JP2008100964A - Biofilm formation inhibitor composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a drug and a composition for inhibiting biofilm formation. <P>SOLUTION: This biofilm formation inhibitor composition contains a compound represented by a component (A) or its salt, and (B) a surfactant, and has a weight ratio of component (A)/component (B) of ≤2. (A) A general formula (1) [R is a 8-14C straight chain or branched alkyl or alkenyl group; EO is ethyleneoxy group; (m) and (n) are each an average addition molar number; and (m)+(n) is a number of 0 to 30]. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a biofilm production inhibitor composition. More specifically, the present invention relates to a biofilm production inhibitor composition for suppressing the production of biofilms composed of microorganisms and microorganism-producing substances in various fields in which microorganisms are involved, and preventing harm resulting therefrom.

  A biofilm is also called a biofilm or slime, and generally refers to an aqueous system in which microorganisms adhere to and grow on the surface of substances to produce macromolecular substances such as polysaccharides and proteins from within microbial cells to form structures. . When a biofilm is formed, harms caused by microorganisms occur and cause problems in various industrial fields. For example, when a biofilm is formed in the piping of a food plant, the biofilm is peeled off and not only leads to contamination of foreign substances in the product, but also causes food poisoning due to microorganism-derived toxins. Furthermore, biofilm formation on the metal surface causes metal corrosion and promotes aging of the equipment.

  Furthermore, microbial control agents such as bactericides and bacteriostatic agents are not sufficiently effective against microorganism aggregates that have formed biofilms compared to microorganisms that are dispersed and suspended in water. There are also many. For example, in the medical field, many cases of nosocomial infections have been reported in recent years due to microorganisms remaining in narrow gaps and holes in medical devices to form biofilms. In the human oral cavity, it is well known that biofilms formed on teeth, so-called dental plaque (dental plaque), cause caries and periodontal disease, and these problems have been studied for a long time.

Until now, in order to suppress biofilms, the idea of not allowing bacteria to grow by giving bactericidal or bacteriostatic action to microorganisms, particularly bacteria, has been generally studied. Patent Document 1 discloses that the number of bacteria can be reduced by using fatty acid, aliphatic alcohol, or the like, and as a result, adhesion of bacteria to the target substance can be prevented. Moreover, in patent document 1, the composition which prepared the emulsion with the antimicrobial oil phase and the emulsifier has shown the bacteria count reduction effect in a comparatively short time, and this is based on that the absolute number of bacteria per unit volume becomes low. This represents the idea of suppressing the adhesion of bacteria to the surface of the target substance.
Patent Document 2 discloses a method of contacting with a specific alkylamine derivative in order to prevent the establishment of pollutant organisms (barnacles, bacterial slime, etc.) on the underwater structure. It did not suppress generation.
Special Table 2002-524257 Special Table 2004-525935

  Patent Document 1 describes the evaluation of bactericidal properties (reducing the number of bacteria by about the fourth power) when microorganisms are sterilized for a relatively short time within 60 minutes or contacted with an antibacterial composition. However, the biofilm problem occurs in units of a long time of several days to several months, and it is practically difficult to link to biofilm production suppression control by a short sterilization evaluation. Fatty acids and fatty alcohols listed as antibacterial oil phases do not have a sufficient bactericidal effect on all microorganisms (bacteria), especially in gram-negative bacteria that often form biofilms and cause problems On the other hand, it does not have the minimum growth inhibitory concentration (MIC), which is an index of long-term bactericidal effect (Science of antiseptic and bactericidal agents; edited by John J. Kabbalah, Fragrance Journal, 1990) . Furthermore, according to the experiments by the inventors, the composition described in Patent Document 1 shows a short-term (up to about 3 hours) bactericidal effect against Pseudomonas aeruginosa and Serratia bacteria among gram-negative bacteria, In the long term (one day or more), it was confirmed that bactericidal and even bacteriostatic effects to suppress bacterial growth were not exhibited, and as a result, a biofilm was formed.

In addition, there are highly bactericidal bactericides such as cationic surfactants with high bactericidal properties and hypochlorite, but bactericidal properties are quickly lost when organic substances are present in the system. As described above, it is difficult to maintain the effect of reducing the number of bacteria over a long period of time.
For these reasons, it has been difficult to fundamentally suppress the production of biofilms from the viewpoint of bactericidal and bacteriostatic.
Accordingly, an object of the present invention is to provide a long-term biofilm formation inhibitor composition.

