WO2013012007A1 - Anti-virus agent composition - Google Patents

Abstract

Provided is an anti-virus agent composition which has low levels of irritation and corrosivity and also has an excellent effect of inactivating viruses, particularly an excellent effect of inactivating non-enveloped viruses such as a norovirus. An anti-virus agent composition comprising the following components (A) and (B): (A) 0.05 to 0.5 vol% inclusive of at least one quaternary ammonium salt selected from a dialkyldimethylammonium salt and an alkylbenzyldimethylammonium salt; and (B) 0.5 to 2 vol% inclusive of benzyl alcohol.

Description

Antiviral composition

The present invention relates to an antiviral composition having a virus inactivating effect.

Viral infectious diseases such as influenza and colds are considered to be one of the important preventive measures to clean fingers and the environment. Among them, viral acute gastroenteritis and diarrhea are not only confirmed as outbreaks in nursing homes, schools, hospitals, etc., but also outbreaks due to food poisoning through contaminated food in restaurants, cooking facilities, etc. There are many reports. These causative viruses include non-enveloped viruses such as norovirus, sapovirus, and rotavirus, and infection of children and the elderly may cause serious symptoms. In addition, hand-foot-and-mouth disease, which is a typical pediatric infection, and foot-and-mouth disease-causing viruses that cause tremendous damage to livestock are also classified as non-enveloped viruses. is there.

In particular, norovirus has been detected not only in Japan but around the world as a causative virus for non-bacterial food poisoning and acute gastroenteritis. As a cause of food poisoning, bacteria such as Staphylococcus aureus and Bacillus cereus have been conventionally known. However, in recent years, food poisoning caused by norovirus causing acute gastroenteritis is about 25% of the number of food poisoning cases, and the number of patients is about about the whole. It accounts for half, is the largest, and is regarded as a problem as a virus that causes mass infection. The infection route is well-known for oral infection caused by eating shellfish such as raw oysters. However, at home and public facilities, the patient's vomit and feces on the fingers and cooking utensils of people who touched contaminated food. A number of secondary infections have been reported from viruses present in or around the patient. In particular, such secondary infection leads to mass infection and spread of infection, and is a major cause of increase in the number of patients.

In order to prevent such food poisoning and infection caused by norovirus, the floor and walls of mass cooking facilities and kitchen spaces such as collective meal facilities and restaurants, food processing equipment, cooking utensils, etc. Implement hygiene measures such as wall virus, cooking utensils, furniture, textile products, etc., and virus removal and virus inactivation of floors and walls of public facilities, equipment, medical equipment, etc., and hand washing Is very important, and it is desirable that these measures be implemented simply with consideration for people and the environment.

Norovirus is an RNA virus that does not have an envelope (membrane-like structure) classified in the Caliciviridae and Norovirus genus, has a strong resistance to acid (gastric acid), and a small amount (about 10 to 100). It is known to be infected with.
At present, there are no vaccines or treatments for norovirus, and the only treatment is to prevent norovirus infection by eliminating viruses or inactivating the virus by cleaning or disinfecting food, cooking utensils, and fingers that may cause it. Is the law. However, since norovirus has strong physicochemical resistance, ethanol and disinfectants including cationic surfactants that are effective against many bacteria are effective against norovirus in general usage. You may not get. Therefore, disinfectants such as chlorine bleach (sodium hypochlorite, etc.), iodine agents (povidone iodine, etc.), aldehyde agents (glutaral, etc.) and peracetic acid preparations are used for inactivating norovirus. However, such disinfectants are highly irritating to humans and corrode metals. In addition, chlorine-based bleaching agent has the effect of bleaching the color pattern of the fiber, and there is a limit in use for textile products and clothing. For this reason, in consideration of the safety of the user, the object to be used, and the usage scene, there are limitations on using these drugs in living environments, fingers, cooking utensils, clothing, and the like.

