In one embodiment, the composition for treating tinea capitis contains a strain of Staphylococcus aureus.

[0041] Staphylococcus humanis strains are coagulase-negative staphylococci (CNS) and Gram-positive bacteria. They are commonly found on the skin. It is hypothesized that Staphylococcus humanis strains exert their bactericidal or bacteriostatic effects by directly or indirectly damaging Trichophyton mentagrophytes, competing for its habitat, or by preventing or inhibiting the proliferation or fixation of Trichophyton mentagrophytes.

[0042] The Staphylococcus aureus strain preferably comprises a Staphylococcus aureus strain with anti-Trichophyton floccosum activity. That is, in a preferred embodiment, a composition for treating tinea capitis containing a Staphylococcus aureus strain with anti-Trichophyton floccosum activity is provided. It should be noted that, in this specification, "anti-Trichophyton floccosum activity" means that, when the anti-Trichophyton floccosum activity is evaluated using the method described in "4. Evaluation of Anti-Trichophyton floccosum Activity" of the examples, the evaluation result is "++" or "+", preferably "++". Therefore, Staphylococcus aureus strains with anti-Trichophyton floccosum activity can treat tinea capitis.

[0043] There are no particular limitations on the strain of Staphylococcus aureus, but strains containing the 16S rDNA containing the base sequence of sequence number 1 or more, or strains containing the 16S rDNA containing the base sequence of sequence number 1 with at least 95% identity and exhibiting anti-tinea capitis activity, are preferred. As shown in the examples, sequence number 1 is the base sequence of the region enclosed by two common sequences shared by 16S rDNA of the genus Staphylococcus. In this case, the common sequence on the 5' side is common sequence 1 (AGCTTGC), and the common sequence on the 3' side is common sequence 2 (AAGCTGG), both of which are included in sequence number 1. Here, for example, "16S rDNA containing the base sequence of sequence number 1" means that the 16S rDNA contains the base sequence of sequence number 1. It should be noted that, in this specification, the base sequences of sequences 1 to 18 can be determined by the methods described in the examples. In addition, if a bacterium has two or more types of 16S rDNA, the 16S rDNA with the highest abundance is considered the 16S rDNA of that bacterium. In cases where there are two or more 16S rDNAs in the same quantity, for Staphylococcus aureus strains, the 16S rDNA with the highest identity to sequence number 1 is used as the 16S rDNA of that bacterium; for Staphylococcus hemolyticus strains, the 16S rDNA with the highest identity to sequence number 10 is used as the 16S rDNA of that bacterium; and for Staphylococcus wortii strains, the 16S rDNA with the highest identity to sequence number 17 is used as the 16S rDNA of that bacterium.

[0044] The base sequence having more than 95% identity with Serial No. 1 is preferably 96% or more, 96.5% or more, 97% or more, 97.5% or more, 98% or more, 98.1% or more, 98.2% or more, 98.3% or more, 98.4% or more, 98.5% or more, 98.6% or more, 98.7% or more, 98.8% or more, 98.9% or more, 99% or more, 99.1% or more, 99.2% or more, 99.3% or more, 99.4% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or 99.9% or more.

[0045] Mutations that have more than 95% identity with the base sequence of sequence number 1 can include substitution, deletion, insertion, addition, and combinations thereof. In a preferred embodiment, the number of substitutions, deletions, insertions, additions, and combinations thereof is preferably 1 to 10, more preferably 1 to 5, and even more preferably 1 to 2.

[0046] As for the aforementioned substitutions, there are no particular limitations, but preferably at least one of the following: guanine at position 45, thymine at position 139, thymine at position 140, adenine at position 309, and cytosine at position 1265. It should be noted that, in one embodiment, guanine at position 45 of sequence number 1 may be substituted with adenine (45G>A (also represented as G45A)), thymine at position 139 may be substituted with cytosine (139T>C), thymine at position 140 may be substituted with cytosine (140T>C), adenine at position 309 may be substituted with cytosine (309A>C), and cytosine at position 1265 may be substituted with thymine (1265C>T).

[0047] As for the above deletion, there are no particular restrictions; the adenine at position 488 of sequence number 1 (488delA (also represented as A488Δ)) can be deleted.

[0048] There are no particular restrictions on the aforementioned insertion; it can be inserted between positions 1182-1183 and between positions 1235-1236 of serial number 1. It should be noted that, in one embodiment, adenine (1182_1183insA (also represented as Δ1183A)) can be inserted between positions 1182-1183 of serial number 1, and guanine (1235_1236insG) can be inserted between positions 1235-1236.

[0049] Furthermore, the base sequence with over 95% identity to sequence number 1 above can be a base sequence preserved by Staphylococcus aureus strains through natural mutation or other means, or a base sequence obtained through artificial mutation treatment. It should be noted that artificial mutation treatments include, for example, gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments can be used alone or in combination of two or more.

[0050] As base sequences that share more than 95% identity with sequence number 1, sequences 2 through 9 can be listed. The mutation sites of sequences 2 through 9 are shown in Table 1 below.

[0051] [Table 1] Serial Number Mutation sites based on sequence number 1 Serial Number 1 - Serial Number 2 139T>C, 140T>C Serial Number 3 1265C>T Serial Number 4 140T>C Serial Number 5 45G>A Serial Number 6 45G>A, 309A>C Serial Number 7 45G>A、488delA Serial Number 8 45G>A、1182_1183insA Serial Number 9 45G>A、1235_1236insG There are no particular restrictions on the Staphylococcus aureus strains that have 16S rDNA containing sequence number 1, and examples include Staphylococcus aureus ATCC27844 and Staphylococcus aureus ATCC27845.

[0052] There are no particular restrictions on the types of Staphylococcus aureus strains that contain 16S rDNA with a base sequence that is more than 95% identical to that of sequence number 1 and have anti-tinea capitis activity. Examples include Staphylococcus aureus ATCC700236 and Staphylococcus aureus A9 (ATCC commissioned number: PTA-125203).

[0053] In one embodiment, there are no particular limitations on the strains of Staphylococcus aureus, and examples include Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus aureus ATCC700236, Staphylococcus aureus FDARGOS_748, Staphylococcus aureus J6, Staphylococcus aureus FDARGOS_136, Staphylococcus aureus A9 (ATCC commissioned number PTA-125203), Staphylococcus aureus C2, Staphylococcus aureus AMT2, Staphylococcus aureus AMT3, Staphylococcus aureus AMT4-C2, Staphylococcus aureus AMT4-G1, Staphylococcus aureus AMT4-D12, etc. In addition to the above, strains belonging to Staphylococcus aureus can be found using "NCBI", "online", saved on February 17, 2022, [searched on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2014 / > The strains described herein. Preferably, the strains belonging to Staphylococcus aureus include at least one strain selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus aureus ATCC700236, Staphylococcus aureus FDARGOS_748, Staphylococcus aureus J6, Staphylococcus aureus FDARGOS_136, and Staphylococcus aureus A9; more preferably, at least one strain selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus aureus ATCC700236, and Staphylococcus aureus A9. It should be noted that the above-mentioned Staphylococcus aureus strains can be used alone or in combination of two or more.

[0054] Staphylococcus aureus strains with anti-Trichophyton floccosum activity can be readily prepared by the following method. Specifically, the method for preparing Staphylococcus aureus strains with anti-Trichophyton floccosum activity includes: (1) collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without symptoms of Trichophyton floccosum); (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and (3) evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine the bacteria with anti-Trichophyton floccosum activity. Each step is described later. The bacterial flora on the ankle and sole of a person usually contains strains of Staphylococcus aureus, which are known to be resident bacteria of the skin. Moreover, according to the above method, strains of Staphylococcus aureus with anti-Trichophyton floccosum activity can be readily prepared.

[0055] The preferred concentration of Staphylococcus aureus strain in the composition for treating tinea capitis is 10. 3 ~10 11 CFU / g, more preferably 10 4 ~10 10 CFU / g, further preferably 10 5 ~10 9 CFU / g. It should be noted that in this specification, "CFU" refers to Colony Forming Unit. Furthermore, in this specification, the concentration of bacterial strains (CFU / g) in the tinea treatment composition is calculated by diluting the tinea treatment composition to an appropriate concentration, applying it to bacterial culture agar medium, and measuring the colonies that appear after incubation (refer to Clinical Microbiology Procedures Handbook Volume 2, Third Edition).

[0056] There are no particular restrictions on the morphology of the Staphylococcus aureus strain in the composition for treating tinea capitis, but it is preferred to be in the form of dried powder (e.g., freeze-dried or spray-dried).

[0057] [Hemolytic Staphylococcus strain] In one embodiment, the composition for treating tinea capitis contains a hemolytic Staphylococcus aureus strain.

[0058] Hemolytic Staphylococcus strains are coagulase-negative staphylococci (CNS) and Gram-positive bacteria. They are commonly found on the skin. It is speculated that hemolytic Staphylococcus strains also inhibit or kill *Trichophyton mentagrophytes* by directly or indirectly damaging it or competing for its habitat.

