Composition for treating tinea

Abstract

Provided is a novel composition for treating ringworm.　This composition for treating ringworm contains at least one selected from the group consisting of a bacterial strain of Staphylococcus hominis, a bacterial strain of Staphylococcus haemolyticus, and a bacterial strain of Staphylococcus warneri.

Description

[Technical Field]

[0001] The present invention relates to a composition for treating tinea. [Background technology]

[0002] Tinea is a skin infection caused by the fungus Trichophyton (also known as tinea fungus). Depending on the site of onset, tinea is classified into tinea pedis (athlete's foot), tinea unguium (nail athlete's foot), tinea manubriata (athlete's foot), tinea corporis (ringworm), tinea cruris (jock itch), and tinea capitis (tinea capitis).

[0003] Tinea is transmitted by the Trichophyton fungus that tinea patients carry. For example, it is known that the main route of infection for the common conditions tinea pedis (athlete's foot) and tinea unguium (tinea unguium) is Trichophyton that is present in swimming pools, public baths, restaurants, slippers, etc.

[0004] Tinea fungus is a non-resident fungus that parasitizes the stratum corneum and subcutaneous tissue of the skin. When it multiplies and invades the stratum corneum, inflammation and blisters occur, causing symptoms such as itching. Tinea fungus feeds on keratin, the main component of keratin.

[0005] Antifungal drugs such as itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, luliconazole, efinaconazole, fosravuconazole, terbinafine, and butenafine have been used to treat tinea.

[0006] For example, Non-Patent Document 1 describes that prepubertal children frequently develop tinea corporis (ringworm) and tinea capitis (tinea capitis), while adolescents and adults frequently develop tinea corporis (ringworm), tinea cruris (jock itch), and tinea pedis (athlete's foot); that because there are diseases that show similar clinical symptoms, it may be preferable to diagnose using potassium hydroxide preparations or culture; and that creams or oral medications should be selected as appropriate for treatment.

[0007] Furthermore, Patent Document 1 describes an invention relating to an external preparation for treating onychomycosis, which contains a) an antifungal active substance, b) a volatile component, c) a medium-chain fatty acid triglyceride, and d) ethyl lactate. According to the external preparation described in Patent Document 1, by using a medium-chain fatty acid triglyceride as a non-volatile component and ethyl lactate as a permeation enhancer, it is described that the preparation has excellent properties in terms of nail permeability, efficacy, and formulation properties.

[0008] Patent Document 1 describes that topical preparations used to treat onychomycosis are superior to oral preparations in that they have fewer systemic side effects and fewer drug-drug interactions. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] International Publication No. 2019 / 088005 [Non-patent literature]

[0010] [Non-Patent Document 1] American Family Physician, Volume 90, Number 10, 702-711. Summary of the Invention [Problem to be solved by the invention]

[0011] In this situation, further development of drugs for treating tinea is underway. [Means for solving the problem]

[0012] The present invention provides, for example, the following compositions for treating tinea.

[0013] [1] A composition for treating ringworm, comprising at least one selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri. [2] The composition for treating tinea described in [1] above, which contains a strain of Staphylococcus hominis. [3] A composition for treating tinea described in [2] above, wherein the Staphylococcus hominis strain has a 16s rDNA containing the base sequence of SEQ ID NO: 1 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 1, and has anti-tinea activity. [4] The composition for treating tinea according to [2] or [3] above, wherein the Staphylococcus hominis strain comprises at least one selected from the group consisting of Staphylococcus hominis ATCC27844, Staphylococcus hominis ATCC27845, Staphylococcus hominis ATCC700236, and Staphylococcus hominis A9 (ATCC accession number: PTA-125203). [5] A composition for treating tinea according to any one of [1] to [4] above, which contains a strain of Staphylococcus haemolyticus. [6] A composition for treating tinea described in [5] above, wherein the Staphylococcus haemolyticus strain has a 16s rDNA containing the base sequence of SEQ ID NO: 10 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 10, and has anti-tinea activity. [7] The composition for treating tinea described in [5] or [6] above, wherein the Staphylococcus haemolyticus strain comprises at least one selected from the group consisting of Staphylococcus haemolyticus ATCC29970, Staphylococcus haemolyticus ATCC700564, and Staphylococcus haemolyticus ATCC29969. [8] A composition for treating tinea according to any one of [1] to [7] above, which contains a strain of Staphylococcus warneri. [9] A composition for treating tinea described in [8] above, wherein the Staphylococcus warneri strain has a 16s rDNA containing the base sequence of SEQ ID NO: 17 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 17, and has anti-tinea activity.

[10] The composition for treating tinea described in [8] or [9] above, wherein the Staphylococcus warneri strain includes Staphylococcus warneri ATCC27836.

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

[10] above, wherein the causative fungus of tinea comprises a fungus of the genus Trichophyton.

[12] The composition for treating tinea described in

[11] above, wherein the causative fungus of tinea comprises at least one fungus selected from the group consisting of a fungal strain of Trichophyton rubrum, a fungal strain of Trichophyton mentagrophytes, and a fungal strain of Trichophyton tonsurans.

[13] The composition for treating tinea according to any one of [1] to

[12] above, which is an external preparation.

[14] A step (1) of collecting and culturing a bacterial flora from at least one of a human ankle and a sole; (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; (3) evaluating the anti-trichophyton activity of the candidate fungus to identify a bacterium having anti-trichophyton activity; (4) a step of preparing a composition for treating tinea, which comprises the bacterium having anti-tinea fungal activity; Including, A method for producing a composition for treating tinea, wherein the bacterium having anti-tinea fungal activity comprises at least one selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri. [Effects of the Invention]

[0014] The present invention provides a novel composition for treating tinea. In a preferred embodiment, the composition can be used in cases where artificial chemical synthesis and extraction of a medicinal compound are difficult or where the medicinal effect is based on the vital reaction of live bacteria, and can have one or more of the following effects: prevention or reduction of side effects, prevention or suppression of skin rash around the affected area, high skin adhesion, maintenance of remission, prevention of recurrence, reduction in the number of administrations, improvement of compliance, and high therapeutic effect. [Brief explanation of the drawings]

[0015] [Figure 1] These are typical examples of evaluation of anti-dermatophyte activity. [Figure 2] FIG. 1 shows the results of the anti-dermatophyte activity of each species of candidate fungi (91 strains) obtained from the ankles and soles. [Figure 3] Phylogenetic tree of known and candidate strains of the genus Staphylococcus. [Figure 4] Phylogenetic tree of known and candidate strains of Staphylococcus hominis. [Figure 5] Phylogenetic tree of known and candidate strains of Staphylococcus haemolyticus. DETAILED DESCRIPTION OF THE INVENTION

[0016] The present invention will be described in detail below.

[0017] <Composition for treating tinea> The composition for treating tinea according to the present invention contains at least one strain selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri. The composition for treating tinea according to the present invention can treat tinea. In this specification, "treatment" means preventing or inhibiting the growth of Trichophyton fungi (so-called tinea fungi), killing them, or stabilizing them.

[0018] It is known that the onset and recurrence of tinea varies from person to person. The present inventors predicted that this individual difference is due to the resident skin flora, and investigated the effects of resident skin flora collected from the ankles and soles of healthy individuals and known strains of Staphylococcus on tinea fungi. As a result, they found that strains of Staphylococcus hominis (hereinafter also referred to as "S. hominis"), Staphylococcus haemolyticus (hereinafter also referred to as "S. haemolyticus"), and Staphylococcus warneri (hereinafter also referred to as "S. warneri"), which are resident skin flora, are effective in treating tinea. According to the present invention, a novel composition for treating tinea can be provided.

[0019] Furthermore, because S. hominis, S. haemolyticus, and S. warneri strains are normal skin flora, they have few or no side effects, and therefore may be effective in preventing or suppressing skin rash around the affected area that can occur with conventional topical preparations.

[0020] Furthermore, because S. hominis, S. haemolyticus, and S. warneri strains are naturally occurring bacteria on the skin, they tend to colonize the skin, which can result in longer-lasting effects on affected areas of the skin and reduced administration frequency compared to conventional topical preparations.

[0021] Furthermore, because S. hominis strains, S. haemolyticus strains, and S. warneri strains are live bacteria that are normally present on the skin, they may have therapeutic effects even when it is difficult to artificially chemically synthesize or extract medicinal compounds, or when the medicinal effects are based on vital reactions derived from live bacteria.

[0022] Therefore, the composition for treating tinea according to the present invention can have advantages such as fewer side effects, better compliance, and higher efficacy compared to conventional topical preparations.

[0023] [Staphylococcus hominis strain] In one embodiment, the composition for treating ringworm comprises a strain of Staphylococcus hominis.

[0024] Staphylococcus hominis strains are coagulase-negative staphylococci (CNS) and gram-positive bacteria. S. hominis strains are normal skin flora. It is presumed that S. hominis strains prevent or inhibit the growth and establishment of tinea fungi, kill tinea fungi, or act as a bacteriostat by directly or indirectly interfering with them or competing for habitat.

