Virus inactivator

Abstract

To provide a virus inactivator that enables the inactivation of viruses present in the environment. [Solution] (A) A virus inactivator comprising maleic acrylate copolymer or a salt thereof as an active ingredient.

Description

[Technical Field]

[0001] The present invention relates to a virus inactivating agent and a virus inactivation method. [Background technology]

[0002] Viral infections, such as viral respiratory infections and infectious gastroenteritis (viral gastroenteritis), primarily spread when infected individuals bring the virus into living spaces. The infection then spreads directly from the infected person or through the environment, including people's hands, clothing, various utensils and materials, walls, air conditioners, and other equipment. Therefore, it is considered effective in preventing the spread of infection by washing and disinfecting hands, clothing, and various utensils and materials that may come into contact with the virus, thereby eliminating or inactivating the virus, as well as inactivating viruses that have been airborne in droplets and viruses that float in the air as aerosols.

[0003] Traditionally, ethanol (high concentration and acidic or alkaline), chlorine-based disinfectants (such as sodium hypochlorite), iodine-based disinfectants, aldehyde-based disinfectants (such as glutaraldehyde), and peracetic acid preparations have been used to inactivate viruses. However, these common disinfectants are highly irritating to mucous membranes and skin, limiting their use due to safety concerns. Furthermore, they require a long time to inactivate viruses, may not be sufficiently effective at room temperature, and their effectiveness may be reduced when used in combination with other bases, such as activators. In addition, spraying chlorine dioxide has been considered as a method to chemically inactivate viruses present in the air, but its effectiveness is not yet certain.

[0004] Acrylic acid-maleic acid copolymer is a type of acrylic acid-based water-soluble polymer that exhibits excellent properties in terms of adsorption to inorganic salts by carboxyl groups and dispersion through ionization. Therefore, it is used in applications such as detergent builders, metal ion chelating agents, and scale inhibitors (calcium carbonate, phosphates, etc.). Meanwhile, Patent Document 1 reports that disinfectant formulations containing acidic polymers with maleic acid as a constituent unit possess both antiviral and antibacterial activity. However, it is completely unknown that acrylate-maleic acid copolymer has virus-inactivating properties. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Special Publication No. 2011-511764 [Overview of the project] [Problems that the invention aims to solve]

[0006] The present invention relates to providing a virus inactivator that enables the inactivation of viruses present in the environment. [Means for solving the problem]

[0007] The inventors have found that maleic acrylate copolymer has the effect of inactivating viruses such as human norovirus, enterovirus, adenovirus, and influenza virus, and that it is useful as a virus inactivator.

[0008] In other words, the present invention relates to a virus inactivator comprising (A) maleic acrylate copolymer or a salt thereof as an active ingredient. The present invention also relates to a method for inactivating viruses, comprising applying (A) a maleic acrylate copolymer or a salt thereof, or a composition containing the same, to an object of concern regarding viral contamination. The present invention also provides a virus inactivation composition comprising (A) maleic acrylate copolymer or a salt thereof as an active ingredient, This invention relates to a virus inactivation composition for use on objects suspected of viral contamination, which is diluted with water to form a diluted solution containing component (A) in an amount of 10 ppm to 15,000 ppm, and applied to the object. The present invention also provides a virus inactivation composition comprising (A) maleic acrylate copolymer or a salt thereof as an active ingredient, This invention relates to a virus inactivation composition in which the concentration of component (A) in the aforementioned composition is 10 ppm or more and 15,000 ppm or less. [Effects of the Invention]

[0009] The virus inactivating agent or virus inactivating composition of the present invention can inactivate viruses attached to hands, hard and soft surfaces in the living environment, viruses dispersed in airborne droplets in living spaces, and prevent or reduce the spread of infection caused by such viruses. [Modes for carrying out the invention]