[0059] The hemolytic staphylococcal strain preferably comprises a hemolytic staphylococcal strain with anti-Trichophyton floccosum activity. That is, in a preferred embodiment, a tinea pedis treatment composition comprising a hemolytic staphylococcal strain with anti-Trichophyton floccosum activity is provided. The hemolytic staphylococcal strain with anti-Trichophyton floccosum activity can treat tinea pedis.

[0060] There are no particular limitations on the hemolytic Staphylococcus strain, but strains containing the 16S rDNA sequence including sequence number 10 or containing a 16S rDNA sequence having at least 95% identity with sequence number 10 and exhibiting anti-tinea capitis activity are preferred. As shown in the example, sequence number 10 is the base sequence of the region enclosed by two common sequences shared by Staphylococcus spp. In this case, the common sequence on the 5' side is common sequence 1 (AGCTTGC), and the common sequence on the 3' side is common sequence 2 (AAGCTGG), both of which are included in sequence number 10. Here, for example, "16S rDNA containing the base sequence of sequence number 10" means that the 16S rDNA sequence contains the base sequence of sequence number 10.

[0061] The base sequence having more than 95% identity with serial number 10 is preferably 96% or more, 96.5% or more, 97% or more, 97.5% or more, 98% or more, 98.1% or more, 98.2% or more, 98.3% or more, 98.4% or more, 98.5% or more, 98.6% or more, 98.7% or more, 98.8% or more, 98.9% or more, 99% or more, 99.1% or more, 99.2% or more, 99.3% or more, 99.4% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or 99.9% or more.

[0062] Mutations that have more than 95% identity with the base sequence of sequence number 10 can include substitutions, deletions, insertions, additions, and combinations thereof. In a preferred embodiment, the number of substitutions, deletions, insertions, additions, and combinations thereof is preferably 1 to 10, more preferably 1 to 5, and even more preferably 1 to 2.

[0063] As for the aforementioned substitutions, there are no particular limitations, but preferably at least one selected from cytosine at position 96, guanine at position 109, adenine at position 398, cytosine at position 952, and cytosine at position 1186 of serial number 10. It should be noted that, in one embodiment, cytosine at position 96 of serial number 10 may be substituted with thymine (96C>T), guanine at position 109 may be substituted with adenine (109G>A), adenine at position 398 may be substituted with guanine (398A>G), and cytosine at position 1186 may be substituted with thymine (1186C>T).

[0064] As for the above deletion, there are no particular restrictions; the adenine at position 1210 (1210delA), position 1219 (1219delA), position 1225 (1225delT), and position 1263 (1263delA) of sequence number 10 may be deleted.

[0065] There are no particular restrictions on the aforementioned insertion; it can be inserted between positions 1117-1118, 1195-1196, and 1234-1235 of sequence number 10. It should be noted that, in one embodiment, guanine (1117-1118insG) can be inserted between positions 1117-1118 of sequence number 10, cytosine (1195-1196insC) can be inserted between positions 1195-1196, and guanine (1234-1235insG) can be inserted between positions 1234-1235.

[0066] The base sequence with over 95% identity to sequence number 10 is also identical to that of the *Staphylococcus aureus* strain. This sequence can be a base sequence retained by other hemolytic *Staphylococcus* strains through natural mutation, or it can be a base sequence obtained through artificial mutation treatment. It should be noted that artificial mutation treatments include, for example, gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments can be used alone or in combination of two or more.

[0067] As base sequences that are more than 95% identical to sequence number 10, sequences 11 to 16 can be listed. The mutation sites of sequences 11 to 16 are shown in Table 2 below.

[0068] [Table 2] Serial Number Mutation sites based on sequence number 10 Serial Number 10 - Serial Number 11 96C>T Serial Number 12 109G>A Serial Number 13 398A>G, 952C>T Serial Number 14 398A>G Serial Number 15 398A>G, 952C>T, 1186C>T Serial Number 16 398A>G, 952C>T, 1117_1118insG, 1195_1196insC, 1210delA, 1219delA, 1225delT, 1234_1235insG, 1263delA There are no particular limitations on hemolytic staphylococcal strains that have a 16S rDNA containing the sequence number 10, and examples include hemolytic staphylococcus ATCC29970 and hemolytic staphylococcus ATCC700564.

[0069] In one embodiment, there are no particular limitations on the hemolytic Staphylococcus strains, and examples include hemolytic Staphylococcus ATCC29970, hemolytic Staphylococcus ATCC700564, hemolytic Staphylococcus ATCC29969, hemolytic Staphylococcus SH_12, and hemolytic Staphylococcus JCSC1435. In addition to the above, hemolytic Staphylococcus strains can be obtained from the internet via NCBI, online, saved on February 17, 2022 [searched on February 17, 2022].<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1141 / > The strains described herein. Preferably, the hemolytic staphylococcal strains include at least one selected from *Staphylococcus ATCC29970*, *Staphylococcus ATCC700564*, *Staphylococcus ATCC29969*, *Staphylococcus SH_12*, and *Staphylococcus JCSC1435*, and more preferably, at least one selected from *Staphylococcus ATCC29970*, *Staphylococcus ATCC700564*, and *Staphylococcus ATCC29969*. It should be noted that the above-mentioned hemolytic staphylococcal strains can be used alone or in combination of two or more.

[0070] The hemolytic staphylococcal strains with anti-Trichophyton floccosum activity are the same as those of Staphylococcus aureus and can be easily prepared by the following method. Specifically, the method for preparing hemolytic staphylococcal strains with anti-Trichophyton floccosum activity includes: (1) collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without symptoms of Trichophyton floccosum); (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and (3) evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine the bacteria with anti-Trichophyton floccosum activity. Hemolytic staphylococcal strains with anti-Trichophyton floccosum activity can also be found in the bacterial flora of the ankle and sole of a person. Therefore, hemolytic staphylococcal strains with anti-Trichophyton floccosum activity can be easily prepared according to the above method.

[0071] The preferred concentration of hemolytic Staphylococcus strains in the composition for treating tinea capitis is 10. 3 ~10 11 CFU / g, more preferably 10 4 ~10 10 CFU / g, further preferably 10 5 ~10 9 CFU / g.

[0072] There are no particular restrictions on the morphology of the hemolytic Staphylococcus strain in the composition for treating tinea capitis, but the preferred form is a dried powder (e.g., freeze-dried or spray-dried).

[0073] [Staphylococcus aureus] In one embodiment, the composition for treating tinea versicolor contains a strain of Staphylococcus aureus.

[0074] Staphylococcus wartii strains are coagulase-negative staphylococci (CNS) and Gram-positive bacteria. They are commonly found on the skin. It is speculated that Staphylococcus wartii strains also exert bactericidal or bacteriostatic effects by directly or indirectly damaging Trichophyton mentagrophytes or competing for its habitat, thus preventing or inhibiting the proliferation of Trichophyton mentagrophytes.

[0075] The *Staphylococcus wartii* strain preferably comprises a *Staphylococcus wartii* strain with anti-*Trichophyton* activity. That is, in a preferred embodiment, a tinea treatment composition comprising a *Staphylococcus wartii* strain with anti-*Trichophyton* activity is provided. The *Staphylococcus wartii* strain with anti-*Trichophyton* activity can treat tinea.

[0076] There are no particular limitations on the *Staphylococcus wortii* strain, but strains containing the 16S rDNA sequence including sequence number 17 or containing a 16S rDNA sequence having at least 95% identity with sequence number 17 and exhibiting anti-tinea capitis activity are preferred. As shown in the examples, sequence number 17 is the base sequence of the region enclosed by two common sequences shared by *Staphylococcus* genus 16S rDNA. In this case, the common sequence on the 5' side is common sequence 1 (AGCTTGC), and the common sequence on the 3' side is common sequence 2 (AAGCTGG), both of which are included in sequence number 17. Here, for example, "16S rDNA containing the base sequence of sequence number 17" means that the 16S rDNA contains the base sequence of sequence number 17.

[0077] The base sequence having more than 95% identity with Serial No. 17 is preferably 96% or more, 96.5% or more, 97% or more, 97.5% or more, 98% or more, 98.1% or more, 98.2% or more, 98.3% or more, 98.4% or more, 98.5% or more, 98.6% or more, 98.7% or more, 98.8% or more, 98.9% or more, 99% or more, 99.1% or more, 99.2% or more, 99.3% or more, 99.4% or more, 99.5% or more, 99.6% or more, 99.7% or more, 99.8% or more, or 99.9% or more.

[0078] Mutations that have more than 95% identity with the base sequence of sequence number 17 can include substitutions, deletions, insertions, additions, and combinations thereof. In a preferred embodiment, the number of substitutions, deletions, insertions, additions, and combinations thereof is preferably 1 to 10, more preferably 1 to 5, and even more preferably 1 to 2.

[0079] As a substitution, there are no particular limitations, but cytosine at position 52 of serial number 17 is preferred. It should be noted that, in one embodiment, cytosine at position 52 of serial number 17 may be substituted with guanine (52C>G).

[0080] Regarding the base sequence that shares over 95% identity with sequence number 17, similar to that of Staphylococcus aureus strains, it can be a base sequence preserved by other Staphylococcus wort strains through natural mutation, or a base sequence obtained through artificial mutation treatment. It should be noted that artificial mutation treatments include, for example, gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments can be used alone or in combination of two or more.