Therefore, in recent years, means that can inactivate viruses that are less irritating and corrosive, or that can be used in textile products are required. For example, a composition containing a lower alcohol, an alkaline substance and a cationic surfactant (Patent Document 1) , A composition containing hydrogen peroxide, benzyl alcohol and a silver component (Patent Document 2), a composition containing a specific amine and / or a quaternary ammonium salt and a specific alkanolamine (Patent Document 3), A composition having a quaternary ammonium salt as an active ingredient and a pH of 5 to 9 (Patent Document 4) or a composition containing a plant extract containing a natural product tannin as an active ingredient (Patent Document 5) has been reported. Yes.

However, even in these compositions, since it takes time to exert the virus inactivating effect, the effect is insufficient for use in hand disinfection and washing operations, or the effect is exhibited in a short time. However, it is still not sufficient in terms of virus inactivation effects such as insufficient reduction of virus infection titer. In addition, there is a problem that it can be applied only to a product formulation that is alkaline or contains a specific component and has an upper composition limit.

JP 2008-189645 A JP 2009-40760 A JP 2005-514427 A JP 2010-53091 A International Publication No. 2008/153077 Pamphlet

Summary of invention

The present invention relates to the following 1) to 5).
1) Antiviral composition containing the following components (A) and (B).
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% 2) A virus inactivation method using a composition containing the following components (A) and (B).
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% 3) Use of a composition containing the following components (A) and (B) for virus inactivation:
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% 4) (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts are used in combination with (B) benzyl alcohol. A method for enhancing the activation effect.
5) (A) Combined with the quaternary ammonium salt of (A) above to enhance the virus inactivating effect of one or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts (B) Use of benzyl alcohol.

TEM photo of NoV VLPs. a) Control, b) Sanizole C: BA = 0.05%: 2.00%, 30 seconds contact

The present invention relates to providing an antiviral composition which has little irritation and corrosivity and has a virus inactivating effect, particularly an excellent inactivating effect against non-enveloped viruses such as Norovirus.

The present inventors have examined components effective for preventing infection of non-enveloped viruses such as Norovirus. When a specific quaternary ammonium salt and benzyl alcohol are used in combination, the quaternary ammonium salt is used. It was found that the virus inactivating effect was greatly increased as compared with the case of using alone, and it was useful as an antiviral composition.

The composition of the present invention has an excellent antiviral action against non-enveloped viruses such as Norovirus, and is less irritating and corrosive, so that it can be used safely and for a short time. Thus, it is possible to inactivate the virus attached to floors and walls of large-scale cooking facilities, kitchen spaces, etc., medical equipment, food processing equipment, cooking utensils and the like. Furthermore, it can be effectively used to inactivate viruses and disinfect hands and fingers attached to floors, walls, furniture, and the like in the dwellings of ordinary households.

Dialkyldimethylammonium salt and alkylbenzyldimethylammonium salt, which are quaternary ammonium salts of component (A), are known compounds as antibacterial cationic surfactants. These may be used alone or in combination of two or more.
In the present invention, the dialkyldimethylammonium salt and the alkylbenzyldimethylammonium salt preferably include a dialkyl (C _8-18 ) dimethylammonium salt and an alkyl (C _12-16 ) benzyldimethylammonium salt, respectively.

That is, the alkyl group in the dialkyldimethylammonium salt includes an alkyl group having 8 to 18 carbon atoms, preferably a linear alkyl group having 8 to 18 carbon atoms. Specific examples include octyl, decyl, dodecyl (lauryl), tetradecyl (myristyl), hexadecyl (cetyl), heptadecyl, octadecyl (stearyl), and more preferably octyl, decyl, and dodecyl.

The alkyl group in the alkylbenzyldimethylammonium salt includes an alkyl group having 12 to 16 carbon atoms, preferably a linear alkyl group having 12 to 16 carbon atoms. Specific examples include dodecyl (lauryl), tetradecyl (myristyl), and hexadecyl (cetyl).