[0025] The S. hominis strain preferably includes an S. hominis strain having anti-trichophyton activity. That is, in a preferred embodiment, a composition for treating tinea is provided that includes an S. hominis strain having anti-trichophyton activity. In this specification, "antitrinobacterial activity" means that when the anti-trichophyton activity is evaluated by the method described in "4. Evaluation of anti-trichophyton activity" in the Examples, the evaluation result is "++" or "+", preferably "++". Therefore, S. hominis strains having anti-trichophyton activity can be used to treat tinea.

[0026] Although there are no particular limitations on the S. hominis strain, it is preferable that the strain has a 16s rDNA containing the nucleotide sequence of SEQ ID NO: 1 or a 16s rDNA containing a nucleotide sequence with 95% or more identity to SEQ ID NO: 1 and has anti-tinea activity. As shown in the Examples, SEQ ID NO: 1 is the nucleotide sequence of the region flanked by two consensus sequences shared by 16s rDNAs of the genus Staphylococcus. Of the two consensus sequences, the consensus sequence on the 5' side is consensus sequence 1 (AGCTTGC), and the consensus sequence on the 3' side is consensus sequence 2 (AAGCTGG), both of which are included in SEQ ID NO: 1. Here, for example, "16s rDNA containing the nucleotide sequence of SEQ ID NO: 1" means that the nucleotide sequence of SEQ ID NO: 1 is contained in the 16s rDNA. In this specification, the nucleotide sequences of SEQ ID NOs: 1 to 18 are identified by the method described in the Examples. Furthermore, if a bacterium has two or more types of 16s rDNA, the most abundant 16s rDNA is designated as the 16s rDNA of that bacterium. If two or more types of 16s rDNA are present in equal numbers, the 16s rDNA with the most identical base sequence (SEQ ID NO: 1 for S. hominis strains, SEQ ID NO: 10 for S. haemolyticus strains, and SEQ ID NO: 17 for S. warneri strains) is designated as the 16s rDNA of that bacterium.

[0027] It is preferred that a base sequence having 95% or more identity to SEQ ID NO: 1 has 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 identity to SEQ ID NO: 1.

[0028] Mutations in a nucleotide sequence having 95% or more identity to SEQ ID NO: 1 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.

[0029] The substitution is not particularly limited, but is preferably at least one substitution selected from the group consisting of guanine at position 45, thymine at position 139, thymine at position 140, adenine at position 309, and cytosine at position 1265 of SEQ ID NO: 1. In one embodiment, guanine at position 45 of SEQ ID NO: 1 can be substituted with adenine (45G>A (also written as G45A)), thymine at position 139 with cytosine (139T>C), thymine at position 140 with cytosine (140T>C), adenine at position 309 with cytosine (309A>C), and cytosine at position 1265 with thymine (1265C>T).

[0030] The deletion is not particularly limited, but may be a deletion of adenine at position 488 of SEQ ID NO: 1 (488delA (also written as A488Δ)).

[0031] The insertion is not particularly limited, and may be between positions 1182 and 1183 and between positions 1235 and 1236 of SEQ ID NO: 1. In one embodiment, an adenine (1182_1183insA (also denoted as Δ1183A)) may be inserted between positions 1182 and 1183 of SEQ ID NO: 1, and a guanine (1235_1236insG) may be inserted between positions 1235 and 1236.

[0032] Furthermore, the base sequence having 95% or more identity to SEQ ID NO: 1 may be one possessed by a S. hominis strain as a result of natural mutation or the like, or may be one obtained by artificial mutation treatment. Examples of artificial mutation treatment include methods such as genetic engineering, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments may be used alone or in combination of two or more types.

[0033] Examples of base sequences having 95% or more identity to SEQ ID NO: 1 include SEQ ID NOs: 2 to 9. The mutation sites in SEQ ID NOs: 2 to 9 are shown in Table 1 below.

[0034] [Table 1]

[0035] Examples of S. hominis strains having 16s rDNA containing the nucleotide sequence of SEQ ID NO: 1 include, but are not limited to, S. hominis ATCC27844, S. hominis ATCC27845, and the like.

[0036] Examples of S. hominis strains that have 16s rDNA containing a base sequence that is 95% or more identical to SEQ ID NO: 1 and have anti-tinea activity include, but are not limited to, S. hominis ATCC700236 and S. hominis A9 (ATCC accession number: PTA-125203).

[0037] In one embodiment, examples of S. hominis strains include, but are not limited to, Staphylococcus hominis ATCC27844, Staphylococcus hominis ATCC27845, Staphylococcus hominis ATCC700236, Staphylococcus hominis FDAARGOS_748, Staphylococcus hominis J6, Staphylococcus hominis FDAARGOS_136, Staphylococcus hominis A9 (ATCC Accession No. PTA-125203), Staphylococcus hominis C2, Staphylococcus hominis AMT2, Staphylococcus hominis AMT3, Staphylococcus hominis AMT4-C2, Staphylococcus hominis AMT4-G1, and Staphylococcus hominis. AMT4-D12, etc. In addition to the above, strains belonging to S. hominis are listed in "NCBI," [online], archived on February 17, 2022, [searched on February 17, 2022], and on the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2014 / > Those described in can be used. Among these, the strain belonging to S. hominis preferably includes at least one selected from the group consisting of S. hominis ATCC27844, S. hominis ATCC27845, S. hominis ATCC700236, S. hominis FDAARGOS_748, S. hominis J6, S. hominis FDAARGOS_136, and S. hominis A9, and more preferably includes at least one selected from the group consisting of S. hominis ATCC27844, S. hominis ATCC27845, S. hominis ATCC700236, and S. hominis A9. The above-mentioned strains belonging to S. hominis may be used alone or in combination of two or more.

[0038] S. hominis strains with anti-trichophyton activity can be easily prepared by the following method. Specifically, the method for preparing S. hominis strains with anti-trichophyton activity includes the steps of (1) collecting and culturing bacterial flora from at least one of the ankle and sole of a human (preferably a healthy individual without tinea symptoms), (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria, and (3) evaluating the anti-trichophyton activity of the candidate bacteria to identify bacteria with anti-trichophyton activity. Each step is described below. The bacterial flora of the human ankle and sole usually includes strains of S. hominis, which are known as normal skin flora. According to the above method, S. hominis strains with anti-trichophyton activity can be easily prepared.

[0039] The concentration of S. hominis strain in the composition for treating ringworm is 10 3 ~10 11 CFU / g is preferred, and 10 4 ~10 10 CFU / g is more preferred, and 10 5 ~10 9 It is more preferably CFU / g. In this specification, "CFU" means colony forming unit. In this specification, the concentration (CFU / g) of the bacterial strain in the composition for treating tinea is calculated by diluting the composition for treating tinea to an appropriate concentration, smearing it on an agar medium for bacterial culture, and counting the colonies that appear after culture (see Clinical Microbiology Procedures Handbook volume 2 THIRD EDITION).

[0040] The form of the S. hominis strain in the composition for treating tinea is not particularly limited, but is preferably in the form of a dry powder (for example, freeze-dried or spray-dried).

[0041] [Staphylococcus haemolyticus strains] In one embodiment, the composition for treating ringworm comprises a strain of Staphylococcus haemolyticus.

[0042] Staphylococcus haemolyticus strains are coagulase-negative staphylococci (CNS) and gram-positive bacteria. S. haemolyticus strains are normal skin flora. It is presumed that S. haemolyticus strains also prevent or inhibit the growth of tinea fungi, kill tinea fungi, or act as bacteriostatic agents by directly or indirectly interfering with the fungus or competing for habitat.

[0043] The S. haemolyticus strain preferably includes an S. haemolyticus strain having anti-trichophytic activity. That is, in one preferred embodiment, a composition for treating tinea is provided, which includes an S. haemolyticus strain having anti-trichophytic activity. The S. haemolyticus strain having anti-trichophytic activity can treat tinea.

[0044] The S. haemolyticus strain is not particularly limited, but is preferably one having a 16s rDNA containing the nucleotide sequence of SEQ ID NO: 10 or a 16s rDNA containing a nucleotide sequence with 95% or more identity to SEQ ID NO: 10 and having anti-tinea activity. As shown in the Examples, SEQ ID NO: 10 is the nucleotide sequence of the region flanked by two consensus sequences shared by 16s rDNAs of the genus Staphylococcus. Of the two consensus sequences, the consensus sequence on the 5' side is consensus sequence 1 (AGCTTGC) and the consensus sequence on the 3' side is consensus sequence 2 (AAGCTGG), both of which are included in SEQ ID NO: 10. Here, for example, "16s rDNA containing the nucleotide sequence of SEQ ID NO: 10" means that the nucleotide sequence of SEQ ID NO: 10 is contained in the nucleotide sequence of the 16s rDNA.