Therefore, the present inventor searched for a composition that suppresses biofilm formation over a long period of time. It was found that biofilm formation was suppressed.
The present invention provides the following component (A)
(A) General formula (1)

(In the formula, R represents a linear or branched alkyl or alkenyl group having 8 to 14 carbon atoms, EO represents an ethyleneoxy group, m and n are average added mole numbers, and m + n is 0 to 30. Or a salt thereof, and (B) a surfactant, and the weight ratio (A) / (B) of the component (A) to the component (B) is 2 or less. A biofilm production inhibitor composition is provided.

  The present invention also provides a method for inhibiting the production of a biofilm by bringing the biofilm production inhibitor composition into contact with a microorganism.

  According to the present invention, biofilm production can be suppressed in the long term.

  The component (A) of the biofilm production inhibitor composition of the present invention has the general formula (1):

Wherein R is a linear or branched alkyl or alkenyl group having 8 to 14 carbon atoms, EO is an ethyleneoxy group, and m and n are the average number of moles added, m + n Is a number from 0 to 30.

  The alkyl group or alkenyl group represented by R may be linear or branched, but preferably has 10 to 12 carbon atoms. The number m + n of the ethyleneoxy group represented by EO is preferably 0-15, particularly preferably 0-5. Examples of the salt of the compound represented by the general formula (1) include mineral acid salts such as hydrochloride, sulfate and phosphate, and organic acid salts such as acetate and citrate.

  The component (A) of the present invention may be present in the system in an amount of 1 ppm or more as a weight concentration capable of exhibiting a long-term biofilm formation inhibitory effect, but is preferably 1 to 10,000 ppm from the viewpoint of economy and effect. -10000 ppm is more preferable, 5-2000 ppm is more preferable, and 10-1000 ppm is particularly preferable.

  Since the component (A) of the present invention has relatively high hydrophobicity and low solubility in water, the present agent can be more effectively used in the aqueous system by using the surfactant to stably exist in the aqueous system. It becomes possible to use.

  Here, “stablely present in the aqueous system” means a state in which the highly hydrophobic component (A) is emulsified / dispersed / solubilized without being separated for a long period of time. A larger amount of component (A) can be emulsified, dispersed and solubilized.

  Although the kind of surfactant which can be used for the biofilm production inhibitor composition of the present invention is not particularly limited, a surfactant capable of stably presenting the component (A) in the aqueous system is desirable. In particular, from the viewpoint of emulsification, dispersion, and solubilization performance, it is preferable to use an anionic surfactant or a nonionic surfactant among the surfactants.

  Examples of the anionic surfactant include lignin sulfonate, alkylbenzene sulfonate, alkyl sulfonate, polyoxyethylene (hereinafter referred to as POE) alkyl sulfonate, POE alkylphenyl ether sulfonate, and POE alkylphenyl. Ether phosphate ester salt, POE arylphenyl ether sulfonate, alkyl sulfate ester salt, POE alkyl sulfate ester salt, POE aryl phenyl ether phosphate ester salt, naphthalene sulfonate, naphthalene sulfonate formalin condensate, POE tribenzylphenyl Ether sulfonate, alkyl phosphate, POE alkyl phosphate, POE tribenzylphenyl ether phosphate ester salt, dialkyl sulfosuccinate, fatty acid salt (soap), POE Al Ether acetates, and among them alkyl sulfates and POE alkyl sulfates, it is more preferable to use a POE alkyl ether acetic acid salt. These anionic surfactants preferably have 10 to 18 alkyl carbon atoms, and the ethylene oxide average addition mole number is preferably 0 to 10, and more preferably 0 to 5.

  Examples of the nonionic surfactant include POE alkyl ether, POE alkyl phenyl ether, polyoxypropylene / POE (block or random) alkyl ether, POE aryl phenyl ether, POE styrenated phenyl ether, POE tribenzyl phenyl ether, and the like. Polyhydric alcohol derivative-type nonionic surfactants; (poly) glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, POE sorbitan fatty acid esters, polyhydric alcohol derivative-type nonionic surfactants such as alkylpolyglycosides, etc. Among them, POE alkyl ether, (poly) glycerin fatty acid ester, alkyl polyglycoside, sorbitan fatty acid ester, and POE sorbitan fatty acid ester are preferable. Alkyl ether. In particular, the HLB of the POE alkyl ether is particularly preferably 10 or more. Further, the POE alkyl ether preferably has 12 to 18 alkyl carbon atoms, and the ethylene oxide average addition mole number is particularly preferably 6 or more.