[0010] In the present invention, (A) maleic acrylate copolymer or a salt thereof is used as an active ingredient for virus inactivation. Maleic acrylate copolymer is a copolymer containing constituent units derived from acrylic acid and constituent units derived from maleic acid. The polymerization mode of maleic acrylate copolymer is not particularly limited and may be random copolymerization, block copolymerization, etc., but random copolymerization is preferred. In the acrylic acid-maleic acid copolymer of the present invention, acrylic acid and maleic acid are essential components as monomers. Maleic acid and acrylic acid may be partially or completely neutralized. Maleic anhydride may also be used as maleic acid. Furthermore, in the present invention, monomers other than acrylic acid (salt) and maleic acid (salt) may be included as monomer components, as long as they do not impair the effects of the present invention. Such monomers can be copolymerized with acrylic acid (salt) or maleic acid (salt), for example, unsaturated monocarboxylic acid monomers such as methacrylic acid and crotonic acid; neutralized products obtained by partially or completely neutralizing the above unsaturated monocarboxylic acid monomers with monovalent metals, divalent metals, ammonia, organic amines, etc.; unsaturated dicarboxylic acid monomers such as fumaric acid, itaconic acid, and citraconic acid; neutralized products obtained by partially or completely neutralizing the above unsaturated dicarboxylic acid monomers with monovalent metals, divalent metals, ammonia, organic amines, etc.; amide monomers such as (meth)acrylamide and t-butyl(meth)acrylamide; (meth) Hydrophobic monomers such as acrylic acid esters, styrene, 2-methylstyrene, and vinyl acetate; unsaturated sulfonic acid monomers such as vinyl sulfonic acid, allyl sulfonic acid, methallyl sulfonic acid, styrene sulfonic acid, 2-acrylamido-2-methylpropanesulfonic acid, 3-alyloxy-2-hydroxypropanesulfonic acid, sulfoethyl (meth)acrylate, sulfopropyl (meth)acrylate, 2-hydroxysulfopropyl (meth)acrylate, and sulfoethylmaleimide; neutralized products obtained by partially or completely neutralizing the above unsaturated sulfonic acid monomers with monovalent metals, divalent metals, ammonia, organic amines, etc.3-Methyl-2-buten-1-ol (prenol), 3-methyl-3-buten-1-ol (isoprenol), 2-methyl-3-buten-2-ol (isoprene alcohol), 2-hydroxyethyl (meth)acrylate, polyethylene glycol mono(meth)acrylate, polypropylene glycol mono(meth)acrylate, polyethylene glycol monoisoprenol ether, polypropylene glycol monoisoprenol ether, polyethylene glycol monoallyl ether, polypropylene glycol monoallyl ether, glycerol monoallyl ether Examples include, but are not limited to, hydroxyl-containing unsaturated monomers such as tel, α-hydroxyacrylic acid, N-methylol(meth)acrylamide, glycerol mono(meth)acrylate, and vinyl alcohol; cationic monomers such as dimethylaminoethyl(meth)acrylate and dimethylaminopropyl(meth)acrylamide; nitrile monomers such as (meth)acrylonitrile; and phosphorus-containing monomers such as (meth)acrylamidemethanephosphonic acid, (meth)acrylamidemethanephosphonic acid methyl ester, and 2-(meth)acrylamide-2-methylpropanephosphonic acid. These monomers may be used individually or in combination of two or more. Examples of salts of maleic acrylate copolymer include alkali metal salts, alkaline earth metal salts, ammonium salts, alkyl or alkenylammonium salts having 1 to 22 carbon atoms, alkyl or alkenyl-substituted pyridinium salts having 1 to 22 carbon atoms, alkanolammonium salts having 1 to 22 carbon atoms, and basic amino acid salts. Preferably, alkali metal salts are used, and more preferably, sodium salts.

[0011] The weight-average molecular weight of the maleic acrylate copolymer or its salt is preferably 1,000 to 1,000,000, from the viewpoint of virus inactivation effect. The weight-average molecular weight can be measured, for example, by gel permeation chromatography (GPC).

[0012] The molar ratio (acrylic acid: maleic acid) of the constituent unit derived from acrylic acid and the constituent unit derived from maleic acid that constitute the acrylic acid-maleic acid copolymer or its salt is preferably 0.20:0.80 to 0.80:0.20 from the viewpoint of the virus inactivating effect.

[0013] A commercially available acrylic acid-maleic acid copolymer or its salt can be used. Also, based on the previously reported information, it is possible to produce a composition that is the same substance as or contains a commercially available product by chemical synthesis.

[0014] In the present invention, the concentration (in terms of acrylic acid-maleic acid copolymer) of (A) acrylic acid-maleic acid copolymer or its salt at the time of applying the virus inactivator to a target (hereinafter also referred to as the treatment target) where virus contamination is a concern is preferably 10 ppm (mass basis) or more, more preferably 30 ppm or more, and even more preferably 40 ppm or more from the viewpoint of further improving the virus inactivating effect. The upper limit is not particularly limited, but it is preferably 15000 ppm or less, more preferably 10000 ppm or less, and preferably 1000 ppm or less, more preferably 500 ppm or less from the viewpoints of the feeling of use and properties when applied to the treatment target. The concentration of the component (A) may be the concentration of the component (A) contained in the virus inactivating composition itself described later, or may be the concentration of the component (A) in the liquid substance at the time of application to the treatment target.

[0015] As shown in the examples described later, the acrylic acid-maleic acid copolymer or its salt exhibits an excellent virus inactivating effect against feline calicivirus, human norovirus, adenovirus, enterovirus, and influenza virus. Therefore, the acrylic acid-maleic acid copolymer or its salt can be a virus inactivator that inactivates viruses, and can also be used for producing a virus inactivator. Also, the acrylic acid-maleic acid copolymer or its salt can be used for inactivating viruses.