[0045] It is preferred that a nucleotide sequence having 95% or more identity to SEQ ID NO: 10 has 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 identity to SEQ ID NO: 10.

[0046] Mutations in a nucleotide sequence having 95% or more identity to SEQ ID NO: 10 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.

[0047] The substitution is not particularly limited, but is preferably at least one selected from the group consisting of cytosine at position 96, guanine at position 109, adenine at position 398, cytosine at position 952, and cytosine at position 1186 of SEQ ID NO: 10. In one embodiment, the cytosine at position 96 of SEQ ID NO: 10 can be substituted with thymine (96C>T), the guanine at position 109 with adenine (109G>A), the adenine at position 398 with guanine (398A>G), the cytosine at position 952 with thymine (952C>T), and the cytosine at position 1186 with thymine (1186C>T).

[0048] The deletion is not particularly limited, but may include deletion of the adenine at position 1210 of SEQ ID NO: 10 (1210delA), the adenine at position 1219 (1219delA), the thymine at position 1225 (1225delT), or the adenine at position 1263 (1263delA).

[0049] The insertion is not particularly limited, and may be between positions 1117 and 1118, between positions 1195 and 1196, or between positions 1234 and 1235 of SEQ ID NO: 10. In one embodiment, a guanine (1117_1118insG) may be inserted between positions 1117 and 1118 of SEQ ID NO: 10, a cytosine (1195_1196insC) may be inserted between positions 1195 and 1196, or a guanine (1234_1235insG) may be inserted between positions 1234 and 1235.

[0050] The base sequence having 95% or more identity to SEQ ID NO: 10 may be one that is possessed by other S. haemolyticus strains as a result of natural mutation or the like, as with S. hominis strains, or may be one that is obtained by artificial mutation treatment. Examples of artificial mutation treatment include genetic engineering, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments may be used alone or in combination of two or more types.

[0051] Examples of base sequences having 95% or more identity with SEQ ID NO: 10 include SEQ ID NOs: 11 to 16. The mutation sites in SEQ ID NOs: 11 to 16 are shown in Table 2 below.

[0052] [Table 2]

[0053] Examples of S. haemolyticus strains having 16s rDNA containing the nucleotide sequence of SEQ ID NO: 10 include, but are not limited to, S. haemolyticus ATCC29970, S. haemolyticus ATCC700564, and the like.

[0054] In one embodiment, the S. haemolyticus strain is not particularly limited, but examples include Staphylococcus haemolyticus ATCC29970, Staphylococcus haemolyticus ATCC700564, Staphylococcus haemolyticus ATCC29969, Staphylococcus haemolyticus SH_12, and Staphylococcus haemolyticus JCSC1435. In addition to the above, other S. haemolyticus strains include those listed in "NCBI," [online], archived February 17, 2022, [searched February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1141 / > can be used. Of these, the S. haemolyticus strain preferably includes at least one selected from the group consisting of S. haemolyticus ATCC29970, S. haemolyticus ATCC700564, S. haemolyticus ATCC29969, S. haemolyticus SH_12, and S. haemolyticus JCSC1435, and more preferably includes at least one selected from the group consisting of Staphylococcus haemolyticus ATCC29970, Staphylococcus haemolyticus ATCC700564, and Staphylococcus haemolyticus ATCC29969. The above-mentioned S. haemolyticus strains may be used alone or in combination of two or more.

[0055] Like S. hominis strains, S. haemolyticus strains with anti-trichophytic activity can be easily prepared by the following method. Specifically, the method for preparing S. haemolyticus strains with anti-trichophytic activity includes the steps of (1) collecting and culturing bacterial flora from at least one of the ankle and sole of a human (preferably a healthy individual without tinea symptoms), (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria, and (3) evaluating the anti-trichophytic activity of the candidate bacteria to identify bacteria with anti-trichophytic activity. S. haemolyticus strains with anti-trichophytic activity can also be contained in the bacterial flora of the human ankle and sole. Therefore, according to the above method, S. haemolyticus strains with anti-trichophytic activity can be easily prepared.

[0056] The concentration of the S. haemolyticus strain in the composition for treating ringworm is 10 3 ~10 11 CFU / g is preferred, and 10 4 ~10 10 CFU / g is more preferred, and 10 5 ~10 9 More preferably, it is CFU / g.

[0057] The form of the S. haemolyticus strain in the composition for treating tinea is not particularly limited, but is preferably in the form of a dry powder (for example, freeze-dried or spray-dried).

[0058] [Staphylococcus warneri] In one embodiment, the composition for treating ringworm comprises a strain of Staphylococcus warneri.

[0059] Staphylococcus warneri strains are coagulase-negative staphylococci (CNS) and gram-positive bacteria. S. warneri strains are normal skin flora. It is presumed that S. warneri strains also prevent or inhibit the growth of tinea fungi, kill tinea fungi, or act as bacteriostatic agents by directly or indirectly interfering with the fungus or competing for habitat.

[0060] The S. warneri strain preferably includes an S. warneri strain having anti-trichophytic activity. That is, in one preferred embodiment, a composition for treating tinea is provided, which includes an S. warneri strain having anti-trichophytic activity. The S. warneri strain having anti-trichophytic activity can treat tinea.

[0061] The S. warneri strain is not particularly limited, but is preferably one that has a 16s rDNA containing the nucleotide sequence of SEQ ID NO: 17 or a 16s rDNA containing a nucleotide sequence with 95% or more identity to SEQ ID NO: 17 and has anti-tinea activity. As shown in the Examples, SEQ ID NO: 17 is the nucleotide sequence of the region flanked by two consensus sequences shared by 16s rDNAs of the genus Staphylococcus. Of the two consensus sequences, the consensus sequence on the 5' side is consensus sequence 1 (AGCTTGC), and the consensus sequence on the 3' side is consensus sequence 2 (AAGCTGG), both of which are included in SEQ ID NO: 17. Here, for example, "16s rDNA containing the nucleotide sequence of SEQ ID NO: 17" means that the nucleotide sequence of SEQ ID NO: 17 is contained in the nucleotide sequence of the 16s rDNA.

[0062] It is preferred that a nucleotide sequence having 95% or more identity to SEQ ID NO: 17 has 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 identity to SEQ ID NO: 17.

[0063] Mutations in a nucleotide sequence having 95% or more identity with SEQ ID NO: 17 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.

[0064] The substitution is not particularly limited, but is preferably a cytosine at position 52 of SEQ ID NO: 17. In one embodiment, the cytosine at position 52 of SEQ ID NO: 17 may be substituted with a guanine (52C>G).

[0065] The base sequence having 95% or more identity to SEQ ID NO: 17 may be one that is possessed by other S. warneri strains as a result of natural mutation or the like, as with S. hominis strains, or may be one that is obtained by artificial mutation treatment. Examples of artificial mutation treatment include genetic engineering, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, and nitrous acid treatment. These mutation treatments may be used alone or in combination of two or more types.

[0066] An example of a base sequence having 95% or more identity with SEQ ID NO: 17 is SEQ ID NO: 18. The mutation sites in SEQ ID NO: 18 are shown in Table 3 below.

[0067] [Table 3]

[0068] Examples of S. warneri strains having 16s rDNA containing the nucleotide sequence of SEQ ID NO: 17 include, but are not limited to, S. warneri ATCC27836.

[0069] In one embodiment, the S. warneri strain is not particularly limited, but includes S. warneri ATCC27836, S. warneri NCTC7291, S. warneri WB224, S. warneri 16A, S. warneri 22.1, etc. In addition to the above, S. warneri strains are also available from "NCBI," [online], archived February 17, 2022, [retrieved February 17, 2022], Internet<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2073 / > can be used. Of these, the S. warneri strain preferably includes at least one selected from the group consisting of Staphylococcus warneri ATCC27836, S. warneri NCTC7291, S. warneri WB224, S. warneri 16A, S. warneri 22.1, and S. warneri WS479, and more preferably includes S. warneri ATCC27836. The above-mentioned S. warneri strains may be used alone or in combination of two or more.

[0070] Like S. hominis strains, S. warneri strains with anti-trichophytic activity can be easily prepared by the following method. Specifically, the method for preparing S. warneri strains with anti-trichophytic activity includes the steps of (1) collecting and culturing bacterial flora from at least one of the ankle and sole of a human (preferably a healthy individual without tinea symptoms), (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria, and (3) evaluating the anti-trichophytic activity of the candidate bacteria to identify bacteria with anti-trichophytic activity. S. warneri strains with anti-trichophytic activity can also be contained in the bacterial flora of the human ankle and sole. Therefore, according to the above method, S. warneri strains with anti-trichophytic activity can be easily prepared.

[0071] The concentration of the S. warneri strain in the composition for treating ringworm is 10 3 ~10 11 CFU / g is preferred, and 10 4 ~10 10 CFU / g is more preferred, and 10 5 ~10 9 More preferably, it is CFU / g.