Suitable dialkyldimethylammonium salts include didecyldimethylammonium salts, and alkylbenzyldimethylammonium salts include dodecylbenzyldimethylammonium salts, tetradecylbenzyldimethylammonium salts, and hexadecylbenzyldimethylammonium salts.

Ammonium salts include salts with halide ions such as F ⁻ , Cl ⁻ , Br ⁻ and I ⁻ , NO ₃ ⁻ , SO ₄ ^2−, etc., and salts with halide ions are preferred and chlorides. More preferred are salts with ions.

More preferable quaternary ammonium salts include dodecylbenzyldimethylammonium chloride, tetradecylbenzyldimethylammonium chloride, hexadecylbenzyldimethylammonium chloride, and didecyldimethylammonium chloride.

The quaternary ammonium salt of the present invention can be produced by a known method, and a commercially available product can also be used. For example, Cotamin D10P (manufactured by Kao Corporation) as a dialkyldimethylammonium salt and Sanizol 08, Sanisole C (manufactured by Kao Corporation), etc. can be purchased and used as an alkylbenzyldimethylammonium salt.

The quaternary ammonium salt alone has an inactivating effect against norovirus (by an alternative experiment with feline calicivirus (FCV)) at a contact time of about 60 minutes alone, but the effect is insufficient in a short time. There is no significant inactivation effect unless used for a long time or at a high concentration.

The component (B) benzyl alcohol can be used by purchasing a commercial product. For example, products from Kanto Chemical Co., Inc. or Tokyo Chemical Industry Co., Ltd. can be purchased and used.
In addition, other organic solvents can be mixed and used with benzyl alcohol as long as the antiviral effect is not affected.

In the composition of the present invention containing the above (A) quaternary ammonium salt and (B) benzyl alcohol, the antiviral activity of the quaternary ammonium salt against norovirus (by an alternative experiment with feline calicivirus (FCV)) is remarkably increased. When a quaternary ammonium salt is used alone, an excellent virus inactivating effect is exhibited even at a concentration and contact time at which antibacterial activity cannot be sufficiently exhibited.

The composition of the present invention may be any composition that exhibits a virus inactivation effect that removes or significantly reduces the ability to infect or proliferate viruses, and may denature or disrupt virus particles and nucleic acids (RNA). As a result, it may be possible to reduce or inhibit the ability of the virus to infect or proliferate by adsorption to the surface of the virus or host cell, binding of the virus with an enzyme, or the like.

Here, the “virus” is preferably a non-enveloped virus, that is, a single-stranded (+) RNA virus having no envelope, a single-stranded (−) RNA virus, a double-stranded RNA virus, a single-stranded virus. Examples include DNA viruses and double-stranded DNA viruses. Examples of such viruses include viruses belonging to the Caliciviridae, Picornaviridae, Parvoviridae, Papillomaviridae, Polyomaviridae, and Adenoviridae families.
Among the above, the composition of the present invention is excellent in inactivating effect against Caliciviridae virus (single-stranded (+) virus), and among the Caliciviridae viruses, the genera Besivirus, Norovirus and Sapovirus It has an excellent inactivation effect on viruses belonging to the group, and in particular has an excellent inactivation effect on viruses belonging to the genus Besivirus and Norovirus.

In the composition of the present invention, the mass ratio of the components (A) and (B) is such that (A) / (B) is 3/1 to 1/500, more preferably 3/1 to 1/40, and further 2/1 to 1/10 is preferable.

The composition of the present invention comprises only other components (A) and (B) or, if necessary, other components such as a chelating agent in order to further improve the antiviral activity and maintain the effect. Along with water-soluble solvents, other surfactants, etc., products and preparations used for cleaning of floors and walls of large-scale cooking facilities, kitchen spaces, medical equipment, food processing equipment, cooking utensils, and various living environments Providing antiviral activity to products and preparations used in foods (eg kitchen detergents, faucet care agents, bath detergents, toilet cleaners, floor and furniture cleaning products, etc.) and clothes and textile products Can be used as a material for.

