JP2020090479A - Biofilm formation inhibitor - Google Patents

Abstract

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 bacteria.

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

Acne bacteria are anaerobic bacteria and therefore live deep inside hair follicles. Acne bacteria grow using sebum accumulated in hair follicles as a nutrient source, and when it deteriorates, it causes inflammation. Due to inflammation, pus accumulates in the hair follicles and becomes a skin rash. Acne known to be involved in Acne bacteria include acne vulgaris that is often seen in adolescents, pustular acne mainly composed of pustules, cystic acne mainly composed of cysts, and acne vulgaris. There are severe cases of acne vulgaris, electric shock acne, etc. For the treatment of such acne, various external preparations and oral preparations have been developed for the purpose of suppressing sebum production, preventing infection, suppressing inflammation, and normalizing keratinization.

Some bacteria form an accumulation of bacterial secretions (polysaccharides, etc.) called a 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. Further, plaques formed between the teeth and between the teeth and the gingiva are considered as a type of biofilm.

Acne bacteria are also known to form biofilms. Since the biofilm accumulates polysaccharides and the like secreted from Acne bacteria, it is difficult for a drug such as an antibacterial agent to penetrate into the biofilm, and the effect is diminished. Therefore, the presence or absence of biofilm formation affects the pathogenicity of various bacterial infections (Non-Patent Document 1). Development of an antibacterial agent that is highly effective against biofilms, plaques, etc. is also underway, and Patent Document 1 discloses an antibacterial agent effective against Staphylococcus aureus, mutans, Escherichia coli, and the like.

Japanese Patent No. 3717960

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

However, the development of a biofilm formation by microorganisms, especially for the acne bacteria that form a biofilm deep inside hair follicles, has not yet been sufficiently developed. Therefore, an object of the present invention is to provide a preparation having a high effect of suppressing such biofilm formation.

The present inventor, as a result of diligent studies to solve the above problems, biofilm formation by biofilm-forming microorganisms, especially acne bacteria which is a causative bacterium of acne, basic amino acids have a particularly remarkable inhibitory effect. I found that it would work. The present invention has been completed by further studies based on such findings.

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

The biofilm formation inhibitor described above can be used 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-amino acids.

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

The biofilm formation inhibitor described above can be used for treating or preventing acne.

The biofilm formation inhibitor can be used for treating or preventing acne on dry skin.

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

The biofilm formation inhibitor described above can be used for the treatment or prevention of acne with inflammation in the hair follicle.

The pH of the biofilm formation inhibitor may be 3.0 to 8.0.

The biofilm formation inhibitor may further contain a surfactant.

The biofilm formation inhibitor described above may be used for acne vulgaris, pustular acne, cystic acne, acne vulgaris or acne blitz.

According to the present invention, it is possible to effectively suppress the formation of biofilm formed by microorganisms, especially acne bacteria.

It is a graph which shows the result of having evaluated biofilm formation suppression rate (%) using the various amino acid compound 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 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 have the meanings commonly 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. The biofilm-forming microorganisms are not particularly limited, for example, acne bacteria, mutans bacteria, Staphylococcus epidermidis, Staphylococcus aureus, bacteria such as Escherichia coli, ringworm, Candida, Malassezia, black mold (for example, , Aspergillus oryzae, Cladosporium bacterium) and the like. From the viewpoint that even higher effects can be expected, the biofilm formation inhibitor of the present invention is preferably used against bacteria, more preferably against Acne or mutans, and particularly preferably Acne. Used against.

As used herein, "inhibition of biofilm formation", whether complete or partial, inhibits microorganisms from forming biofilms, or the growth rate of biofilms formed. It means to reduce. When detecting biofilm formation inhibition of Acne bacteria, the method is not limited as long as it is a known method, but if it is the method described in the test example below, the biofilm formation inhibition rate is at least 20% or more. Is more preferable, 30% or more is more preferable, 40% or more is still more preferable, 50% or more is still more preferable, and 60% or more is particularly preferable.

As used herein, the term “acne bacterium” refers to Propionibacterium acnes (P. acnes), a genetic analogue thereof, or a mutant thereof.