[0072] The form of the S. warneri strain in the composition for treating tinea is not particularly limited, but is preferably in the form of a dry powder (for example, freeze-dried or spray-dried).

[0073] [Other Staphylococcus strains] The composition for treating ringworm may further contain other strains of the genus Staphylococcus, where "other strains of the genus Staphylococcus" means strains of the genus Staphylococcus other than S. hominis, S. haemolyticus, and S. warneri.

[0074] Other strains of the genus Staphylococcus include, but are not limited to, bacterial species that have been reported to be isolated from humans, such as strains of Staphylococcus saprophyticus, Staphylococcus kloosii, Staphylococcus lugdunensis, Staphylococcus capitis, Staphylococcus caprae, Staphylococcus epidermidis (Staphylococcus epidermidis), and Staphylococcus aureus (Staphylococcus aureus).

[0075] S. saprophyticus strains are available from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1350 / > Examples include those described in. As for S. kloosii strains, "NCBI," [online], archived February 17, 2022, [retrieved February 17, 2022], Internet<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 66884 / > Examples include those described in. S. lugdunensis strains are available from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2548 / > Examples include those described in. S. capitis strains are available from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2054 / > Examples include those described in. S. caprae strains include those from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1971 / > Examples include those described in. As for strains of S. epidermidis (Staphylococcus epidermidis), "NCBI," [online], archived February 17, 2022, [retrieved February 17, 2022], Internet<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 155 / > Examples include those described in. As for S. aureus strains, "NCBI," [online], archived on February 17, 2022, [searched on February 17, 2022], Internet<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 154 / > Examples include those described in.

[0076] From the viewpoint of achieving a higher therapeutic effect against tinea, the other Staphylococcus strains preferably include at least one selected from the group consisting of S. capitis, S. caprae, S. epidermidis (Staphylococcus epidermidis), Staphylococcus kloosii, Staphylococcus lugdunensis, and Staphylococcus saprophyticus, more preferably at least one selected from the group consisting of S. capitis and S. caprae strains, and particularly preferably at least one of S. capitis ATCC27840 and S. caprae ATCC35538. Furthermore, from the viewpoint of adjusting the balance of normal skin flora at the site to which the composition for treating tinea is applied, the other strains of the genus Staphylococcus preferably include strains of S. epidermidis (Staphylococcus epidermidis), and more preferably include at least one of S. epidermidis ATCC12228 and S. epidermidis ATCCC14990. These other strains of the genus Staphylococcus may be used alone or in combination of two or more.

[0077] The concentration of other Staphylococcus strains in the composition for treating tinea is not particularly limited, but may be 10 3 ~10 11 CFU / g is preferred, and 10 4 ~10 10 CFU / g is more preferred, and 10 5 ~10 9 It is more preferable that the CFU / g is within the above range. When two or more other strains of the genus Staphylococcus are contained in combination, it is preferable that each of the two or more other strains of the genus Staphylococcus falls within the above range for each species.

[0078] The form of the other Staphylococcus strain in the composition for treating tinea is not particularly limited, but is preferably in the form of a dry powder (e.g., freeze-dried or spray-dried).The form of the other Staphylococcus strain is preferably the same as that of at least one selected from the group consisting of the above-mentioned S. hominis strain, S. haemolyticus strain, and S. warneri strain.

[0079] [Strains of other genera] The composition for treating ringworm may further contain a strain of another genera, where "strain of another genera" means a bacterial strain other than a strain of the genus Staphylococcus.

[0080] The strains of other genera are not particularly limited, but are preferably normal skin bacteria (i.e., normal skin bacteria other than Staphylococcus bacteria), and examples thereof include strains of Cutibacterium acnes, Streptococcus pyogenes (group A β-hemolytic streptococcus), Pseudomonas aeruginosa, Escherichia coli, Bacillus cereus, Enterococcus faecalis, and Moraxella osloensis.

[0081] As for the strain of Cutibacterium acnes, "NCBI", [online], archived on February 17, 2022, [searched on February 17, 2022], Internet<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1140 / > Examples include those described in. Streptococcus pyogenes (group A beta-hemolytic streptococcus) strains are available from NCBI, [online], archived on February 17, 2022, [searched on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 175 / > Examples include those described in. Pseudomonas aeruginosa strains are available from NCBI, [online], archived on February 17, 2022, [searched on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 187 / > Examples include those described in. For Escherichia coli strains, see "NCBI," [online], archived February 17, 2022, [searched February 17, 2022], Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 167 / > Examples include those described in. Bacillus cereus strains are available from NCBI, [online], archived on February 17, 2022, [searched on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 157 / > Examples include those described in. Enterococcus faecalis strains are available from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 808 / > Examples include those described in. Moraxella osloensis strains are available from NCBI, [online], archived February 17, 2022, [retrieved February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 12468 / > Examples include those described in.

[0082] Of these, from the viewpoint of obtaining a higher therapeutic effect against tinea, it is preferable that the strain of other genera includes at least one selected from the group consisting of Pseudomonas aeruginosa strains, Escherichia coli strains, Bacillus cereus strains, Enterococcus faecalis strains, and Moraxella osloensis strains, it is more preferable that the strain includes at least one selected from the group consisting of Pseudomonas aeruginosa strains, Escherichia coli strains, and Enterococcus faecalis strains, and it is even more preferable that the strain includes at least one selected from the group consisting of Pseudomonas aeruginosa ATCC27853, Escherichia coli ATCC25922, and Enterococcus faecalis ATCC29212. Furthermore, from the viewpoint of adjusting the balance of normal skin bacteria in the affected area to which the composition for treating tinea is applied, the strains of other genera preferably include at least one selected from the group consisting of strains of Cutibacterium acnes, strains of Streptococcus pyogenes (group A beta-hemolytic streptococcus), and strains of Pseudomonas aeruginosa, more preferably strains of Cutibacterium acnes, and even more preferably strains of Cutibacterium acnes ATCC 6919. These strains of other genera may be used alone or in combination of two or more.

[0083] The concentration of the strain of other genera in the composition for treating tinea is not particularly limited, but may be 103 ~10 11 CFU / g is preferred, and 10 4 ~10 10 CFU / g is more preferred, and 10 5 ~10 9 It is more preferable that the CFU / g is 0.5 CFU / g. When two or more strains of other genera are contained in combination, it is preferable that each of the two or more strains of other genera falls within the above range for each species.

[0084] The form of the strain of another genera in the composition for treating tinea is not particularly limited, but is preferably in the form of a dry powder (e.g., freeze-dried or spray-dried).The form of the strain of another genera is preferably the same as that of at least one selected from the group consisting of the above-mentioned S. hominis strain, S. haemolyticus strain, and S. warneri strain.

[0085] [Antifungal agent] The composition for treating tinea may further contain an antifungal agent. By including an antifungal agent, the therapeutic effect of tinea may be enhanced.

[0086] Antifungal agents include, but are not limited to, itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, luliconazole, efinaconazole, fosravuconazole, terbinafine, butenafine, etc. These antifungal agents may be used alone or in combination of two or more.

[0087] The concentration of the antifungal agent in the composition for treating tinea is not particularly limited, but is preferably 0.1 to 30% by mass, and more preferably 0.5 to 15% by mass, relative to the total mass of the composition for treating tinea.

[0088] [Additives] The composition for treating tinea may contain additives, including, but not limited to, bases, humectants, thickeners, emulsifiers, emulsifier aids, preservatives, stabilizers, pH adjusters, etc.

[0089] (Base) Examples of the base include a hydrophobic base, a hydrophilic base, and water. In this specification, the term "base" 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.

[0090] The hydrophobic base is not particularly limited, and examples thereof include higher hydrocarbons such as squalane, liquid paraffin, light liquid paraffin, petrolatum, ceresin wax, microcrystalline wax, squalene, and gelled hydrocarbons; oils and fats such as olive oil, jojoba oil, sesame oil, soybean oil, cacao butter, camellia oil, peanut oil, beef tallow, lard, triacetin, and hardened castor oil; waxes such as beeswax, white beeswax, carnauba wax, and lanolin; fatty acids such as stearic acid and oleic acid; higher alcohols such as lanolin alcohol, myristyl alcohol, cetanol (cetyl alcohol), stearyl alcohol, cetostearyl alcohol, and cholesterol; and fatty acid esters such as isopropyl myristate, stearyl myristate, and medium-chain fatty acid triglycerides.

[0091] The hydrophilic base is not particularly limited, but examples thereof include lower alcohols such as ethanol, propanol, and isopropanol; polyhydric alcohols such as glycerin, 1,3-butylene glycol, and propylene glycol; sugar alcohols such as sorbitol and mannitol; and macrogol.

[0092] The above-mentioned bases may be used alone or in combination of two or more.

[0093] (wetting agent) The humectant keeps the skin moist when the ringworm treatment composition is applied. The humectant is not particularly limited, but examples thereof include petrolatum, glycerin, propylene glycol, and 1,3-butylene glycol. The above-mentioned wetting agents may be used alone or in combination of two or more kinds.

