Composition for treating ringworm

Abstract

Provided is a novel composition for treating tinea capitis. The composition for treating white ringworm contains at least one selected from the group consisting of a human staphylococcus strain, a hemolytic staphylococcus strain, and a Staphylococcus vorus strain.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202610581522.8 (22) Application Date 2023.05.15 (30) Priority Data 2022-080165 2022.05.16 JP (62) Divisional Application Data 202380040262.8 2023.05.15 (71) Applicant Maruhiro Co., Ltd. Address Osaka City, Osaka Prefecture, Japan (72) Inventors Naoki Hayashi, Shoji Kanayama (74) Patent Agency China Patent Agency (Hong Kong) Limited 72001 Patent Attorney Yue Xinchen, Peng Chang (51) Int.Cl. A61K 35 / 74 (2015.01) A61P 31 / 10 (2006.01) C12N 1 / 20 (2026.01) C12N 1 / 205 (2026.01) C12R 1 / 44 (2006.01) (54) Invention Title: Composition for Treating Tinea (57) Abstract: A novel composition for treating tinea is provided. The composition for treating tinea contains at least one bacterium selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains. Claims: 2 pages Description: 29 pages Sequence List (electronic publication) Drawings: 3 pages CN 122297526 A 2026.06.30 CN 1 22 29 75 26 A 1. A composition for treating tinea, comprising at least one bacterium with anti-tinea activity selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains, wherein the Staphylococcus wartii strain comprises Staphylococcus wartii ATCC27836. 2. The tinea capitis treatment composition of claim 1, comprising a Staphylococcus aureus strain. 3. The tinea capitis treatment composition of claim 2, wherein the Staphylococcus aureus strain has 16S rDNA comprising the base sequence of Serial Number 1 or 16S rDNA comprising a base sequence having at least 95% identity with Serial Number 1, and has anti-tinea capitis activity. 4. The tinea capitis treatment composition of 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 capitis treatment composition of claim 1, comprising a hemolytic Staphylococcus aureus strain. 6. The tinea capitis treatment composition of claim 5, wherein the hemolytic Staphylococcus aureus strain has 16S rDNA comprising the base sequence of Serial Number 10 or 16S rDNA comprising a base sequence having at least 95% identity with Serial Number 10, and has anti-tinea capitis activity.7. The tinea 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 treatment composition according to claim 1, comprising a Staphylococcus aureus strain. 9. The tinea treatment composition according to any one of claims 1 to 8, wherein the pathogenic fungus of tinea comprises Trichophyton spp. 10. The tinea treatment composition according to claim 9, wherein the pathogenic fungus of tinea comprises at least one selected from Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans fungi. 11. The tinea treatment composition 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 unguium, comprising: a step (1) collecting and culturing bacterial flora from at least one location on the ankle and sole of a person; a step (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; a step (3) evaluating the anti-tinea unguium activity of the candidate bacteria and identifying bacteria with anti-tinea unguium activity; and a step (4) preparing a composition for treating tinea unguium containing bacteria with said anti-tinea unguium activity; wherein the bacteria with said anti-tinea unguium activity comprises at least one selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains, wherein the Staphylococcus wartii strain comprises Staphylococcus wartii ATCC27836. 13. Use of at least one bacterium with anti-tinea unguium activity selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains in the manufacture of a tinea unguium treatment, wherein the Staphylococcus wartii strain comprises Staphylococcus wartii ATCC27836. 14. The use according to claim 13, comprising a Staphylococcus aureus strain. 15. The use according to claim 14, wherein the Staphylococcus aureus strain has 16S rDNA comprising the base sequence of sequence number 1 or 16S rDNA comprising 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. Claims 1 / 2 Page 2 CN 122297526 A 17. The use according to claim 13, comprising a hemolytic Staphylococcus aureus strain. 18. The use according to claim 17, wherein the hemolytic Staphylococcus aureus strain has 16S rDNA comprising the base sequence of sequence number 10 or 16S rDNA comprising 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, comprising a strain of Staphylococcus aureus. 21. The use according to any one of claims 13-20, wherein the pathogenic fungus of tinea capitis comprises a fungus of the genus Trichophyton. 22. The use according to claim 21, wherein the pathogenic fungus of tinea capitis comprises at least one selected from a fungal strain of Trichophyton rubrum, a fungal strain of Trichophyton mentagrophytes, and a fungal strain of Trichophyton tonsurans. 23. The use according to any one of claims 13-20, wherein it is a topical agent. Claims 2 / 2 Page 3 CN 122297526 A Tinea capitis Treatment Composition Technical Field

[0001] The present invention relates to tinea capitis treatment compositions, etc. Background Art

[0002] Tinea capitis is a skin infection caused by fungi of the genus Trichophyton (so-called Trichophyton). Based on the location of the infection, tinea capitis is classified as tinea pedis (athlete's foot), onychomycosis (nail fungus), tinea manuum (hand fungus), tinea corporis (body fungus), tinea cruris (groin fungus), tinea capitis (scalp fungus), etc.

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

[0004] Trichophyton fungi are transient fungi that parasitize the keratin or subcutaneous tissue of the skin. If Trichophyton fungi proliferate and erode into the stratum corneum, inflammation or blisters will occur, resulting in symptoms such as itching. It should be noted that Trichophyton fungi use keratin, a major component of keratin, as a nutrient source.

[0005] Conventionally, antifungal drugs such as itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, iriconazole, flusaconazole, terbinafine, and butenafine have been used as treatments for tinea unguium.

[0006] For example, Non-Patent Document 1 describes the following: Tinea corporis (tinea unguium) and tinea capitis (tinea capitis) are more common in pre-adolescent children. On the other hand, tinea corporis (tinea unguium), tinea cruris (tinea cruris), and tinea pedis (tinea pedis) are more common in young adults and adults. Since there are diseases showing the same clinical symptoms, it is sometimes preferred to make a diagnosis by potassium hydroxide preparation or culture. The treatment drugs can be appropriately selected as creams or oral preparations.

[0007] In addition, Patent Document 1 describes an invention relating to a topical preparation for the treatment of onychomycosis containing a) an antifungal active substance, b) a volatile component, c) a medium-chain fatty acid triglyceride, and d) ethyl lactate. According to Patent Document 1The topical formulation describes superior properties from the viewpoints of nail permeability, efficacy, and formulation properties by using medium-chain fatty acid triglycerides as non-volatile components and ethyl lactate as a permeation enhancer.

[0008] It should be noted that, compared with oral formulations, the topical formulation for treating onychomycosis described in Patent Document 1 is superior in terms of fewer systemic side effects and drug interactions.

[0009] Prior Art Documents Patent Document 1: International Publication No. 2019 / 088005.

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

[0011] Problem to be Solved by the Invention In this case, further development of a tinea pedis treatment drug is underway.

[0012] Means for Solving the Problem The present invention provides, for example, a tinea pedis treatment composition with the following formulation.

[0013] [1] A tinea treatment composition containing at least one strain selected from Staphylococcus hominis strain (page 1 / 29, CN 122297526 A), Staphylococcus haemolyticus strain, and Staphylococcus warneri strain.

[0014] [2] The tinea treatment composition according to [1] above contains a Staphylococcus hominis strain.

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

[0016] [4] The tinea 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 commissioned number: PTA-125203).

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

[0018] [6] The tinea treatment composition according to [5] above, wherein the hemolytic Staphylococcus aureus 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]According to the tinea treatment composition of [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] According to any one of [1] to [7] above, the tinea treatment composition contains a Staphylococcus aureus strain.

[0021] [9] According to the tinea treatment composition of [8] above, wherein the Staphylococcus aureus strain has a 16S rDNA containing a base sequence containing 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] According to the tinea treatment composition of [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 a fungus of the genus Trichophyton.