In the present specification, the “mutans bacterium” refers to Streptococcus mutans (S. mutans) or a genetic analogue thereof or a mutant thereof.

As used herein, the term "staphylococcus epidermidis" refers to Staphylococcus epidermidis (S. epidermidis) or its genetic analogs or mutants thereof.

As used herein, the term "Staphylococcus aureus" refers to Staphylococcus aureus (S. aureus) or its genetic analogs or mutants thereof.

As used herein, the term “Escherichia coli” refers to Escherichia coli (E. coli) or a genetic analogue thereof or a mutant thereof. The type of surface antigen (O antigen and H antigen) is not limited as long as it is a biofilm-forming Escherichia coli.

In the present specification, the “trichophyton fungus” means a Trichophyton rubrum (T. rubrum), Trichophyton mentagrophytes (T. mentagrophytes), Trichophyton tonsurans (T. tonsurans), etc., or a genetic analogue thereof. Alternatively, it means a mutant thereof.

In the present specification, the “Candida bacterium” refers to a fungus of the genus Candida including Candida albicans (C. albicans) and the like, genetic analogues thereof or mutants thereof.

In the present specification, the term “malassezia fungus” means 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) and the like, or a genetic analogue thereof or a mutant thereof.

As used herein, the term “A. oryzae” refers to Aspergillus oryzae (A. oryzae)-containing Aspergillus fungi or a genetic analogue thereof or a mutant thereof.

In the present specification, the term "cladosporium fungus" means Cladosporium cladosporioides (C. cladosporioides), Cladosporium sphaerospermum (C. sphaerospermum), etc., and their genetic analogues or mutations 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, but are not limited to, 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.), sulfoalkylalkylenediamine (N,N'-bis(3-sulfopropyl)ethylenediamine etc.), epsilon aminocaproic acid, 2-amino-2-methyl-1-propanol (AMP), 2-amino Examples thereof include 2-methyl-1,3-propanediol, organic amines such as meglumine, urea and creatinine, and pharmaceutically acceptable salts thereof. In the present invention, the basic amino acids may be either L-form and/or D-form, but L-form is preferably used.

As the basic amino acids, from the viewpoint of effectively suppressing biofilm formation (particularly, biofilm formation of Acne bacteria), basic amino acids are preferable, and at least selected from the group consisting of arginine, histidine, lysine, and ornithine. One or more components are more preferable, arginine and/or histidine are further preferable, and arginine is particularly preferable. The basic amino acids may be synthesized by known methods and used, or commercially available products may be obtained and used. The basic amino acids may be used alone or in any combination of two or more. In addition, since basic amino acids in biofilm formation inhibitors are not antibiotics, they are effective against microorganisms that are clinically partially or completely resistant to one or more antibiotics. is there. In addition, the acne bacterium is originally a resident bacterium of the skin and has a role of suppressing the invasion and reproduction of other pathogenic bacteria by keeping the skin weakly acidic. Therefore, excessive sterilization/suppression of Acne bacteria by a strong drug such as an antibiotic may rather cause a burden on the skin. By suppressing the biofilm formation of Acne bacteria, it is possible to enhance the penetrating power of various drugs, and to effectively sterilize and suppress Acne bacteria without using strong drugs. Alternatively, by suppressing the biofilm formation of acne bacteria, various drugs (for example, antibacterial agents including anti-acne bacteria agents described below, keratin/sebum-dissolving agents, anti-inflammatory agents, antibiotics, antipruritic agents, topical agents). Anesthetics, vitamins, wound healing agents, etc.) can be used in small doses.

In the present specification, the pharmaceutically acceptable salts of basic amino acids include, for example, alkali metal salts, alkaline earth metal salts, salts with organic bases, and the like, sodium, potassium, calcium, magnesium, Examples thereof include salts with ammonium, diethanolamine, ethylenediamine and the like. 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 and L-glucamine; and salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and 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 salts with acidic amino acids such as aspartic acid and glutamic acid. The "pharmaceutically acceptable salt" may include a solvate or a hydrate of the salt.

