JP2016210751A - Biofilm formation inhibitor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide formulations that can inhibit biofilm formation by microorganisms, such as acne bacteria.SOLUTION: A biofilm formation inhibitor that effectively inhibits biofilm formation by microorganisms contains basic amino acids, which consists of at least one or more selected from arginine, histidine, lysine, and ornithine. Particularly, the biofilm formation inhibitor is used for the treatment or prevention of repetitive acne and further acne associate with inflammation in the hair follicle.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a preparation capable of suppressing biofilm formation by microorganisms such as acne.

  Acne is a skin disease that causes comedones, papules, nodules and pustules, sometimes accompanied by inflammation of the hair follicles, and is mainly found on the face, chest and back. Acne is known to be caused mainly by acne bacteria (Propionibacterium acnes, P. acnes), which are resident in the skin.

  Acne is an anaerobic bacterium that lives deep inside the hair follicle. Acne bacteria proliferate using sebum accumulated in the hair follicle as a nutrient source, and when it gets worse, it causes inflammation. Due to inflammation, pus accumulates in the hair follicle and becomes a skin rash. Acne that is known to be associated with acne bacteria includes acne vulgaris that is easily seen in adolescence, pustular acne mainly composed of pustules, cystic acne mainly composed of cysts, and acne vulgaris. There are congestive acne and electric shock acne that are severe cases. For the treatment of acne, various external preparations and oral preparations have been developed for the purpose of suppressing sebum production, preventing infection, suppressing inflammation, normalizing keratinization, and the like.

  Some bacteria form an accumulation of bacterial secretions (polysaccharides, etc.) called biofilm and continue to adhere to the adherend with viscosity. As a familiar example, it is known that the slime of the sink is a biofilm formed by bacteria. In addition, plaque that occurs between teeth or at the boundary between teeth and gums is considered as a kind of biofilm.

  Acne bacteria are also known to form biofilms. Since the biofilm is accumulated with polysaccharides secreted from acne bacteria, it is difficult for drugs such as antibacterial agents to penetrate inside, and the effect is reduced. Therefore, the presence or absence of biofilm formation affects the pathogenicity of various bacterial infections (Non-patent Document 1). Development of antibacterial agents that are highly effective against biofilms, plaques, and the like is also underway. Patent Document 1 discloses antibacterial preparations effective against Staphylococcus aureus, mutans bacteria, Escherichia coli, and the like.

Japanese Patent No. 3717960

Tom Coenye et al., Research in Microbiology 158 (2007) 386-392

  However, the development of biofilm formation by microorganisms, particularly against acne bacteria that form a biofilm deep in the hair follicle, has not been sufficiently developed yet. Therefore, an object of this invention is to provide the formulation with the high effect which suppresses such biofilm formation.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have shown that basic amino acids have a particularly remarkable inhibitory effect on biofilm formation by microorganisms that form biofilms, particularly acne that is the cause of acne. I found out that it works. The present invention has been completed by further studies based on such knowledge.

  That is, the present invention relates to a biofilm formation inhibitor containing basic amino acids.

  Said biofilm formation inhibitor may be used in order to suppress biofilm formation of acne bacteria.

  The basic amino acids may be at least one selected from the group consisting of arginine, histidine, lysine and ornithine.

  The basic amino acids may be L-form amino acids.

  The biofilm formation inhibitor can be applied to the skin or the oral cavity.

  The biofilm formation inhibitor can be used for the treatment or prevention of acne.

  The biofilm formation inhibitor can be used for the treatment or prevention of acne in dry skin.

  The biofilm formation inhibitor can be used for the treatment or prevention of acne that tends to be repeated.

  The biofilm formation inhibitor can be used for the treatment or prevention of acne accompanied by inflammation in the hair follicle.

  The biofilm formation inhibitor may have a pH of 3.0 to 8.0.

  The biofilm formation inhibitor may further contain a surfactant.

  The biofilm formation inhibitor described above can be used against acne vulgaris, pustular acne, cystic acne, concentrated acne or electric acne.

  According to the present invention, biofilm formation formed by microorganisms, particularly acne bacteria, can be effectively suppressed.

It is a graph which shows the result of having evaluated the biofilm formation suppression rate (%) using the various amino acid compounding preparations adjusted to pH7. It is a graph which shows the result of having evaluated the biofilm formation suppression rate (%) using various amino acid compound combination preparations adjusted to pH5.

  Hereinafter, the present invention will be described in detail. It should be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified.

  The present invention relates to a biofilm formation inhibitor containing basic amino acids.

  The biofilm formation inhibitor of the present invention can be used to suppress biofilm formation by various microorganisms. Although it does not specifically limit as microorganisms which form a biofilm, For example, bacteria, such as acne bacteria, mutans bacteria, S. epidermidis, Staphylococcus aureus, Escherichia coli, ringworm, candida, malassezia, black mold (for example, And fungi such as Aspergillus oryzae and Cladosporium). From the viewpoint that a higher effect can be expected, the biofilm formation inhibitor of the present invention is preferably used against bacteria, more preferably used against acne or mutans, and particularly preferably acne. Used for.

  In the present specification, “inhibition of biofilm formation” refers to the inhibition of the formation of biofilm by microorganisms, whether complete or partial, or the growth rate of the formed biofilm. It means to reduce. In the case of detecting the suppression of acne biofilm formation, the method is not limited as long as it is a known method, but if it is the method described in the test examples described later, the biofilm formation inhibition rate is at least 20% or more. It is preferably 30% or more, more preferably 40% or more, still more preferably 50% or more, and particularly preferably 60% or more.

  As used herein, “Acne fungus” refers to Propionibacterium acnes (P. acnes), a genetic analog thereof, or a mutant thereof.