[0024]

[12] The tinea treatment composition according to

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

[0025]

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

[12] above is a topical application.

[0026]

[14] A method for manufacturing a composition for treating tinea pedis, comprising: a step (1) of collecting and culturing bacterial flora from at least one location on the ankle and sole of a person; a step (2) of isolating and culturing the cultured bacterial flora to obtain candidate bacteria; a step (3) of evaluating the anti-tinea pedis activity of the candidate bacteria and identifying bacteria with anti-tinea pedis activity; and a step (4) of preparing a composition for treating tinea pedis containing bacteria with said anti-tinea pedis activity; wherein the bacteria with said anti-tinea pedis activity contain at least one selected from Staphylococcus aureus strains, Staphylococcus hemolyticus strains, and Staphylococcus wartii strains.

[0027] Description of the effects of the invention 2 / 29 pages 5 CN 122297526 A According to the present invention, a new composition for treating tinea pedis is provided. According to a preferred embodiment of the present invention, it can be used even when the artificial chemical synthesis and extraction of the medicamentous compound is difficult or the medicamentous activity is based on the life reaction derived from live bacteria, and can have one or more of the following effects: prevention or reduction of side effects, prevention or inhibition of skin periphery of the affected area.Skin rashes, improve skin adhesion, maintain relief, prevent recurrence, reduce the number of applications, improve compliance, and achieve higher therapeutic effects.

[0028] [Figure 1] Typical examples of various evaluations of anti-Trichophyton floccosum activity.

[0029] [Figure 2] A graph showing the anti-Trichophyton floccosum activity results of various species of candidate bacteria (91 strains) obtained from the ankle and sole.

[0030] [Figure 3] Phylogenetic tree of known and candidate strains of Staphylococcus.

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

[0032] [Figure 5] Phylogenetic tree of known and candidate strains of Staphylococcus hemolyticus. Detailed Description

[0033] Hereinafter, the present invention will be described in detail.

[0034] <Composition for Treating Trichophyton Floccosum> The composition for treating trichophyton floccosum according to the present invention contains at least one strain selected from Staphylococcus aureus, Staphylococcus hemolyticus, and Staphylococcus variabilis. According to the present invention, a therapeutic composition for tinea can be used to treat tinea. It should be noted that, in this specification, "treatment" means preventing or inhibiting the proliferation of Trichophyton spp. fungi (so-called Trichophyton) by sterilization or bacteriostasis.

[0035] It is known that there are individual differences in the onset and recurrence of tinea. The inventors predicted that individual differences lie in the resident skin flora and studied the effects of resident skin flora and known strains of Staphylococcus spp. collected from the ankles and soles of healthy individuals on Trichophyton. The results showed that Staphylococcus hominis strains, Staphylococcus haemolyticus strains, and Staphylococcus warneri strains, which are resident skin flora, are effective in treating tinea. According to the present invention, a new therapeutic composition for tinea can be provided.

[0036] It should be noted that since Staphylococcus aureus strains, Staphylococcus hemolyticus strains, and Staphylococcus wartii strains are resident bacteria of the skin, 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 occur with conventional topical agents.

[0037] In addition, since Staphylococcus aureus strains, Staphylococcus hemolyticus strains, and Staphylococcus wartii strains are resident bacteria of the skin, they easily adhere to the skin. Therefore, for example, 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 number of applications.

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

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

[0040] [Staphylococcus aureus strain] Specification 3 / 29 pages 6 CN 122297526 A In one embodiment, the tinea treatment composition contains a Staphylococcus aureus strain.

[0041] The Staphylococcus aureus strain is a coagulase-negative staphylococcus (CNS) and is a Gram-positive bacterium. Staphylococcus aureus strains are resident bacteria of the skin. It is presumed that Staphylococcus aureus strains prevent or inhibit the proliferation or fixation of Trichophyton mentagrophytes by directly or indirectly damaging them, competing for their living space, etc., thereby killing or inhibiting them.

[0042] The Staphylococcus aureus strain preferably contains a Staphylococcus aureus strain with anti-Trichophyton mentagrophyte activity. That is, in a preferred embodiment, a tinea treatment composition containing a Staphylococcus aureus strain with anti-Trichophyton mentagrophyte 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 tinea.

[0043] There are no particular limitations on the Staphylococcus aureus strains, but strains with anti-Trichophyton floccosum activity are preferred, as they have a 16S rDNA containing the base sequence of sequence number 1 or a 16S rDNA containing a base sequence that has more than 95% identity with sequence number 1. As shown in the examples, sequence number 1 is the base sequence of the region between two common sequences common to the 16S rDNA of Staphylococcus genus. In this case, among the two common sequences, 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), 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 sequence numbers 1 to 18 can be determined by the methods described in the examples. Furthermore, when 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. When there are two or more types of 16S rDNA in equal quantities, for Staphylococcus aureus strains, the 16S rDNA with the base sequence having the highest identity with sequence number 1 is considered the 16S rDNA of that bacterium; for Staphylococcus hemolyticus strains, the 16S rDNA with the base sequence having the highest identity with sequence number 10 is considered the 16S rDNA of that bacterium; for Staphylococcus wartii strains, the 16S rDNA with the base sequence having the highest identity with sequence number 10 is considered the 16S rDNA of that bacterium; and for Staphylococcus aureus strains, the 16S rDNA with the base sequence having the highest identity with sequence number 10 is considered the 16S rDNA of that bacterium.The 16S rDNA with the highest identity to sequence number 17 is used as the 16S rDNA of this bacterium.

[0044] The base sequence with more than 95% identity to sequence number 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] As mutations in the base sequence having 95% or more identity with Serial No. 1, examples 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 above-mentioned substitutions, there are no particular limitations, but it is preferred to be a substitution of 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, the guanine at position 45 of sequence number 1 can be replaced by adenine (45G>A (also represented as G45A)), the thymine at position 139 can be replaced by cytosine (139T>C), the thymine at position 140 can be replaced by cytosine (140T>C), the adenine at position 309 can be replaced by cytosine (309A>C), and the cytosine at position 1265 can be replaced by thymine (1265C>T).

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

[0048] As for the above insertion, there is no particular limitation, it can be inserted between positions 1182 and 1183, and between positions 1235 and 1236 of sequence number 1. It should be noted that, in one embodiment, adenine (1182_1183insA (also represented as Δ1183A)) can be inserted between positions 1182 and 1183 of sequence number 1, and guanine (1235_...

[0049] (Page 4 / 29, Specification CN 122297526 A 1236insG) can be inserted between positions 1235 and 1236). Furthermore, the base sequence having more than 95% identity with the above-mentioned sequence number 1 can be a base sequence preserved by Staphylococcus aureus strains through natural mutation, or a base sequence obtained through artificial mutation treatment.It should be noted that artificial mutation treatments can include, for example, gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, nitrous acid treatment, etc. These mutation treatments can be used alone or in combination of two or more.

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

[0051] [Table 1] Mutation sites when the sequence number is based on sequence number 1 Sequence number 1 - Sequence number 2 139T>C, 140T>C Sequence number 3 1265C>T Sequence number 4 140T>C Sequence number 5 45G>A Sequence number 6 45G>A, 309A>C Sequence number 7 45G>A, 488delA Sequence number 8 45G>A, 1182_1183insA Sequence number 9 45G>A, 1235_1236insG As a Staphylococcus aureus strain with 16S rDNA containing the base sequence of sequence number 1, there are no particular restrictions, and Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, etc. can be listed.

