JP2010215598A - Virucidal agent composition for calicivirus and method of application thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a virucidal agent composition for calicivirus which has an excellent virucidal effect and is highly safe for handling, and a method for application of the same. <P>SOLUTION: The virucidal agent for calicivirus includes 1,4-bis(3,3'-(1-decyl pyridinium)methyloxy)butane dibromide and an ethylenediamine tetraacetate as effective components. The virucidal method for calicivirus comprises bringing 1,4-bis(3,3'-(1-decyl pyridinium)methyloxy)butane dibromide into contact with caliciviruses in the presence of the ethylenediamine tetraacetate. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a calicidal virus composition and a method of using the same.

  One of the reasons for struggling with hygiene management measures at food production and cooking facilities and medical facilities is food poisoning caused by norovirus of the Caliciviridae family. In FY2006, the incidence of food poisoning due to norovirus was the highest in both the number of cases and the number of patients, the number of occurrences was about 500, and the number of patients was very high at about 27600, resulting from its strong infectivity. This has led to the occurrence of large-scale food poisoning. In addition, it was said that the cause of food poisoning due to norovirus was to eat bivalves such as oysters, but recently, norovirus contained in food, feces and vomit is infected from person to person, Furthermore, it has been found that secondary contamination, such as spreading through contact with a clean surface via contaminated food or water, is the main cause. To prevent this secondary contamination, thorough hand washing, sterilization of cooking utensils and tools such as cutting boards and knives, sterilization of cooking tables and work tables, toilet door knobs and toilet seats, cleaning of medical equipment and treatment of vomiting, etc. Becomes important.

  Therefore, a drug used for sanitation in food production facilities, cooking facilities, and medical facilities is desired to be effective against calicivirus, safe, less corrosive, and economical.

  By the way, “Norovirus” was formerly called Norwalk virus or small spherical virus, but it was determined by the International Virological Society as the Norovirus genus of the Caliciviridae family. In other words, norovirus is an RNA-type virus that has only RNA, and is surrounded by a protein shell called capsid and does not have a membrane made of sugar and lipid called envelope.

  In general, a virus having an envelope can be inactivated because the envelope is easily destroyed by a drug and cannot bind to a host cell receptor. However, because norovirus does not have this envelope, it is resistant to drugs.

As a test for norovirus, there is a method in which norovirus is collected from human stool, contacted with a drug, DNA destruction of norovirus is confirmed by PCR method, and viability of norovirus is discriminated. Since norovirus has not been successfully infected to cultured cells and animals, it is used in this way collected from human-derived feces. However, since this has a problem that it is not quantitative and expensive, feline calicivirus F9 strain, which is the same calicivirus, is currently used as an alternative virus, and EPA and CEN norovirus test methods Then, feline calicivirus F9 strain is used as an alternative virus.

  Because this feline calicivirus test method can be handled safely without human infection and can be quantified by culture, feline calicivirus is commonly used as a substitute for norovirus in Japan and overseas. .

  As a method for inactivating norovirus, a method using a sodium hypochlorite solution and a method using hot water disinfection are known (see Non-Patent Document 1). However, in hot water disinfection, it is desirable to maintain a temperature of 85 ° C. or higher. However, if hot water is applied to the object to be disinfected, the temperature decreases due to contact with the object to be disinfected, and the desired temperature can be maintained. It is difficult and there is a risk of burns, making it unsafe.

  Moreover, in the disinfection method using a sodium hypochlorite solution, rust and corrosion may occur when the object to be disinfected is a metal. Furthermore, depending on the disinfectant, there are safety problems such as bleaching due to the bleaching action / effect of sodium hypochlorite and the odor of chlorine filling the kitchen / workplace.

  Because of these problems, alcohol is usually used for the sterilization of cooking utensils such as cutting boards and kitchen knives. However, alcohol inactivates the feline calicivirus F9 strain, which is a substitute for norovirus. (See Non-Patent Documents 2 and 3).