[0016] The viruses targeted by the virus inactivator in this invention include all types of viruses, regardless of the type of nucleic acid (RNA, DNA) and whether or not they have an envelope. Enveloped viruses include those with RNA as their nucleic acid, such as influenza virus, coronavirus, SARS coronavirus, SARS coronavirus-2, RSV, mumps virus, lassa virus, dengue virus, rubella virus, and human immunodeficiency virus, and those with DNA as their nucleic acid, such as human herpesvirus, vaccinia virus, and hepatitis B virus. Furthermore, examples of viruses that do not have an envelope include those with RNA as their nucleic acid, such as norovirus, poliovirus, echovirus, hepatitis A virus, hepatitis E virus, rhinovirus, astrovirus, rotavirus, coxsackievirus, enterovirus, and sapovirus, and those with DNA as their nucleic acid, such as adenovirus, B19 virus, papovavirus, and human papillomavirus. SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is a SARS-related coronavirus that causes acute respiratory illness (COVID-19).

[0017] Of these, influenza virus, norovirus, adenovirus, and enterovirus are preferred, with norovirus being more preferred. Norovirus includes HNV (human norovirus), FCV (feline calicivirus), MNV (mouse norovirus), and others. Noroviruses are divided into seven gene groups (genogroups, GI to GVII) based on the homology of their genome base sequences. The HNVs that infect humans are GI9 types (GI.1, GI.2, GI.3, GI.4, GI.5, GI.6, GI.7, GI.8, GI.9), GII19 types (GII.1, GII.2, GII.3, GII.4, GII.5, GII.6, GII.7, GII.8, GII.9, GII.10, GII.12, GII.13, GII.14, GII.15, GII.16, GII.17, GII.20, GII.21, GII.22), and GIV1 type (GIV.1). However, the HNV in this invention may be any of these genotypes.

[0018] In the present invention, virus inactivation means reducing or eliminating the activity of the virus and thereby eliminating its ability to infect host cells. The virus inactivation effect can be confirmed, for example, by bringing the test product into contact with the virus, infecting host cells with the virus, and measuring the viral infectivity titer. Here, the host cell can be any cell in which the target virus can proliferate. For example, for feline calicivirus, cat kidney-derived cell lines (CRFK) can be used; for adenovirus, human laryngeal cancer-derived cells (HEp-2) or human lung epithelial cell-derived cells (A549) can be used; for enterovirus, African green monkey kidney epithelial cells (Vero) can be used; for influenza virus, canine kidney cells (MDCK), African green monkey kidney epithelial cells (Vero), or duck embryonic stem cell-derived cell lines (EB66); and for human coronavirus, human ileocecal adenocarcinoma cells (HCT-8), African green monkey kidney epithelial cells (VeroE6), or human liver cancer-derived cell lines (Huh7) can be used. Furthermore, the human norovirus inactivation effect can be evaluated by a system in which human intestinal organoids are cultured in three dimensions on an extracellular matrix, and then differentiated to obtain intestinal epithelial cells, which are then infected with HNV (see Examples).

[0019] The virus inactivator in the present invention may be in the form of maleic acrylate copolymer or a salt thereof used alone, or it may be in the form of a composition containing maleic acrylate copolymer or a salt thereof (for example, a virus inactivation composition). That is, the virus inactivator in the present invention may be a virus inactivation composition that exhibits a virus inactivation effect, or it may be a material or formulation for incorporating into such a composition. The virus inactivation composition may be in an undiluted form or a diluted form. The undiluted form is applied to the target object without dilution and used for virus inactivation. The diluted form is diluted with a suitable medium such as water to achieve an active ingredient concentration in which the virus inactivation effect is exerted by the acrylic acid maleic acid copolymer or its salt, and used for virus inactivation of the target object. The preferred active ingredient concentration for which the virus inactivation effect is exerted is as described above.

[0020] Acrylates maleic acid copolymer or its salts can be used in either a liquid or gaseous state, but from the viewpoint of virus inactivation effect, it is preferable to use it in a liquid state.