  As used herein, “mutans bacterium” refers to Streptococcus mutans (S. mutans), a genetic analog thereof, or a mutant thereof.

  As used herein, “Staphylococcus epidermidis” refers to Staphylococcus epidermidis (S. epidermidis), a genetic analog thereof, or a mutant thereof.

  As used herein, “S. aureus” refers to Staphylococcus aureus (S. aureus), a genetic analog thereof, or a mutant thereof.

  As used herein, “Escherichia coli” refers to Escherichia coli (E. coli), a genetic analog thereof, or a mutant thereof. As long as E. coli forms a biofilm, the types of surface antigens (O antigen and H antigen) are not limited.

  In the present specification, “Rhizobium fungi” refers to Trichophyton rubrum (T. rubrum), Trichophyton mentagrophytes (T. mentagrophytes), Trichophyton tonsurans (T. tonsurans) and the like, or genetic analogs thereof Or it refers to those mutants.

  As used herein, “Candida” refers to Candida fungi including Candida albicans (C. albicans) and the like, their genetic analogs, or mutants thereof.

In the present specification, “Malassezia” refers to Malassezia furfur (M. furfur)
, Malassezia globosa (M. globosa), Malassezia restricta (M. restricta), Malassezia pachydermatis (M. pachydermatis), Malassezia dermatis (M. dermatis), Malassezia japonica (M. japonica), Malassezia yamatoensis (M.yamatoensis) It refers to the genus Malassezia or its genetic analogues or mutants thereof.

  In the present specification, “Astragalus fungi” refers to Aspergillus oryzae (A. oryzae) and the like, fungi belonging to the genus Aspergillus, genetic analogs thereof, or mutants thereof.

  In the present specification, “Cladosporia” refers to Cladosporium cladosporioides (C. cladosporioides), Cladosporium sphaerospermum (C. sphaerospermum), etc., or a genetic analog thereof or a mutation thereof. Refers to the body.

  In the present specification, the basic amino acids are not limited as long as they have an inhibitory effect on biofilm formation. Arginine, lysine, histidine, ornithine, δ-hydroxylysine, alkanolamine (trometamol, Monoethanolamine, diethanolamine, triethanolamine, etc.), sulfoalkylpiperazine (HEPES, 1,4-bis (2-sulfoethyl) piperazine, 1,4-bis (3-sulfopropyl) piperazine, 1,4-bis (4 -Sulfobutyl) piperazine etc.), sulfoalkylalkylene diamine (N, N'-bis (3-sulfopropyl) ethylenediamine etc.), epsilon aminocaproic acid, 2-amino-2-methyl-1-propanol (AMP), 2-amino -2-methyl-1,3-propyl Propanediol, organic amines such as meglumine, urea and creatinine, or the like of these pharmaceutically acceptable salts, and the like. In the present invention, as the basic amino acids, either L-form and / or D-form may be used, but L-form is preferably used.

  As the basic amino acids, basic amino acids are preferable from the viewpoint of effectively suppressing biofilm formation (in particular, biofilm formation of acne), and at least selected from the group consisting of arginine, histidine, lysine and ornithine. One or more components are more preferred, arginine and / or histidine are more preferred, and arginine is particularly preferred. Basic amino acids may be synthesized by a known method or may be obtained and used commercially. Moreover, basic amino acids may be used alone or in any combination of two or more. In addition, since the basic amino acids in the biofilm formation inhibitor are not antibiotics, they are also effective against clinically resistant microorganisms that are partially or completely resistant to one or more antibiotics. is there. Acne bacteria are inherently resident bacteria of the skin, and have a role of suppressing the invasion and propagation of other pathogenic bacteria by keeping the skin weakly acidic. Therefore, excessively sterilizing and suppressing acne bacteria with a powerful drug such as antibiotics may cause a burden on the skin. By suppressing the biofilm formation of acne bacteria, it is possible to increase the penetration of various drugs and effectively sterilize and inhibit acne bacteria without using a powerful drug. Or, by suppressing the biofilm formation of acne bacteria, various drugs (for example, antibacterial agents including anti-acne bacteria as described later, keratin / sebony dissolving agents, anti-inflammatory agents, antibiotics, antipruritic agents, topical Anesthetics, vitamins, wound healing agents, etc.) can be used in small doses.

  In the present specification, examples of the pharmaceutically acceptable salt of basic amino acids include salts with alkali metal salts, alkaline earth metal salts, organic bases, and the like, such as sodium, potassium, calcium, magnesium, A salt with ammonium or diethanolamine, ethylenediamine and the like can be mentioned. Furthermore, ammonia, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris (hydroxymethyl) aminomethane, N, N-bis (hydroxyethyl) piperazine, 2-amino-2-methyl-1-propanol, ethanolamine, Salts of amines such as N-methylglucamine, L-glucamine; and salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; methanesulfonic acid, benzenesulfonic acid, paratoluenesulfone Acid, acetic acid, propionic acid, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid , Salts with organic acids such as salicylic acid; or aspartic acid, glutamic acid, etc. And salts with sexual amino acid may also be mentioned. “Pharmaceutically acceptable salt” may include solvates or hydrates of salts.

  The content of the basic amino acids can be 0.0001% by weight or more based on the total amount of the preparation, as an example, from the viewpoint of more effectively suppressing biofilm formation. It is preferably at least wt%, more preferably at least 0.005 wt%, even more preferably at least 0.01 wt%, particularly preferably at least 0.05 wt%. Most preferably, it is 1% by weight or more. The content of basic amino acids can be, for example, 30% by weight or less, preferably 25% by weight or less, more preferably 20% by weight or less, based on the total amount of the preparation. It is preferably 15% by weight or less, particularly preferably 10% by weight or less. The content of basic amino acids can be, for example, 0.0001% to 30% by weight, preferably 0.001% to 25% by weight, based on the total amount of the formulation. More preferably, the content is 0.005 wt% to 20 wt%, particularly preferably 0.01 wt% to 15 wt%, and most preferably 0.1 wt% to 10 wt%.