Here, as the chelating agent, (1) tripolyphosphoric acid, pyrophosphoric acid, orthophosphoric acid, hexametaphosphoric acid and alkali metal salts thereof, (2) ethylenediaminetetraacetic acid, hydroxyiminodiacetic acid, dihydroxyethylglycine, nitrilotriacetic acid, hydroxyethylenediamine Triacetic acid, diethylenetriaminepentaacetic acid, triethylenetetraminehexaacetic acid and alkali metal salts or alkaline earth metal salts thereof, (3) aminotrimethylenephosphonic acid, 1-hydroxyethylidene-1,1-diphosphonic acid, ethylenediaminetetramethylenephosphone Acid, diethylenetriaminepentamethylenephosphonic acid, N-oxide of aminotrimethylenephosphonic acid and alkali metal salts or alkaline earth metal salts thereof, (4) acrylic acid and methacrylic acid? Monomers or copolymers of selected monomers, acrylic acid-maleic acid copolymer, poly α-hydroxyacrylic acid and alkali metal salts thereof, (5) citric acid, succinic acid, malic acid, fumaric acid, tartaric acid, One or more polyvalent carboxylic acids selected from malonic acid and maleic acid and alkali metal salts thereof; (6) alkylglycine-N, N-diacetic acid, aspartic acid-N, N-diacetic acid, serine- Examples thereof include N, N-diacetic acid, glutamic acid diacetic acid, ethylenediamine disuccinic acid, and salts thereof.

In addition, the composition of the present invention may further contain, for example, additives such as oil, higher fatty acids, silicones, pH adjusters, thickeners, suspending agents, powder components, natural extracts, pigments, and fragrances as appropriate. It can be used to produce an antiviral composition that is blended and stirred and dispersed or dissolved.

Here, as pH adjusters, acid agents such as inorganic acids such as hydrochloric acid and sulfuric acid, organic acids such as citric acid, succinic acid, malic acid, fumaric acid, tartaric acid, malonic acid, maleic acid, sodium hydroxide and water Alkaline agents such as sodium carbonate and potassium carbonate such as potassium oxide, ammonia and derivatives thereof, amine salts such as monoethanolamine, diethanolamine, and triethanolamine may be used alone or in combination. In addition, these acid agents and alkali agents may be used in combination as a buffer system.
The above composition is preferably adjusted to have a pH of 2 to 12 at 20 ° C. with the above pH adjuster, and particularly to have a pH of 4 to 11 in terms of effects on humans and substrates. To preferred.

The content of the quaternary ammonium salt of the component (A) in the composition of the present invention is 0.05 vol% or more and 0.5 vol% or less, more preferably 0.1 vol% or more from the viewpoint of antiviral activity. More preferably, it is 0.2 vol% or more, and 0.5 vol% or less, 0.2 vol% or less, or 0.1 vol% or less is mentioned from the viewpoint of blending into a product or the like. For example, 0.05 to 0.5 vol%, 0.1 to 0.5 vol%, 0.2 to 0.5 vol%, 0.05 to 0.2 vol%, 0.05 to 0.1 vol%, 0.1 to 0.2 vol% etc. are mentioned.
The content of benzyl alcohol as component (B) is 0.5 to 2 vol%, but from the viewpoint of antiviral activity, it is more preferably 1 vol% or more, and even more preferably 1.5 vol% or more. From the viewpoint of property and blending, 2 vol% or less, 1.5 vol% or less, or 1.0 vol% or less can be mentioned. Examples thereof include 0.5 to 2 vol%, 0.5 to 1 vol%, 0.5 to 1.5 vol%, and 1 to 2 vol%.