[0081] As a base sequence that has more than 95% identity with sequence number 17, sequence number 18 can be listed. The mutation sites of sequence number 18 are shown in Table 3 below.

[0082] [Table 3] Serial Number Mutation sites based on sequence number 17 Serial Number 17 - Serial Number 18 52C>G There are no particular limitations on the type of Staphylococcus aureus strain that has a 16S rDNA containing sequence number 17, such as Staphylococcus aureus ATCC27836.

[0083] In one embodiment, there are no particular limitations on the strain of *Staphylococcus vulgaris*, and examples include *Staphylococcus vulgaris* ATCC27836, *Staphylococcus vulgaris* NCTC7291, *Staphylococcus vulgaris* WB224, *Staphylococcus vulgaris* 16A, and *Staphylococcus vulgaris* 22.1. In addition to the above, *Staphylococcus vulgaris* strains can be found using "NCBI," "online," saved on February 17, 2022, [searched on February 17, 2022], or on the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2073 / > The strains described herein. Preferably, the *Staphylococcus variabilis* strain includes at least one selected from *Staphylococcus variabilis* ATCC27836, *Staphylococcus variabilis* NCTC7291, *Staphylococcus variabilis* WB224, *Staphylococcus variabilis* 16A, *Staphylococcus variabilis* 22.1, and *Staphylococcus variabilis* WS479, and more preferably includes *Staphylococcus variabilis* ATCC27836. It should be noted that the above-mentioned *Staphylococcus variabilis* strains can be used alone or in combination of two or more.

[0084] The *Staphylococcus wartii* strain with anti-Trichophyton floccosum activity is the same as the *Staphylococcus aureus* strain and can be easily prepared by the following method. Specifically, the method for preparing the *Staphylococcus wartii* strain with anti-Trichophyton floccosum activity includes: (1) collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without symptoms of Trichophyton floccosum); (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and (3) evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine the bacteria with anti-Trichophyton floccosum activity. The *Staphylococcus wartii* strain with anti-Trichophyton floccosum activity can also be found in the bacterial flora of the ankle and sole of a person. Therefore, according to the above method, the *Staphylococcus wartii* strain with anti-Trichophyton floccosum activity can be easily prepared.

[0085] The preferred concentration of Staphylococcus aureus strain in the composition for treating tinea capitis is 10. 3 ~10 11 CFU / g, more preferably 10 4 ~10 10 CFU / g, further preferably 10 5 ~10 9 CFU / g.

[0086] There are no particular restrictions on the morphology of Staphylococcus aureus strains in the composition for treating tinea capitis, but dry powder form (e.g., freeze-dried form, spray-dried form) is preferred.

[0087] [Other Staphylococcus strains] The composition for treating tinea may further contain other Staphylococcus strains. Here, "Staphylococcus strains" means Staphylococcus strains other than Staphylococcus aureus, hemolytic Staphylococcus, and Staphylococcus wartii.

[0088] As for other Staphylococcus strains, there are no particular restrictions; saprophytic Staphylococcus can be listed as an example. Staphylococcus saprophyticus ) strain, Staphylococcus krill ( Staphylococcus kloosii ) strain, Staphylococcus ludensii ( Staphylococcus lugdunensis ) strains, Staphylococcus aureus ( Staphylococcus capitis ) strain, Staphylococcus aureus ( Staphylococcus caprae ) strains, Staphylococcus epidermidis ( Staphylococcus epidermidis ) strain, Staphylococcus aureus ( Staphylococcus aureus ) strains and other bacterial species with human isolation reports.

[0089] As saprophytic Staphylococcus ( S . saprophyticus The strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1350 / > The strains recorded in the literature.

[0090] As Staphylococcus kurizinus ( S . kloosii The strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 66884 / > The strains recorded in the literature.

[0091] As Staphylococcus ludenum ( S . lugdunensis The strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2548 / > The strains recorded in the literature.

[0092] As cephalosporin ( S . capitisThe strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2054 / > The strains recorded in the literature.

[0093] As Staphylococcus aureus ( S . caprae The strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1971 / > The strains recorded in the literature.

[0094] As Staphylococcus epidermidis ( S . epidermidis The strain can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 155 / > The strains recorded in the literature.

[0095] As Staphylococcus aureus ( S . aureus The strain can be listed as "NCBI" or "online," saved on February 17, 2022, [retrieved on February 17, 2022], from the internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 154 / > The strains recorded in the literature.

[0096] From the viewpoint of achieving better therapeutic effects in treating tinea capitis, other Staphylococcus strains are preferably selected from Staphylococcus capitis, Staphylococcus membranaceus, and Staphylococcus epidermidis. S . epidermidis The composition contains at least one strain of Staphylococcus aureus, Staphylococcus krusei, Staphylococcus ludens, and Staphylococcus saprophyticus, more preferably at least one strain selected from Staphylococcus cephalosporin and Staphylococcus capsulatum, and particularly preferably at least one strain selected from Staphylococcus cephalosporin ATCC27840 and Staphylococcus capsulatum ATCC35538. Furthermore, from the viewpoint of adjusting the balance of resident skin flora at the application site of the tinea treatment composition, other Staphylococcus strains preferably include Staphylococcus epidermidis (S. aureus). S . epidermidisThe strains are further preferably composed of at least one of Staphylococcus epidermidis ATCC12228 and Staphylococcus epidermidis ATCCC14990. It should be noted that these other Staphylococcus strains can be used alone or in combination of two or more.

[0097] There are no particular limitations on the concentration of other Staphylococcus strains in the composition for treating tinea capitis, but 10 is preferred. 3 ~10 11 CFU / g, more preferably 10 4 ~10 10 CFU / g, further preferably 10 5 ~10 9 CFU / g. It should be noted that when the product contains two or more other Staphylococcus strains, it is preferable that each of the two or more other Staphylococcus strains is within the range described above.

[0098] The morphology of other Staphylococcus strains in the composition for treating tinea capitis is not particularly limited, but dry powder form (e.g., freeze-dried or spray-dried) is preferred. It should be noted that the morphology of the other Staphylococcus strains is preferably the same as at least one of the strains selected from the above-mentioned Staphylococcus humanis, Staphylococcus hemolyticus, and Staphylococcus wartii strains.

[0099] [Other genera and strains] The composition for treating tinea may further contain bacterial strains of other genera. Here, "other genera" refers to bacterial strains other than Staphylococcus.

[0100] There are no particular restrictions on other genera and strains, but resident skin bacteria (i.e., resident skin bacteria other than Staphylococcus) are preferred, such as *Dermatobacterium acnes*. Cutibacterium acnes (Acne bacteria) strains, Streptococcus pyogenes ( Streptococcus pyogenes (Group A) β Hemolytic streptococci) strains, Pseudomonas aeruginosa ( Pseudomonas aeruginosa (Pseudomonas aeruginosa) strain, Escherichia coli ( Escherichia coli ) strains, Bacillus cereus ( Bacillus cereus ) strains, Enterococcus faecalis ( Enterococcus faecalis ) strain, Moraxella osloi Moraxella osloensis ) strains, etc.

[0101] As a strain of *Propionibacterium acnes* (acne fungus), it can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1140 / > The strains recorded in the literature.

[0102] As a group A streptococcus pyogenes β Hemolytic streptococcus strains, listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 175 / > The strains recorded in the literature.

[0103] As a strain of Pseudomonas aeruginosa, it can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 187 / > The strains recorded in the literature.

[0104] As for the *E. coli* strain, the following can be listed: "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 167 / > The strains recorded in the literature.

[0105] As a strain of Bacillus cereus, it can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 157 / > The strains recorded in the literature.

[0106] As a strain of Enterococcus faecalis, it can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 808 / > The strains recorded in the literature.

[0107] As an Oslo Moraxella strain, it can be listed as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 12468 / > The strains recorded in the literature.

[0108] Among these strains, from the viewpoint of achieving a higher therapeutic effect for tinea versicolor, it is preferable to include at least one strain selected from *Pseudomonas aeruginosa*, *Escherichia coli*, *Bacillus cereus*, *Enterococcus faecalis*, and *Moraxella osloensis*. More preferably, it includes at least one strain selected from *Pseudomonas aeruginosa*, *Escherichia coli*, and *Enterococcus faecalis*. Even more preferably, it includes at least one strain selected from *Pseudomonas aeruginosa* ATCC27853, *Escherichia coli* ATCC25922, and *Enterococcus faecalis* ATCC29212. Furthermore, from the viewpoint of adjusting the balance of resident skin flora at the application site of the tinea versicolor treatment composition, it is preferable to include strains selected from *Propionibacterium acnes* (acne bacteria) and *Streptococcus pyogenes* (group A). β The bacteria contain at least one strain of hemolytic streptococcus and a strain of Pseudomonas aeruginosa, more preferably including a strain of Propionibacterium acnes, and even more preferably including Propionibacterium acnes ATCC6919. It should be noted that these other genera and strains can be used alone or in combination of two or more.