  In the present invention, the pH of the biofilm formation inhibitor is appropriately set according to the type and content of the ingredients, the application, the formulation form, the method of use, and the like. As an example, when used as a biofilm formation inhibitor against acne bacteria, the pH is not limited from the viewpoint that basic amino acids can more effectively inhibit the biofilm of acne bacteria, but the pH is 3.0. To 8.0, preferably 3.1 to 7.8, more preferably 3.2 to 7.6, and even more preferably 3.3 to 7.4. Preferably, the pH is 3.4 to 7.2, particularly preferably 3.5 to 7.

  In this invention, the viscosity of a biofilm formation inhibitor is suitably set according to the kind and content of a compounding component, a use, a formulation form, a usage method, etc. The viscosity of the biofilm formation inhibitor of acne is not limited from the viewpoint that the active ingredient penetrates into the biofilm and the preparation remains in the affected area, but is preferably 0.001 mPa · s to 500000 mPa · s. More preferably 0.005 mPa · s to 450,000 mPa · s, still more preferably 0.01 mPa · s to 400000 mPa · s, and particularly preferably 0.05 mPa · s to 350,000 mPa · s. And most preferably 0.1 mPa · s to 300,000 mPa · s. The viscosity is not particularly limited as long as it is a known method, but the viscosity at 20 ° C. measured with a rotational viscometer (RB-80H viscometer, manufactured by Toki Sangyo Co., Ltd., rotor: H7, rotation speed: 10 rpm). It can be.

  In the present specification, the biofilm formation inhibitor is not particularly limited as long as it is a known form as a pharmaceutical, a quasi-drug, or a cosmetic, but it can be used as an external preparation. Such external preparations impregnated biofilm formation inhibitors in sheets such as liquids, suspensions, emulsions, creams, ointments, gels, liniments, lotions, aerosols, powders, and non-woven fabrics. Depending on the form of the sheet or the like, it can be formulated by a known method. Preferably, it is in the form of a solution, suspension, emulsion, or cream, and more preferably in the form of a solution, emulsion, or cream. By formulating in such a form, biofilm formation can be effectively suppressed.

  In the present specification, the biofilm formation inhibitor can be used for topical administration of the skin surface such as the face, perilip, scalp, neck, chest, back, abdomen, arms, oral cavity, buttocks, buttocks, pubic area, etc. . When used as an inhibitor of acne biofilm formation, it can be administered locally for various acne conditions, such as microcomedones (corn plug generation, early plugging of the horn plugs) and so-called pimples (adults) It can be used as a prophylactic and / or therapeutic agent for acne), white acne (white papules), black acne (open comedones), red acne with inflammation (red papules), or yellow acne (purulent acne). The acne biofilm formation inhibitor should be used as a preventive and / or therapeutic agent for acne vulgaris, pustular acne, cystic acne, congestive acne, and electric acne. Can do. Acne is sometimes called open or occlusive depending on whether the hair follicles are opened or closed by sebum or keratin plugs. Can also be used. The biofilm formation inhibitor of Acne can also be used for acne caused by atopic dermatitis. Acne bacteria are also known as causative bacteria for sarcoidosis, and therefore can be used as a preventive and / or therapeutic agent in affected areas of the skin where sarcoidosis granulomatous lesions have occurred. In this specification, the biofilm formation inhibitor of an acne bacterium is used effectively when the acne bacterium grows in or around the hair follicle to form a biofilm.

  Dry skin, often seen in adult acne, is prone to excessive sebum secretion and clogged plugs, and changes in the surrounding environment and stress may increase biofilm formation by acne bacteria. Moreover, when biofilm formation increases, there is a concern that acne bacteria protected by biofilm may easily develop acne symptoms. It is known that the increase in biofilm formation leads to an increase in lipase activity, and lipase decomposes triglycerides in sebum to produce free fatty acids, causing inflammation in the hair follicle. Therefore, the biofilm formation inhibitor of the present invention can treat and / or prevent acne in such dry skin, treat and / or prevent acne that tends to be repeated, treat and / or prevent acne with inflammation in the hair follicle. It can be used effectively for prevention. The biofilm inhibitor of acne bacteria can be used as a pharmaceutical, a quasi-drug, or a cosmetic for the purpose of such prevention and / or treatment.

  When a biofilm formation inhibitor is administered locally, it can be administered directly from the container containing the biofilm formation inhibitor to the affected area, or the necessary amount can be taken to the fingertip and administered to the affected area. It is. From the viewpoint of hygiene, it is also possible to take a necessary amount of a biofilm formation inhibitor in a clean sponge, cotton swab, cloth, tissue, skin patch, etc. and administer it to the affected area.

In the present specification, the effective dosage of the biofilm formation inhibitor can be appropriately set according to the type and content of the ingredients, the application and the formulation, but the face, perilip, scalp, neck, chest, back, When applied to the skin surface such as the abdomen, arm, mouth, buttocks, buttocks, pubic area, etc., once per day (for example, about 1 to 3 times) or when symptoms such as itching and pain appear, 1 per unit area Times, about 5 to 100 mg / cm ² . The timing of use can be set as appropriate. For example, it is preferable to use it on clean skin after washing the affected area (for example, after washing the face).