The composition of the present invention is a place where there is a risk of virus attachment, for example, kitchen (kitchen floor, kitchen space such as walls), sink, refrigerator, etc. Tableware such as dishes, refrigerators, toilets, baths, toilets, living rooms such as living rooms and bedrooms, textile products such as sofas, cushions, bedding and curtains can be used by contacting them. Here, the contact is not particularly limited, and examples include direct application of the composition to the target surface, filling in a spraying device such as a trigger or aerosol, and spraying.

There is no restriction | limiting in particular in the usage form at the time of making the composition of this invention contact. For example, it can be liquid, gel, or paste.
In the case of a liquid, it can be used as a spray, lotion or the like. For example, a known spray container such as a trigger spray container (direct pressure or pressure accumulation type) or a dispenser type pump spray container can be filled, and the spray amount can be adjusted to spray the virus in a place where it is or is thought to exist. And so on.

Also, it can be used as a wet wipe (such as a wet napkin). For example, the composition of the present invention, solubilized on a base material comprising a natural fiber, a regenerated fiber such as rayon, a synthetic fiber such as polyethylene, polypropylene, polyester, or a woven or non-woven web of a mixture of natural fiber and synthetic fiber It can be used by containing a carrier and various additives as required. Specifically, it can be used for sterilization of skin conditioners, pet care products, surfaces of household kitchens and bathrooms, surfaces of exercise equipment, and the like.

When it is made into a gel using a water-soluble gel or oil-based gel, or in a paste form, it is used as a medical product by directly applying it to the human body, pets, etc., or carrying it on paper or non-woven fabric, It can also be used as a filter for an air purifier.

Furthermore, the composition of the present invention can be used by kneading it with a resin. Examples of resins that can be kneaded include natural, petroleum, synthetic waxes, rosin resins, ethylene-vinyl acetate copolymers, ethylene-vinyl alcohol copolymers, polyesters, polyolefins, acrylic resins, etc. Is mentioned.
The kneaded material can be used as it is, or can be supported on a porous carrier, formed into a sheet, or used as a laminate. Examples of the porous carrier include a natural polymer such as cellulose and chitosan, the above synthetic resin, an inorganic porous material such as calcium silicate, and the like in an arbitrary shape such as a granular shape and a sheet shape. The above-mentioned kneaded product or laminate can be used, for example, by installing it in an air conditioner, toilet, bathroom, living room, hospital room, hospital waiting room, dust box, etc. and gradually evaporating the antiviral composition.

With respect to the above-described embodiment, the following aspects are disclosed in the present invention.
<1> An antiviral composition containing the following components (A) and (B).
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% <2 > A virus inactivation method using a composition containing the following components (A) and (B).
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% <3 > The method of <2>, wherein the composition is brought into contact with a place where there is a risk of virus attachment.
<4> Use of a composition containing the following components (A) and (B) for virus inactivation.
(A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol% <5 > (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts, and (B) benzyl alcohol is used in combination with the quaternary ammonium salt virus. A method for enhancing the inactivation effect.
<6> (A) the quaternary ammonium salt of (A) above for enhancing the virus inactivating effect of one or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts; (B) Use of benzyl alcohol in combination.
<7> In the above items <1> to <6>, the component (A) is more preferably 0.1 vol% or more, more preferably 0.2 vol% or more, and 0.5% or less, 0.2 vol%. Hereinafter, it is 0.1 vol% or less.
<8> In the above items <1> to <6>, the component (B) is more preferably 1 vol% or more, more preferably 1.5 vol% or more, and 2 vol% or less, 1.5 vol% or less, or 1 0.0 vol% or less.
<9> In the above <1> to <6>, the mass ratio of the components (A) and (B) is preferably (A) / (B) of 3/1 to 1/500, more preferably Is 3/1 to 1/40, more preferably 2/1 to 1/10.
<10> In the above items <1> to <9>, the quaternary ammonium salt (A) is preferably one or more selected from octylbenzyldimethylammonium chloride, dodecylbenzyldimethylammonium chloride and didecyldimethylammonium chloride. It is.
<11> In the above <1> to <9>, the virus to be suitably applied is a non-enveloped virus.
<12> In the above items <1> to <9>, a virus that is preferably applied is a virus belonging to the genus Norovirus.