[0109] There are no particular limitations on the concentration of other strains in the composition for treating tinea capitis, but 10 is preferred. 3 ~10 11 CFU / g, more preferably 10 4 ~10 10 CFU / g, further preferably 10 5 ~10 9 CFU / g. It should be noted that when two or more strains from other genera are present, it is preferable that each species of the two or more strains from other genera be within the range described above.

[0110] There are no particular restrictions on the morphology of other strains in the composition for treating tinea capitis, but dry powder form (e.g., freeze-dried or spray-dried) is preferred. It should be noted that the morphology of other strains is preferably the same as at least one of the strains selected from the above-mentioned Staphylococcus aureus, Staphylococcus hemolyticus, and Staphylococcus wartii strains.

[0111] [Antifungal agents] The composition for treating tinea capitis may further contain an antifungal agent. The presence of an antifungal agent further enhances the therapeutic effect of tinea capitis.

[0112] There are no particular limitations on antifungal agents, and examples include itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, ifluconazole, flusaconazole, terbinafine, and butenafine. These antifungal agents can be used alone or in combination of two or more.

[0113] There is no particular limitation on the concentration of the antifungal agent in the tinea pedis treatment composition, but it is preferably 0.1 to 30% by mass, more preferably 0.5 to 15% by mass, relative to the total mass of the tinea pedis treatment composition.

[0114] [additive] Compositions for treating tinea may also contain additives. There are no particular restrictions on additives, but examples include bases, wetting agents, thickeners, emulsifiers, co-emulsifiers, preservatives, stabilizers, and pH adjusters.

[0115] (Matrix) Examples of matrices include hydrophobic matrices, hydrophilic matrices, and water. It should be noted that, in this specification, "matrix" refers to an additive having a content of 40% by mass or more, preferably 50% to 99% by mass, relative to the total mass of the composition for treating tinea capitis.

[0116] There are no particular limitations on the hydrophobic matrix mentioned above. Examples include squalane, liquid paraffin, light liquid paraffin, petrolatum, ceresin, microcrystalline wax, squalene, gelled hydrocarbons, and other higher hydrocarbons; olive oil, jojoba oil, sesame oil, soybean oil, cocoa butter, camellia oil, peanut oil, tallow, lard, triacetyl glycerol, hydrogenated castor oil, and other oils; beeswax, white beeswax, carnauba wax, lanolin, and other waxes; stearic acid, oleic acid, and other fatty acids; lanolin alcohol, myristol, cetyl alcohol, stearyl alcohol, cetearyl alcohol, cholesterol, and other higher alcohols; and isopropyl myristate, stearyl myristate, medium-chain triglycerides, and other fatty acid esters.

[0117] There are no particular limitations on the hydrophilic matrix mentioned above, and examples include lower alcohols such as ethanol, propanol, and isopropanol; polyols such as glycerol, 1,3-butanediol, and propylene glycol; sugar alcohols such as sorbitol and mannitol; and polyethylene glycol.

[0118] The above-mentioned substrates can be used alone or in combination of two or more.

[0119] (Wetting agent) The wetting agent keeps the skin moist when applying the composition for treating tinea capitis.

[0120] There are no particular restrictions on what can be used as a wetting agent; examples include petrolatum, glycerin, propylene glycol, and 1,3-butanediol.

[0121] The above wetting agents can be used alone or in combination of two or more.

[0122] (Thickener) Thickeners increase the viscosity or gel the composition used to treat tinea capitis.

[0123] There are no particular restrictions on thickeners, and examples include gelatin, agar, carrageenan, gum arabic, tragacanth, sodium alginate, propylene glycol alginate, acrylic polymers, polyvinyl alcohol (partial saponification), sodium alginate, methylcellulose, carboxymethylcellulose, water-soluble cellulose derivatives (hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, and hydrophobic hydroxypropylmethylcellulose, etc.), sodium polyacrylate, glyceryl monooleate, pectin, xanthan gum, etc.

[0124] The above thickeners can be used alone or in combination of two or more.

[0125] (Emulsifier) Emulsifiers emulsify the compositions for treating tinea capitis into oil-in-water (o / w) or water-in-oil (w / o) emulsions.

[0126] As emulsifiers, there are no particular limitations, and examples include cationic surfactants such as alkylamine salts, alkylamine polyoxyethylene adducts, fatty acid triethanolamine monoester salts, acylaminoethyl diethylamine salts, and fatty acid polyamine condensates; anionic surfactants such as polyoxyethylene alkyl ether phosphates, aminosulfates, saturated higher fatty acid salts, and N-acyl amino acid salts; nonionic surfactants such as sucrose fatty acid esters, glycerol fatty acid esters, polyglycerol fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene dehydrated sorbitol fatty acid esters, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, polyethylene glycol, polyethylene glycol fatty acid esters, polyoxyethylene polyoxypropylene glycol, and polyoxyethylene alkyl ethers; and amphoteric surfactants such as alkyl betaine, alkylamide betaine, alkyl sulfobetaine, imidazoline, lauryl dimethylaminoacetic acid betaine, and alkyl diaminoethyl glycine.

[0127] The above emulsifiers can be used alone or in combination of two or more.

[0128] (Emulsifier) Emulsifiers stabilize the emulsion when the composition for treating tinea capitis is emulsified into an oil-in-water (o / w) or water-in-oil (w / o) form.

[0129] There are no particular restrictions on what can be used as a co-emulsifier; examples include cetyl alcohol, stearyl alcohol, oleyl alcohol, and isostearyl alcohol.

[0130] The above-mentioned emulsifiers can be used alone or in combination of two or more.

[0131] (preservative) Preservatives prevent or inhibit contamination and decomposition of the composition for treating tinea capitis caused by foreign microorganisms.

[0132] As preservatives, there are no particular restrictions, and examples include p-hydroxybenzoic acid, methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, sodium benzoate, chlorobutanol, chlorocresol, benzyl alcohol, salicylic acid, phenoxyethanol, thymol, butylated hydroxytoluene, sodium edetate hydrate, sodium dehydroacetate, sorbic acid, potassium sorbate, benzalkonium chloride, etc.

[0133] The above-mentioned preservatives can be used alone or in combination of two or more.

[0134] (Stabilizer) Stabilizers prevent or inhibit the death, decomposition, and physical changes of components contained in compositions used to treat tinea capitis.

[0135] As stabilizers, there are no particular restrictions, and examples include sodium bisulfite, sodium metabisulfite, ascorbic acid, tocopherol, butylated hydroxytoluene, sodium edetate (EDTA), and benzotriazole.

[0136] The above stabilizers can be used alone or in combination of two or more.

[0137] (pH adjuster) pH adjusters maintain the stability of the components contained in the tinea pedis treatment composition and prevent or reduce the irritation of the tinea pedis treatment composition to organisms.

[0138] As pH adjusters, there are no particular limitations, and examples include lactic acid, acetic acid, acetates (such as sodium acetate), citric acid, citrates (such as sodium citrate), phosphoric acid, phosphates (such as sodium phosphate), diisopropanolamine, triisopropanolamine, triethanolamine, potassium hydroxide, and sodium hydroxide.

[0139] The above-mentioned pH adjusters can be used alone or in combination of two or more.

[0140] [Dosage Form] There are no particular limitations on the dosage form of the tinea pedis treatment composition, but a topical formulation is preferred. By using the tinea pedis treatment composition as a topical formulation, at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii can be attached to the skin, effectively preventing or inhibiting the proliferation or attachment of tinea pedis, thereby achieving bactericidal or bacteriostatic effects.

[0141] There are no particular restrictions on the specific form of topical agents; examples include ointments, creams, gels, and lotions.

[0142] It should be noted that ointments are semi-solid topical preparations applied to the skin, in which the active ingredient is dissolved or dispersed in a base. Examples include oily ointments and water-soluble ointments. Creams are semi-solid topical preparations applied to the skin, emulsified into an oil-in-water (o / w) or water-in-oil (w / o) form. Gels are gel-like topical preparations applied to the skin, including aqueous gels and oily gels. Lotions are liquid topical preparations in which the active ingredient (such as Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, or Staphylococcus wartii strains) is dissolved or dispersed in an aqueous solution.

[0143] [Trichophyton floccosum] The causative agent of tinea capitis is Trichophyton mentagrophytes, specifically Trichophyton mentagrophytes (…). Trichophyton (This refers to a fungus.) Among the aforementioned Trichophyton species are fungal strains of Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, etc.

[0144] Examples of fungal strains of the aforementioned Trichophyton rubrum include Trichophyton rubrum ATCC28188 and Trichophyton rubrum ATCC22402.

[0145] Examples of fungal strains of the aforementioned Trichophyton mentagrophytes include Trichophyton mentagrophytes TIMM2789 and Trichophyton mentagrophytes ATCCMYA-4439.

[0146] Examples of fungal strains of *Trichophyton tonsurans* include *Trichophyton tonsurans* ATCC56186 and *Trichophyton tonsurans* ATCC28942.

[0147] The pathogens causing tinea capitis preferably include fungi of the genus Trichophyton, more preferably at least one fungal strain selected from Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans, further preferably including fungal strains of Trichophyton rubrum, and particularly preferably including Trichophyton rubrum ATCC28188. It should be noted that the pathogens causing tinea capitis can be one or more species.