  Surfactants can be used alone or in combination of two or more in order to enhance the emulsification / dispersion / solubilization performance.

The weight ratio (A) / (B) of the component (A) to the component (B) in the biofilm production inhibitor composition is preferably 2/1 to 1/100 from the viewpoint of long-term biofilm production inhibition effect. 2/1 to 1/50 is more preferable, 2/1 to 1/20 is more preferable, and 1/1 to 1/10 is particularly preferable.
The biofilm production inhibitor composition preferably contains 0.01 to 10% by weight of component (A), more preferably 0.1 to 10% by weight, particularly preferably 0.5 to 5% by weight, and The component (B) is preferably contained in an amount of 0.1 to 30% by weight, more preferably 1 to 20% by weight.

  The biofilm production inhibitor composition of the present invention can be used in combination with a bactericidal agent or an antibacterial agent. Generally, when a biofilm is formed, a situation occurs in which the bactericidal agent is hardly effective. However, when the biofilm formation is suppressed by the biofilm formation inhibitor of the present invention, the bactericidal agent can be sufficiently effective.

  The above bactericides and antibacterials include benzalkonium chloride, benzethonium chloride, didecyldimethylammonium chloride, cetylpyridinium chloride, chlorhexidine hydrochloride, chlorhexidine gluconate, polyhexamethylenebiguanidine hydrochloride, triclosan, isopropyl Examples include methylphenol, trichlorocarbanilide, ethanol, isopropyl alcohol and the like.

  In the biofilm production inhibitor composition of the present invention, it is possible to use a thickener in order to increase the viscosity and improve the adhesion to an object.

  Furthermore, a chelating agent may be added to the biofilm production inhibitor composition of the present invention. Examples of the chelating agent include ethylenediaminetetraacetic acid (EDTA), succinic acid, salicylic acid, oxalic acid, malic acid, lactic acid, fumaric acid, tartaric acid, citric acid, gluconic acid, tripolyphosphoric acid, 1-hydroxyethane-1,1-diphosphone. Examples include acids, polyacrylic acid, acrylic acid / maleic acid copolymers, and salts thereof.

  The biofilm production inhibitor composition of the present invention can take various forms such as liquids, pastes, powders, tablets and the like depending on applications. The biofilm production inhibitor composition may be a one-component type in which all components are mixed, but it may be divided into several divided packages depending on ease of use.

The biofilm production inhibitor composition of the present invention is effective when used in a water dilution system. A fixed amount of the diluted water of the composition is accumulated and the object is immersed for use. When the object covers a wide range, the mist may be sprayed using a spray device, or the foamed material may be sprayed using a foaming machine. Further, the composition may be applied by pouring a water-diluted solution or brushing. Alternatively, the object may be wiped off by impregnating the towel with a water-diluted solution. If the conditions for contact with microorganisms are satisfied, it is possible to attach or apply a water dilution of the composition to the surface where microorganisms may be present. The water dilution of the composition preferably has a weight concentration of component (A) of 1 to 10,000 ppm, more preferably 5 to 10,000 ppm.
In addition, depending on the object, it is possible to spread it in the form of cream or ointment without using a water dilution system. In this case, the component (A) is provided in a form dissolved, dispersed, and emulsified in an appropriate solvent, and the weight concentration of the component (A) at the time of use is preferably 1 to 10,000 ppm, and preferably 5 to 10,000 ppm. More preferably.

  The present invention also provides a method for inhibiting biofilm production by bringing a biofilm production inhibitor composition into contact with a microorganism. Here, the contact between the biofilm formation inhibitor composition and the microorganism is preferably performed continuously.

  The biofilm production inhibitor composition of the present invention can be used in a wide range of fields in which biofilms are a concern. For example, the present invention can be applied to cleaning agents for food or beverage production plants with a high risk of bacterial contamination. In addition, the present invention can be applied to cleaning agents such as medical devices that easily form biofilms, such as endoscopes and artificial dialysis machines. Furthermore, since it has high safety, it can be used as a cleaning agent, dentifrice, oral care agent, denture care agent and the like for human subjects.