  In this specification, a biofilm formation inhibitor can be used before and after a symptom appears. For example, the biofilm formation inhibitor of acne bacteria can be administered before and after the symptoms caused by acne bacteria appear. By administering the biofilm formation inhibitor of acne bacteria to the affected area before the symptoms caused by acne bacteria appear, it becomes possible to inhibit the biofilm formation of acne bacteria. Moreover, it becomes possible to reduce the growth rate of the biofilm formed by administering the biofilm formation inhibitor of an acne bacterium to an affected part after the symptom resulting from an acne bacterium appears.

  In the present specification, the biofilm inhibitor can further contain a surfactant.

  Examples of the surfactant include sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, diglycerol sorbitan penta-2-ethylhexylate, and diglycerol sorbitan tetra-2-ethylhexylate. Sorbitan fatty acid esters; propylene glycol fatty acid esters such as propylene glycol monostearate; polyoxyethylene hydrogenated castor oil 40 (HCO-40), polyoxyethylene hydrogenated castor oil 50 (HCO-50), polyoxyethylene hardened Hardened castor oil derivatives such as castor oil 60 (HCO-60), polyoxyethylene hydrogenated castor oil 80; polyoxyethylene (20) sorbitan (polysorbate 20) monolaurate, monostearate Polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene (20) sorbitan (polysorbate 60), polyoxyethylene monooleate (20) sorbitan (polysorbate 80), polyoxyethylene (20) sorbitan isostearate; Oxyethylene monococonut oil fatty acid glyceryl; glycerin alkyl ether; alkyl glucoside; polyoxyalkylene such as polyoxyethylene lauryl ether, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, polyoxyethylene behenyl ether Alkyl ethers; amines such as stearylamine and oleylamine; polyoxyethylene / methylpolysiloxane copolymer; Silicone surfactants such as uril PEG-9 polydimethylsiloxyethyl dimethicone, PEG-9 polydimethylsiloxyethyl dimethicone; laurate, palmitate, cocoyl glutamate, coconut oil methylalanine salt, acylmethyl taurate, Examples include anionic surfactants such as polyoxyethylene lauryl sulfate, amphoteric surfactants such as lauryl diaminoethyl glycine salt, coconut oil fatty acid betaine salt, and nonionic surfactants such as polyoxyethylene lauryl alcohol ether. It is done. Preferred are polyoxyethylene sorbitan fatty acid esters and polyoxyethylene alkylene alkyl ethers. The surfactant may be synthesized and used by a known method, or a commercially available product may be obtained and used. Moreover, surfactant may be used individually by 1 type and may be used in combination of 2 or more types arbitrarily.

  When the biofilm formation inhibitor contains a surfactant, the content of the surfactant can be appropriately set according to the type of surfactant, the type and amount of other components, and the like. As an example, the content of the surfactant can be 0.0001% by weight or more, preferably 0.001% by weight or more, and 0.002% by weight or more based on the total amount of the preparation. More preferably, it is more preferably 0.005% by weight or more, particularly preferably 0.01% by weight or more, and most preferably 0.05% by weight or more. Further, as an example, the content of the surfactant can be 30% by weight or less, preferably 25% by weight or less, more preferably 20% by weight or less, based on the total amount of the preparation. 15 wt% or less is particularly preferable, and 10 wt% or less is most preferable. As an example, the content of the surfactant can be 0.0001 wt% to 30 wt%, preferably 0.001 wt% to 25 wt%, based on the total amount of the preparation. More preferably, the content is 002 wt% to 20 wt%, particularly preferably 0.01 wt% to 15 wt%, and most preferably 0.05 wt% to 10 wt%.

  In this specification, the biofilm formation inhibitor of an acne bacterium can contain a known acne therapeutic agent component, or can be used in combination with a known acne therapeutic agent as a separate preparation. Known acne treatment agents include, but are not limited to, antibacterial agents including anti-acne antibacterial agents, keratin and sebum solubilizers, anti-inflammatory agents, antibiotics, antipruritic agents, local anesthetics, vitamins, wound treatment agents, etc. It is done. In the present invention, a known acne therapeutic agent can be blended in the biofilm formation inhibitor to exert the effect together. In addition, the administration method in the case of using an acne biofilm formation inhibitor in combination with a known acne treatment agent is not limited as long as the effect of the present invention is exhibited, but the biofilm formation inhibitor of acne bacteria By inhibiting the film and then administering a known acne therapeutic agent, penetration of the known acne therapeutic agent can also be enhanced.

  Antibacterial agents including anti-acne antibacterial agents include, for example, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, sulfur, benzoyl peroxide, piroctone olamine, zinc pyrithione, crimbazole, rilopirox, ketoconazole, itraconazole, guaiazulene, camazulene, chlorhexylene Benzalkonium chloride, acrinol, ethanol, benzethonium chloride, cresol, gluconic acid and its derivatives, popidone iodine, potassium iodide, iodine, isopropylmethylphenol, triclocarban, triclosan, photosensitizer 101, photosensitizer 201, paraben, Phenoxyethanol, 1,2-pentanediol, alkyldiaminoglycine hydrochloride, pyroctoolamine, miconazole, nadifloxacin Homosurufamin, sulfadiazine, (also referred to as resorcinol) resorcinol, and the like.

  Examples of the keratolytic agent include salicylic acid, glycolic acid, lactic acid, malic acid, resorcin (also referred to as resorcinol), and the like.