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

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

In the present invention, the viscosity of the biofilm formation inhibitor is appropriately set according to the type and content of the blended components, the application, the formulation form, the method of use, and the like. The viscosity of the acne bacterium biofilm formation inhibitor is not limited from the viewpoint that the active ingredient penetrates into the biofilm and the formulation remains in the affected area, but is preferably 0.001 mPa·s to 500,000 mPa·s. Yes, more preferably 0.005 mPa·s to 450,000 mPa·s, further 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. Viscosity is not particularly limited as long as it is a known method, but viscosity at 20° C. measured by a rotational viscometer (RB-80H type viscometer, manufactured by Toki Sangyo Co., Ltd., rotor; H7, rotational speed; 10 rpm). Can be

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

In the present specification, the biofilm formation inhibitor can be used for local administration of the skin surface such as face, perilipse, scalp, neck, chest, back, abdomen, arm, oral cavity, armpit, buttocks, genital area. .. When used as a biofilm formation inhibitor for Acne bacteria, it can be administered locally to various acne conditions, such as microcomedones (horn plug formation, early plugging of the corner plug) and so-called pimples (adults). Acne), white acne (white papule), black acne (open comedone), and red acne (red papule) or yellow acne (purulent acne) with inflammation can be used as a prophylactic and/or therapeutic agent. In addition, the acne bacterium biofilm formation inhibitor should be used as a prophylactic and/or therapeutic agent for acne vulgaris, pustular acne, cystic acne, acne vulgaris, and acne acne. You can Acne is sometimes referred to as open or obstructive, depending on whether the hair follicle is open or closed due to sebum or keratin plugs. Can also be used for. The acne bacterium biofilm formation inhibitor can also be used for acne caused by atopic dermatitis. In addition, since the acne bacterium is also known as a causative bacterium of sarcoidosis, it can be used as a preventive and/or therapeutic agent even in an affected area of the skin where granulomatous lesions of sarcoidosis are caused. In the present specification, the biofilm formation inhibitor for Acne bacteria is effectively used when Acne bacteria grow in or around hair follicles to form biofilms.

Dry skin, which is often seen in adult acne, is prone to excessive sebum secretion and clogging of the keratin plug, and such changes in the surrounding environment and stress may increase biofilm formation by acne bacteria. Further, there is concern that acne bacteria protected by the biofilm may repeatedly develop acne symptoms as biofilm formation increases. It is known that an increase in biofilm formation leads to an increase in lipase activity, the lipase decomposes triglyceride in sebum to produce free fatty acid, and causes inflammation in the hair follicle. Therefore, the biofilm formation inhibitor of the present invention is a treatment and/or prevention of acne in such dry skin, a treatment and/or prevention of acne that tends to be repeated, a treatment of acne with inflammation in the hair follicle and/or It can be used effectively for prevention. The acne bacterium biofilm inhibitor can be used as a drug, a quasi drug, or a cosmetic for the purpose of such prevention and/or treatment.

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

In the present specification, the effective dose of the biofilm formation inhibitor may be appropriately set depending on the type and content of the compounding ingredients, the application and the form of the formulation, but the face, the periphery of the lips, the scalp, the neck, the chest, the back, When applied to the skin surface of the abdomen, arms, oral cavity, armpit, buttocks, genital area, etc., several times a day (for example, about 1 to 3 times) or 1 times per unit area when symptoms such as itching and pain appear. The dose is about 5 to 100 mg/cm ² . The timing of use can be set as appropriate, but it is preferably used for clean skin after washing the affected area (for example, after washing the face when using it on the face).

In the present specification, the biofilm formation inhibitor can be used before and after the appearance of symptoms. For example, the biofilm formation inhibitor of Acne bacteria can be administered before and after the appearance of symptoms caused by Acne bacteria. 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. In addition, by administering a biofilm formation inhibitor of Acne bacteria to the affected area after the appearance of symptoms caused by Acne bacteria, the growth rate of the biofilm formed can be reduced.