[0148] Tinea capitis is caused by the aforementioned Trichophyton fungus. It should be noted that, based on the affected area, tinea capitis is classified into tinea pedis (athlete's foot), onychomycosis (nail fungus), tinea manuum (hand fungus), tinea corporis (body fungus), tinea cruris (groin fungus), and tinea capitis (scalp fungus), etc. Since the skin penetration of topical agents varies depending on the affected area, it is preferable to adjust the composition of the topical agent appropriately according to the affected area.

[0149] [Application Method] There are no particular restrictions on the method of application of the composition for treating tinea capitis. When the composition for treating tinea capitis is a topical agent, it is applied to the affected area.

[0150] There is no particular limitation on the number of applications; it can be any of the following: 3 times a day, 2 times a day, once a day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, or once every 7 days. Preferably, the number of applications is once a day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, or once every 7 days. The tinea capitis treatment composition of this invention contains at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii, which exhibit excellent skin adhesion. Therefore, compared to conventional tinea capitis treatments, it can maintain remission or prevent recurrence through the sustained effect of live bacteria adhesion. Furthermore, it can reduce the number of applications or improve patient compliance.

[0151] When used topically, the preferred application rate of the composition for treating tinea capitis is 0.1~180 mg / cm³. 2 More preferably 1~10 mg / cm 2 At this time, the amount of at least one of the Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains selected from the tinea pedis treatment composition applied per application is preferably 10. 0 ~10 8 CFU / cm 2 More preferably 10 1 ~10 7 CFU / cm 2 More preferably 10 2 ~10 6 CFU / cm 2 It should be noted that when the composition for treating tinea capitis contains two or more strains of Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii, the total amount thereof is preferably within the range described above.

[0152] There are no particular limitations on the affected area to which the tinea treatment composition is applied; it is generally used for affected areas exhibiting tinea symptoms. It should be noted that the tinea treatment composition of this invention contains at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus, and Staphylococcus wartii, which are resident bacteria of the skin. Therefore, compared to conventional topical agents, it reduces the frequency of application due to its high skin adhesion and does not cause, or hardly causes, rashes on the skin surrounding the affected area, thus improving patient compliance.

[0153] The above-mentioned tinea pedis treatment composition can be used alone to exert its therapeutic effect, or it can be combined with other therapeutic agents.

[0154] Other therapeutic agents mentioned above may include oral and topical formulations. In this case, the oral or topical formulation contains at least one antifungal drug selected from itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, ilfluconazole, flusaconazole, terbinafine, and butenafine. Two or more of the other therapeutic agents may also be combined.

[0155] That is, according to one embodiment of the present invention, a tinea pedis treatment composition and a combination thereof with other therapeutic agents are provided. In this case, when the tinea pedis treatment composition is a topical agent, the other therapeutic agents are preferably oral agents. Furthermore, the tinea pedis treatment composition and the other therapeutic agents may be applied simultaneously or sequentially.

[0156] <Method for manufacturing composition for treating tinea capitis> According to one embodiment of the present invention, a method for manufacturing a composition for treating tinea capitis is provided. The manufacturing method includes: a step (1) collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without tinea capitis symptoms); a step (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; a step (3) evaluating the anti-tinea capitis activity of the candidate bacteria to identify bacteria with anti-tinea capitis activity; and a step (4) preparing a composition for treating tinea capitis containing bacteria with the aforementioned anti-tinea capitis activity. In this case, the bacteria with the aforementioned anti-tinea capitis activity contain at least one selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains. It should be noted that the Staphylococcus aureus strain is preferably a Staphylococcus aureus strain with anti-tinea capitis activity. Furthermore, the hemolytic Staphylococcus aureus strain is preferably a hemolytic Staphylococcus aureus strain with anti-tinea capitis activity. Additionally, the Staphylococcus wartii strain is preferably a Staphylococcus wartii strain with anti-tinea capitis activity. It should be noted that specific examples of Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus variabilis strains are as described above.

[0157] [Process (1)] Step (1) involves collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without symptoms of tinea pedis). Bacterial flora are collected from the ankle and / or sole using a swab or similar means and smeared onto a culture medium for incubation. Each bacterial strain then forms a colony.

[0158] [Process (2)] Step (2) involves isolating and culturing the bacterial flora cultured in step (1) to obtain candidate bacteria. Colonies formed during the culture in step (1) are collected using platinum rings or similar methods and spread onto a culture medium for isolation and culture. This allows for the isolation of a large number of bacteria contained in the bacterial flora of the human ankle and / or sole. It should be noted that the cultured bacteria are candidate bacteria, preferably suspended in a bacterial stock solution and preserved as a skin bacterial library at low temperatures (e.g., -100 to -50°C, preferably -90 to -70°C).

[0159] [Process (3)] Step (3) is the process of evaluating the anti-Trichophyton floccosum activity of the candidate bacteria obtained in step (2) to identify bacteria with anti-Trichophyton floccosum activity.

[0160] In step (3), firstly, it is evaluated which candidate bacteria isolated from the ankle and / or sole of a person has anti-Trichophyton floccosum activity. At this time, the evaluation of the anti-Trichophyton floccosum activity of the candidate bacteria is carried out by the method described in "4. Evaluation of Anti-Trichophyton Floccosum Activity" of the example.

[0161] Next, bacteria with antifungal activity against Trichophyton mentagrophytes are selected and identified. There are no particular limitations on the identification method; methods such as comparing 16S rDNA sequences and constructing phylogenetic trees can be listed. For example, in the method of comparing 16S rDNA sequences, the target bacteria's 16S rDNA sequence can be analyzed, and the DNA sequence data can be used for NCBI BLAST search to identify the bacteria. Alternatively, in the method of constructing phylogenetic trees, the bacterial 16S rDNA sequence can be analyzed, and the 16S rDNA sequence data of known bacteria can be combined for analysis to construct a phylogenetic tree, which can then be used to identify the target bacteria. It should be noted that in step (3), when identifying the target bacteria as Staphylococcus humanis strains, hemolytic Staphylococcus aureus strains, or Staphylococcus wartii strains, steps may include calculating the identity of the target bacteria's 16S rDNA sequence with sequence numbers 1, 10, 17, etc., and confirming the DNA sequence identity of the target bacteria.

[0162] The bacteria with anti-Trichophyton floccosum activity identified through step (3) may contain at least one strain selected from Staphylococcus aureus with anti-Trichophyton floccosum activity, hemolytic Staphylococcus aureus with anti-Trichophyton floccosum activity, and Staphylococcus wartii with anti-Trichophyton floccosum activity. Therefore, by extracting at least one strain selected from the target Staphylococcus aureus with anti-Trichophyton floccosum activity, hemolytic Staphylococcus aureus with anti-Trichophyton floccosum activity, and Staphylococcus wartii with anti-Trichophyton floccosum activity from the preserved skin bacterial library, strains of Staphylococcus aureus with anti-Trichophyton floccosum activity, hemolytic Staphylococcus aureus with anti-Trichophyton floccosum activity, and Staphylococcus wartii with anti-Trichophyton floccosum activity can be obtained.

[0163] [Process (4)] Step (4) is the process of preparing a tinea therapeutic composition containing bacteria with anti-Trichophyton floccosum activity. The bacteria with anti-Trichophyton floccosum activity identified in step (3) are appropriately cultured, purified, freeze-dried, etc., to prepare a tinea therapeutic composition containing bacteria with anti-Trichophyton floccosum activity. It should be noted that the bacteria with the above-mentioned anti-Trichophyton floccosum activity contain at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii strains, preferably at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii strains with anti-Trichophyton floccosum activity.

[0164] Treatment methods According to one aspect of the present invention, a treatment method for tinea capitis is provided. In this method, the treatment involves applying the aforementioned tinea capitis treatment composition to the affected area. The tinea capitis treatment composition and its method of application are as described above. Example

[0165] The present invention will be illustrated below with examples, but the present invention is not limited thereto.

[0166] 1. Collection of candidate bacteria Bacterial flora were collected from one site each of the ankle and sole of five healthy individuals using swabs. The swabs containing bacterial flora were streaked onto Anero Columbia RS blood agar and incubated aerobically at 35°C for one day. After incubation, colonies representative of morphology, color, and dryness were collected using platinum rings.

[0167] Platinum rings with collected bacterial colonies attached were smeared onto Anero Columbia RS blood agar medium and cultured aerobically at 35°C for 1 day.

[0168] After culturing, the developed bacterial colonies were recovered using a platinum ring, suspended in a 25% glycerol bacterial stock solution, and stored at -80°C. It should be noted that a total of 91 candidate strains were collected. At this point, 52 candidate strains were obtained from the ankle, and 39 candidate strains were obtained from the sole.

[0169] 2. Preparation of the known bacteria As a known bacterium, the following 32 strains of bacteria were prepared.