  Further, it is known that a disinfectant containing a cationic surfactant system exhibits an effect on food poisoning bacteria but has no inactivation effect on viruses (Non-patent Document 3). And 4).

  On the other hand, there is also a report that virus inactivation is observed when a cationic surfactant is used and an alkali agent is added (see Non-Patent Documents 5 and 6).

  In addition, there are reports that the combination of ethanol, alkaline substances and cationic fungicides is effective in inactivating feline calicivirus and can be expected to be applied to disinfection of hard surfaces contaminated with microorganisms containing viruses (non- (See Patent Document 7).

  Japanese Patent Application Laid-Open No. 2007-45732 discloses a disinfecting solution containing 0.05 to 0.5% by weight of a polyhexamethylene biguanide compound and having a pH in the range of 9 to 12, wherein the disinfecting solution contains alcohol. What contains 40-80 weight% is described. Furthermore, it can contain other antibacterial components such as benzalkonium chloride, cetylviridinium chloride, phenoxyethanol, chlorohexidine gluconate, lauryl sulfate, etc. An article for disinfection is disclosed (see Patent Document 1).

  The method of inactivating a virus by adding an alkali agent to the cationic surfactant or polyhexamethylene biguanide compound as described above has a problem that it is difficult to handle and safe because it has a high pH. It was.

JP 2007-45732 A

“Norovirus Food Poisoning Countermeasures (Proposal)” Food and Drug Hygiene Council Food Hygiene Division Food Poisoning Department (October 12, 2007) “Inactivation of Caliciviruses”, American Society for Microbiology, Vol. 70, no. 8, 4538-4543 (2004) “Inactivation of feline calicirus, a Norwalk virus surrogate”, J. Am. Hosp. Infect, 41, 51-57 (1999) “Virtual Disinfectants and Feline Viruses”, Am. J. et al. Vet. Res. , Vol. 41, no. 3, 410-414 (1980) "Efficacy of common used discovered for the inactivation of calicirus on strawberry, lettuce, and food-contact surface", J. Am. Food. Prot. 64, 1430-1434 (2001) “Effects of quarterly ammonium compounds with 0.1% sodium hydroxide on sine vesicular disease virus”, J. Am. Vet. Med. Sci. , 59, 323-328 (2007) “Development of norovirus-substituted feline calicivirus and ethanol preparation effective for various microorganisms”, antibacterial and antifungal, Vol. 35, no. 11, 725-732 (2007)

  However, the technical contents and compositions described in these documents only satisfy specific performances, and it is a fact that comprehensively satisfactory products are not obtained.

  The present invention has been made in view of such circumstances, and an object thereof is to provide a calicidal virus composition having an excellent calicidal virus effect and high safety in handling, and a method for using the same.

  1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide is widely known as a bactericidal agent and has little effect on calicivirus. However, as a result of intensive studies to achieve the above object, the present inventors have found 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide to ethylenediaminetetraacetic acid. It has been found that the use of salt has an excellent virucidal effect against calicivirus while maintaining the bactericidal effect.

  Such a result was completely unpredictable because both components alone had little calicidal virus effect.

  That is, the present invention relates to (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide in an amount of 0.01% by mass or more, and (B) ethylenediaminetetraacetate in an amount of 0.0% by mass. The first gist is a calicivirus composition containing 000625% by mass or more.

  The present invention also provides 0.01% by mass or more of (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide and 0.000625% by mass of (B) A second aspect is a calicivirus method characterized by contacting with a calicivirus in the presence of ethylenediaminetetraacetate.

  According to the present invention, by combining (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide and (B) ethylenediaminetetraacetate, the effect of calicivirus is achieved. Can be obtained. In addition, the calicidal virus composition of the present invention can be used in a sanitary manner, and since it does not use an alkaline agent, the handling safety is high.

  Next, the best mode for carrying out the present invention will be described in detail.