[0052] There are no particular limitations on the Staphylococcus aureus strains that have anti-tinea capitis activity and contain a 16S rDNA containing a base sequence that has more than 95% identity with sequence number 1. 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, [retrieved on February 17, 2022], Internet.<https: / / www .ncbi .nlm .nih .gov / genome / browse / #! / prokaryotes / 2014 / > The strains described herein. Among them, strains belonging to Staphylococcus aureus preferably include those selected from Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus aureus ATCC700236, and Staphylococcus aureus.The strain comprises at least one of the following: FDAARGOS_748, Staphylococcus aureus J6, FDAARGOS_136, and Staphylococcus aureus A9; more preferably, it comprises at least one of the following: ATCC27844, ATCC27845, ATCC700236, and 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 easily prepared by the following method. Specifically, the method for preparing Staphylococcus aureus strains with anti-Trichophyton floccosum activity includes: a step (1) of 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); a step (2) of isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and a step (3) of evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine bacteria with anti-Trichophyton floccosum activity. Each step is described later. The bacterial flora on the ankle and sole of a person usually includes 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 easily prepared.

[0055] The concentration of Staphylococcus aureus strains in the composition for treating Trichophyton floccosum is preferably 10³ to 10¹¹ CFU / g, more preferably 10⁴ to 10¹⁰ CFU / g, and even more preferably 10⁵ to 10⁹ 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 culturing (refer to Clinical Microbiology Procedures Handbook Volume 2, Third Edition).

[0056] The morphology of the Staphylococcus aureus strain in the tinea treatment composition is not particularly limited, but is preferably in dry powder form (e.g., freeze-dried form, spray-dried form).

[0057] [Hemolytic Staphylococcus strains] In one embodiment, the tinea treatment composition contains hemolytic Staphylococcus strains.

[0058] The hemolytic Staphylococcus strains are coagulation-negative staphylococci (CNS) and are Gram-positive bacteria. Hemolytic Staphylococcus strains are resident bacteria of the skin. It is speculated that hemolytic Staphylococcus strains also prevent or inhibit the growth of Trichophyton mentagrophytes through direct or indirect damage or competition for its habitat.The proliferation of Trichophyton floccosum is inhibited by bactericidal or bacteriostatic agents.

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

[0060] There are no particular limitations on the hemolytic Staphylococcus strain, but it is preferred to have a 16S rDNA containing the base sequence containing sequence number 10 or a 16S rDNA containing a base sequence having more than 95% identity with sequence number 10 and having anti-Trichophyton floccosum activity. As shown in the example, sequence number 10 is the base sequence of the region between two common sequences common to the 16S rDNA of the genus Staphylococcus. At this point, of the two shared sequences mentioned above, the shared sequence on the 5' side is shared sequence 1 (AGCTTGC), and the shared sequence on the 3' side is shared sequence 2 (AAGCTGG), both of which are contained in sequence number 10. Here, for example, "16S rDNA containing the base sequence of sequence number 10" means that the 16S rDNA has the base sequence of sequence number 10 in its base sequence.

[0061] The base sequence having 95% or more identity with Serial No. 10 preferably 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 with Serial No. 10.

[0062] As mutations of the base sequence having 95% or more identity with Serial No. 10, substitution, deletion, insertion, addition, and combinations thereof can be listed. 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 above-mentioned substitutions, there are no particular limitations, but preferably at least one of 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-mentioned deletion, there are no particular restrictions; adenine at position 1210 (1210delA), adenine at position 1219 (1219delA), thymine at position 1225 (1225delT), and adenine at position 1263 (1263delA) of sequence number 10 may be deleted.

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

[0066] The base sequence having more than 95% identity with the above-mentioned sequence number 10 is also the same as that of the Staphylococcus aureus strain. It can be a base sequence retained by other hemolytic Staphylococcus strains through natural mutation, etc., or it can be a base sequence obtained through artificial mutation treatment. It should be noted that, as an example of artificial mutation treatment, methods such as gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, nitrous acid treatment, etc. can be listed. These mutation treatments can be used alone or in combination of two or more.

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

[0068] [Table 2] Mutation sites when the sequence number is based on sequence number 10: Sequence number 10 - Sequence number 11 96C>T Sequence number 12 109G>A Sequence number 13 398A>G, 952C>T Sequence number 14 398A>G Sequence number 15 398A>G, 952C>T, 1186C>T Sequence number 16 398A>G, 952C>T, 1117-1118insG, 1195-1196insC, 1210delA, 1219delA, 1225delT, 1234-1235insG, 1263delA As a 16S sequence containing the base sequence of sequence number 10 The hemolytic staphylococcal strains that can be used to generate rDNA are not particularly limited, and examples include hemolytic staphylococci ATCC29970 and hemolytic staphylococci ATCC700564.

[0069] In one embodiment, the hemolytic staphylococcal strain is not particularly limited, and examples include hemolytic staphylococci ATCC29970 and hemolytic staphylococci ATCC700564.Staphylococcus ATCC29970, hemolytic Staphylococcus ATCC700564, hemolytic Staphylococcus ATCC29969, hemolytic Staphylococcus SH_12, hemolytic Staphylococcus JCSC1435, etc. In addition to the above, as hemolytic Staphylococcus strains, they can be found using "NCBI," "online," saved on February 17, 2022, [searched on February 17, 2022], on the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1141 / > The strains described herein. Preferably, the hemolytic staphylococcal strain includes at least one selected from hemolytic staphylococcal ATCC29970, hemolytic staphylococcal ATCC700564, hemolytic staphylococcal ATCC29969, hemolytic staphylococcal SH_12, and hemolytic staphylococcal JCSC1435; more preferably, it includes at least one selected from hemolytic staphylococcal ATCC29970, hemolytic staphylococcal ATCC700564, and hemolytic staphylococcal 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 human staphylococcal strains and can be easily prepared by the following methods. Specifically, the method for preparing a hemolytic staphylococcal strain with anti-Trichophyton floccosum activity includes: a step (1) of 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); a step (2) of isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and a step (3) of evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine bacteria with anti-Trichophyton floccosum activity. The hemolytic staphylococcal strain with anti-Trichophyton floccosum activity may also be included in the bacterial flora of the ankle and sole of a person. Therefore, according to the above method, a hemolytic staphylococcal strain with anti-Trichophyton floccosum activity can be easily prepared.

[0071] The concentration of the hemolytic staphylococcal strain in the composition for treating Trichophyton floccosum is preferably 10³ to 10¹¹ CFU / g, more preferably 10⁴ to 10¹⁰ CFU / g, and even more preferably 10⁵ to 10⁹ CFU / g.

[0072] The morphology of the hemolytic staphylococcus strain in the tinea pedis treatment composition is not particularly limited, but is preferably in dry powder form (e.g., freeze-dried form, spray-dried form).

[0073] [Staphylococcus wartii] In one embodiment, the tinea pedis treatment composition contains a Staphylococcus wartii strain.

[0074] The Staphylococcus wartii strain is a coagulase-negative staphylococcus (coagulasenegat...Staphylococcus ivermectina (CNS) is a Gram-positive bacterium. Staphylococcus ivermectina strains are resident bacteria of the skin. It is speculated that Staphylococcus ivermectina strains also prevent or inhibit the proliferation of Trichophyton mentagrophytes through direct or indirect damage, competition for habitat, etc., thereby killing or inhibiting the bacteria.

[0075] The Staphylococcus ivermectina strain preferably includes a strain with anti-Trichophyton mentagrophyte activity. That is, in a preferred embodiment, a composition for treating tinea tinea containing a Staphylococcus ivermectina strain with anti-Trichophyton mentagrophyte activity is provided. The Staphylococcus ivermectina strain with anti-Trichophyton mentagrophyte activity can treat tinea tinea.