[0094] (thickener) The thickener increases the viscosity or causes the composition for treating ringworm to become gelatinous. Examples of thickeners include, but are not limited to, gelatin, agar, carrageenan, gum arabic, tragacanth, sodium alginate, propylene glycol alginate, carboxyvinyl polymer, polyvinyl alcohol (partially saponified), sodium alginate, methylcellulose, carboxymethylcellulose, water-soluble cellulose derivatives (hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydrophobized hydroxypropyl methylcellulose, etc.), sodium polyacrylate, glycerin monooleate, pectin, xanthan gum, etc. The above-mentioned thickeners may be used alone or in combination of two or more kinds.

[0095] (emulsifier) The emulsifier emulsifies the composition for treating tinea into an oil-in-water (o / w) type or a water-in-oil (w / o) type. The emulsifier is not particularly limited, but examples thereof 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, alkyl sulfates, saturated higher fatty acid salts, and N-acylamino acid salts; nonionic surfactants such as sucrose fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene sorbitan 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 zwitterionic surfactants such as alkyl betaines, alkylamido betaines, alkyl sulfobetaines, imidazolines, lauryl dimethylaminoacetic acid betaine, and alkyldiaminoethyl glycines. The above-mentioned emulsifiers may be used alone or in combination of two or more kinds.

[0096] (emulsifying aid) The co-emulsifier stabilizes the emulsion when the composition for treating tinea is emulsified into an oil-in-water (o / w) type or a water-in-oil (w / o) type. The emulsifying aid is not particularly limited, but examples thereof include cetanol (cetyl alcohol), stearyl alcohol, oleyl alcohol, and isostearyl alcohol. The above-mentioned emulsifying aids may be used alone or in combination of two or more kinds.

[0097] (preservative) The preservative prevents or inhibits contamination and degradation of the composition for treating ringworm by foreign microorganisms. The preservative is not particularly limited, but examples thereof include parahydroxybenzoic acid, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, sodium benzoate, chlorobutanol, chlorocresol, benzyl alcohol, salicylic acid, phenoxyethanol, thymol, dibutylhydroxytoluene, sodium edetate hydrate, sodium dehydroacetate, sorbic acid, potassium sorbate, and benzalkonium chloride. The above preservatives may be used alone or in combination of two or more.

[0098] (stabilizer) The stabilizer prevents or inhibits the death, decomposition, and physical changes of the ingredients contained in the composition for treating ringworm. The stabilizer is not particularly limited, but examples thereof include sodium hydrogen sulfite, sodium pyrosulfite, ascorbic acid, tocopherol, dibutylhydroxytoluene, sodium edetate (EDTA), and benzotriazole. The above-mentioned stabilizers may be used alone or in combination of two or more kinds.

[0099] (pH adjuster) The pH adjuster maintains the stability of the ingredients contained in the composition for treating tinea and prevents or reduces irritation of the composition for treating tinea to the living body. The pH adjuster is not particularly limited, but examples thereof 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. The above pH adjusters may be used alone or in combination of two or more.

[0100] [Dosage form] The dosage form of the composition for treating tinea is not particularly limited, but is preferably an external preparation. By using the composition for treating tinea as an external preparation, at least one selected from the group consisting of S. hominis strains, S. haemolyticus strains, and S. warneri strains can be established on the skin, thereby effectively preventing or suppressing the growth and establishment of tinea fungi, and sterilizing or bacteriostatic.

[0101] The specific form of the topical preparation is not particularly limited, but examples include ointments, creams, gels, lotions, and the like.

[0102] Ointments are semi-solid topical preparations in which the active ingredient to be applied to the skin is dissolved or dispersed in a base, and examples include oily ointments and water-soluble ointments. Creams are semi-solid topical preparations to be applied to the skin that are emulsified as oil-in-water (o / w) or water-in-oil (w / o) emulsions. Gels are gel-type topical preparations to be applied to the skin, and examples include aqueous gels and oily gels. Lotions are liquid topical preparations in which the active ingredient (e.g., S. hominis strain, S. haemolyticus strain, S. warneri strain) is dissolved or dispersed in an aqueous solution.

[0103] [Tinea fungus] The causative fungus of tinea is tinea fungus, specifically, a fungus of the genus Trichophyton. Examples of the tinea fungus include fungal strains of Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

[0104] Examples of the fungal strain of Trichophyton rubrum include Trichophyton rubrum ATCC28188 and Trichophyton rubrum ATCC22402.

[0105] Examples of the fungal strain of Trichophyton mentagrophytes include Trichophyton mentagrophytes TIMM2789 and Trichophyton mentagrophytes ATCCMYA-4439.

[0106] Examples of the fungal strain of Trichophyton tonsurans include Trichophyton tonsurans ATCC56186 and Trichophyton tonsurans ATCC28942.

[0107] The causative fungus of ringworm preferably includes a fungus of the genus Trichophyton, more preferably includes at least one selected from the group consisting of a fungal strain of Trichophyton rubrum, a fungal strain of Trichophyton mentagrophytes, and a fungal strain of Trichophyton tonsurans, further preferably includes a fungal strain of Trichophyton rubrum, and particularly preferably includes Trichophyton rubrum ATCC 28188. The causative fungus of ringworm may be one type or two or more types.

[0108] Tinea is caused by the Trichophyton fungus. Tinea is classified into tinea pedis (athlete's foot), tinea unguium (nail athlete's foot), tinea manubriata (athlete's foot), tinea corporis (ringworm), tinea cruris (jock itch), tinea capitis (tinea capitis), etc. depending on the site of onset. Since the skin penetration of topical preparations varies depending on the site of onset, it is preferable to appropriately adjust the composition of the topical preparation depending on the site of onset.

[0109] [Administration method] The method of administering the composition for treating tinea is not particularly limited, but when the composition for treating tinea is an external preparation, it is applied to the affected area.

[0110] The number of administrations (applications) is not particularly limited and may be three times a day, twice a day, once a day, once every two days, once every three days, once every four days, once every five days, once every six days, or once every seven days, etc. Of these, the number of administrations is preferably once a day, once every two days, once every three days, once every four days, once every five days, once every six days, or once every seven days. The composition for treating tinea according to the present invention contains at least one strain selected from the group consisting of S. hominis, S. haemolyticus, and S. warneri, which have excellent skin-adhering properties. Therefore, compared to conventional tinea therapeutic agents, the composition exhibits sustained effects associated with the adhesion of live bacteria, enabling maintenance of remission and prevention of recurrence, etc., and also enabling a reduction in the number of administrations and increased patient compliance.

[0111] When the tinea therapeutic composition is used as a topical agent, the amount of application is 0.1 to 180 mg / cm 2 is preferably 1 to 10 mg / cm 2 In this case, the amount of application of at least one selected from the group consisting of S. hominis strain, S. haemolyticus strain, and S. warneri strain in the composition for treating tinea is more preferably 10 0 ~10 8 CFU / cm 2 Preferably, 10 1 ~10 7 CFU / cm 2 It is more preferable that 2~10 6 CFU / cm 2 When the composition for treating tinea contains two or more strains of S. hominis, S. haemolyticus, and S. warneri, it is preferable that the total amount thereof be within the above range.

[0112] The affected area to which the composition for treating tinea is applied is not particularly limited, but is usually an area with symptoms of tinea. Note that the composition for treating tinea according to the present invention contains at least one selected from the group consisting of strains of S. hominis, S. haemolyticus, and S. warneri, which are normal skin bacteria, and therefore has high skin adhesion, allowing for a reduction in the number of administrations compared to conventional topical preparations, and causing no or almost no skin rash problems around the affected area, thereby increasing patient compliance.

[0113] The above-mentioned composition for treating tinea exhibits a therapeutic effect when used alone, but may also be used in combination with other therapeutic agents.

[0114] The other therapeutic agents include oral and topical agents. In this case, the oral or topical agent contains at least one antifungal agent such as itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, luliconazole, efinaconazole, fosravuconazole, terbinafine, and butenafine. Two or more of the above-mentioned other therapeutic agents may be combined.

[0115] That is, according to one aspect of the present invention, a combination of a tinea therapeutic composition and another therapeutic agent is provided. In this case, when the tinea therapeutic composition is an external preparation, the other therapeutic agent is preferably an oral preparation. Furthermore, the tinea therapeutic composition and the other therapeutic agent may be administered simultaneously or sequentially.

[0116] <Method for producing a composition for treating tinea> One aspect of the present invention provides a method for producing a composition for treating tinea. The method includes the steps of (1) collecting and culturing a bacterial flora from at least one of the ankle and sole of a human (preferably a healthy individual without tinea symptoms), (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria, (3) evaluating the anti-tinea fungal activity of the candidate bacteria to identify bacteria with anti-tinea fungal activity, and (4) preparing a composition for treating tinea containing the bacteria with anti-tinea fungal activity. The bacteria with anti-tinea fungal activity include at least one selected from the group consisting of S. hominis strains, S. haemolyticus strains, and S. warneri strains. The S. hominis strain is preferably an S. hominis strain with anti-tinea fungal activity. The S. haemolyticus strain is preferably an S. haemolyticus strain with anti-tinea fungal activity. Furthermore, the S. warneri strain is preferably an S. warneri strain having an anti-trichophytic activity. Specific examples of S. hominis strains, S. haemolyticus strains, and S. warneri strains are as described above.