  In the present invention, (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide is 0.01 mass% or more, and (B) ethylenediaminetetraacetate is 0.000625 mass. % Or more of (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane having a mass of 0.01% by mass or more. It is characterized by contacting dibromide with calicivirus in the presence of 0.000625% by mass or more of (B) ethylenediaminetetraacetate.

  (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide used in the present invention is commercially available. For example, trade name: “Hygenia” (manufactured by Tama Chemical Co., Ltd.) ) Etc.

  (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide has almost no calicivirus effect even if it acts on calicivirus alone, but surprisingly (B) When (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide is allowed to act in the presence of ethylenediaminetetraacetate, an extremely high calicicidal virus effect is obtained. It has been found that it can be obtained. The mechanism of action is unknown, but at present it is thought to be due to the synergistic effect of ethylenediaminetetraacetate and 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide. Yes.

  Since the calicidal virus composition of the present invention is effective against not only bacteria but also calicivirus, it can be used hygienically. In addition, since an alkaline agent is not used, the handling safety is high. Therefore, the calicidal virus composition of the present invention is useful as a calicidal virus agent in food production, cooking facilities and medical facilities.

  (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide, which is an active ingredient of the calicidal virus composition of the present invention, is used as an active ingredient when used. It is used in the range of 0.01% by mass or more in the virus agent composition or diluent. That is, if it is less than 0.01% by mass, the desired calicivirus performance cannot be obtained, which is not preferable.

  Moreover, (B) ethylenediaminetetraacetate is used at 0.000625 mass% or more. That is, if it is less than 0.000625 mass%, the desired calicidal virus performance cannot be obtained, which is not preferable.

  In order to use the calicidal virus method of the present invention safely in handling, preferably the calicidal virus composition is used as a stock solution at room temperature or diluted 10-fold to 1000-fold with water, and the pH (JIS Z-8802) is used. : 1984 “pH measurement method”) is preferably brought into contact with calicivirus at 25 ° C. in the range of 4-8.

  Furthermore, (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide and (B) ethylenediaminetetraacetate solution should be prepared separately and mixed for use. You can also.

  In the present invention, an anionic surfactant, a nonionic surfactant, a cationic surfactant, and an amphoteric surfactant can be blended as necessary.

  Examples of the anionic surfactant include those represented by the following general formulas (1) to (4):

(1)

(2)

(3)

[In general formula (1)-(3), R is a C3-C20 alkyl group or alkenyl group, R 'is a C1-C4 alkyl group, n is an integer of 1-8, m is 1-2. An integer represents M, hydrogen, an alkali metal, an amine or an alkanolamine, and AO represents ethylene oxide, propylene oxide or butylene oxide, respectively. ]

(4)

[In General Formula (4), R represents a C3 to C20 alkyl group or alkenyl group, m and n each represent an integer of 1 to 2, and M represents hydrogen, an alkali metal, an amine, or an alkanolamine. ]

  Nonionic surfactants include, for example, polyoxyalkylene alkyl ethers, polyoxyalkylene fatty acid esters, polyoxyethylene sorbitan fatty acid esters, alkyl polyglycosides, sucrose fatty acid esters, alkyl polyglycerin ethers, fatty acid alkanolamides, alkylamine oxides, Examples include block copolymers of polyoxyethylene and polyoxypropylene and polyoxyalkylene styrenated phenyl ethers.