  As the anti-inflammatory agent, either a steroidal anti-inflammatory agent or a non-steroidal anti-inflammatory agent can be used. Specifically, allantoin, glycyrrhizic acid, glycyrrhetinic acid or a pharmaceutically acceptable salt thereof, dexamethasone acetate valerate, dexamethasone, prodonisolone valerate acetate (prednisolone valerate acetate ester), prodonisolone acetate, prodonisolone, hydrocortisone acetate, hydrocortisone , Ufenamate, bufexamac, ibuprofenpiconol, and glycol salicylate are exemplified, but not limited thereto. Furthermore, for example, indomethacin, diclofenac, piroxicam, epsilon-aminocaproic acid, berberine, lysozyme, azulene, bromelain, serrapeptase, semi-alkaline proteinase, and pharmaceutically acceptable salts thereof are exemplified.

  As the antibiotic, for example, any of macrolide antibiotics, lincomycin antibiotics, aminoglycoside antibiotics, chloramphenicol antibiotics and the like can be used. Examples of macrolide antibiotics include erythromycin, azithromycin, clarithromycin, tilmicosin, tylosin, etc., examples of lincomycin antibiotics include clindamycin, lincomycin, etc., and aminoglycoside antibiotics Examples include gentamicin sulfate, and chloramphenicol antibiotics include chloramphenicol.

  Examples of the antipruritic agent include d-camphor, dl-camphor, menthol, phenol, crotamiton and the like.

  Examples of the local anesthetic include dibucaine hydrochloride, isothipenzil hydrochloride, lidocaine hydrochloride, thecaine, dibucaine, lidocaine, ethyl paraaminobenzoate and the like.

  As vitamins, vitamin A such as retinol, retinal, retinoic acid and derivatives thereof (for example, retinol palmitate, tretinoin); nicotinic acid, nicotinamide, dl-α-tocopherol nicotinate, benzyl nicotinate, nicotine Nicotinic acids such as methyl acid, β-butoxyethyl nicotinate, 1- (4-methylphenyl) ethyl nicotinate; pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-panthecine, D-panthetin, Pantothenic acids such as coenzyme A and pantothenyl ethyl ether; vitamin E such as dl-α-tocopherol, dl-α-tocopherol acetate, dl-α-tocopherol succinate, dl-α-tocopherol calcium succinate; ascorbic acid A Ascorbic acid derivatives such as sodium corbate, dehydroascorbic acid, sodium ascorbate phosphate, magnesium ascorbate phosphate, magnesium, ascorbigen-A, ascorbyl stearate, ascorbyl palmitate, dipalmitate L-ascorbyl Some vitamin C: thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate phosphate, thiamine Monophosphate, thiamine diphosphate, thiamine diphosphate hydrochloride, thiamine triphosphate, thiamine triphosphate monophosphate, dibenzoylthia Vitamins such as riboflavin, flavin mononucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, riboflavin 5'-phosphate sodium, riboflavin tetranicotinate B2: Vitamin B6 such as pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, 5′-phosphate pyridoxal hydrochloride and pyridoxamine hydrochloride; biotins such as biotin and bioticin; vitamin B12 such as cyanocobalamin, hydroxocobalamin and deoxyadenosylcobalamin; Folic acid such as folic acid, pteroylglutamic acid; vitamin D such as methyl hesperidin, ergocalciferol, cholecalciferol; phylloquinone, Vitamin Ks such as farnoquinone; vitamin-like action factors such as adapalene (vitamin A-like action factor), carnitine, ferulic acid, α-lipoic acid, orotic acid and the like.

(Examples of combinations with known acne treatments)
In the present invention, it is preferable to use a combination of a known acne therapeutic agent together with basic amino acids having the ability to suppress biofilm formation. Known acne treatments that can be used in combination in this way are not particularly limited, but preferably antibacterial agents (for example, salicylic acid, sulfur, isopropylmethylphenol, ethanol, homosulfamine, sulfadiazine, nadifloxacin, benzoyl peroxide, Resorcin), keratolytic agents (eg salicylic acid, glycolic acid, resorcin), antibiotics (eg clindamycin), vitamins (eg adapalene).

  In this specification, when used as a pharmaceutical, quasi-drug, cosmetic, etc., the biofilm formation inhibitor is mixed with a known base or carrier to the extent that it does not impair the effects of the present invention. Can do. Other biofilm formation inhibitors include, for example, surfactants, oils, alcohols, thickeners, preservatives, antioxidants, antioxidants, preservatives, chelating agents, pH adjusters, stabilizers. Additives such as solubilizing agents, suspending agents, tonicity agents, buffers, soothing agents, dispersants, fragrances, coloring agents, pigments, water and the like can be blended. These additives can be used alone or in combination of two or more.

  Bases or carriers include liquid paraffin, squalane, gelled hydrocarbons (such as plastibase), ozokerite, α-olefin oligomers, hydrocarbons such as light liquid paraffin; methyl polysiloxane, cross-linked methyl polysiloxane, highly polymerized methyl Polysiloxane, Cyclic silicone, Alkyl-modified silicone, Cross-linked alkyl-modified silicone, Amino-modified silicone, Polyether-modified silicone, Polyglycerin-modified silicone, Cross-linked polyether-modified silicone, Cross-linked alkyl polyether-modified silicone, Silicone / alkyl chain copolymer Modified polyether-modified silicone, silicone / alkyl chain co-modified polyglycerin-modified silicone, polyether-modified branched silicone, polyglycerin-modified branched silicone, acrylic silicone, Silicone oil such as phenyl-modified silicone, silicone resin; polyethylene glycol; dioxane; isopropyl myristate, octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, pentaerythritol tetra-2-ethylhexanoate Esters; lower alcohols such as ethanol and isopropanol; glycol ethers such as diethylene glycol monomethyl ether and diethylene glycol monoethyl ether; polyhydric alcohols such as polyethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin and isoprene glycol; water An aqueous base such as

  A base or a support | carrier can be used individually by 1 type or in combination of 2 or more types.