[0117] [Process (1)] Step (1) involves collecting and culturing bacterial flora from at least one of the ankle and sole of a human (preferably a healthy individual without tinea symptoms). Bacterial flora is collected from the ankle and / or sole of a human using a swab or the like, and then smeared on a medium and cultured. This allows colonies to form for each strain.

[0118] [Process (2)] Step (2) is a step of isolating and culturing the bacterial flora cultured in step (1) to obtain candidate bacteria. The colonies formed by the culture in step (1) are each collected using a platinum loop or the like, smeared on a medium, and isolated and cultured. This allows the isolation of a large number of bacteria contained in the bacterial flora contained in the human ankle and / or sole. The cultured bacteria are candidate bacteria, and are preferably suspended in a bacterial stock solution and stored at low temperatures (e.g., −100 to −50°C, preferably −90 to −70°C) as a skin bacterial library.

[0119] [Process (3)] Step (3) is a step of evaluating the anti-trichophyton activity of the candidate fungus obtained in step (2) to identify a bacterium having anti-trichophyton activity.

[0120] In step (3), first, the candidate fungi isolated from the human ankle and / or sole are evaluated for their anti-trichophyton activity by the method described in "4. Evaluation of anti-trichophyton activity" in the Examples.

[0121] Next, bacteria with anti-trichophyton activity are selected and identified. Identification methods include, but are not limited to, methods based on 16s rDNA sequence comparison and methods for creating a phylogenetic tree. For example, in methods based on 16s rDNA sequence comparison, the 16s rDNA sequence of the target bacterium is analyzed, and the bacterium can be identified by performing a BLAST search on NCBI using the DNA sequence data. In methods for creating a phylogenetic tree, the 16s rDNA sequence of the bacterium is analyzed in combination with 16s rDNA sequence data of known bacteria to create a phylogenetic tree, and the target bacterium can be identified from the phylogenetic tree. Note that, if the target bacterium is identified as a strain of S. hominis, S. haemolyticus, or S. warneri, step (3) may include a step of calculating the identity of the 16s rDNA sequence of the target bacterium to SEQ ID NOs: 1, 10, 17, etc., and a step of confirming the DNA sequence identity of the target bacterium.

[0122] The bacteria having anti-trichophyton activity identified in step (3) may include at least one selected from the group consisting of an S. hominis strain having anti-trichophyton activity, an S. haemolyticus strain having anti-trichophyton activity, and an S. warneri strain having anti-trichophyton activity. Therefore, by extracting at least one selected from the group consisting of a target S. hominis strain having anti-trichophyton activity, an S. haemolyticus strain having anti-trichophyton activity, and an S. warneri strain having anti-trichophyton activity from the stored skin bacteria library, it is possible to obtain an S. hominis strain having anti-trichophyton activity, an S. haemolyticus strain having anti-trichophyton activity, or an S. warneri strain having anti-trichophyton activity.

[0123] [Process (4)] Step (4) is a step of preparing a composition for treating tinea containing bacteria having anti-tinea fungal activity. The bacteria having anti-tinea fungal activity identified in step (3) are appropriately cultured, purified, lyophilized, etc. to prepare a composition for treating tinea containing bacteria having anti-tinea fungal activity. The bacteria having anti-tinea fungal activity include at least one selected from the group consisting of S. hominis strains, S. haemolyticus strains, and S. warneri strains, and preferably at least one selected from the group consisting of S. hominis strains having anti-tinea fungal activity, S. haemolyticus strains having anti-tinea fungal activity, and S. warneri strains.

[0124] <Treatment method> According to one aspect of the present invention, there is provided a method for treating tinea, which comprises applying the above-described composition for treating tinea to the affected area. The composition for treating tinea and its administration method are as described above. [Example]

[0125] The present invention will be specifically explained below with reference to examples, but the present invention is not limited to these examples.

[0126] 1.Collection of candidate bacteria Bacterial flora was collected using swabs from one ankle and one sole of the foot of five healthy volunteers. The swabs containing the bacterial flora were streaked onto AnaeroColumbia RS blood agar medium and cultured at 35°C under aerobic conditions for one day. After culture, colonies characteristic of morphology, color, and dryness were collected using a platinum loop.

[0127] The platinum loop containing the collected colonies was smeared on an AnaeroColumbia RS blood agar medium and cultured for isolation at 35°C under aerobic conditions for one day.

[0128] After cultivation, the grown colonies were collected with a platinum loop, suspended in a 25% glycerol bacterial stock solution, and stored at -80°C. A total of 91 candidate strains were collected, including 52 strains from the ankle and 39 strains from the sole.

[0129] 2. Preparation of Known Bacteria The following 32 strains of bacteria were prepared as known bacteria. (1)Acinetobacter baumannii ATCC17978 (2)Corynebacterium glaucum JCM12208 (3)Corynebacterium jeikeium ATCC43734 (4)Corynebacterium striatum NBRC15291 (5)Enterococcus faecalis ATCC29212 (6) Escherichia coli ATCC25922 (7)Micrococcus luteus ATCC4698 (8)Moraxella atlantae NBRC14588 (9)Neisseria gonorrhoeae ATCC19424 (10)Pseudomonas aeruginosa ATCC27853 (11) Staphylococcus aureus ATCC29213 (12) Staphylococcus epidermidis ATCC12228 (13) Staphylococcus haemolyticus ATCC29970 (14)Staphylococcus hominis ATCC27844 (15) Streptococcus agalactiae ATCC13813 (16)Streptococcus mitis JCM12971 (17) Streptococcus pyogenes ATCC12344 (18) Staphylococcus aureus ATCC12600 (19) Staphylococcus epidermidis ATCC14990 (20) Staphylococcus capitis ATCC27840 (21)Staphylococcus caprae ATCC35538 (22) Staphylococcus cohnii ATCC29974 (23) Staphylococcus haemolyticus ATCC700564 (24) Staphylococcus haemolyticus ATCC29969 (25)Staphylococcus hominis ATCC27845 (26)Staphylococcus hominis ATCC700236 (27) Staphylococcus kloosii ATCC43959 (28)Staphylococcus lentus ATCC700403 (29)Staphylococcus lugdunensis ATCC49576 (30)Staphylococcus lugdunensis ATCC43809 (31)Staphylococcus warneri ATCC27836 (32)Staphylococcus saprophyticus ATCC15305

[0130] 3. Preparation of evaluation medium To evaluate the anti-trichophyton activity of candidate fungi and known fungi, a fungal pour medium containing trichophyton was prepared as an evaluation medium.

[0131] Specifically, one bead of Trichophyton rubrum ATCC28188 stored in a microbank was taken, smeared on a potato dextrose agar (PDA) slant medium, and statically cultured at 30°C under aerobic conditions.

[0132] After the cultivation, phosphate-buffered saline (PBS) containing 0.05% polysorbate 80 was added to the PDA agar slant medium to prepare a fungal inoculum.

[0133] PDA was dissolved in ultrapure water to a concentration of 39.0 g / L, and the resulting solution was heated in an autoclave (121 °C, 20 min). After cooling, the PDA solution was mixed with the fungal inoculum solution, poured into petri dishes, and solidified to prepare fungal pour medium.

[0134] 4. Evaluation of anti-tinea fungus activity The anti-tinea activity of candidate fungi (91 strains) and known fungi (32 strains) was evaluated using a fungal pour medium as an evaluation medium.

[0135] Specifically, one bead containing a candidate or known bacterium stored in a microbank was taken, smeared on an Anaerobic Columbia RS blood agar medium, and cultured at 35°C under aerobic conditions for 1 to 2 days. The resulting bacteria were then collected using a platinum loop.

[0136] The collected bacteria were streaked onto a fungal pour medium and incubated at 30°C under aerobic conditions until the growth of Trichophytons caused turbidity in the agar medium, which could be visually confirmed. After incubation, the growth inhibition zone formed on the fungal pour medium was observed, and the anti-trichophyton activity of the candidate or known fungus was evaluated according to the size of the growth inhibition zone according to the following criteria. Typical examples of each evaluation are shown in Figure 1.

[0137] ++: A clear area of ​​inhibition of tinea fungus growth was observed +: No clear growth inhibition area was observed, but growth of tinea fungus was inhibited. -: No inhibition of tinea fungus growth was observed

[0138] First, the results obtained for known bacteria (32 strains) are shown in Table 4 below.

[0139] [Table 4]

[0140] The results in Table 4 show that of the known bacteria (32 strains), 9 strains were "++", 10 strains were "+", and 13 strains were "-".