Hereinafter, the present invention will be described in more detail based on examples.
Example 1
(1) Preparation of antiviral agent As the quaternary ammonium salt of (a), alkylbenzyldimethylammonium chloride (Sanisol C (manufactured by Kao), linear alkyl (mixture of 12 to 16 carbon atoms) benzyldimethylammonium chloride) and di Decyldimethylammonium chloride (manufactured by Kao) was prepared using benzyl alcohol (BA, (WAKO)) as (b) and diluted with sterilized water to the concentrations shown in Tables 1 to 3 below. .

(2) Preparation of test virus and virus suspension Feline calicivirus F-9 ATCC VR-782 (feline calicivirus (FCV)) was used as the test virus. Since norovirus culture systems have not been established, feline calicivirus classified into the same caliciviridae from morphology and genetic information is used as an alternative virus for norovirus, and a method for confirming virus infectivity is common. (Journal of Hospital Infection, 41, p51-57, 1999).
The cells used were CRFK (Crandell Rees feline kidney) cells (Dainippon Sumitomo Pharma Co., Ltd.). The medium used was a cell growth medium prepared by adding 10% fetal calf serum to Eagle MEM “Nissui” (1) (Nissui Pharmaceutical). As the cell maintenance medium, Eagle MEM “Nissui” (1) to which 2% fetal calf serum was added was used.

CRFK cells were monolayer cultured in tissue culture flasks using cell growth media. After monolayer culture, the cell growth medium was removed from the flask and inoculated with the test virus. Next, cell maintenance medium was added and cultured in a carbon dioxide incubator (CO ₂ concentration: 5%) at 37 ° C. ± 1 ° C. for 1 to 5 days. After culturing, cell morphology was observed using an inverted phase contrast microscope to confirm that cytopathic effect (CPE) occurred. Next, the culture solution was centrifuged (3,000 rpm, 10 minutes), and the resulting supernatant was used as a virus suspension.

(3) Virus inactivation test 0.1 ml of the virus suspension was added to 1 ml of each specimen shown in Tables 1 to 3 and mixed to obtain a working solution. Acting at room temperature, test 1 was diluted with cell maintenance medium after 5, 10, 20 and 30 minutes. Test 2 was similarly diluted after 0.5, 1 and 5 minutes. However, dilution was performed at a dilution concentration confirmed by a preliminary test. In addition, it tested similarly using purified water as a control, and measured the virus infectious titer at the start.

(4) Method of measuring virus infection titer Using cell growth medium, the cells used were cultured in a monolayer in a tissue culture microplate (96 well), and then the cell growth medium was removed and 0.1 ml of cell maintenance medium was added. . Next, the working fluid was diluted 10-fold using a cell maintenance medium. 0.1 ml of each working solution was inoculated into 4 wells and cultured in a carbon dioxide incubator (CO ₂ concentration: 5%) at 37 ° C. ± 1 ° C. for 4-7 days. After culturing, the presence or absence of CPE was observed using an inverted phase contrast microscope, and 50% tissue culture infectious dose (TCID50) was obtained by the Reed-Muench method (The American journal of hygiene, 27, 493-497, 1938). ) Was calculated and converted to the virus infectivity per ml of working fluid.