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

Examples of the surfactant include sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, penta-2-ethylhexyl diglycerol sorbitan, and tetra-2-ethylhexyl diglycerol sorbitan. 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 hydrogenated Castor oil derivatives such as castor oil 60 (HCO-60) and polyoxyethylene hydrogenated castor oil 80; polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monostearate (polysorbate) 60), polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene (20) sorbitan monooleate (polysorbate 80) and polyoxyethylene (20) sorbitan isostearate; polyoxyethylene monococonut oil fatty acid glyceryl; glycerin alkyl ether Alkyloxysides; polyoxyethylene lauryl ether, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, polyoxyethylene behenyl ether, and other polyoxyalkylene alkyl ethers; stearylamine, oleylamine, etc. Amines; Silicone-based surfactants such as polyoxyethylene/methylpolysiloxane copolymer, lauryl PEG-9 polydimethylsiloxyethyl dimethicone, PEG-9 polydimethylsiloxyethyl dimethicone; laurate, palmitate, cocoyl Anionic surfactants such as glutamate, coconut oil methylalanine salt, acylmethyltaurine salt, polyoxyethylene lauryl sulfate, amphoteric surfactants such as lauryl diaminoethylglycine salt, coconut oil fatty acid betaine salt, polyoxyethylene Examples include nonionic surfactants such as lauryl alcohol ether. Preferred are polyoxyethylene sorbitan fatty acid esters and polyoxyethylene alkylene alkyl ethers. The surfactant may be synthesized by a known method and used, or a commercially available product may be obtained and used. In addition, the surfactant may be used alone or in any combination of two or more.

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. The content of the surfactant can be, for example, 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, further preferably 0.01% by weight or more, most preferably 0.05% by weight or more. The content of the surfactant can be, for example, 30% by weight or less, preferably 25% by weight or less, and more preferably 20% by weight or less, based on the total amount of the preparation. 15% by weight or less is particularly preferable, and 10% by weight or less is most preferable. As an example, the content of the surfactant may be 0.0001% by weight to 30% by weight, preferably 0.001% by weight to 25% by weight, based on the total amount of the preparation, and is preferably 0. It is more preferably 002 to 20% by weight, particularly preferably 0.01 to 15% by weight, and most preferably 0.05 to 10% by weight.

In the present specification, the acne bacterium biofilm formation inhibitor may contain a known acne therapeutic drug component, or may be used as a separate formulation in combination with a known acne therapeutic drug. Known acne therapeutic agents include, but are not limited to, antibacterial agents including anti-acne bacteria agents, keratin/sebum-dissolving agents, anti-inflammatory agents, antibiotics, antipruritic agents, local anesthetics, vitamins, and wound healing agents. Be done. In the present invention, a known acne treatment drug may be incorporated into the biofilm formation inhibitor so that the effect can be exerted together. In addition, the administration method when using a biofilm formation inhibitor of Acne bacteria in combination with a known acne therapeutic agent is not limited as long as the effect of the present invention is exhibited, but biofilm formation inhibitor of Acne bacteria bio It is also possible to increase the penetration of the known acne therapeutic agent by administering the known acne therapeutic agent after suppressing the film.

Examples of the antibacterial agent containing an anti-acne agent include, for example, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, sulfur, benzoyl peroxide, piroctone olamine, zinc pyrithione, climbazole, lillopirox, ketoconazole, itraconazole, guaiazulene, camazulene, chlorhexidine. , Benzalkonium chloride, acrinol, ethanol, benzethonium chloride, cresol, gluconic acid and its derivatives, povidone iodine, potassium iodide, iodine, isopropylmethylphenol, triclocarban, triclosan, photosensitizer No. 101, photosensitizer No. 201, paraben, Phenoxyethanol, 1,2-pentanediol, alkyldiaminoglycine hydrochloride, pyroctoolamine, miconazole, nadifloxacin, homosulfamine, sulfadiazine, resorcinol (also called resorcinol) and the like can be mentioned.