Example 1: Composition of biofilm production inhibitor composition and assay component for biofilm production inhibition ability (A) [General formula (1): EO represents ethyleneoxy group, m and n represent average number of moles added ]

(A-1) C8 amine [Farmin 08D, manufactured by Kao Corporation, R = C8 alkyl, m + n = 0]
(A-2) C10 amine [decylamine, manufactured by Wako Pure Chemical Industries, Ltd., R = C10 alkyl, m + n = 0]
(A-3) C12 amine [Farmin 20D, manufactured by Kao Corporation, R = C12 alkyl, m + n = 0]
(A-4) C12 amine hydrochloride [dodecylamine hydrochloride, manufactured by Wako Pure Chemical Industries, Ltd., R = C12, m + n = 0]
(A-5) Cocoamine [Farmin CS, manufactured by Kao Corporation, R = C8-14 alkyl (coconut composition), m + n = 0]
(A-6) Cocoamine acetate [Acetamine 24, manufactured by Kao Corporation, R = C8-14 alkyl (coconut composition), m + n = 0]
(A-7) Cocoamine ethylene oxide 2 mol adduct [Amete 102, manufactured by Kao Corporation, R = C8-14 alkyl (coconut composition), m + n = 2]
(A-8) Cocoamine ethylene oxide 5 mol adduct [Amate 105, manufactured by Kao Corporation, R = C8-14 alkyl (coconut composition), m + n = 5]

Component (A ′) [General formula (2): EO represents an ethyleneoxy group, and m and n represent the average number of moles added]

(A′-1) C3 amine [propylamine, manufactured by Wako Pure Chemical Industries, Ltd., R ′ = C3 alkyl, m + n = 0]
(A′-2) C3 amine hydrochloride [propylamine hydrochloride, manufactured by Wako Pure Chemical Industries, Ltd., R ′ = C3 alkyl, m + n = 0]
(A′-3) C6 amine [hexylamine, manufactured by Wako Pure Chemical Industries, Ltd., R ′ = C6 alkyl, m + n = 0]
(A′-4) C6 amine hydrochloride [hexylamine hydrochloride, manufactured by Wako Pure Chemical Industries, Ltd., R ′ = C6 alkyl, m + n = 0]
(A′-5) Cured Tallowamine [Farmin 86T, manufactured by Kao Corporation, R ′ = C16,18 alkyl, m + n = 0]
(A′-6) C18 amine [Farmin 80, manufactured by Kao Corporation, R ′ = C18 alkyl, m + n = 0]
(A′-7) Oleylamine [Farmin O, manufactured by Kao Corporation, R ′ = C18 alkenyl, m + n = 0]
(A′-8) Cured Tallowamine Ethylene Oxide 2-Mole Adduct [Amit 302, manufactured by Kao Corporation, R ′ = C16,18 alkyl, m + n = 2]
(A′-9) Cured Tallowamine Ethylene oxide 20 mol adduct [Amete 320, manufactured by Kao Corporation, R ′ = C16,18 alkyl, m + n = 20]

Component (B) Surfactant [Numbers in parentheses indicate the average number of moles of ethylene oxide added. ]
<Anionic surfactant>
(B-1) Sodium lauryl sulfate [Emar 0, manufactured by Kao Corporation]
(B-2) Polyoxyethylene (2) Sodium lauryl ether sulfate [Emar 20C, manufactured by Kao Corporation; effective content 25% by weight]
(B-3) Polyoxyethylene (4.5) sodium lauryl ether acetate [Kao Akipo RLM45-NV, manufactured by Kao Corporation; effective 24% by weight]
<Nonionic surfactant>
(B-4) Polyoxyethylene (6) lauryl ether [Emulgen 106, manufactured by Kao Corporation]
(B-5) Polyoxyethylene (12) lauryl ether [Emulgen 120, manufactured by Kao Corporation]
(B-6) Lauryl glycoside [Mydol 12, manufactured by Kao Corporation]
(B-7) Decylglycerin monocaprylate [SY Glister MCA750, manufactured by Sakamoto Pharmaceutical Co., Ltd.]
(B-8) Sorbitan monolaurate [Leodol SP-L10, manufactured by Kao Corporation]
(B-9) Polyoxyethylene (6) sorbitan monolaurate [Leodol TW-L106, manufactured by Kao Corporation]
(B-10) Polyoxyethylene (20) sorbitan monolaurate [Leodol TW-L120, manufactured by Kao Corporation]