[0141] Furthermore, of the candidate bacteria (91 strains), 23 strains were "++", 32 strains were "+", and 36 strains were "-".

[0142] 5. Identification of Candidate Bacteria To analyze the relationship between the anti-dermatophyte activity of the candidate fungi and the type of the candidate fungi, the candidate fungi were identified by comparing their 16s rDNA sequences.

[0143] Specifically, total DNA was extracted and purified from candidate bacteria cultured on AnaeroColumbia RS blood agar medium according to the NucleoSpin Microbial DNA manual, and the DNA concentration and purity were measured using the NanoDrop ultra-microspectrophotometer.

[0144] Next, according to the instructions in the Bacterial 16S rDNA PCR Kit, the purified total DNA was used as a template to amplify the 16S rDNA sequence region 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 the NucleoSpin Gel and PCR Clean-up kit, and the DNA concentration and purity were measured using a NanoDrop ultra-microspectrophotometer. Agarose gel electrophoresis confirmed that the DNA fragment size had been amplified as expected.

[0145] The amplified DNA fragment (16S rDNA template sample) was mixed with primers (F1 (SEQ ID NO: 19), F2 (SEQ ID NO: 21), or R2 (SEQ ID NO: 20)) included in the Bacterial 16S rDNA PCR Kit, and the target sequence was subjected to Sanger sequencing. After assembling the resulting sequences, the candidate bacterial species was identified by performing a NCBI BLAST search on the region between consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG), which are shared by Staphylococcus 16S rDNA (approximately 1,240-1,280 bp). The target region of 16S rDNA (approximately 1,240-1,280 bp) is part of the amplified DNA fragment (approximately 1,500 bp) and contains consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG). Furthermore, if consensus sequence 1 (AGCTTGC) was not present in the obtained sequence, it was replaced with GGTTGG and TGCTTGC in that order, and if consensus sequence 2 (AAGCTGG) was not present, it was replaced with GAAGTTGG, AAGTCGG, GAAGTCGA, AAGTCAG, GAGCTGG, AAGGGGGG, AAGTCTGG, CAGTCAG, and CCGCTGG in that order. The candidate bacterial species were identified by performing a BLAST search on NCBI for the base sequence of the region flanked by each consensus sequence (approximately 1,240 to 1,280 bp). If the obtained sequence did not contain either or both of consensus sequence 1 (AGCTTGC) or its alternative sequences (GGTTGG, TGCTTGC), and consensus sequence 2 (AAGCTGG) or its alternative sequences (GAAGTTGG, AAGTCGG, GAAGTCGA, AAGTCAG, GAGCTGG, AAGGGGGG, AAGTCTGG, CAGTCAG, CCGCTGG), the candidate bacterial species was identified by performing a BLAST search on NCBI for the full-length base sequence of the obtained sequence (amplified DNA fragment (approximately 1,500 bp)).

[0146] To identify the candidate bacterial species, we selected those with 99% or more identity from the top 10 sequences found in the BLAST search. If the top 10 selected bacterial species included multiple results from the same genus but different species, we used the genus name (Staphylococcus sp. and Micrococcus sp.). If no species meeting the criteria was identified, the species was deemed unknown.

[0147] The identified candidate fungi and the results of their anti-trichophyton activity are shown in Tables 5 and 6 below.

[0148] [Table 5]

[0149] [Table 6]

[0150] The identified candidate bacteria are organized by species in Tables 5 and 6, and the percentage of anti-tinea fungal activity exhibited by each species is analyzed and shown in Figure 2. The results in Figure 2 show that 100% and 86% of Staphylococcus hominis and Staphylococcus haemolyticus, respectively, showed a "++" result.

[0151] Furthermore, of the 92 candidate strains, the results of the anti-dermatophyte activity by isolation site for 52 candidate strains obtained from the ankle and 39 candidate strains obtained from the sole of the foot are shown in Table 7 below.

[0152] [Table 7]

[0153] The results in Table 7 show that of the 52 strains from the ankle, 33% were "++", 31% were "+", and 37% were "-", while of the 39 strains from the sole, 15% were "++", 41% were "+", and 44% were "-". Compared to the sole, the proportion of strains showing "++" was found to be approximately twice as high in the ankle.

[0154] 6. Construction of a phylogenetic tree of the Staphylococcus genus A phylogenetic tree was created for 27 known strains of the genus Staphylococcus (Table 8 below) and 45 candidate strains identified as belonging to the genus Staphylococcus (Table 9 below, excerpts from Tables 5 and 6) based on the phylogenetic tree for the 16S rDNA sequences of 27 species of the genus Staphylococcus in a 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). The created phylogenetic tree is shown in Figure 3. The phylogenetic tree was created using CLC sequence viewer (ver. 8.0).

[0155] [Table 8]

[0156] [Table 9]

[0157] Comparing the phylogenetic tree in Figure 3 with the results of the anti-dermatophyte activity of the candidate fungi, the clusters were roughly divided into three clusters: Cluster 1, which mainly showed "-" results; Cluster 2, which mainly showed "++" results; and Cluster 3, which mainly showed "+" results. Of these, Cluster 2 consisted of S. hominis and S. haemolyticus.

[0158] Therefore, we created phylogenetic trees for six known strains of S. hominis and S. haemolyticus bacteria (Table 10 below) and 15 candidate strains identified as S. hominis or S. haemolyticus (Table 11 below, excerpts from Table 9). The created phylogenetic trees for S. hominis and S. haemolyticus are shown in Figures 4 and 5.

[0159] [Table 10]

[0160] [Table 11]

[0161] 7.16s rDNA sequence (1) 16s rDNA sequence of Staphylococcus hominis strains The 16s rDNA sequence of the known bacterium Staphylococcus hominis ATCC27844 was confirmed in the ATCC database, and the nucleotide sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. As a result, the 16s rDNA sequence of Staphylococcus hominis ATCC27844 had the nucleotide sequence of SEQ ID NO: 1 (Staphylococcus hominis ATCC27844 genome positions: 716544-717811). Note that SEQ ID NO: 1 contains consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG), and therefore the 5' end of the nucleotide sequence of SEQ ID NO: 1 is consensus sequence 1 (AGCTTGC) and the 3' end is consensus sequence 2 (AAGCTGG).

[0162] Furthermore, the 16S rDNA sequences of candidate strains Nos. 2, 3, 37, 40, 45, 62, 64, and 74, which were identified as Staphylococcus hominis strains, were confirmed for the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) (approximately 1,240-1,280 bp) using the same method as described above in "5. Identification of Candidate Bacteria." As a result, the 16S rDNA sequences of candidate strains Nos. 3, 40, 45, 64, and 74 had the nucleotide sequence of SEQ ID NO: 5, the 16S rDNA sequence of candidate strain No. 2 had the nucleotide sequence of SEQ ID NO: 7, the 16S rDNA sequence of candidate strain No. 37 had the nucleotide sequence of SEQ ID NO: 8, and the 16S rDNA sequence of candidate strain No. 62 had the nucleotide sequence of SEQ ID NO: 9.

[0163] Furthermore, the 16s rDNA sequences of the known bacteria Staphylococcus hominis ATCC27845 and Staphylococcus hominis ATCC700236 were confirmed in the ATCC database, and the base sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. Also, the 16s rDNA sequence of Staphylococcus hominis A9 (ATCC accession number: PTA-125203) was confirmed in the PATRIC database, and the base sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. As a result, the 16s rDNA sequence of Staphylococcus hominis ATCC27845 had the base sequence of SEQ ID NO: 1 (Staphylococcus hominis ATCC27845 genome position: 761822-763089), and the 16s rDNA sequences of Staphylococcus hominis ATCC700236 and Staphylococcus hominis A9 (ATCC accession number: PTA-125203) had the base sequence of SEQ ID NO: 5 (Staphylococcus hominis ATCC700236 genome position: 849942-851209, Staphylococcus hominis A9 genome position: 185-1452).

[0164] (2) 16s rDNA sequences of Staphylococcus haemolyticus strains The 16s rDNA sequence of the known bacterium Staphylococcus haemolyticus ATCC29970 was confirmed in the ATCC database, and the nucleotide sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. As a result, the 16s rDNA sequence of Staphylococcus haemolyticus ATCC29970 had the nucleotide sequence of SEQ ID NO: 10 (Staphylococcus haemolyticus ATCC29970 genome positions: 807302 to 808569).