Examples of the keratolytic agent include salicylic acid, glycolic acid, lactic acid, malic acid, resorcin (also called 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, glycyrrhizinic acid, glycyrrhetinic acid or a pharmaceutically acceptable salt thereof, dexamethasone acetate valerate, dexamethasone, prodnisolone valerate acetate (prednisolone valerate acetate), prodnisolone acetate, prodnisolone acetate, hydrocortisone acetate, hydrocortisone Ufenamate, bufexamac, ibuprofen piconol, glycol salicylate are exemplified, but are 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 the macrolide antibiotics include erythromycin, azithromycin, clarithromycin, tilmicosin, tylosin, and the like.Examples of the lincomycin antibiotics include clindamycin, lincomycin and the like, and the aminoglycoside antibiotics include Examples thereof include gentamicin sulfate, and examples of the chloramphenicol antibiotic 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, isothipendyl hydrochloride, lidocaine hydrochloride, thecaine, dibucaine, lidocaine, ethyl paraaminobenzoate and the like.

As vitamins, vitamins A such as retinol, retinal, retinoic acid and their derivatives (eg, retinol palmitate, tretinoin); nicotinic acid, nicotinic acid amide, nicotinic acid dl-α-tocopherol, benzyl nicotinate, nicotine. Nicotinic acids such as methyl acidate, β-butoxyethyl nicotinate, 1-(4-methylphenyl)ethyl nicotinate; pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-panthesine, D-panthetine, Pantothenic acids such as coenzyme A and pantothenyl ethyl ether; vitamin Es such as dl-α-tocopherol, dl-α-tocopherol acetate, dl-α-tocopherol succinate, and dl-α-tocopherol succinate; ascorbic acid Ascorbic acid derivatives such as sodium ascorbate, dehydroascorbic acid, sodium ascorbate phosphate, magnesium ascorbate phosphate, ascorbogen-A, ascorbyl stearate, ascorbyl palmitate, dipalmitate L-ascorbyl Vitamin Cs; thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate ester phosphate, Vitamin B1s such as thiamine monophosphate ester, thiamine diphosphate ester, thiamine diphosphate ester hydrochloride, thiamine triphosphate ester, thiamine triphosphate ester monophosphate salt, dibenzoyl thiamine, dibenzoyl thiamine hydrochloride salt; riboflavin, flavin mononucleotide , Flavin adenine dinucleotide, riboflavin butyric acid ester, riboflavin tetrabutyric acid ester, riboflavin 5'-phosphate sodium salt, riboflavin tetranicotinic acid ester, and other vitamin B2s; pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, 5'-pyridoxal phosphate , Vitamin B6s such as pyridoxamine hydrochloride; biotins such as biotin and bioticin; vitamin B12s such as cyanocobalamin, hydroxocobalamin, deoxyadenosylcobalamin; folic acid such as folic acid and pteroylglutamic acid; methylhesperidin, ergocalciferol, and chole. Vitamin Ds such as calciferol; phylloquino And vitamin Ks such as farnoquinone; adapalene (vitamin A-like acting factor), vitamins such as carnitine, ferulic acid, α-lipoic acid, orotic acid and the like.

(Example of combination with known acne remedy)
In the present invention, it is preferable to use a known acne remedy in combination with a basic amino acid having biofilm formation inhibiting ability. Known acne therapeutic agents that can be used in combination in such a manner 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 the present specification, the biofilm formation inhibitor, when used as a drug, a quasi drug, a cosmetic, etc., should be mixed with a known base or carrier to prepare a formulation as long as the effect of the present invention is not impaired. You can In addition, 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, isotonic agents, buffering agents, soothing agents, dispersants, fragrances, coloring agents, pigments and water can be added. These additives may be used alone or in combination of two or more.