[0076] There are no particular limitations on the Staphylococcus ivermectina strain, but strains having a 16S rDNA containing a base sequence including sequence number 17 or more, or a 16S rDNA containing a base sequence having 95% or more identity with sequence number 17 and having anti-Trichophyton mentagrophyte activity, are preferred. As shown in the example, sequence number 17 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 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 95% or more 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] As mutations of the base sequence having 95% or more identity with Serial No. 17, substitution, deletion, insertion, addition, and combinations thereof can be listed. 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 the above-mentioned substitutions, there are no particular limitations, but cytosine at position 52 of sequence number 17 is preferred. It should be noted that, in one embodiment, cytosine at position 52 of sequence number 17 may be substituted with guanine (52C>G).

[0080] The base sequence having more than 95% identity with the above-mentioned sequence number 17 is also the same as that of Staphylococcus aureus strains.The base sequence can be one that is preserved by other Staphylococcus aureus strains through natural mutation, or it can be a base sequence obtained through artificial mutation treatment. It should be noted that, for example, artificial mutation treatments include gene manipulation, genome editing, ultraviolet irradiation, radiation irradiation, ethyl methanesulfonate (MES) treatment, N-methyl-N-nitrosoguanidine (NTG) treatment, nitrous acid treatment, etc. 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 site of sequence number 18 is shown in Table 3 below.

[0082] [Table 3] Mutation sites with sequence number 17 as the reference Description 8 / 29 pages 11 CN 122297526 A Sequence number 17 - Sequence number 18 52C>G As a *Staphylococcus aureus* strain having a 16S rDNA containing the base sequence of sequence number 17, there are no particular limitations; *Staphylococcus aureus* ATCC27836 can be listed.

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

[0084] The *Staphylococcus vulgaris* strains with anti-Trichophyton floccosum activity are the same as those of *Staphylococcus aureus* strains and can be easily prepared by the following methods. Specifically, the method for preparing a *Staphylococcus variabilis* strain with anti-Trichophyton floccosum activity includes: a step (1) of 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); a step (2) of isolating and culturing the cultured bacterial flora to obtain candidate bacteria; and a step (3) of evaluating the anti-Trichophyton floccosum activity of the candidate bacteria to determine the bacteria with anti-Trichophyton floccosum activity. Regarding *Staphylococcus variabilis* strains with anti-Trichophyton floccosum activity...Coccal strains may also be present in the bacterial flora of the ankle and sole of the foot. Therefore, according to the above method, Staphylococcus aureus strains with anti-Trichophyton floccosum activity can be easily prepared.

[0085] The concentration of Staphylococcus aureus strains in the tinea pedis treatment composition is preferably 10³ to 10¹¹ CFU / g, more preferably 10⁴ to 10¹⁰ CFU / g, and even more preferably 10⁵ to 10⁹ CFU / g.

[0086] The morphology of Staphylococcus aureus strains in the tinea pedis treatment composition is not particularly limited, but is preferably in dry powder form (e.g., freeze-dried form, spray-dried form).

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

[0088] Other Staphylococcus strains are not particularly limited, and can be listed as strains with human isolation reports such as *Staphylococcus saprophyticus*, *Staphylococcus kloosii*, *Staphylococcus lugdunensis*, *Staphylococcus capitis*, *Staphylococcus caprae*, *Staphylococcus epidermidis*, and *Staphylococcus aureus*.

[0089] As for *Staphylococcus saprophyticus* strains, 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 / 1350 / > The strains described in

[0090] include Staphylococcus kloosii strains such as those listed in "NCBI", "Online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 66884 / > The strains described in

[0091] include Staphylococcus lugdunensis strains such as "NCBI", "Online", February 17, 2022.Instruction manual, page 9 / 29, 12 CN 122297526 A, dated [retrieved on February 17, 2022], Internet.<https: / / www .ncbi .nlm .nih .gov / genome / browse / #! / prokaryotes / 2548 / > The strains recorded in

[0092] include Staphylococcus capitis strains such as those listed in "NCBI", "Online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 2054 / > The strains recorded in

[0093] include: Staphylococcus caprae strains such as those listed in NCBI, online, saved on February 17, 2022, [retrieved on February 17, 2022], and available on the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 1971 / > The strains recorded in

[0094] include Staphylococcus epidermidis, which 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

[0095] include Staphylococcus aureus strains such as "NCBI", "online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www .ncbi .nlm .nih .gov / genome / browse / #! / prokaryotes / 154 / > The strains described herein.

[0096] As other Staphylococcus strains, from the viewpoint of obtaining a higher therapeutic effect on tinea capitis, it is preferable to include at least one strain selected from Staphylococcus cephalosporin, Staphylococcus capitis, Staphylococcus epidermidis, Staphylococcus krillii, Staphylococcus ludensii, and Staphylococcus saprophyticus. More preferably, it includes at least one strain selected from Staphylococcus cephalosporin and Staphylococcus capitis. Particularly preferred is at least one strain selected from Staphylococcus cephalosporin and Staphylococcus capitis.The composition for treating tinea versicolor contains at least one of *Staphylococcus epidermidis* strains, preferably 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] The concentration of other *Staphylococcus* strains in the composition for treating tinea versicolor is not particularly limited, but is preferably 10³ to 10¹¹ CFU / g, more preferably 10⁴ to 10¹⁰ CFU / g, and even more preferably 10⁵ to 10⁹ CFU / g. It should be noted that when two or more other *Staphylococcus* strains are included, it is preferable that each species of the two or more other *Staphylococcus* strains falls within the above-mentioned range.

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

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

[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. Examples include Cutibacterium acnes (acne bacteria), Streptococcus pyogenes (group A beta-hemolytic streptococci), Pseudomonas aeruginosa (aeruginosa), Escherichia coli, Bacillus cereus, Enterococcus faecalis, Moraxella osloensis, etc. Instruction manual, page 10 / 29, 13 CN 122297526 A

[0101] As a strain of *Propionibacterium acnes* (acne bacteria), 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 / 1140 / > The strains described in

[0102] include Streptococcus pyogenes (Group A β-hemolytic streptococci). Examples of such strains include "NCBI", "Online", and 2022.Saved on the 17th of the month, [retrieved on February 17, 2022], Internet<https: / / www .ncbi .nlm .nih .gov / genome / browse / #! / prokaryotes / 175 / > The strains recorded in

[0103] include Pseudomonas aeruginosa strains such as those listed in "NCBI", "Online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 187 / > The strains recorded in

[0104] include Escherichia coli strains such as those from NCBI, online, saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 167 / > The strains recorded in

[0105] include Bacillus cereus strains such as those listed in "NCBI", "Online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 157 / > The strains recorded in

[0106] include Enterococcus faecalis strains such as those listed in "NCBI", "Online", saved on February 17, 2022, [retrieved on February 17, 2022], and the Internet.<https: / / www.ncbi.nlm.nih.gov / genome / browse / #! / prokaryotes / 808 / > The strains recorded in

[0107] include: "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 described herein.

[0108] Among them, from the viewpoint of obtaining a higher therapeutic effect on tinea versicolor, it is preferable that the strains include at least one strain selected from Pseudomonas aeruginosa (Pseudomonas aeruginosa), Escherichia coli, Bacillus cereus, Enterococcus faecalis, and Moraxella osloi. More preferably, the strains include at least one strain selected from Pseudomonas aeruginosa (Pseudomonas aeruginosa), Escherichia coli, and Enterococcus faecalis. Even more preferably, the strains include Pseudomonas aeruginosa ATCC27853 and Escherichia coli ATCC25922.At least one species of Enterococcus faecalis ATCC29212. Additionally, from the viewpoint of adjusting the balance of resident skin flora at the application site of the tinea treatment composition, it is preferable to include at least one species selected from Propionibacterium acnes (acne bacteria), Streptococcus pyogenes (group A beta-hemolytic streptococci), and Pseudomonas aeruginosa (Pseudomonas aeruginosa) strains, more preferably Propionibacterium acnes (acne bacteria) strains, and even more preferably Propionibacterium acnes ATCC6919. It should be noted that these other species can be used alone or in combination of two or more.