(5) Test result It was confirmed that BA alone did not reduce the infectious titer of FCV by contact for 5 minutes (Table 1). Alkylbenzyldimethylammonium chloride alone (0.05%) reduces the infectivity only by 2.2 log TCID ₅₀ after 5 minutes of FCV contact, compared to 1.0% for alkylbenzyldimethylammonium chloride (0.05%). % And 2.0% of BA reduces the 4 log TCID ₅₀ infectivity in 5 minutes and 0.5 minutes of contact, respectively, and the combination of alkylbenzyldimethylammonium chloride and BA provides a high antiviral effect in a short time. It was confirmed that this was exhibited (Table 2). Similarly, didecyldimethylammonium chloride alone (0.05%) reduces the infectious value only by 2.4 log TCID ₅₀ in 5 minutes, whereas didecyldimethylammonium chloride (0.05%) contains 0.5% BA. When 5% was added, a decrease of 4log TCID ₅₀ was confirmed in 0.5 minutes when 2% BA was added (Table 3). From this, it was shown that the high FCV inactivation effect (norovirus inactivation effect) can be exhibited in a short time by using the cationic surfactant of the present invention and BA together.

Example 2
(1) Preparation of test solution Alkylbenzyldimethylammonium chloride (Sanisol C (manufactured by Kao), linear alkyl (mixture of 12 to 16 carbon atoms) benzyldimethylammonium chloride) and didecyl as the quaternary ammonium salt of (a) Dimethylammonium chloride (DDAC; manufactured by Kao) was used as benzyl alcohol (BA, (WAKO)) as (b).
(2) Novirus virus-like particles (NoV VLPs) Shape test method NoV VLPs are expressed and purified in large quantities of capsid proteins that compose norovirus. I'm taking it. It has been confirmed that morphologically and immunologically it is similar to norovirus except that it does not have RNA as a genetic factor (Hansman, GS, et al. Genetic and antigenic diversity among noroviruses. J Gen. Virol. 87: 909-919, 2006).
NoV VLPs were produced using a recombinant protein mass expression system using baculovirus according to the method of Hansman, G. S. et al. (Non-patent Document 1). That is, the gene region encoding the structural protein (accession No. AB365435) encoding the structural protein constituting NoV VLPs in the genome of baculovirus is recombined into a vector to produce a recombinant baculovirus and infect insect cells. As a result, the recombinant protein was expressed in large quantities, and NoV VLPs were recovered and purified.
Test solutions having a concentration twice the values shown in Tables 4, 5 and 6 were prepared and mixed in equal amounts with NoV VLPs solution. After contact for 0.5, 1, 5 minutes, 30 μL was extracted and the test solution was removed. Specifically, 30 μL extracted was brought into contact with 360 μL SM-2 Bio-beads suspension (50% w / v) dispensed in a microtube, stirred for 2 minutes, and then 10 μL of the supernatant was subjected to TEM observation. SM-2 bio beads were suspended in 10 mM HEPES so as to be 50 (w / v)% and autoclaved.
The NoV VLPs after the contact test were used for TEM (Transmission Electron Microscope) observation, and the anti-norovirus effect of each test solution was predicted by capturing changes in the particle structure.

(3) Negative staining method The grid for TEM observation is 400 mesh (carbon deposition reinforcement) with a copper collodion film (Nisshin EM), and in the cleaning mode by ion sputtering (E-1030: Hitachi) immediately before use. Glow discharge (1 mA, 30 sec) was performed to make the grid surface field hydrophilic. 10 μL of the sample supernatant was quickly dropped on the grit, and after about 3 minutes, excess water on the grid was slowly sucked off using a Kim towel. The sample was dried for about 1 minute, and then the grid surface on which the sample supernatant was adsorbed was brought into contact with a staining solution (2% phosphotungstic acid solution: pH 6.8) dripped on the parafilm. Using a Kim towel, the excess staining solution on the grid was slowly blotted out. Thereafter, 10 μL of ion-exchanged water passed through a 0.45 μm filter was dropped onto the grid, washed for 30 seconds, and then excess water on the grid was blotted using a Kim towel and dried sufficiently.

(4) TEM observation method A dyed sample was obtained by using a transmission electron microscope H7650 (Hitachi), a pressurization voltage of 10 kV, an emission current of 10 μA, a condenser lens scale No. 2, objective lens scale No. Observation was made promptly under the condition of 3.