As a base or carrier, liquid paraffin, squalane, gelled hydrocarbon (plastibase, etc.), ozokerite, α-olefin oligomer, hydrocarbon such as light liquid paraffin; methyl polysiloxane, cross-linked methyl polysiloxane, highly polymerized methyl Polysiloxane, cyclic silicone, alkyl modified silicone, crosslinked alkyl modified silicone, amino modified silicone, polyether modified silicone, polyglycerin modified silicone, crosslinked polyether modified silicone, crosslinked alkylpolyether modified silicone, silicone/alkyl chain Modified polyether modified silicone, silicone/alkyl chain co-modified polyglycerin modified silicone, polyether modified branched silicone, polyglycerin modified branched silicone, acrylic silicone, phenyl modified silicone, silicone oil such as silicone resin; polyethylene glycol; dioxane; myristin Esters such as isopropyl acid, octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, pentaerythritol tetra-2-ethylhexanoate; lower alcohols such as ethanol and isopropanol; diethylene glycol monomethyl ether, diethylene glycol mono Glycol ethers such as ethyl ether; polyhydric alcohols such as polyethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin and isoprene glycol; water-based bases such as water.

The base or carrier may be used alone or in combination of two or more.

Examples of the oil component include natural animal and vegetable oils and fats, hydrocarbon oils, ester oils, silicone oils, higher alcohols, higher fatty acids, animal and vegetable and synthetic essential oils, and the like.

Examples of natural animal and vegetable oils and fats include avocado oil, linseed oil, almond oil, olive oil, cocoa oil, beef tallow, tung 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, mokuro, coconut oil , Hardened coconut oil, peanut oil, lanolin, egg yolk oil, rosehip oil and the like.

As the hydrocarbon oil, paraffinic hydrocarbons and olefinic hydrocarbons are used, and examples thereof include squalane, squalene, ceresin, paraffin, pristane, microcrystalline wax, liquid paraffin and petrolatum.

As the ester oil, 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, isoprypir myristate, octyl myristate. Dodecyl, cetyl myristate, myristyl myristate, octyl palmitate, 2-ethylhexyl palmitate, 2-hexyldecyl palmitate, 2-heptylundecyl palmitate, cholesteryl 12-hydroxystearate, phytosteryl oleate, diisomalate. Stearyl, paramethoxycinnamic acid ester, tetrarosinic acid pentaerythritol and the like can be mentioned.

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

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

As the higher fatty acid, a saturated or unsaturated straight chain or branched chain 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 thereof include myristic acid, lauric acid, lanolin acid, linoleic acid and linolenic acid.

Examples of the thickener include guar gum, locust bean gum, carrageenan, xanthan gum, carboxymethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydrophobized hydroxypropyl methyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer. , Alkyl acrylate methacrylic acid copolymer, polyethylene glycol, bentonite, (hydroxyethyl acrylate/acryloyl dimethyl taurine Na) copolymer, (acryloyl dimethyl taurine ammonium/vinyl pyrrolidone) copolymer and the like.

Preferable preservatives include, for example, paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like.

Preferable examples of the antioxidant include sulfite, ascorbic acid and the like.

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, paraoxy. Examples thereof include methyl benzoate and phenoxyethanol.

Examples of the chelating agent include EDTA.sodium salt and EDTA.calcium.sodium salt.

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.) ), organic bases (triethanolamine, diisopropanolamine, triisopropanolamine, etc.) and the like.

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

Preferable examples of the solubilizing agent 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, for example, stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, surfactants such as glyceryl monostearate; for example, polyvinyl alcohol, Examples thereof include hydrophilic polymers such as polyvinylpyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose.

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

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

Preferable examples of soothing agents include benzyl alcohol and the like.

Examples of the dispersant include sodium pyrophosphate, sodium hexametaphosphate, polyvinyl alcohol, polyvinylpyrrolidone, methyl vinyl ether/maleic anhydride cross-linked copolymer, organic acid and the like.

Examples of the colorant include inorganic pigments and natural pigments.

In the present specification, the biofilm formation inhibitor can contain other active ingredients as long as the effects of the present invention are not impaired. Specific examples of the active ingredient include, for example, a moisturizing component, a pearly luster imparting agent, a conditioning agent, a blood circulation promoting component, an astringent component, an ultraviolet absorbing component, an ultraviolet scattering component, a washing component, a peptide or its derivative, an amino acid or its derivative, and a cell. An activating component etc. are mentioned.