[0170] (1) Acinetobacter baumannii ( Acinetobacter baumannii ATCC17978 (2) Corynebacterium bluegrass ( Corynebacterium glaucum JCM12208 (3) Corynebacterium jejuni ( Corynebacterium jeikeium ATCC43734 (4) Corynebacterium bandingense ( Corynebacterium striatum ) NBRC15291 (5) Enterococcus faecalis ATCC29212 (6) Escherichia coli ATCC25922 (7) Micrococcus luteus ( Micrococcus luteus ATCC4698 (8) Moraxella tarda ( Moraxella atlantae ) NBRC14588 (9) Neisseria gonorrhoeae ( Neisseria gonorrhoeae ATCC19424 (10) Pseudomonas aeruginosa ATCC27853 (11) Staphylococcus aureus ATCC29213 (12) Staphylococcus epidermidis ATCC12228 (13) Hemolytic Staphylococcus aureus ATCC29970 (14) Staphylococcus aureus ATCC27844 (15) Agalactiae Streptococcus ( Streptococcus agalactiae ATCC13813 (16) Streptococcus pyogenes ( Streptococcus mitis JCM12971 (17) Streptococcus pyogenes ATCC12344 (18) Staphylococcus aureus ATCC12600 (19) Staphylococcus epidermidis ATCC14990 (20) Staphylococcus aureus ATCC27840 (21) Staphylococcus aureus ATCC35538 (22) Staphylococcus aureus ( Staphylococcus cohnii ATCC29974 (23) Hemolytic Staphylococcus aureus ATCC700564 (24) Hemolytic Staphylococcus aureus ATCC29969 (25) Staphylococcus aureus ATCC27845 (26) Staphylococcus aureus ATCC700236 (27) Staphylococcus krill ATCC43959 (28) Staphylococcus aureus ( Staphylococcus lentus ATCC700403 (29) Staphylococcus ludens ATCC49576 (30) Staphylococcus ludensburgii ATCC43809 (31) Staphylococcus aureus ATCC27836 (32) Staphylococcus saprophyticus ATCC15305 3. Evaluation of the preparation of the culture medium. To evaluate the antifungal activity of candidate and known fungi against Trichophyton mentagrophytes, a fungal mixed dilution medium containing Trichophyton mentagrophytes was prepared as an evaluation medium.

[0171] Specifically, take one bead of Trichophyton rubrum (ATCC28188) preserved in Microbank, spread it on potato dextrose agar (PDA) slant agar medium, and incubate it statically at 30°C under aerobic conditions.

[0172] After cultivation, the fungal inoculum was prepared by adding phosphate-buffered saline (PBS) containing 0.05% polysorbate 80 to PDA slant agar medium.

[0173] PDA was dissolved in ultrapure water to achieve a concentration of 39.0 g / L. The resulting solution was heated in an autoclave (121°C, 20 minutes). After air cooling, the fungal inoculum was mixed and diluted in the PDA solution, and then dispensed into petri dishes for solidification, thus preparing a fungal mixed dilution culture medium.

[0174] 4. Evaluation of antifungal activity against Trichophyton mentagrophytes The anti-tinea pedis activity of candidate strains (91 strains) and known strains (32 strains) was evaluated using fungal mixed dilution medium as the evaluation medium.

[0175] Specifically, take one bead containing candidate or known bacteria stored in a microbank, spread it on Anero Columbia RS blood agar medium, and incubate it under aerobic conditions at 35°C for 1-2 days. Collect the obtained bacteria using a platinum ring.

[0176] Platinum rings with collected bacteria attached were streaked onto a fungal mixed dilution medium and incubated aerobically at 30°C until turbidity in the agar medium caused by the proliferation of Trichophyton mentagrophytes was visually confirmed. After incubation, the proliferation-inhibiting zone formed in the fungal mixed dilution medium was observed. Based on the size of the proliferation-inhibiting zone, the anti-Trichophyton mentagrophyte activity of candidate or known bacteria was evaluated according to the following criteria. It should be noted that typical examples of each evaluation are shown below. Figure 1 middle.

[0177] ++: A clearly defined region of inhibited proliferation was observed. +: No clearly defined areas of proliferation inhibition were observed, but inhibition of Trichophyton mentagrophytes proliferation was observed. - No inhibition of Trichophyton mentagrophyte proliferation was observed. First, the results obtained from the known bacteria (32 strains) are shown in Table 4 below.

[0178] [Table 4] serial number strains strain result 1 Acinetobacter baumannii ATCC17978 - 2 Blue-gray corynebacterium JCM12208 - 3 Corynebacterium jejuni ATCC43734 - 4 Corynebacterium banding NBRC15291 - 5 Enterococcus faecalis ATCC29212 ++ 6 E. coli ATCC25922 ++ 7 Micrococcus luteus ATCC4698 - 8 Atlanta Moraxella NBRC14588 - 9 Neisseria gonorrhoeae ATCC19424 - 10 Pseudomonas aeruginosa ATCC27853 ++ 11 Staphylococcus aureus ATCC29213 - 12 Staphylococcus epidermidis ATCC12228 + 13 hemolytic staphylococci ATCC29970 + 14 Staphylococcus aureus ATCC27844 ++ 15 agalactococcus ATCC13813 - 16 Streptococcus suis JCM12971 - 17 Streptococcus pyogenes ATCC12344 - 18 Staphylococcus aureus ATCC12600 + 19 Staphylococcus epidermidis ATCC14990 + 20 Staphylococcus aureus ATCC27840 + 21 Staphylococcus aureus ATCC35538 + 22 Staphylococcus coli ATCC29974 - 23 hemolytic staphylococci ATCC700564 ++ 24 hemolytic staphylococci ATCC29969 ++ 25 Staphylococcus aureus ATCC27845 ++ 26 Staphylococcus aureus ATCC700236 ++ 27 Staphylococcus krill ATCC43959 + 28 Staphylococcus aureus ATCC700403 - 29 Staphylococcus ludenum ATCC49576 + 30 Staphylococcus ludenum ATCC43809 + 31 Staphylococcus aureus ATCC27836 ++ 32 Staphylococcus saprophyticus ATCC15305 + As shown in Table 4, among the known bacteria (32 strains), 9 strains were "++", 10 strains were "+", and 13 strains were "-".

[0179] In addition, among the candidate bacteria (91 strains), the results were: 23 strains were "++", 32 strains were "+", and 36 strains were "-".

[0180] 5. Identification of candidate bacteria To analyze the relationship between the antifungal activity of candidate bacteria and the species of candidate bacteria, candidate bacteria were identified. Identification of candidate bacteria was performed by comparing 16S rDNA sequences.

[0181] Specifically, following the instructions for NucleoSpin Microbial DNA, total DNA was extracted and purified from candidate bacteria cultured in Anero Columbia RS blood agar medium, and the DNA concentration and purity were determined using NanoDrop as an ultra-micro spectrophotometer.

[0182] Next, following the instructions of the Bacterial 16S rDNA PCR Kit, using purified total DNA as a template, the 16S rDNA sequence region was amplified by PCR using the included primers F1 (SEQ ID NO: 19) and R2 (SEQ ID NO: 20). The amplified DNA fragment (approximately 1,500 bp) was purified using a NucleoSpin gel and a PCR cleaning kit, and the DNA concentration and purity were determined using a NanoDrop spectrophotometer. Agarose gel electrophoresis confirmed that the amplified DNA fragment size was as expected.

[0183] The amplified DNA fragment (16S rDNA template sample) was mixed with the primers (F1 (serial number 19), F2 (serial number 21), or R2 (serial number 20)) provided with the Bacterial 16S rDNA PCR Kit, and the target sequence was analyzed using Sanger sequencing. After assembling the obtained sequencing sequence, the candidate bacterial species were identified by performing an NCBI BLAST search targeting the region between the common sequence 1 (AGCTTGC) and common sequence 2 (AAGCTGG) shared by Staphylococcus spp. At this point, the target region of the 16S rDNA (approximately 1,240–1,280 bp) is part of the amplified DNA fragment (approximately 1,500 bp), containing common sequence 1 (AGCTTGC) and common sequence 2 (AAGCTGG). In addition, in the obtained sequencing sequences, in the absence of common sequence 1 (AGCTTGC), the sequences were replaced with GGTTGG and TGCTTGC in that order. In the absence of common sequence 2 (AAGCTGG), the sequences were replaced with GAGTTGG, AAGTGCGG, GAAGTCGA, AAGTCAG, GAGTGG, AAGGGGGG, AAGCTGG, CAGTCAG, and CCGCTGG in that order. The candidate bacterial species were identified by performing NCBI BLAST search targeting the base sequence (approximately 1,240–1,280 bp) of the regions sandwiched by each common sequence. It should be noted that, in cases where the obtained sequencing sequence does not contain any or both of the following: common sequence 1 (AGCTTGC) or its alternative sequences (GGTTGG, TGCTTGC) and common sequence 2 (AAGCTGG) or its alternative sequences (GAAGTTGG, AAGTGCGG, GAAGTCGA, AAGTCAG, GAGTGG, AAGGGGGG, AAGTCAG, CAGTCAG, CCGCTGG), the candidate bacterial species are identified by performing a BLAST search on NCBI targeting the full-length base sequence of the obtained sequence (the amplified DNA fragment (approximately 1,500 bp)).