Examples of the cationic surfactant include alkyl dimethyl benzyl ammonium chloride,
Alkyldimethylethylbenzylammonium chloride,
Alkylpyridinium ammonium chloride,
Dialkylmethylbenzylammonium chloride,
Didecyldimethylammonium chloride,
Didecyldimethylammonium adipate,
Didecyldimethylammonium gluconate,
Didecyldimethylammonium propionate,
Didecyldimethylammonium bromide,
Didecyl monomethylhydroxyethylammonium chloride,
Didecyl monomethylhydroxyethylammonium adipate,
Didecyl monomethylhydroxyethylammonium gluconate,
Didecyl monomethylhydroxyethylammonium sulfonate,
Didecyl monomethylhydroxyethylammonium bromide,
N, N′-hexamethylenebis (4-carbamoyl-1-decylpyridinium bromide), alkyldimethylhydroxyethylammonium chloride,
Alkyldimethylhydroxyethylammonium adipate,
Alkyldimethylhydroxyethylammonium gluconate,
Alkyldimethylhydroxyethylammonium bromide,
Alkyltrimethylammonium chloride,
Alkyltrimethylammonium adipate,
Alkyltrimethylammonium gluconate,
Alkyltrimethylammonium bromide,
Dioctyldimethylammonium chloride,
Dioctyldimethylammonium adipate,
Dioctyldimethylammonium gluconate,
Dioctyldimethylammonium bromide,
Octyldecyldimethylammonium chloride,
Octyldecyldimethylammonium adipate,
Octyldecyldimethylammonium gluconate,
Octyldecyldimethylammonium bromide,
Didecylmethyl polyoxyethylene ammonium propionate,
A quaternary ammonium salt selected from methylbenzethonium chloride,
Polyhexamethylene biguanide hydrochloride,
Polyhexamethylene guanidine phosphate,
Polyhexamethylene guanidine chloride,
Guanidine cationic surfactant of chlorhexidine gluconate,
It is at least one selected from hexadecyltributylphosphonium, alkyldiamine or a salt thereof. These may be used alone or in combination of two or more.

  Examples of the amphoteric surfactant include at least one selected from alkylaminocarboxylic amphoteric surfactants, glycine amphoteric surfactants, alkylpolyaminoethylglycine hydrochloride, and sodium alkyldiaminoethylglycine.

  In addition, this invention can also contain another microbicide, an antifoamer, a cleaning builder, etc. as needed. Among these, as a disinfectant, for example, 5-chloro-2-methyl-4-isothiazolin-3-one, benzoisothiazolin-3-one, as an antifoaming agent, for example, as a silicon compound, a cleaning builder, Examples thereof include nitrilotriacetic acid, citric acid, malic acid, gluconic acid, iminodiacetic acid, and alkali metal salts thereof, ammonium salts, and alkanolamines such as monoethanolamine, triethanolamine, and monoisopropanolamine.

    Hereinafter, examples will be described together with comparative examples. The present invention is not limited to the examples.

  As the calicivirus composition of the present invention, test calicivirus compositions having the compositions of Examples 1 to 31 and Comparative Examples 1 to 13 shown in Tables 1 to 4 below are prepared, and norovirus and cat are prepared. The virucidal effect against calicivirus and the sterilizing and cleaning properties against general bacteria were evaluated. The numerical values in each table are due to the presence of each component, the unit is mass%, and the active ingredient concentration is shown in ppm unit as necessary. The test methods and evaluation criteria for each item are as shown below.

<Effects against calicivirus>
Calidicidal virus test 1: Effect on norovirus [test method]
100 μl of human stool-derived norovirus suspension was mixed with 400 μl of the reagent agent and stirred with a vortex mixer. RNA extraction is started 15 minutes after standing and DNase treatment is performed. Thereafter, a norovirus-derived DNA band was amplified by RT-PCR (reverse transcriptase-polymerase chain reaction).
Next, the DNA band was confirmed by 2.5% agarose gel electrophoresis, and the effect was confirmed.
[Criteria]
If the DNA band is negative in the above test, there is an effect,
If the DNA band is positive in the above test, there is no effect,
It was.
Calidicidal virus test 2: Effect on feline calicivirus [Test method]
270 μL of the reagent stock solution and 30 μL of feline calicivirus F9 strain (FCV) were contacted for 10 minutes, then diluted 10-fold with α-Minimum Essential Medium (α-MEM) (manufactured by Wako Pure Chemical Industries, Ltd.), and stepwise A 10-fold dilution was performed. 50 μL of α-MEM was added to cat kidney cells (CRFK) previously cultured in a 96-well microplate, and the previously diluted solution was added thereto. The cells were cultured for 4 days at 37 ° C. in the presence of 5% CO ₂ to confirm the presence or absence of cell degeneration.
A blank using ultrapure water instead of the reagent was used, and the Log Reduction of FCV by the reagent was calculated from these differences, and evaluated according to the following criteria.
[Criteria]
○: Log Reduction is 4 or more,
Δ: Log Reduction is 2 or more and less than 4,
X: Log Reduction is less than 2,
Δ and ○ were determined to be practical.