  Examples of the oil include natural animal and plant oils, hydrocarbon oils, ester oils, silicone oils, higher alcohols, higher fatty acids, animals and plants, and synthetic essential oils.

  Examples of natural animal and plant oils and fats include avocado oil, linseed oil, almond oil, olive oil, cacao oil, beef tallow, giraffe oil, wheat germ oil, sesame oil, rice germ oil, rice bran oil, safflower oil, soybean oil, evening primrose oil , Camellia oil, corn oil, rapeseed oil, horse fat, persic oil, palm oil, palm kernel oil, castor oil, sunflower oil, pork fat, grape oil, jojoba oil, macadamia nut oil, mink oil, cottonseed oil, molasses, palm oil , Hardened coconut oil, peanut oil, lanolin, egg yolk oil, rosehip oil and the like.

  Examples of the hydrocarbon oil include paraffinic hydrocarbons and olefinic hydrocarbons, and examples include squalane, squalene, ceresin, paraffin, pristane, microcrystalline wax, liquid paraffin, and petroleum jelly.

  As ester oils, synthetic esters, esters of higher alcohols and higher fatty acids are used, for example, diisobutyl adipate, 2-hexyldecyl adipate, di-2-heptylundecyl adipate, isostearyl isostearate, Trimethylolpropane triisostearate, cetyl 2-ethylhexanoate, neopentyl glycol di-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, pentaerythritol tetra-2-ethylhexanoate, cetyl octanoate, Oleyl oleate, octyldodecyl oleate, decyl oleate, neopentyl glycol dicaprate, 2-ethylhexyl succinate, isocetyl stearate, butyl stearate, diisopropyl sebacate, cetyl lactate Tetradecyl lactate, isopril pyr myristate, octyldodecyl myristate, cetyl myristate, myristyl myristate, octyl palmitate, 2-ethylhexyl palmitate, 2-hexyldecyl palmitate, 2-heptylundecyl palmitate, 12-hydroxystearin Examples thereof include cholesteryl acid, phytosteryl oleate, diisostearyl malate, paramethoxycinnamate, and pentarosyl tetrarosinate.

  Examples of the silicone oil include dimethylpolysiloxane, highly polymerized methylpolysiloxane, methylphenylpolysiloxane, methylhydrogenpolysiloxane, octamethylcyclotetrasiloxane, octamethylcyclopentasiloxane, decamethylcyclohexasiloxane, stearoxysilicone, etc. Higher alkoxy-modified silicone, alkyl-modified silicone, higher fatty acid ester-modified silicone, and the like.

  Examples of the higher alcohol include octyldodecanol, isostearyl alcohol, oleyl alcohol, stearyl alcohol, cetanol, and behenyl alcohol.

  As the higher fatty acid, a saturated or unsaturated linear or branched fatty acid having 12 to 22 carbon atoms can be used. For example, isostearic acid, oxystearic acid, oleic acid, stearic acid, palmitic acid, behenic acid, Examples include myristic acid, lauric acid, lanolinic acid, linoleic acid, and linolenic acid.

  Examples of the thickener include guar gum, locust bean gum, carrageenan, xanthan gum, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydrophobized hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer. , Alkyl methacrylate copolymer, polyethylene glycol, bentonite, (hydroxyethyl acrylate / acryloyldimethyltaurine Na) copolymer, (acryloyldimethyltaurine ammonium / vinylpyrrolidone) copolymer, and the like.

  Preferable examples of the preservative include paraoxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.

  Preferable examples of the antioxidant include sulfite and ascorbic acid.

  Examples of the antioxidant include dibutylhydroxytoluene (BHT), butylhydroxyanisole, sorbic acid, sodium sulfite, ascorbic acid, erythorbic acid, L-cysteine hydrochloride and the like.

  Preservatives include benzoic acid, sodium benzoate, dehydroacetic acid, sodium dehydroacetate, isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, butyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, benzyl paraoxybenzoate, paraoxybenzoate Examples include methyl benzoate and phenoxyethanol.

  Examples of the chelating agent include EDTA / disodium salt, EDTA / calcium disodium salt, and the like.

  Examples of pH adjusters include inorganic acids (hydrochloric acid, sulfuric acid, etc.), organic acids (lactic acid, sodium lactate, citric acid, sodium citrate, succinic acid, sodium succinate, etc.), inorganic bases (potassium hydroxide, sodium hydroxide, etc.) ) And organic bases (such as triethanolamine, diisopropanolamine, and triisopropanolamine).

  Examples of the stabilizer include sodium polyacrylate, dibutylhydroxytoluene, butylhydroxyanisole and the like.

  Preferable examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like.

  Suitable examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate; for example, polyvinyl alcohol, Examples thereof include hydrophilic polymers such as polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.

  Preferable examples of the isotonic agent include sodium chloride, glycerin, D-mannitol and the like.

  Preferable examples of the buffer include buffer solutions such as phosphate, acetate, carbonate, citrate and the like.

  Preferable examples of the soothing agent include benzyl alcohol.

  Examples of the dispersant include sodium pyrophosphate, sodium hexametaphosphate, polyvinyl alcohol, polyvinyl pyrrolidone, methyl vinyl ether / maleic anhydride crosslinked copolymer, and organic acid.

  Examples of the colorant include inorganic pigments and natural pigments.

  In this specification, a biofilm formation inhibitor can contain another active ingredient in the range which does not impair the effect of this invention. Specific examples of the active ingredient include, for example, a moisturizing ingredient, a pearl luster imparting agent, a conditioning agent, a blood circulation promoting ingredient, an astringent ingredient, an ultraviolet absorbing ingredient, an ultraviolet scattering ingredient, a cleaning ingredient, a peptide or a derivative thereof, an amino acid or a derivative thereof, a cell An activation component etc. are mentioned.