Moisturizing ingredients include polyhydric alcohols such as glycerin, 1,3-butylene glycol, propylene glycol, polyethylene glycol and diglycerin; saccharides such as trehalose, xylitol and oligosaccharides; sodium hyaluronate, heparin analogues, 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 Lipids such as lipids; chamomile extract, hamamelis extract, tea extract, perilla extract, plant extract such as Yokuinin extract and the like.

Examples of the pearly 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. , Vinylimidazolium trichloride/vinylpyrrolidone copolymer, hydroxyethylcellulose/dimethyldiallylammonium chloride copolymer, vinylpyrrolidone/quaternized dimethylaminoethylmethacrylate copolymer, polyvinylpyrrolidone/alkylaminoacrylate copolymer, polyvinylpyrrolidone /Alkylaminoacrylate/vinylcaprolactam copolymer, vinylpyrrolidone/methacrylamidopropyltrimethylammonium chloride copolymer, alkylacrylamide/acrylate/alkylaminoalkylacrylamide/polyethylene glycol methacrylate copolymer, adipic acid/dimethylaminohydroxypropylethylenetriamine Examples thereof include copolymers.

Examples of blood circulation promoters include acetylcholine, ictamol, caffeine, capsaicin, cantharisin tincture, gamma-oryzanol, ginger oriental tincture, zingerone, cepharanthin, senbri extract, tannic acid, capsicum tincture, tolazoline, tocopherol nicotinate, and benzyl nicotinate. Esters and the like can be mentioned.

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

As the ultraviolet absorbing component, octyltriazone, hexyl diethylaminohydroxybenzoylbenzoate, octyl dimethoxybenzylidene dioxoimidazolidine propionate, 2-ethylhexyl paramethoxycinnamate, t-butylmethoxydibenzoylmethane, phenylbenzimidazole sulfonic acid, Examples include octyl methoxycinnamate and ethylhexyl methoxycinnamate.

As the ultraviolet scattering component, hydrous silicic acid, zinc silicate, cerium silicate, titanium silicate, zinc oxide, zirconium oxide, cerium oxide, titanium oxide, iron oxide, inorganic compounds such as silicic acid anhydride, those inorganic compounds With hydrous silicic acid, aluminum hydroxide, inorganic powder such as mica or talc, or composite with resin powder such as polyamide, polyethylene, polyester, polystyrene, nylon, etc. Examples include processed products.

As a cleaning component, an alkali metal salt such as potassium laurate, potassium myristate, potassium palmitate or potassium stearate, a soap such as an alkanolamide salt or an amino acid salt, an amino acid-based interface 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 alkys sulfosuccinate ester, coconut oil fatty acid monoethanolamide, coconut oil fatty acid ethanolamide Fatty acid alkanolamides such as, sodium lauryl phosphate, monoalkyl phosphate ester salts such as sodium polyoxyethylene lauryl ether phosphate, coconut oil fatty acid amide propyldimethylaminoacetic acid betaine, lauryldimethylaminoacetic acid betaine, 2-alkyl-N- Betaine-type amphoteric surfactants such as carboxymethyl-N-hydroxyethyl imidazolinium betaine, lauryl hydroxysulfobetaine and lauroylamidoethyl hydroxyethyl carboxymethyl betaine hydroxypropyl sodium phosphate, amino acid-type amphoteric surfactants such as sodium lauryl aminopropionate Activator etc. are mentioned.

Peptides or derivatives thereof, 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, hydrolyzing conchiolin, silk proteolytic peptide, hydrolyzing silk, sodium lauroyl hydrolyzing silk, soybean proteolytic peptide, hydrolyzing soybean protein, wheat protein, wheat proteolytic peptide, hydrolyzing wheat protein, casein decomposing peptide , Acylated peptides (palmitoyl oligopeptide, palmitoyl pentapeptide, palmitoyl tetrapeptide, etc.) and the like.

As the amino acid or its derivative, 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-activating components include γ-aminobutyric acid, amino acids such as ε-aminocaproic acid, retinol, thiamine, riboflavin, pyridoxine hydrochloride, vitamins such as pantothenic acids, glycolic acid, α-hydroxy acids such as lactic acid, tannin, Examples include flavonoid, saponin, allantoin, and Photosensitizer No. 301.