[0109] The concentration of other species in the tinea treatment composition is not particularly limited, but is preferably 10³ to 10¹¹ CFU / g, more preferably 10⁴ to 10¹⁰ CFU / g, and even more preferably 10⁵ to 10⁹ CFU / g. It should be noted that when two or more other species are included, it is preferable that each species of the two or more other species falls within the above-mentioned range.

[0110] The morphology of other strains in the tinea pedis treatment composition is not particularly limited, but is preferably in the form of dried powder (e.g., freeze-dried or spray-dried). 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 agent] The tinea pedis treatment composition may further contain an antifungal agent. By containing an antifungal agent, the therapeutic effect of tinea pedis can be further improved. Specification 11 / 29 pages 14 CN 122297526 A

[0112] As an antifungal agent, there are no particular limitations, and examples include itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, iriconazole, flusaconazole, terbinafine, butenafine, etc. These antifungal agents can be used alone or in combination of two or more.

[0113] The concentration of the antifungal agent in the tinea pedis treatment composition is not particularly limited, but 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] [Additives] The tinea pedis treatment composition may also contain additives. There are no particular limitations on additives, and examples include matrix, wetting agent, thickener, emulsifier, co-emulsifier, preservative, stabilizer, pH adjuster, etc.

[0115] (Matrix) Examples of matrix include hydrophobic matrix, hydrophilic matrix, 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 tinea pedis treatment composition.

[0116] There are no particular limitations on the above-mentioned hydrophobic matrix, and examples include squalane, liquid paraffin, light liquid paraffin, etc.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; isopropyl myristate, stearyl myristate, medium-chain triglycerides and other fatty acid esters.

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

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

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

[0120] There are no particular limitations on the wetting agent, and examples include petrolatum, glycerin, propylene glycol, 1,3-butanediol, etc.

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

[0122] (Thickening Agent) The thickening agent increases the viscosity or gels the composition for treating tinea capitis.

[0123] There are no particular limitations on the thickening agent, and examples include gelatin, agar, carrageenan, gum arabic, tragacanth gum, sodium alginate, propylene glycol alginate, acrylic polymer, polyvinyl alcohol (partially saponified), 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) ​​The emulsifier emulsifies the composition for treating tinea capitis into an oil-in-water (o / w) or water-in-oil (w / o) emulsion.

[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 alkyl betaine, alkyl amide betaine, etc.Alkali, alkyl sulfobetaine, imidazoline, lauryl dimethylaminoacetic acid betaine, alkyl diaminoethyl glycine, and other amphoteric surfactants, etc.

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

[0128] (Co-emulsifier) ​​The co-emulsifier stabilizes 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 limitations on the co-emulsifier, and examples include cetyl alcohol, stearyl alcohol, oleyl alcohol, isostearyl alcohol, etc.

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

[0131] (Preservative) The preservative prevents or inhibits contamination and decomposition of the composition for treating tinea capitis caused by foreign microorganisms.

[0132] As preservatives, there are no particular limitations, and examples include p-hydroxybenzoic acid, methylparaben, ethylparaben, propylparaben, butylparaben, 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 preservatives can be used alone or in combination of two or more.

[0134] (Stabilizer) The stabilizer prevents or inhibits the death, decomposition, and physical changes of the components contained in the composition for treating tinea capitis.

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

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

[0137] (pH adjuster) The pH adjuster maintains the stability of the components contained in the tinea pedis treatment composition and prevents or reduces the irritation of the tinea pedis treatment composition to organisms.

[0138] There are no particular limitations on the pH adjuster, and examples include lactic acid, acetic acid, acetate (sodium acetate, etc.), citric acid, citrate (sodium citrate, etc.), phosphoric acid, phosphate (sodium phosphate, etc.), diisopropanolamine, triisopropanolamine, triethanolamine, potassium hydroxide, sodium hydroxide, etc.

[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 corporis treatment composition as a topical agent, 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 corporis, and achieving bactericidal or bacteriostatic effects.

[0141] There are no particular limitations on the specific form of the topical agent, and ointments, creams, gels, lotions, etc. can be listed.

[0142] It should be noted that an ointment is a semi-solubilized preparation in which the active ingredient is dissolved or dispersed in a matrix and applied to the skin.Solid topical agents include oily ointments and water-soluble ointments. Creams are semi-solid topical agents that are emulsified into an oil-in-water (O / W) or water-in-oil (W / O) form and applied to the skin. Gels are gel-like topical agents applied to the skin, including aqueous gels and oily gels. Lotions are liquid topical agents made by dissolving or dispersing the active ingredient (Staphylococcus aureus strain, hemolytic Staphylococcus aureus strain, Staphylococcus vulgaris strain, etc.) in an aqueous solution.

[0143] [Trichophyton] The pathogen of Trichophyton is Trichophyton, specifically a fungus of the genus Trichophyton. Examples of Trichophyton include fungal strains of Trichophyton rubrum, Trichophyton mentagrophytes, and Trichophyton tonsurans.

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

[0145] Examples of fungal strains of *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 pathogenic fungus of *T. rubrum* preferably includes fungi of the genus *Trichophyton*, more preferably includes at least one fungal strain selected from *Trichophyton rubrum*, *Trichophyton mentagrophytes*, and *Trichophyton tonsurans*, further preferably includes a fungal strain of *Trichophyton rubrum*, and particularly preferably includes *Trichophyton rubrum* ATCC28188. It should be noted that the pathogenic fungus of *T. rubrum* 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 as tinea pedis (athlete's foot), onychomycosis (nail fungus), tinea manuum (hand fungus), tinea corporis (body fungus), tinea cruris (groin fungus), tinea capitis (scalp fungus), etc. Since the skin permeability 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 limitations on the application method of the tinea capitis treatment composition. When the tinea capitis treatment composition is a topical agent, it is applied to the affected area.

[0150] There are no particular limitations on the number of applications (times of application). It can be any one of the following: 3 times a day, 2 times a day, 1 time ... The tinea capitis treatment composition of the present invention contains at least one strain selected from Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii, which have excellent skin adhesion.Therefore, compared with conventional tinea treatments, the sustained effect of live bacteria fixation can maintain remission or prevent recurrence, and can also reduce the number of applications or improve patient compliance.

[0151] When the tinea treatment composition is for external use, the application amount is preferably 0.1 to 180 mg / cm2, more preferably 1 to 10 mg / cm2. In this case, the application amount of at least one strain of Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii in each application is preferably 100 to 108 CFU / cm2, more preferably 101 to 107 CFU / cm2, and even more preferably 102 to 106 CFU / cm2. It should be noted that when the tinea treatment composition contains two or more strains of Staphylococcus aureus, hemolytic Staphylococcus aureus, and Staphylococcus wartii, the total amount is preferably within the above range.

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

[0153] The above-mentioned tinea treatment composition can exert its therapeutic effect by use alone or in combination with other therapeutic agents.

[0154] Examples of the above-mentioned other therapeutic agents include oral and topical formulations. At this time, the instructions for the above-mentioned oral or topical preparation (page 14 / 29, CN 122297526 A) contain at least one of the following antifungal drugs: itraconazole, miconazole, clotrimazole, ketoconazole, bifonazole, lanoconazole, ruliconazole, iriconazole, flusaconazole, terbinafine, butenafine, etc. Two or more of the above-mentioned 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 with other therapeutic agents are provided. At this time, when the tinea pedis treatment composition is a topical preparation, the other therapeutic agents are preferably oral preparations. Furthermore, the tinea pedis treatment composition and other therapeutic agents may be applied simultaneously or sequentially.