  As moisturizing ingredients, polyhydric alcohols such as glycerin, 1,3-butylene glycol, propylene glycol, polyethylene glycol and diglycerin; sugars such as trehalose, xylitol and oligosaccharides; sodium hyaluronate, heparin analog, chondroitin sulfate High molecular compounds such as sodium, collagen, elastin, keratin, chitin, chitosan; amino acids such as glycine, aspartic acid, arginine; natural moisturizing factors such as sodium lactate, urea, sodium pyrrolidonecarboxylate; ceramide, cholesterol, phosphorus Examples include lipids such as lipids; plant extract extracts such as chamomile extract, chamamelis extract, tea extract, perilla extract, and yokuinin extract.

  Examples of the pearl luster imparting agent include ethylene glycol distearate, ethylene glycol monostearate, and triethylene glycol distearate.

  Examples of the conditioning agent include cationized cellulose, cationized starch, cationized fenugreek gum, cationized guar gum, cationized tara gum, cationized locust bean gum, cationized xanthan gum, diallyl quaternary ammonium salt / acrylamide copolymer, polyquaternium , Vinyl imidazolium trichloride / vinyl pyrrolidone copolymer, hydroxyethyl cellulose / dimethyldiallylammonium chloride copolymer, vinyl pyrrolidone / quaternized dimethylaminoethyl methacrylate copolymer, polyvinyl pyrrolidone / alkylamino acrylate copolymer, polyvinyl pyrrolidone / Alkylamino acrylate / vinyl caprolactam copolymer, vinyl pyrrolidone / methacrylamidopropyl chloride Trimethyl ammonium copolymer, alkyl acrylamide / acrylate / alkylaminoalkyl acrylamide / polyethylene glycol methacrylate copolymers, adipic acid / dimethylaminohydroxypropyl ethylene triamine copolymer.

  Examples of the blood circulation promoter include acetylcholine, ictamol, caffeine, capsaicin, cantalis tincture, gamma oryzanol, ginger tincture, gingeron, cephalanthin, assembly extract, tannic acid, red pepper tincture, trazoline, tocopherol nicotinate, benzyl nicotinate Examples include esters.

  Examples of the astringent component include zinc oxide, zinc sulfate, allantoin hydroxyaluminum, aluminum chloride, zinc sulfocolate and tannic acid.

  Examples of the ultraviolet absorbing component include octyl triazone, diethylaminohydroxybenzoyl hexyl benzoate, dimethoxybenzylidene dioxoimidazolidine propionate octyl, paramethoxycinnamate 2-ethylhexyl, t-butylmethoxydibenzoylmethane, phenylbenzimidazolesulfonic acid, Examples include octyl methoxycinnamate and ethylhexyl methoxycinnamate.

  Examples of ultraviolet scattering components include hydrous silicic acid, zinc silicate, cerium silicate, titanium silicate, zinc oxide, zirconium oxide, cerium oxide, titanium oxide, iron oxide, silicic anhydride, and other inorganic compounds. Coated with inorganic powders such as hydrous silicic acid, aluminum hydroxide, mica and talc, compounded with resin powders such as polyamide, polyethylene, polyester, polystyrene and nylon, and with silicon oil and fatty acid aluminum salts, etc. Processed ones are listed.

  Detergents include soaps such as alkali metal salts such as potassium laurate, potassium myristate, potassium palmitate or potassium stearate, alkanolamide salts or amino acid salts, amino acid-based interfaces such as sodium cocoyl glutamate, sodium cocoyl methyl taurate Activator, ether sulfate ester salt such as sodium laureth sulfate, ether carboxylate salt such as sodium lauryl ether acetate, sulfosuccinate ester salt such as sodium alkylsulfosuccinate ester, coconut oil fatty acid monoethanolamide, coconut oil fatty acid ethanolamide Fatty acid alkanolamides, monoalkyl phosphates such as sodium lauryl phosphate, sodium polyoxyethylene lauryl ether phosphate, coconut oil Acid amidopropyldimethylaminoacetic acid betaine, lauryldimethylaminoacetic acid betaine, 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, laurylhydroxysulfobetaine and lauroylamidoethylhydroxyethylcarboxymethylbetaine hydroxypropyl phosphate sodium Betaine type amphoteric surfactants such as amino acid type amphoteric surfactants such as sodium laurylaminopropionate.

  Peptides or derivatives thereof include keratin-degrading peptide, hydrolyzed keratin, collagen, fish-derived collagen, atelocollagen, gelatin, elastin, elastin-degrading peptide, collagen-degrading peptide, hydrolyzed collagen, hydroxypropylammonium chloride hydrolyzed collagen, elastin-degrading peptide , Conchiolin degrading peptide, hydrolyzed conchiolin, silk proteolytic peptide, hydrolyzed silk, lauroyl hydrolyzed silk sodium, soy proteolytic peptide, hydrolyzed soy protein, wheat protein, wheat proteolytic peptide, hydrolyzed wheat protein, casein degrading peptide Acylated peptides (palmitoyl oligopeptide, palmitoyl pentapeptide, palmitoyl tetrapeptide, etc.).

  Amino acids or derivatives thereof include betaine (trimethylglycine), proline, hydroxyproline, serine, glycine, alanine, phenylalanine, β-alanine, threonine, glutamic acid, glutamine, asparagine, aspartic acid, cysteine, cystine, methionine, leucine, isoleucine , Valine, taurine, γ-aminobutyric acid, γ-amino-β-hydroxybutyric acid, carnitine, carnosine, creatine and the like.