<Bactericidal test: Bactericidal effect on general bacteria>
〔Test method〕
Based on the European Standard Test Method EN1040, the following evaluation criteria were used.
〔Evaluation criteria〕
○: Decrease in the number of bacteria over 5 Log reduction △: Decrease in the number of bacteria with Log reduction of 4 or more and less than 5 ×: Decrease in the number of bacteria less than 4 Log reduction,
Δ and ○ were determined to be practical.
[Test strain]
Pseudomonas aeruginosa (Pseudomonas aeruginosa) NBRC 13736 Initial number of bacteria: 4.0 × 10 ⁷ cfu / ml,
Staphylococcus aureus subsp. aureus (Staphylococcus aureus) NBRC 13276 Initial number of bacteria: 3.0 × 10 ⁷ cfu / ml,
Escherichia coli (E. coli) NBRC 12734 Initial number of bacteria: 3.8 × 10 ⁷ cfu / ml
[Contact time / temperature] 5 minutes / 20 ° C
[Use dilution water] Purified water

<PH measurement>
Using a pH meter (model: pH METER F-12, manufactured by HORIBA, Ltd.), the measurement was performed according to JIS Z-8802: 1984 “pH measurement method”.

<Detergency>
〔Test method〕
A test piece was prepared by attaching oil and fat stains (JIS-K3362: 1998-9.2 model stains) to a stainless steel piece (5 cm × 8 cm) at 500 mg / sheet and drying at 25 ° C. for 1 hour. The test detergent aqueous solution (40 ° C.) was subjected to a detergency test (250 rpm, 10 minutes) using a linac type tester, then rinsed with running water and allowed to dry naturally. Then, based on the weight change before and after cleaning, the cleaning efficiency (%) was calculated from the following formula and evaluated according to the following criteria.
Washing efficiency (%) = [weight before washing (g) −weight after washing (g)] / [weight before washing (g) −weight of clean stainless steel piece (g)] × 100
〔Evaluation criteria〕
○: Cleaning rate of 80% or more (excellent in cleaning)
Δ: Cleaning rate of 50% or more to less than 80% (slightly superior in cleaning properties)
X: Cleaning rate less than 50% (inferior in cleaning properties)
And the evaluation criteria of ○ and Δ were determined to be practical.

The components used in the examples and comparative examples are shown below.
Component (A): 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide Product name: “Hygenia”, 100% by mass of active ingredient manufactured by Tama Chemical Industry Co., Ltd.

(B) Component chelating agent 1
Ethylenediaminetetraacetate Product name: “Chillest 2B”, manufactured by Cylest (active ingredient 100% by mass)

(Other ingredients)
Chelating agent 2
L-glutamic acid diacetic acid tetrasodium Product name: “GLDA 4Na-R”, manufactured by Showa Denko KK (active ingredient 40% by mass)
Chelating agent 3
Nitrilotriacetate sodium product name: “Chillest NTA”, manufactured by Cylest Chemical Co. (active ingredient 98% by mass)
Chelating agent 4
Methyl glycine diacetic acid Product name: “Trilon M” manufactured by BASF (active ingredient 40% by mass)