[0184] It should be noted that when identifying candidate bacterial species, species showing more than 99% identity with the top 10 sequences found in the BLAST search were selected. In cases where the selected top 10 species included multiple results from the same genus but different species, identification by genus name was used (Staphylococcus species and Micrococcus species...). Micrococcus sp If no suitable bacterial species were identified, the result is considered unknown.

[0185] The results of the identification of candidate bacteria and antifungal activity against Trichophyton mentagrophytes are shown in Tables 5 and 6 below.

[0186] Tables 5 and 6 present the identified candidate bacteria organized by species, and the results obtained from analyzing the proportion of antifungal activity shown by each species are displayed. Figure 2 In the middle. By Figure 2 The results showed that Staphylococcus aureus and hemolytic Staphylococcus aureus accounted for 100% and 86% respectively, showing a "++" result.

[0187] In addition, the results of the anti-Trichophyton mentagrophyte activity of 52 candidate bacteria obtained from the ankle and 39 candidate bacteria obtained from the sole, based on the isolation site, are shown in Table 7 below.

[0188] [Table 7] According to the results in Table 7, among the 52 strains on the ankle, 33% were "++", 31% were "+", and 37% were "-". Among the 39 strains on the sole, 15% were "++", 41% were "+", and 44% were "-". It can be seen that the proportion of strains showing "++" on the ankle is approximately twice that on the sole.

[0189] 6. Construction of a phylogenetic tree for the genus Staphylococcus. Referring to the phylogenetic tree of 27 Staphylococcus species based on the 16S rDNA sequences of 27 Staphylococcus species in the paper by Ghebremedhin B et al. (J Clin Microbiol, 2008; 46: 1019-1025: Genetic classification and distinguishing of Staphylococcus species based on different partial gap, 16S rRNA, hsp60, rpoB, sodA, and tuf gene sequences), a phylogenetic tree was constructed targeting the 27 known Staphylococcus strains (Table 8 below) and 45 Staphylococcus strains identified as Staphylococcus from the candidate strains (bacteria extracted from Tables 5 and 6 are shown in Table 9 below). The constructed phylogenetic tree is shown in... Figure 3 It should be noted that the CLC sequence viewer (ver. 8.0) was used in the creation of the phylogenetic tree.

[0190] [Table 8] serial number strains strain 1 Staphylococcus alright ATCC43957 2 Staphylococcus aureus ATCC12600 3 Staphylococcus aureus ATCC33753 4 Staphylococcus aureus ATCC27840 5 Staphylococcus aureus ATCC35538 6 Staphylococcus aureus () CIP103274 7 Staphylococcus chromogenicus CBCC1462 8 Staphylococcus coli GH137 9 Staphylococcus dolphinus () ATCC49171 10 Staphylococcus epidermidis ATCC14990 11 Staphylococcus aureus () ATCC43958 12 Staphylococcus feli () GD521 13 Staphylococcus aureus () VIII1 14 hemolytic staphylococci ATCC29970 15 Staphylococcus aureus ATCC27844 16 Staphylococcus suis () ATCC11249 17 Staphylococcus intermedia () H11 / 68 18 Staphylococcus krill ATCC43959 19 Staphylococcus aureus JCM2426 20 Staphylococcus ludenum ATCC43809 21 Staphylococcus aureus () MB4 22 Staphylococcus saprophyticus ATCC15305 23 Staphylococcus stearothermia DSM4807 24 Staphylococcus aureus () DSM20345 25 Staphylococcus pseudosporins MK148 26 Staphylococcus aureus ATCC27836 27 Staphylococcus xylose () JCM2418 [Table 9] Will Figure 3 The phylogenetic tree was compared with the antifungal activity results of candidate bacteria, and the results were roughly divided into cluster 1, which mainly showed "-" results; cluster 2, which mainly showed "++" results; and cluster 3, which mainly showed "+" results. Cluster 2 consisted of Staphylococcus aureus and hemolytic Staphylococcus aureus. cocci constitute.

[0191] Therefore, a phylogenetic tree was constructed for six known Staphylococcus humanis and Staphylococcus hemolyticus strains (Table 10 below) and 15 strains of bacteria identified as Staphylococcus humanis or Staphylococcus hemolyticus from the candidate strains (the bacteria extracted from Table 9 are shown in Table 11 below). The constructed phylogenetic tree for Staphylococcus humanis and Staphylococcus hemolyticus is shown in... Figure 4 and Figure 5 middle.

[0192] [Table 10] serial number strains strain 1 Staphylococcus aureus ATCC27844 2 Staphylococcus aureus ATCC27845 3 Staphylococcus aureus ATCC700236 4 Staphylococcus aureus A9 5 hemolytic staphylococci ATCC29970 6 hemolytic staphylococci ATCC700564 [Table 11] serial number strains result 2 Staphylococcus aureus ++ 3 Staphylococcus aureus ++ 37 Staphylococcus aureus ++ 40 Staphylococcus aureus ++ 45 Staphylococcus aureus ++ 62 Staphylococcus aureus ++ 64 Staphylococcus aureus ++ 74 Staphylococcus aureus ++ 20 hemolytic staphylococci ++ 22 hemolytic staphylococci ++ 23 hemolytic staphylococci ++ 24 hemolytic staphylococci ++ 25 hemolytic staphylococci ++ 33 hemolytic staphylococci ++ 35 hemolytic staphylococci + 7. 16S rDNA sequence (1) 16S rDNA sequence of Staphylococcus aureus strain The 16S rDNA sequence of Staphylococcus aureus ATCC27844, a known bacterium, was confirmed using the ATCC database. The base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was also confirmed. As a result, the 16S rDNA sequence of Staphylococcus aureus ATCC27844 contains the base sequence of sequence number 1 (genomic location of Staphylococcus aureus ATCC27844: 716544~717811). It should be noted that because sequence number 1 contains shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG), the 5' end of the sequence in sequence number 1 is shared sequence 1 (AGCTTGC), and the 3' end is shared sequence 2 (AAGCTGG).

[0193] In addition, the 16S rDNA sequences of candidate strains identified as Staphylococcus aureus strains numbered 2, 3, 37, 40, 45, 62, 64, and 74 were confirmed using the same method as described in "5. Identification of Candidate Strains" (approximately 1,240–1,280 bp) in the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG). The results showed that the 16S rDNA sequences of candidate strains numbered 3, 40, 45, 64, and 74 contained the sequence number 5; the 16S rDNA sequence of candidate strain number 2 contained the sequence number 7; the 16S rDNA sequence of candidate strain number 37 contained the sequence number 8; and the 16S rDNA sequence of candidate strain number 62 contained the sequence number 9.

[0194] Furthermore, the 16S rDNA sequences of Staphylococcus aureus ATCC27845 and ATCC700236 (both well-known strains) were confirmed using the ATCC database, and the base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. Additionally, the 16S rDNA sequence of Staphylococcus aureus A9 (ATCC commission number: PTA-125203) was confirmed using the PATRIC database, and the base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. As a result, the 16S rDNA sequence of Staphylococcus aureus ATCC27845 has the sequence number 1 (genome location of Staphylococcus aureus ATCC27845: 761822~763089), and the 16S rDNA sequences of Staphylococcus aureus ATCC700236 and Staphylococcus aureus A9 (ATCC commissioned number: PTA-125203) have the sequence number 5 (genome location of Staphylococcus aureus ATCC700236: 849942~851209, and genome location of Staphylococcus aureus A9: 185~1452).

[0195] (2) 16S rDNA sequence of hemolytic Staphylococcus strains The 16S rDNA sequence of *Staphylococcus hemolyticus* ATCC29970, a known bacterium, was confirmed using the ATCC database. The base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was also confirmed. As a result, the 16S rDNA sequence of *Staphylococcus hemolyticus* ATCC29970 has the base sequence of sequence number 10 (genomic location of *Staphylococcus hemolyticus* ATCC29970: 807302~808569).

[0196] Furthermore, the 16S rDNA sequences of candidate strains identified as hemolytic Staphylococcus aureus (strains 22-25, 33, and 35) were confirmed using the same method as described in "5. Identification of Candidate Strains," identifying the base sequence (approximately 1,240-1,280 bp) between the shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG). As a result, the 16S rDNA sequences of candidate strains 22, 24, 25, and 33 contained the base sequence at sequence number 13; the 16S rDNA sequence of candidate strain 35 contained the base sequence at sequence number 14; and the 16S rDNA sequence of candidate strain 23 contained the base sequence at sequence number 16. The 16S rDNA sequence of strain 20, identified as hemolytic Staphylococcus aureus, contained shared sequence 1 (AGCTTGC) but not shared sequence 2 (AAGCTGG). Because the 16S rDNA sequence of candidate bacterium number 20 has an alternative sequence (GAGCTGG) to the common sequence 2, the base sequence (approximately 1,267 bp) of the region sandwiched between the common sequence 1 (AGCTTGC) and the alternative sequence (GAGCTGG) to the common sequence 2 was confirmed. As a result, the 16S rDNA sequence of candidate bacterium number 20 has the base sequence of sequence number 15.