[0156] <Method for Manufacturing a Tinea Pediatric Treatment Composition> According to one embodiment of the present invention, a method for manufacturing a tinea pedis treatment composition is provided. The above 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 symptoms of tinea corporis); a step (2) isolating and culturing the cultured bacterial flora to obtain candidate bacteria; a step (3) evaluating the antifungal activity of the candidate bacteria to identify bacteria with antifungal activity; and a step (4) preparing a mixture containing the above-mentioned antifungal activity.Step (4) of the composition for treating tinea pedis with bacteria that have anti-tinea pedis activity. At this time, the bacteria having the above-mentioned anti-tinea pedis activity contain at least one selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains. It should be noted that the above-mentioned Staphylococcus aureus strains are preferably Staphylococcus aureus strains with anti-tinea pedis activity. In addition, the above-mentioned hemolytic Staphylococcus aureus strains are preferably hemolytic Staphylococcus aureus strains with anti-tinea pedis activity. In addition, the above-mentioned Staphylococcus wartii strains are preferably Staphylococcus wartii strains with anti-tinea pedis activity. It should be noted that specific examples of Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains are as described above.

[0157] [Step (1)] Step (1) is a step of collecting bacterial flora from at least one location on the ankle and sole of a person (preferably a healthy person without tinea pedis symptoms) and culturing it. Bacterial flora are collected from the ankle and / or sole of a person using a swab or the like, and smeared on a culture medium for culturing. As a result, each strain forms a colony.

[0158] [Step (2)] Step (2) is a step of isolating and culturing the bacterial flora cultured in Step (1) to obtain candidate bacteria. Colonies formed by the culture in Step (1) are collected using platinum rings or the like and spread on a culture medium for isolation and culture. Thus, a large number of bacteria contained in the bacterial flora of the human ankle and / or sole can be isolated. It should be noted that the various bacteria cultured are candidate bacteria, preferably suspended in a bacterial stock solution and preserved as a skin bacterial library at low temperature (e.g., -100~-50°C, preferably -90~-70°C).

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

[0160] In Step (3), firstly, it is evaluated which candidate bacteria among the candidate bacteria isolated from the human ankle and / or sole have anti-Trichophyton floccosum activity. At this point, 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 anti-Trichophyton floccosum activity are selected and identified. There are no particular limitations on the identification method, and methods such as comparing 16S rDNA sequences and constructing a phylogenetic tree can be listed. For example, in the method of comparing 16S rDNA sequences, the bacteria can be identified by analyzing the 16S rDNA sequence of the target bacteria and using the DNA sequence data for NCBI BLAST search. In addition, in the method of constructing a phylogenetic tree, the bacteria can be identified by analyzing the 16S rDNA sequence of the bacteria and combining the 16S rDNA sequence data of known bacteria for analysis, and the target bacteria can be identified by the phylogenetic tree. It should be noted that in step (3), when identifying the target bacteria, Staphylococcus aureus strain, hemolytic Staphylococcus aureus strain, or Staphylococcus vulgaris strain, the bacteria can be identified by comparing the 16S rDNA sequence of the target bacteria with the 16S rDNA sequence data of known bacteria.In the case of Staphylococcus strains, the process may include calculating the 16S rDNA sequence of the target bacteria and confirming the identity of the DNA sequence of the target bacteria, etc.

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

[0163] [Step (4)] Step (4) is a step of preparing a tinea treatment 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 treatment 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 selected from Staphylococcus aureus strains, hemolytic Staphylococcus aureus strains, and Staphylococcus wartii strains, preferably containing at least one selected from Staphylococcus aureus strains with anti-Trichophyton floccosum activity, hemolytic Staphylococcus aureus strains with anti-Trichophyton floccosum activity, and Staphylococcus wartii strains.

[0164] <Treatment Method> According to one aspect of the present invention, a treatment method for tinea is provided. In this case, the above-mentioned treatment method includes applying the above-mentioned tinea treatment composition to the affected area. The tinea treatment composition and its application method are as described above. Examples

[0165] Hereinafter, examples are given to specifically illustrate the present invention, 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 medium and incubated aerobically at 35°C for one day. After incubation, representative colonies in terms of morphology, color, and dryness were collected using platinum rings.

[0167] Platinum rings containing the collected colonies were smeared onto Anero Columbia RS blood agar medium and isolated and incubated aerobically at 35°C for one day.

[0168] After incubation, the developed colonies were recovered using platinum rings, suspended in 25% glycerol bacterial stock solution, and stored at -80°C. It should be noted that a total of 91 candidate bacteria were collected. At this time, 52 candidate bacteria were obtained from the ankle, and [the remaining 52 were obtained from the sole].The number of candidate bacteria obtained was 39.

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

[0170] (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 hemolyticus 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 cephalosporin ATCC27840 (21) Staphylococcus capsulatum ATCC35538 ​​(22) 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 lentus ATCC700403 (29) Staphylococcus ludens ATCC49576 (30) Staphylococcus ludens ATCC43809 (31) Staphylococcus wortii ATCC27836 (32)Staphylococcus saprophyticus ATCC15305 3. Preparation of Evaluation Culture Medium In order to evaluate the anti-Trichophyton floccosum activity of the candidate and known strains, a fungal mixed dilution culture medium containing Trichophyton floccosum was prepared as the evaluation culture medium.

[0171] Specifically, one bead of Trichophyton floccosum (Trichophyton rubrum ATCC28188) stored in Microbank was spread onto potato dextrose agar (PDA) slant agar and incubated aerobically at 30°C.

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

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

[0174] 4. Evaluation of anti-Trichophyton spp. The anti-Trichophyton spp. activity was evaluated using the fungal mixed dilution medium as the evaluation medium.

[0175] Specifically, one bead containing candidate or known bacteria stored in a Microbank was taken and spread onto Anero Columbia RS blood agar medium. The bead was incubated at 35°C under aerobic conditions for 1-2 days. Bacteria were collected using a platinum ring (page 17 / 29 of the manual, 20 CN 122297526 A).

[0176] The platinum ring with the collected bacteria was streaked onto the fungal mixed dilution medium and incubated at 30°C under aerobic conditions until turbidity in the agar medium caused by Trichophyton spp. proliferation could be visually confirmed. After culturing, the proliferation-inhibiting regions formed in the fungal mixed dilution medium were observed. Based on the size of the proliferation-inhibiting regions, the anti-Trichophyton mentagrophytes activity of the candidate or known strains was evaluated according to the following criteria. It should be noted that typical examples of each evaluation are shown in Figure 1.

[0177] ++: Clear proliferation-inhibiting regions were observed +: No clear proliferation-inhibiting regions were observed, but proliferation inhibition of Trichophyton mentagrophytes was observed -: No proliferation inhibition of Trichophyton mentagrophytes was observed First, the results obtained for known strains (32 strains) are shown in Table 4 below.