Component (A) or (A ′) is fixed at 1% by weight, component (B) is at a concentration selected from 1% by weight, 3% by weight, 6% by weight and 10% by weight, and the remainder is effective with ion-exchanged water. Mixed weight. The compound was diluted to 100 ppm as a component (A) or (A ′) using Mueller Hinton medium [manufactured by Nippon Becton Dickinson Co., Ltd.] into a 24-well microplate (manufactured by Asahi Techno Glass Co., Ltd.). Weighed 2 mL.
Pseudomonas aeruginosa NBRC13275, Serratia marcescens NBRC12648, Klebsiella pneumoniae ATCC13883, soybean-Casein Digest Agar [SCD agar medium] Was used to inoculate the test solution in the microplate using a sterilized bamboo skewer from a colony formed by pre-culturing at 37 ° C. for 24 hours. The culture solution was discarded after culturing at 37 ° C. for 48 hours, and the formation state of the biofilm adhered to the microplate wall was visually observed. The state of the biofilm is ◎ when the biofilm covers 0 to less than 20% of the plate wall surface, ◯ when it covers 20% or more and less than 40%, △ or 60% when it covers 40% or more but less than 60% What covered the above was made into x.
The results are shown in Tables 1-1 to 1-4.

Example 2: Biofilm production reduction test in a large-capacity plastic cup Pseudomonas aeruginosa NBRC13275 was used at 37 ° C for 24 hours using Soybean-Casein Digest Agar [SCD agar medium: manufactured by Nippon Pharmaceutical Co., Ltd.] Pre-cultured. The colonies grown on the medium are scraped off and suspended in 10 mM sterile phosphate buffer (pH 7.2), then washed twice by centrifugation at 5,000 × g for 15 minutes at 10 ° C., and again with 10 mM sterile phosphorus. A bacterial solution was prepared by suspending in an acid buffer (pH 7.2) and adjusting the bacterial concentration to 1.0 (OD ₆₀₀ = 1.0) at ₆₀₀ nm absorbance. Thereafter, 100 mL of Mueller Hinton medium (manufactured by Nippon Becton Dickinson Co., Ltd.) and 15 kinds of test agents selected from Example 1 may be put into a 200 mL sterilized screw cup (Eiken Equipment Co., Ltd.). After mixing and adjusting the concentration as the component (A) or (A ′) to 5 ppm, 10 ppm, 50 ppm, 100 ppm or 500 ppm, 0.1 mL of the aforementioned prepared bacterial solution was inoculated. In addition, as a control, a test group in which bacteria were inoculated in the same manner as described above on a Mueller Hinton medium to which no drug was added was simultaneously provided.
These were statically cultured at 37 ° C., and after 1 day, 2 days, 3 days, and 5 days, the number of bacteria in the culture was measured and the biofilm formed in the cup was visually observed. The precipitate was taken out by centrifugation at 5,000 × g for 30 minutes at 5 ° C., dried in a vacuum desiccator for 24 hours, and weighed to obtain the weight of the biofilm produced in the culture solution. The biofilm production state is ○ when the biofilm is not confirmed in the cup, △ when the biofilm starts to form at the gas-liquid interface in the cup, and the biofilm enters the culture solution from the gas-liquid interface. The state that has been generated up to now was determined as x.
The results are shown in Tables 2-1 to 2-3.

  From the above results, it can be seen that the product of the present invention can effectively suppress the production of biofilm.

Claims (6)

Next ingredient (A)
(A) General formula (1)

(In the formula, R represents a linear or branched alkyl or alkenyl group having 8 to 14 carbon atoms, EO represents an ethyleneoxy group, m and n are average added mole numbers, and m + n is 0 to 30. Or a salt thereof, and (B) a surfactant, and the weight ratio (A) / (B) of the component (A) to the component (B) is 2 or less. Biofilm production inhibitor composition.   (B) The biofilm production inhibitor composition according to claim 1, wherein the surfactant is at least one selected from the group consisting of an anionic surfactant and a nonionic surfactant.   (B) The biofilm production inhibitor composition according to claim 1, wherein the surfactant is a nonionic surfactant.   The biofilm production | generation inhibitor composition of any one of Claims 1-3 is made to contact microorganisms, and the method of suppressing the production | generation of a biofilm.   The method for inhibiting biofilm production according to claim 4, wherein the biofilm production inhibitor composition and the microorganism are continuously contacted.   The method for suppressing the production of a biofilm according to claim 4 or 5, wherein the weight concentration of the component (A) during use of the biofilm production inhibitor composition is 1 to 10,000 ppm.