[0197] Furthermore, the 16S rDNA sequence of *Staphylococcus hemolyticus* ATCC700564, a known bacterium, was confirmed using the ATCC database. The base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was also confirmed. As a result, the 16S rDNA sequence of *Staphylococcus hemolyticus* ATCC700564 has the base sequence of sequence number 10 (genomic location of *Staphylococcus hemolyticus* ATCC700564: 214068~215335).

[0198] The results obtained in (1) and (2) above are shown in Tables 12 and 13 below.

[0199] [Table 12] strains strain Serial Number Evaluation results of antifungal activity against Trichophyton mentagrophytes Staphylococcus aureus ATCC27844 Serial Number 1 ++ Staphylococcus aureus Number 2 Serial Number 7 ++ Staphylococcus aureus Number 3 Serial Number 5 ++ Staphylococcus aureus Number 37 Serial Number 8 ++ Staphylococcus aureus Number 40 Serial Number 5 ++ Staphylococcus aureus Number 45 Serial Number 5 ++ Staphylococcus aureus Number 62 Serial Number 9 ++ Staphylococcus aureus Number 64 Serial Number 5 ++ Staphylococcus aureus Number 74 Serial Number 5 ++ Staphylococcus aureus ATCC27845 Serial Number 1 ++ Staphylococcus aureus ATCC700236 Serial Number 5 ++ Staphylococcus aureus A9 Serial Number 5 ND hemolytic staphylococci ATCC29970 Serial Number 10 + hemolytic staphylococci Number 20 Serial Number 15 ++ hemolytic staphylococci Number 22 Serial Number 13 ++ hemolytic staphylococci Number 23 Serial Number 16 ++ hemolytic staphylococci Number 24 Serial Number 13 ++ hemolytic staphylococci Number 25 Serial Number 13 ++ hemolytic staphylococci Number 33 Serial Number 13 ++ hemolytic staphylococci Number 35 Serial Number 14 + hemolytic staphylococci ATCC700564 Serial Number 10 ++ *ND: No data [Table 13] It should be noted that in Table 13, serial numbers 5 and 7-9 are based on serial number 1, and serial numbers 13-16 are based on serial number 10. Additionally, for example, "45G>A" in serial number 5 means that the G (guanine) at position 45 of serial number 1 is mutated to A (adenine). Furthermore, for example, "488delA" in serial number 7 means that the A (adenine) at position 488 of serial number 1 is deleted. Moreover, for example, "1182_1183insA" in serial number 8 means that A (adenine) is inserted between positions 1182 and 1183 of serial number 1.

[0200] (3) 16S rDNA sequence of Staphylococcus aureus strain The 16S rDNA sequence of *Staphylococcus aureus* ATCC27836, a well-known bacterium, was confirmed using the ATCC database. The base sequence of the region enclosed by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was also confirmed. As a result, the 16S rDNA sequence of *Staphylococcus aureus* ATCC27836 has sequence number 17 (genomic location of *Staphylococcus aureus* ATCC27836: 725369~726636).

[0201] As can be seen from the results of 7 (1) to (3) above, Staphylococcus aureus with 16S rDNA containing sequence number 1 or having more than 95% identity with sequence number 1, Staphylococcus hemolyticus with 16S rDNA containing sequence number 10 or having more than 95% identity with sequence number 10, and Staphylococcus variabilis with 16S rDNA containing sequence number 17 or having more than 95% identity with sequence number 17 have anti-Trichophyton mentagrophytes activity.

[0202] 8. Effects against other Trichophyton species In sections 4-7 above, a fungal mixed dilution medium containing *Trichophyton rubrum* ATCC28188 (a type of Trichophyton) was used as the evaluation medium (see "3. Preparation of Evaluation Medium" above). Here, the effects on other Trichophyton species (*Trichophyton rubrum*) were evaluated. T . rubrum The effect of ATCC28188 and other Trichophyton species.

[0203] (1) Preparation of a mixed dilution culture medium containing other Trichophyton species Except for using beads of Trichophyton rubrum ATCC22402, Trichophyton mentagrophytes TIMM2789, Trichophyton mentagrophytes ATCCMYA-4439, Trichophyton tonsurans ATCC56186, or Trichophyton tonsurans ATCC28942 instead of Trichophyton rubrum ATCC28188 beads, five mixed dilution culture media containing Trichophyton floccosum were prepared using the same method as described in "3. Preparation of Evaluation Culture Media" above.

[0204] (2) Evaluation of antifungal activity against other Trichophyton species For the anti-Trichophyton floccosum activity of well-known fungi such as Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus hemolyticus ATCC29969, Staphylococcus hemolyticus ATCC700564, and Staphylococcus variabilis ATCC27836, which have been evaluated against Trichophyton rubrum ATCC28188, the anti-Trichophyton floccosum activity was evaluated using a mixed dilution culture medium containing the above five fungi.

[0205] The evaluation method was the same as that described in "4. Evaluation of antifungal activity against Trichophyton mentagrophytes". The results obtained are presented together with the results of antifungal activity against Trichophyton rubrum ATCC28188 in Table 13 below.

[0206] As shown in Table 14, the five other Trichophyton species also exhibited the same antifungal activity as Trichophyton rubrum ATCC28188.

Claims

1. A composition for treating tinea capitis, comprising at least one bacterium selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains that has anti-tinea capitis activity. The *Staphylococcus variabilis* strain mentioned therein includes *Staphylococcus variabilis* ATCC27836.

2. The tinea pedis treatment composition according to claim 1, comprising a strain of Staphylococcus aureus.

3. The tinea pedis treatment composition according to claim 2, wherein the Staphylococcus aureus strain has 16S rDNA containing the base sequence of sequence number 1 or 16S rDNA containing a base sequence having more than 95% identity with sequence number 1, and has anti-tinea pedis activity.

4. The tinea pedis treatment composition according to claim 2, wherein the Staphylococcus aureus strain comprises at least one selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845 and Staphylococcus aureus ATCC700236.

5. The tinea pedis treatment composition according to claim 1, wherein it contains a hemolytic staphylococcal strain.

6. The tinea pedis treatment composition according to claim 5, wherein the hemolytic staphylococcal strain has 16S rDNA containing the base sequence of sequence number 10 or 16S rDNA containing a base sequence having more than 95% identity with sequence number 10, and has anti-tinea pedis activity.

7. The tinea pedis treatment composition according to claim 5, wherein the hemolytic staphylococcal strain comprises at least one selected from hemolytic staphylococcus ATCC29970, hemolytic staphylococcus ATCC700564 and hemolytic staphylococcus ATCC29969.

8. The tinea pedis treatment composition according to claim 1, wherein it contains Staphylococcus aureus strain.

9. The tinea pedis treatment composition according to any one of claims 1 to 8, wherein the pathogenic fungus of tinea pedis comprises Trichophyton spp.

10. The tinea pedis treatment composition according to claim 9, wherein the pathogenic fungus of tinea pedis comprises at least one fungal strain selected from Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

11. The composition for treating tinea capitis according to any one of claims 1 to 8, wherein it is a topical application.

12. A method for manufacturing a composition for treating tinea capitis, comprising: The process of collecting and culturing bacterial flora from at least one location on the ankle and sole of a person (1), Step (2) involves isolating and culturing the cultured bacterial flora to obtain candidate bacteria. The steps (3) for evaluating the antifungal activity of the candidate bacteria and identifying bacteria with antifungal activity, and Step (4) of preparing a tinea treatment composition containing bacteria having the anti-tinea fungal activity; The bacteria possessing the aforementioned anti-Trichophyton floccosum activity include at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii. The *Staphylococcus variabilis* strain mentioned therein includes *Staphylococcus variabilis* ATCC27836.

13. Use of at least one bacterium with anti-tinea capitis activity, selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains, in the manufacture of tinea capitis treatment drugs. The *Staphylococcus variabilis* strain mentioned therein includes *Staphylococcus variabilis* ATCC27836.

14. The use according to claim 13, wherein it comprises a human staphylococcal strain.

15. The use according to claim 14, wherein the Staphylococcus aureus strain has 16S rDNA containing the base sequence of sequence number 1 or 16S rDNA containing a base sequence having more than 95% identity with sequence number 1, and has anti-tinea capitis activity.

16. The use according to claim 14, wherein the Staphylococcus aureus strain comprises at least one selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845 and Staphylococcus aureus ATCC700236.

17. The use according to claim 13, wherein it contains a hemolytic staphylococcal strain.

18. The use according to claim 17, wherein the hemolytic staphylococcal strain has a 16S rDNA containing the base sequence of sequence number 10 or a 16S rDNA containing a base sequence having more than 95% identity with sequence number 10, and has anti-tinea capitis activity.

19. The use according to claim 17, wherein the hemolytic staphylococcal strain comprises at least one selected from hemolytic staphylococcal ATCC29970, hemolytic staphylococcal ATCC700564 and hemolytic staphylococcal ATCC29969.

20. The use according to claim 13, wherein it contains a strain of Staphylococcus aureus.

21. The use according to any one of claims 13 to 20, wherein the pathogenic fungus of tinea capitis comprises Trichophyton spp.

22. The use according to claim 21, wherein the pathogenic fungus of tinea corporis comprises at least one fungal strain selected from Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

23. The use according to any one of claims 13 to 20, wherein it is a topical agent.

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