[0178] [Table 4] Results of Bacterial Species and Strains: 1 Acinetobacter baumannii ATCC17978 - 2 Corynebacterium blueii JCM12208 - 3 Corynebacterium jejuni ATCC43734 - 4 Corynebacterium striatum NBRC15291 - 5 Enterococcus faecalis ATCC29212 ++ 6 Escherichia coli ATCC25922 ++ 7 Micrococcus luteus ATCC4698 - 8 Moraxella tarda NBRC14588 - 9 Neisseria gonorrhoeaeATCC19424 - 10 *Pseudomonas aeruginosa* ATCC27853 ++ 11 *Staphylococcus aureus* ATCC29213 - 12 *Staphylococcus epidermidis* ATCC12228 + 13 *Staphylococcus hemolyticus* ATCC29970 + 14 *Staphylococcus hominis* ATCC27844 ++ 15 *Streptococcus agalactiae* ATCC13813 - 16 *Streptococcus pyogenes* JCM12971 - 17 *Streptococcus pyogenes* ATCC12344 - 18 *Staphylococcus aureus* ATCC12600 + 19 *Staphylococcus epidermidis* ATCC14990 + 20 *Staphylococcus cephalosporin* ATCC27840 + 21 *Staphylococcus capsulatum* ATCC35538 ​​+ 22 *Staphylococcus coli* ATCC29974 - 23 Hemolytic Staphylococcus ATCC700564 ++ 24 Hemolytic Staphylococcus ATCC29969 ++ 25 Staphylococcus aureus ATCC27845 ++ 26 Staphylococcus aureus ATCC700236 ++ 27 Staphylococcus krill ATCC43959 + 28 Staphylococcus stoichioides ATCC700403 - Instructions for use, pages 18 / 29, 21 CN 122297526 A 29 Staphylococcus ludens ATCC49576 + 30 Staphylococcus ludens ATCC43809 + 31 Staphylococcus variabilis ATCC27836 ++ 32 Staphylococcus saprophyticus ATCC15305 + As shown in Table 4, among the known bacteria (32 strains), the results are: 9 strains are "++", 10 strains are "+", and 13 strains are "-".

[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 In order to analyze the relationship between the anti-Trichophyton mentagrophyte effect of the candidate bacteria and the species of the candidate bacteria, the candidate bacteria were identified. The identification of candidate bacteria was performed by comparing the 16S rDNA sequences.

[0181] Specifically, according to the instructions of NucleoSpin Microbial DNA, total DNA was extracted and purified from the 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, according to the instructions of the Bacterial 16S rDNA PCR kit, using the purified total DNA as a template, the 16S rDNA sequence region was amplified by PCR using the included primers F1 (serial number 19) and R2 (serial number 20). The amplified DNA fragment (approximately 1,500 μL) was...The DNA fragments (16S rDNA template samples) were purified using NucleoSpin gel and a PCR cleaning kit, and the DNA concentration and purity were determined using NanoDrop as an ultra-micro spectrophotometer. The amplified DNA fragment size as theoretically expected was confirmed by agarose gel electrophoresis.

[0183] The amplified DNA fragments (16S rDNA template samples) were mixed with 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 by Sanger sequencing. After assembling the obtained sequencing sequences, the candidate bacterial species were identified by NCBI BLAST search targeting the base sequence (approximately 1,240–1,280 bp) between the common sequence 1 (AGCTTGC) and common sequence 2 (AAGCTGG) of Staphylococcus genus 16S rDNA. 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). Furthermore, in the obtained sequencing sequences, where common sequence 1 (AGCTTGC) is absent, it is replaced with the sequence GGTTGG and TGCTTGC; where common sequence 2 (AAGCTGG) is absent, it is replaced with the sequence GAGTTGG, AAGTCGG, GAAGTCGA, AAGTCAG, GAGGCTGG, AAGGGGGG, AAGCTGG, CAGTCAG, and CCGCTGG. Candidate bacterial species are identified by performing a BLAST search on the regions containing each common sequence (approximately 1,240–1,280 bp) using NCBI. It should be noted that, in cases where the obtained sequencing sequences do 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, GAGCTGG, AAGGGGGG, AAGCTGG, 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 hit in the BLAST search are selected. Here, if the selected top 10 species include multiple results from the same genus but different species, the results are not considered.In this case, identification was performed by genus name (Staphylococcus species and Micrococcus sp.). The case where no species meeting the criteria was identified is considered unknown.

[0185] The identified candidate bacteria and the results of their anti-Trichophyton floccosum activity are shown in Tables 5 and 6 below. Specification 19 / 29 pages 22 CN 122297526 A Specification 20 / 29 pages 23 CN 122297526 A

[0186] In Tables 5 and 6, the identified candidate bacteria are organized by species, and the results obtained by analyzing the proportion of anti-Trichophyton floccosum activity shown by each species are shown in Figure 2. As shown in Figure 2, Staphylococcus aureus and hemolytic Staphylococcus showed "++" results of 100% and 86%, respectively.

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

[0188] [Table 7] Specification 21 / 29 pages 24 CN 122297526 A According to the results in Table 7, among the 52 strains from the ankle, 33% were "++", 31% were "+", and 37% were "-", while among the 39 strains from the sole, 15% were "++", 41% were "+", and 44% were "-". It can be seen that the proportion of strains showing "++" on the ankle is about twice that on the sole.

[0189] 6. The phylogenetic tree of Staphylococcus was constructed with reference to 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 16S rDNA sequences of 27 Staphylococcus species. The phylogenetic tree was constructed for 27 known Staphylococcus strains (Table 8 below) and 45 strains of Staphylococcus identified 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 CLC sequence viewer (ver. 8.0) was used in the construction of the phylogenetic tree.

[0190] [Table 8] Numbered Bacterial Species and Strains 1 Staphylococcus arlettae ATCC43957 2 Staphylococcus aureus ATCC12600 3 Staphylococcus auriculata4. Staphylococcus auricularis (ATCC33753) 5. Staphylococcus capitis (ATCC27840) 6. Staphylococcus goatii (ATCC35538) 7. Staphylococcus carnosus (CIP103274) 8. Staphylococcus chromogenes (CBCC1462) 9. Staphylococcus coli (GH137) 10. Staphylococcus delphini (ATCC49171) 11. Staphylococcus epidermidis (ATCC14990) (Instructions for use, pages 22 / 29, CN 122297526 A) 12. Staphylococcus equorum (ATCC43958) 13. Staphylococcus felis (GD521) 14. Staphylococcus gallinarum (VIII1) 15. Hemolytic Staphylococcus ATCC29970 16. Staphylococcus hominis ATCC27844 17. Staphylococcus hyicus ATCC11249 18. Staphylococcus intermedius H11 / 68 19. Staphylococcus krill ATCC43959 20. Staphylococcus sluggish JCM2426 21. Staphylococcus ludens ATCC43809 22. Staphylococcus muscae MB4 23. Staphylococcus saprophyticus ATCC15305 24. Staphylococcus schleiferi DSM4807 25. Staphylococcus sciuri DSM20345 26. Staphylococcus simulans MK148 26 Staphylococcus aureus ATCC27836 27 Staphylococcus xylosus JCM2418 [Table 9] Specification 23 / 29 pages 26 CN 122297526 A The phylogenetic tree in Figure 3 is compared with the anti-Trichophyton floccosum activity results of the candidate bacteria, and they are roughly divided into cluster 1, which mainly shows "-" results, cluster 2, which mainly shows "++" results, and cluster 3, which mainly shows "+" results. Among them, cluster 2 consists of Staphylococcus aureus and hemolytic Staphylococcus.

[0191] Therefore, 6 strains of known Staphylococcus aureus and hemolytic Staphylococcus were prepared (Table 10 below) and candidate bacteria.A phylogenetic tree of 15 strains of bacteria identified as Staphylococcus humanis or Staphylococcus hemolyticus (the bacteria extracted from Table 9 are shown in Table 11 below) was prepared. The phylogenetic trees of Staphylococcus humanis and Staphylococcus hemolyticus are shown in Figures 4 and 5.