(5) Results of particle shape evaluation by TEM observation-: No effect can be expected (no change in particle structure)
-/ +: Little effect expected (with slightly deformed particles)
+: Slight effect can be expected (about half of the particles are deformed)
++: An effect can be expected (the majority of deformed particle shapes)
+++: Significant effect can be expected (only deformed particles)

It was confirmed that the particle structure of NoV VLPs did not change at all by contact with BA alone for 5 minutes (Table 4). Alkylbenzyldimethylammonium chloride alone (0.05%) shows slight deformation of the particle structure of NoV VLPs after 5 minutes of contact, whereas alkylbenzyldimethylammonium chloride (0.05%) has 0.5% %, 1.0%, and 2.0% of BA added to confirm the change in particle shape to the extent that the effect can be expected by contact for 5, 1 and 0.5 minutes, respectively. It was suggested that the antiviral effect was exhibited in a short time against Norovirus (Table 5). Similarly, with didecyldimethylammonium chloride (DDAC) alone (0.05%), only a slight change in particle shape was observed, whereas BA was added to DDAC (0.05%). When 0.5% was added, 1 minute was observed, and when 2.0% BA was further added, a change in the particle shape was observed to such an extent that an effect could be expected in 0.5 minutes (Table 6). This suggests that the anti-norovirus effect can be exhibited in a short time by using a cationic surfactant and BA in combination.
FIG. 1 shows a TEM photograph when Sanisole C (0.05%) and BA (2.00%) were added for 0.5 minutes. From FIG. 1, it was confirmed that the particle structure of Norovirus virus like particles (NoV VLPs) composed only of norovirus structural proteins was deformed by the combined action of alkylbenzyldimethylammonium salt and benzyl alcohol. .

Claims (14)

1. An antiviral composition containing the following components (A) and (B).
   (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol%
2. The quaternary ammonium salt of (A) is at least one selected from dodecylbenzyldimethylammonium chloride, tetradecylbenzyldimethylammonium chloride, hexadecylbenzyldimethylammonium chloride and didecyldimethylammonium chloride. Composition.
3. The composition according to claim 1 or 2, wherein the virus is a non-enveloped virus.
4. The composition according to claim 1 or 2, wherein the virus is a virus belonging to the genus Norovirus.
5. A virus inactivation method using a composition containing the following components (A) and (B).
   (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol%
6. The quaternary ammonium salt of (A) is at least one selected from dodecylbenzyldimethylammonium chloride, tetradecylbenzyldimethylammonium chloride, hexadecylbenzyldimethylammonium chloride and didecyldimethylammonium chloride. Method.
7. The method according to claim 5 or 6, wherein the virus is a non-enveloped virus.
8. The method according to claim 5 or 6, wherein the virus is a virus belonging to the genus Norovirus.
9. Use of a composition containing the following components (A) and (B) for virus inactivation:
   (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts 0.05 vol% to 0.5 vol% (B) benzyl alcohol 0.5 vol% to 2 vol%
10. The quaternary ammonium salt of (A) is at least one selected from dodecylbenzyldimethylammonium chloride, tetradecylbenzyldimethylammonium chloride, hexadecylbenzyldimethylammonium chloride and didecyldimethylammonium chloride. use.
11. The use according to claim 9 or 10, wherein the virus is a non-enveloped virus.
12. The use according to claim 9 or 10, wherein the virus is a virus belonging to the genus Norovirus.
13. (A) One or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts are used in combination with (B) benzyl alcohol. A method for enhancing the activation effect.
14. (A) In combination with the quaternary ammonium salt of (A) above for enhancing the virus inactivating effect of one or more quaternary ammonium salts selected from dialkyldimethylammonium salts and alkylbenzyldimethylammonium salts ( B) Use of benzyl alcohol.

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