  Cell activation components include amino acids such as γ-aminobutyric acid and ε-aminocaproic acid, vitamins such as retinol, thiamine, riboflavin, pyridoxine hydrochloride, pantothenic acids, α-hydroxy acids such as glycolic acid and lactic acid, tannins, Examples include flavonoids, saponins, allantoin, and photosensitizer 301.

  Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited thereto.

(Preparation of formulation)
A biofilm formation inhibitor having the composition shown in Table 1 or 2 was prepared according to a conventional method. Specifically, each component was added to physiological saline (manufactured by Otsuka Pharmaceutical Co., Ltd.) at the concentrations shown in Table 1 or 2. The mixture was stirred at room temperature and adjusted to pH 7 or pH 5 with hydrochloric acid or sodium hydroxide aqueous solution to obtain a uniform preparation.

(Acne bacteria biofilm formation inhibition test)
In this test, L-histidine (manufactured by Wako Pure Chemical Industries, Ltd.), L-arginine (manufactured by Wako Pure Chemical Industries, Ltd.), glycine (manufactured by Wako Pure Chemical Industries, Ltd.), L-glutamic acid (manufactured by Wako Pure Chemical Industries, Ltd.) Implemented. Specifically, each test solution inoculated into a 96-well round bottom plate so that the P. acnes suspension becomes 10 ⁸ (CFU / mL) bacterial solution is dispensed at 100 μL per well. Thereafter, anaerobic culture was performed at 36 ° C. for 4 days. After incubation, each well was washed twice with 200 μL of purified water and stained with 100 μL of 0.5% crystal violet for 20 minutes. Each well was washed 3 times with 200 μL of purified water, and then 160 μL of 33% acetic acid was used to dissolve crystal violet adhering to the plate. Using a microplate reader (Molecular Devices, Versa max), the absorbance at 590 nm at which crystal violet exhibits absorbance was measured to obtain the amount of biofilm formed. A physiological saline adjusted to pH 7 or 5 (manufactured by Otsuka Pharmaceutical Co., Ltd.) was prepared as a control, and the biofilm formation inhibition rate was calculated by the following formula using the ratio to the biofilm formation amount. For the test of statistical significance, P value was calculated using Dunnett test (n = 6). The results are shown in Tables 1 and 2 and FIGS.

[Equation 1]
Biofilm formation inhibition rate (%) = 100−absorbance of residual biofilm in each test solution / absorbance of control × 100 (%)

  As shown in the above results, at pH 7, the formation of biofilms of acne was significantly suppressed by basic amino acids such as arginine and histidine. On the other hand, the neutral amino acid glycine, which is not a basic amino acid, and the acidic amino acid glutamic acid, did not show an inhibitory effect on acne biofilms. Further, even at pH 5, the biofilm of acne was remarkably suppressed by basic amino acids. Based on this result, for example, when basic amino acids are used in combination with other known acne therapeutic agents, it is possible to increase the penetration of the known acne therapeutic agents into acne bacteria and enhance the therapeutic effect.

(Reference test example: Antibacterial test)
This test was performed using L-arginine (manufactured by Wako Pure Chemical Industries, Ltd.) and L-histidine (manufactured by Wako Pure Chemical Industries, Ltd.). Each test solution was prepared in the same manner as in the above example (pH 7), and then inoculated to a 96-well round bottom plate with 10 ⁸ (CFU / mL) suspension of P. acnes suspension. 100 μL of the test solution was dispensed per well, and anaerobic culture was performed at 36 ° C. for 4 days. After culture, 100 μL (n = 3) of a 96-well round bottom plate preparation was collected, diluted 10-fold to 1000-fold with physiological saline (Otsuka Pharmaceutical), seeded on GAM agar medium (Nissui Pharmaceutical), and agar plate The number of viable bacteria was determined by surface smearing. In addition, physiological saline adjusted to pH 7 (manufactured by Otsuka Pharmaceutical Co., Ltd.) was prepared as a control, and the number of viable bacteria was determined by the same treatment as in the case of using the test solution. The results are shown in Table 3.

  As shown in the above results, antibacterial activity against acne bacteria was not recognized in basic amino acids such as arginine and histidine. Therefore, it was shown that the biofilm formation inhibitor of the present invention does not exhibit bactericidal ability but has biofilm formation inhibiting ability.

(Formulation example)
Hereinafter, the formulation example of the biofilm formation inhibitor of this invention is shown. Formulation examples 1 to 4 are lotions, and formulation examples 5 to 8 are examples of creams (oil-in-water type). All are prepared according to conventional methods. These formulation examples can be used to suppress biofilm formation by microorganisms such as acne.

Claims (10)

  A biofilm formation inhibitor containing basic amino acids.   The biofilm formation inhibitor of Claim 1 which suppresses the biofilm formation of an acne microbe.   The biofilm formation inhibitor according to claim 1 or 2, wherein the basic amino acids are at least one selected from the group consisting of arginine, histidine, lysine and ornithine.   The biofilm formation inhibitor according to any one of claims 1 to 3, wherein the basic amino acids are L-form amino acids.   The biofilm formation inhibitor as described in any one of Claims 1-4 applied to skin or an intraoral area.   The biofilm formation inhibitor according to any one of claims 1 to 5, which is used for treatment or prevention of acne.   The biofilm formation inhibitor as described in any one of Claims 1-6 used for the treatment or prevention of acne in dry skin.   The biofilm formation inhibitor according to any one of claims 1 to 7, which is used for treatment or prevention of acne that tends to repeat.   The biofilm formation inhibitor according to any one of claims 1 to 8, which is used for treatment or prevention of acne accompanied by inflammation in a hair follicle.   The biofilm formation inhibitor according to any one of claims 1 to 9, wherein the biofilm formation inhibitor has a pH of 3.0 to 8.0.

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