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

(Preparation of formulation)
The 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 a concentration 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.

(Biofilm formation inhibition test of Acne bacteria)
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.), and L-glutamic acid (manufactured by Wako Pure Chemical Industries, Ltd.) It carried out using. Specifically, each test solution inoculated into a 96-well round-bottom plate so that the P. acnes suspension became 10 ⁸ (CFU/mL) bacterial solution was dispensed at 100 μL per well. Then, anaerobic culture was carried out at 36° C. for 4 days. After culturing, each well was washed twice with 200 μL of purified water and stained with 100 μL of 0.5% crystal violet for 20 minutes. After washing each well with 200 μL of purified water three times, the crystal violet attached to the plate was dissolved using 160 μL of 33% acetic acid. Using a microplate reader (Molecular Devices, Versa max), the absorbance at 590 nm at which crystal violet exhibits an absorbance was measured to determine the amount of biofilm formation. A physiological saline solution (manufactured by Otsuka Pharmaceutical Co., Ltd.) adjusted to pH 7 or 5 was prepared as a control, and the biofilm formation inhibition rate was calculated by the following formula using the ratio to this biofilm formation amount. For the test of statistically significant difference, P value was calculated using Dunnett test (n=6). The results are shown in Tables 1-2 and FIGS.

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

As shown in the above results, at pH 7, basic amino acids such as arginine and histidine significantly suppressed the biofilm formation of P. acnes. On the other hand, glycine, which is a neutral amino acid that is not a basic amino acid, and glutamic acid, which is an acidic amino acid, were not effective in suppressing the biofilm of Acne bacteria. In addition, even at pH 5, the basic amino acids significantly suppressed the biofilm of P. acnes. From this result, for example, when the basic amino acids are used in combination with other known acne therapeutic agents, it becomes possible to enhance the penetration of the known acne therapeutic agents into the acne bacterium 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.). After preparing each test solution (pH 7) in the same manner as in the above-mentioned example, a 96-well round bottom plate was inoculated with 10 ⁸ (CFU/mL) bacterial solution 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 culturing, 100 μL (n=3) of the preparation in a 96-well round bottom plate was collected, diluted 10- to 1000-fold with physiological saline (Otsuka Pharmaceutical Co., Ltd.), seeded on GAM agar medium (Nissui Pharmaceutical Co., Ltd.), and agar plate. The number of viable bacteria was determined by the surface smear method. In addition, physiological saline adjusted to pH 7 (manufactured by Otsuka Pharmaceutical Co., Ltd.) was prepared as a control, and treated in the same manner as in the case of using the test solution to determine the viable cell count. The results are shown in Table 3.

As shown in the above results, basic amino acids such as arginine and histidine did not show antibacterial action against P. acnes. Therefore, it was shown that the biofilm formation inhibitor of the present invention does not exhibit bactericidal activity but has biofilm formation inhibitory activity.

(Formulation example)
The formulation examples of the biofilm formation inhibitor of the present invention are shown below. Prescription examples 1 to 4 are lotions, and prescription examples 5 to 8 are creams (oil-in-water type). Both are prepared according to conventional methods. These formulation examples can be used to suppress biofilm formation by microorganisms such as Acne bacteria.

Claims (10)

A biofilm formation inhibitor containing basic amino acids. The biofilm formation inhibitor according to claim 1, which suppresses biofilm formation of Acne bacteria. 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 according to any one of claims 1 to 4, which is applied to the skin or the oral cavity. The biofilm formation inhibitor according to any one of claims 1 to 5, which is used for treating or preventing acne. The biofilm formation inhibitor according to any one of claims 1 to 6, which is used for treating or preventing acne on dry skin. The biofilm formation inhibitor according to any one of claims 1 to 7, which is used for treating or preventing acne that tends to be repeated. The biofilm formation inhibitor according to any one of claims 1 to 8, which is used for treating or preventing acne with 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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