(Nonionic surfactant)
Nonionic surfactant 1
Polyoxyalkylene alkyl ether Product name: “Adekatol LB83”, manufactured by ADEKA (active ingredient 100% by mass)
Nonionic surfactant 2
Polyoxyalkylene fatty acid ester Product name: “EMALEX 6300M-ST”, manufactured by Nippon Emulsion Co., Ltd. (active ingredient 100% by mass)
Nonionic surfactant 3
Polyoxyalkylene alkyl ether Product name: “Neugen XL60”, manufactured by Daiichi Kogyo Seiyaku Co., Ltd. (active ingredient 100% by mass)
Nonionic surfactant 4
Coconut oil fatty acid diethanolamide (1: 1 type)
Product name: “Adekasol COA” manufactured by ADEKA (active ingredient 100% by mass)

(Amphoteric surfactant)
Amphoteric surfactant 1
Sodium Lauraminopropionate Product name: “Levon APL” manufactured by Sanyo Chemical Industries (active ingredient 30% by mass)
Amphoteric surfactant 2
Sodium alkyldiaminoethylglycine
Product name: “Levon S” manufactured by Sanyo Chemical Industries (active ingredient 30% by mass)

(Anionic surfactant)
Anionic surfactant 1
Polyoxyethylene alkyl ether phosphoric acid product name: “Phosphanol RA-600”, manufactured by Toho Chemical Industry Co., Ltd. (active ingredient 100% by mass)
Anionic surfactant 2
Sodium polyoxyethylene lauryl ether acetate Product name: “Bulite LCA30D”, manufactured by Sanyo Chemical Industries (active ingredient 29% by mass)

(Cationic surfactant)
Cationic surfactant 1
Alkyl chloride (C12) benzalkonium Product name: “Cation G50” manufactured by Sanyo Chemical Industries, Ltd. (active ingredient 50% by mass)
Cationic surfactant 2
Didecyldimethylammonium chloride Product name: “Burdac 2280” manufactured by Lonza Japan (active ingredient 80% by mass)
Cationic surfactant 3
Polyhexamethylene biguanide hydrochloride Product name: “Proxel IB” manufactured by Arch Chemical Co., Ltd. (active ingredient 20% by mass)

(Solubilizer)
Solubilizer 1
Propylene glycol Product name: “ADEKA PROPYLENE GLYCOL (PG)”, manufactured by Asahi Denka Kogyo Co., Ltd. (active ingredient 100% by mass)
Solubilizer 2
Sodium benzoate Product name: “Sodium benzoate”, manufactured by Toagosei Co., Ltd. (active ingredient 100% by mass)
Solubilizer 3
Meta-xylene sulfonic acid soda Product name: “Meta-xylene sulfonic acid soda”, manufactured by Mitsubishi Gas Chemical Company (active ingredient 100% by mass)

  From the results of Tables 1 to 4, all of Examples 1 to 31 of the composition of the present invention have excellent performance in all items in terms of virucidal activity against feline calicivirus and sterilization against general bacteria. You can see that Moreover, it turns out that Example goods 1 and 2 have the outstanding virucidal performance also with respect to norovirus.

  From the above results, it was confirmed that the product of this example had effective virucidal performance and sterilization performance against calicivirus.

According to the present invention, by combining (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide and (B) ethylenediaminetetraacetate, the effect of calicivirus is achieved. Can be obtained. In addition, the calicidal virus composition of the present invention can be used in a sanitary manner, and since it does not use an alkaline agent, the handling safety is high.

Claims (2)

(A) 0.01% by mass or more of 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butanedibromide,
(B) 0.000625 mass% or more of ethylenediaminetetraacetate,
A calicicidal virus composition comprising: 0.01% by mass or more of (A) 1,4-bis (3,3 ′-(1-decylpyridinium) methyloxy) butane dibromide was added to 0.000625% by mass of (B) ethylenediaminetetraacetate A method of killing calicivirus comprising contacting with a calicivirus in the presence.