[0192] [Table 10] Number of Bacterial Species / Strain 1 Staphylococcus aureus ATCC27844 Instruction Manual 24 / 29 Page 27 CN 122297526 A 2 Staphylococcus aureus ATCC27845 3 Staphylococcus aureus ATCC700236 4 Staphylococcus aureus A9 5 Hemolytic Staphylococcus aureus ATCC29970 6 Hemolytic Staphylococcus aureus ATCC700564 [Table 11] Number of Bacterial Species / Results 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 Staphylococcus aureus ++ 22 Hemolytic Staphylococcus aureus ++ 23 Hemolytic Staphylococcus aureus ++ 24 Hemolytic Staphylococcus aureus ++ 25 Hemolytic Staphylococcus ++ 33 Hemolytic Staphylococcus ++ 35 Hemolytic Staphylococcus + 7. 16S rDNA sequence (1) The 16S rDNA sequence of Staphylococcus aureus strain was confirmed by the ATCC database. The 16S rDNA sequence of Staphylococcus aureus ATCC27844, a well-known bacterium, was confirmed, and the base sequence of the region sandwiched between shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. As a result, the 16S rDNA sequence of Staphylococcus aureus ATCC27844 has the base sequence of sequence number 1 (genome location of Staphylococcus aureus ATCC27844: 716544~717811). It should be noted that since sequence number 1 contains shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG), the 5' end of the base sequence of sequence number 1 is shared sequence 1 (AGCTTGC), and the 3' end is shared sequence 2 (AAGCTGG).

[0193] Furthermore, for the 16S rDNA sequences of candidate strains identified as Staphylococcus aureus strains numbered 2, 3, 37, 40, 45, 62, 64, and 74, the base sequences (approximately 1,240~1,280 bp) of the region sandwiched between shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) were confirmed using the same method as described in "5. Identification of Candidate Strains" above. As a result, the 16S rDNA sequences of candidate strains numbered 3, 40, 45, 64, and 74 have the base sequence of sequence number 5, and the 16S rDNA sequences of candidate strain number 2 have the base sequence of sequence number 5.The rDNA sequence has the base sequence of sequence number 7, the 16S rDNA sequence of candidate bacterium number 37 has the base sequence of sequence number 8, and the 16S rDNA sequence of candidate bacterium number 62 has the base sequence of sequence number 9.

[0194] In addition, the 16S rDNA sequences of Staphylococcus aureus ATCC27845 and Staphylococcus aureus ATCC700236, which are known bacteria, were confirmed by the ATCC database, and the base sequence of the region sandwiched by shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. In addition, the 16S rDNA sequence of Staphylococcus aureus A9 (ATCC commission number: Specification 25 / 29 pages 28 CN 122297526 A PTA-125203) was confirmed by the PATRIC database, and the base sequence of the region sandwiched 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) The 16S rDNA sequence of the hemolytic staphylococcus strain was confirmed using the ATCC database. The 16S rDNA sequence of hemolytic staphylococcus ATCC29970, a known bacterium, was confirmed, and the base sequence of the region sandwiched between shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. As a result, the 16S rDNA sequence of hemolytic staphylococcus ATCC29970 has the base sequence of sequence number 10 (genomic location of hemolytic staphylococcus ATCC29970: 807302~808569).

[0196] Furthermore, the 16S rDNA sequences of candidate strains 22-25, 33, and 35, identified as hemolytic Staphylococcus strains, were confirmed using the same method as described in "5. Identification of Candidate Strains" above, identifying the base sequence (approximately 1,240-1,280 bp) of the region enclosed by 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 of sequence number 13, and the 16S rDNA sequence of candidate strain 35...The 16S rDNA sequence of candidate bacterium number 23 has the base sequence of sequence number 14. The 16S rDNA sequence of candidate bacterium number 20, identified as a hemolytic staphylococcal strain, has a common sequence 1 (AGCTTGC) but lacks a common sequence 2 (AAGCTGG). Because the 16S rDNA sequence of candidate bacterium number 20 has a substitution sequence (GAGCTGG) for common sequence 2, the base sequence (approximately 1,267 bp) of the region enclosed by common sequence 1 (AGCTTGC) and the substitution sequence (GAGCTGG) for 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, and the base sequence of the region sandwiched between shared sequence 1 (AGCTTGC) and shared sequence 2 (AAGCTGG) was confirmed. As a result, the 16S rDNA sequence of Staphylococcus hemolyticus ATCC700564 has the base sequence of sequence number 10 (genome 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] Evaluation results of antifungal activity against Trichophyton mentagrophytes: Staphylococcus aureus ATCC27844 Serial No. 1 ++ Staphylococcus aureus Serial No. 2 Serial No. 7 ++ Staphylococcus aureus Serial No. 3 Serial No. 5 ++ Staphylococcus aureus Serial No. 37 Serial No. 8 ++ Staphylococcus aureus Serial No. 40 Serial No. 5 ++ Staphylococcus aureus Serial No. 45 Serial No. 5 ++ Staphylococcus aureus Serial No. 62 Serial No. 9 ++ Staphylococcus aureus Serial No. 64 Serial No. 5 ++ Staphylococcus aureus Serial No. 74 Serial No. 5 ++ Staphylococcus aureus ATCC27845 Serial No. 1 ++ Staphylococcus aureus ATCC700236 Serial No. 5 ++ Instructions for use 26 / 29 pages 29 CN 122297526 A Staphylococcus aureus A9 Serial No. 5 ND Hemolytic Staphylococcus aureus ATCC29970 Serial No. 10 + Hemolytic Staphylococcus aureus, serial number 20, sequence number 15, ++; Hemolytic Staphylococcus aureus, serial number 22, sequence number 13, ++; Hemolytic Staphylococcus aureus, serial number 23, sequence number 16, ++; Hemolytic Staphylococcus aureus, serial number 24, sequence number 13, ++; Hemolytic Staphylococcus aureus, serial number 25, sequence number 13, ++; Hemolytic Staphylococcus aureus, serial number 33, sequence number 13, ++; Hemolytic Staphylococcus aureus, serial number 35, sequence number 14, +; Hemolytic Staphylococcus aureus 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" recorded in serial number 5 means that the G (guanine) at position 45 of serial number 1 is mutated to A (adenine). Additionally, for example, "488delA" recorded in serial number 7 means that the A (adenine) at position 488 of serial number 1 is deleted. Furthermore, for example, "1182_1183insA" recorded in serial number 8 means that A (adenine) is inserted between positions 1182 and 1183 of serial number 1.

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

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

[0202] 8. Effects against other Trichophyton floccosum In 4 to 7 above, a fungal mixed dilution medium containing Trichophyton rubrum ATCC28188, which is Trichophyton floccosum, was used as the evaluation medium (refer to “3. Preparation of evaluation medium” above). Here, the effects on other Trichophyton species (Trichophyton rubrum ATCC28188) were evaluated.

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

[0204] (2)Evaluation of antifungal activity against other Trichophyton species: Staphylococcus aureus ATCC27844, Staphylococcus aureus ATCC27845, Staphylococcus hemolyticus ATCC29969, Staphylococcus hemolyticus ATCC700564, and Staphylococcus wartii ATCC27836, which have been previously evaluated for their antifungal activity against Trichophyton rubrum ATCC28188, were evaluated using a mixed dilution culture medium containing the aforementioned five fungi.

[0205] The evaluation method was performed using the same method as described in "4. Evaluation of antifungal activity against Trichophyton rubrum". The results obtained are presented together with the results of the antifungal activity against Trichophyton rubrum ATCC28188 in Table 13 below. Instruction manual, pages 28 / 29, 31, CN 122297526 A

[0206] As shown in Table 14, it also exhibits the same antifungal activity as *Trichophyton rubrum* ATCC28188 among five other *Trichophyton* species. Instruction manual, pages 29 / 29, 32, CN 122297526 A, Figure 1, Figure 2, Appendix 1 / 3, page 33, CN 122297526 A, Figure 3, Appendix 2 / 3, page 34, CN 122297526 A, Figure 4, Figure 5, Appendix 3 / 3, page 35, CN 122297526 A Abstract A novel composition for treating ringworm is provided. The composition for treating ringworm contains 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*.

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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