[0165] Furthermore, the 16s rDNA sequences of candidate bacteria Nos. 22 to 25, 33, and 35, identified as Staphylococcus haemolyticus strains, were confirmed for the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) (approximately 1,240 to 1,280 bp) using the same method as described above in "5. Identification of Candidate Bacteria." The results showed that the 16s rDNA sequences of candidate bacteria Nos. 22, 24, 25, and 33 had the nucleotide sequence of SEQ ID NO: 13, the 16s rDNA sequence of candidate bacteria No. 35 had the nucleotide sequence of SEQ ID NO: 14, and the 16s rDNA sequence of candidate bacteria No. 23 had the nucleotide sequence of SEQ ID NO: 16. The 16s rDNA sequence of candidate bacteria No. 20, identified as a Staphylococcus haemolyticus strain, contained consensus sequence 1 (AGCTTGC) but not consensus sequence 2 (AAGCTGG). The 16s rDNA sequence of No. 20 contained an alternative sequence (GAGCTGG) to consensus sequence 2, so the base sequence (approximately 1,267 bp) of the region flanked by consensus sequence 1 (AGCTTGC) and the alternative sequence (GAGCTGG) to consensus sequence 2 was confirmed. As a result, the 16s rDNA sequence of candidate bacterium No. 20 contained the base sequence of SEQ ID NO: 15.

[0166] Furthermore, the 16s rDNA sequence of the known bacterium Staphylococcus haemolyticus ATCC700564 was checked against the ATCC database, and the nucleotide sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. As a result, the 16s rDNA sequence of Staphylococcus haemolyticus ATCC700564 had the nucleotide sequence of SEQ ID NO: 10 (Staphylococcus haemolyticus ATCC700564 genome position: 214068-215335).

[0167] The results obtained in (1) and (2) above are shown in Tables 12 and 13 below.

[0168] [Table 12]

[0169] [Table 13]

[0170] In Table 13, SEQ ID NOs: 5 and 7 to 9 are based on SEQ ID NO: 1, and SEQ ID NOs: 13 to 16 are based on SEQ ID NO: 10. For example, "45G>A" in SEQ ID NO: 5 means that G (guanine) at position 45 of SEQ ID NO: 1 has been mutated to A (adenine). For example, "488delA" in SEQ ID NO: 7 means that A (adenine) at position 488 of SEQ ID NO: 1 has been deleted. For example, "1182_1183insA" in SEQ ID NO: 8 means that A (adenine) has been inserted between positions 1182 and 1183 of SEQ ID NO: 1.

[0171] (3) 16s rDNA sequence of Staphylococcus warneri strains The 16s rDNA sequence of the known bacterium Staphylococcus warneri ATCC27836 was confirmed in the ATCC database, and the nucleotide sequence of the region flanked by consensus sequence 1 (AGCTTGC) and consensus sequence 2 (AAGCTGG) was confirmed. As a result, the 16s rDNA sequence of Staphylococcus warneri ATCC27836 had the nucleotide sequence of SEQ ID NO: 17 (Staphylococcus warneri ATCC27836 genome position: 725369-726636).

[0172] Based on the results of 7(1) to (3) above, S. hominis having a 16s rDNA containing SEQ ID NO: 1 or a base sequence having 95% or more identity to SEQ ID NO: 1, S. haemolyticus having a 16s rDNA containing SEQ ID NO: 10 or a base sequence having 95% or more identity to SEQ ID NO: 10, and S. warneri having a 16s rDNA containing SEQ ID NO: 17 or a base sequence having 95% or more identity to SEQ ID NO: 17 are considered to have anti-tinea fungal activity.

[0173] 8. Effective against other tinea fungi In the above sections 4 to 7, a fungal pour medium containing the tinea fungus Trichophyton rubrum ATCC28188 was used as the evaluation medium (see "3. Preparation of evaluation medium" above). Here, the effects on other tinea fungi (tinea fungi other than T. rubrum ATCC28188) were evaluated.

[0174] (1) Preparation of fungal pour medium containing other dermatophytes Fungal pour medium containing five species of dermatophytes was prepared in the same manner as in "3. Preparation of evaluation medium" above, except that beads of Trichophyton rubrum ATCC22402, Trichophyton mentagrophytes TIMM2789, Trichophyton mentagrophytes ATCCMYA-4439, Trichophyton tonsurans ATCC56186, or Trichophyton tonsurans ATCC28942 were used instead of beads of Trichophyton rubrum ATCC28188.

[0175] (2) Evaluation of anti-dermatophytic activity against other dermatophytes The anti-tinea fungal activities of the known fungi S. hominis ATCC27844, S. hominis ATCC27845, S. haemolyticus ATCC29969, S. haemolyticus ATCC700564, and S. warneri ATCC27836, which were evaluated for their anti-tinea fungal activity against Trichophyton rubrum ATCC28188, were also evaluated using fungal pour medium containing the above five species of tinea fungi.

[0176] The evaluation method was the same as in "4. Evaluation of anti-trichophyton activity" above. The results obtained are shown in Table 13 below, along with the results of the anti-trichophyton activity against T. rubrum ATCC28188.

[0177] [Table 14]

[0178] The results in Table 14 show that the same anti-dermatophytic activity as that of T. rubrum ATCC28188 was also exhibited against five other species of dermatophytes.

Claims

1. A composition for treating tinea, comprising at least one bacterium having anti-tinea activity selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri, A composition for treating tinea, wherein the Staphylococcus warneri strain comprises Staphylococcus warneri ATCC27836.

2. The composition for treating ringworm according to claim 1, comprising a strain of Staphylococcus hominis.

3. 3. The composition for treating tinea according to claim 2, wherein the Staphylococcus hominis strain has a 16s rDNA containing the base sequence of SEQ ID NO: 1 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 1, and has anti-tinea activity.

4. The composition for treating tinea according to claim 2, wherein the Staphylococcus hominis strain comprises at least one selected from the group consisting of Staphylococcus hominis ATCC27844, Staphylococcus hominis ATCC27845, and Staphylococcus hominis ATCC700236.

5. The composition for treating tinea according to claim 1, comprising a strain of Staphylococcus haemolyticus.

6. 6. The composition for treating tinea according to claim 5, wherein the Staphylococcus haemolyticus strain has a 16s rDNA containing the base sequence of SEQ ID NO: 10 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 10, and has anti-tinea activity.

7. 6. The composition for treating tinea according to claim 5, wherein the Staphylococcus haemolyticus strain comprises at least one selected from the group consisting of Staphylococcus haemolyticus ATCC29970, Staphylococcus haemolyticus ATCC700564, and Staphylococcus haemolyticus ATCC29969.

8. The composition for treating ringworm according to claim 1, comprising a strain of Staphylococcus warneri.

9. The composition for treating tinea according to any one of claims 1 to 8, wherein the causative fungus of tinea comprises a fungus of the genus Trichophyton.

10. The composition for treating tinea according to claim 9, wherein the causative fungus of tinea comprises at least one selected from the group consisting of fungal strains of Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

11. The composition for treating tinea according to any one of claims 1 to 8, which is an external preparation.

12. (1) collecting and culturing bacterial flora from at least one of a human ankle and a human foot sole; (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; (3) evaluating the anti-trichophyton activity of the candidate fungus to identify a bacterium having anti-trichophyton activity; (4) a step of preparing a composition for treating tinea, which comprises the bacterium having anti-tinea fungal activity; Including, A method for producing a composition for treating tinea, wherein the bacterium having anti-tinea fungal activity comprises at least one selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri, The method for producing the Staphylococcus warneri includes Staphylococcus warneri ATCC27836.

13. 1. Use of at least one bacterium having anti-tinea activity selected from the group consisting of a strain of Staphylococcus hominis, a strain of Staphylococcus haemolyticus, and a strain of Staphylococcus warneri in the manufacture of a therapeutic agent for tinea, The Staphylococcus warneri strain includes Staphylococcus warneri ATCC27836.

14. 14. The use according to claim 13, comprising a strain of Staphylococcus hominis.

15. The use described in claim 14, wherein the Staphylococcus hominis strain has a 16s rDNA containing the base sequence of SEQ ID NO: 1 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 1, and has anti-tinea activity.

16. The use of claim 14, wherein the Staphylococcus hominis strain comprises at least one selected from the group consisting of Staphylococcus hominis ATCC27844, Staphylococcus hominis ATCC27845, and Staphylococcus hominis ATCC700236.

17. 14. The use according to claim 13, comprising a strain of Staphylococcus haemolyticus.

18. The use described in claim 17, wherein the Staphylococcus haemolyticus strain has a 16s rDNA containing the base sequence of SEQ ID NO: 10 or a 16s rDNA containing a base sequence having 95% or more identity to SEQ ID NO: 10, and has anti-tinea activity.

19. The use of claim 17, wherein the Staphylococcus haemolyticus strain comprises at least one selected from the group consisting of Staphylococcus haemolyticus ATCC29970, Staphylococcus haemolyticus ATCC700564, and Staphylococcus haemolyticus ATCC29969.

20. 14. The use according to claim 13, comprising a strain of Staphylococcus warneri.

21. The use according to any one of claims 13 to 20, wherein the causative fungus of ringworm comprises a fungus of the genus Trichophyton.

22. 22. The use of claim 21, wherein the causative fungus of ringworm comprises at least one selected from the group consisting of fungal strains of Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

23. The use according to any one of claims 13 to 20, which is an external preparation.

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