[0021] The above-mentioned virus inactivation compositions include those used in liquid or gaseous form. The virus inactivation composition used in the liquid phase may contain, in addition to acrylic acid maleic acid copolymer or its salts, antimicrobial substances such as bases, hypochlorous acid, hydrogen peroxide, and silver ion compounds, as well as cationic antimicrobial agents (such as benzethonium chloride), bactericides (such as triclosan and isopropylmethylphenol), and surfactants. Furthermore, the virus inactivation composition is prepared by appropriately blending additives such as chelating agents, humectants, lubricants, builders, buffers, abrasives, electrolytes, bleaches, fragrances, dyes, foaming control agents, corrosion inhibitors, essential oils, thickeners, pigments, gloss enhancers, enzymes, detergents, solvents, dispersants, polymers, silicones, and hydrophobic substances. The form of such a composition may be, but is not limited to, liquid, emulsion, cream, lotion, paste, gel, sheet (base-supported), or oil. The virus inactivation composition can be appropriately incorporated into various cleaning agents (laundry detergents, household cleaning agents, dishwashing detergents, hair washes, hand washes, body washes, etc.), disinfectants, sanitary product compositions, etc. When using the virus inactivation composition of the present invention in laundry detergents, dishwashing detergents, etc., the virus inactivation composition can be used after diluting with water. In this case, the dilution ratio is preferably 10 times by mass or more, more preferably 100 times by mass or more, even more preferably 800 times by mass or more, and also preferably 5000 times by mass or less, and more preferably 4000 times by mass or less. Since the virus inactivation effect is achieved even with such dilution, the concentration of the active ingredient in the virus inactivation composition can be appropriately designed.

[0022] Examples of the above-mentioned sanitary product compositions include lotions, creams, shampoos, hair conditioners, hand soaps, body washes, facial cleansers, bath additives, foams, antiperspirants, deodorants, underarm odor preventatives, and oral hygiene products (mouthwash, toothpaste, mouth fresheners, gargles, etc.). The composition can be prepared by conventional methods by appropriately combining carriers that are acceptable as cosmetics, etc. (for example, diluents, dispersants, buffers, pH adjusters, emulsifiers, surfactants, preservatives, stabilizers, antioxidants, colorants, humectants, thickeners, disinfectants, fragrances, etc.).

[0023] A virus inactivation composition used in the gas phase (for example, a spatial virus inactivation composition) can be prepared by blending a base and various additives (polyols (dipropylene glycol, propylene glycol, etc.), surfactants, UV absorbers, antioxidants, preservatives, deodorants, natural extracts, silicones, thickeners, dyes, pigments, colorants, oils, fragrances, etc.) with a maleic acrylate copolymer or a salt thereof. Such a composition may take the form of a liquid or gel, but a liquid form is preferred.

[0024] In this specification, examples of bases include conventionally known substances, whether oily or aqueous, such as water, methanol, ethanol, propanol, isopropanol, butanol, isobutanol, 3-methyl-3-methoxybutanol, propylene glycol, triethylene glycol, dimethyl ether, liquid propane, petrolatum, lanolin, castor oil, and paraffinic hydrocarbons (e.g., liquid paraffin). These bases can be used individually or in combination of two or more. Furthermore, when the virus inactivation composition is to be prepared as a gel, it can be prepared by appropriately adding a natural gelling agent or a synthetic gelling agent, such as a water-soluble gelling agent like carrageenan or gellan gum, or an oil-soluble gelling agent like metal soap or aluminum octylate, according to conventionally known methods.

[0025] The content of the active ingredient in the embodiment in which the virus inactivating agent of the present invention is used as a composition can be appropriately determined depending on the form of the composition. For example, the content of maleic acrylate copolymer or its salt relative to the total amount of the composition is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, even more preferably 0.05% by mass or more, and even more preferably 0.5% by mass or more. Furthermore, there is no particular upper limit, for example, preferably 99.999% by mass or less, and even more preferably 10% by mass or less.

[0026] The virus inactivator in the present invention is active under acidic, neutral, and alkaline conditions. However, from the viewpoint of safety and virus inactivation effect, the pH (at 20°C) of the composition in which the virus inactivator is used as a composition is preferably 4 to 11, more preferably 8 to 11, and even more preferably 10 to 11. The pH shall be measured using a pH meter after adjusting the temperature to 20°C. The virus inactivator in the present invention has high activity under alkaline conditions, and the pH (at 20°C) of the composition in which the virus inactivator is used as a composition, or when applied, is preferably 8 to 12, more preferably 9 to 11, and more preferably 10 to 11. In the present invention, the virus inactivating agent is preferably a composition in which the virus inactivating agent is used, or the pH (at 20°C) at the time of application is preferably between pH 5 and 8, from the viewpoint of application to neutral detergents, tolerance for the incorporation of general-purpose ingredients, and achieving both a mild feel and low reactivity to the skin and application object, and a high virus inactivation effect.

[0027] The virus inactivator of the present invention makes it possible to inactivate viruses in objects where viral contamination is a concern. Objects where viral contamination is a concern to which acrylic acid maleic acid copolymer or its salts, or compositions containing the same, can be applied include, for example, the skin or mucous membranes of animals to which viruses adhere, hard or soft surfaces of inanimate objects, objects such as waste, and living spaces where viruses are airborne or dispersed. Here, examples of surfaces of inanimate objects include hard surfaces in homes and business facilities such as counters, sinks, restrooms, toilets, bathtubs, showers, floors, windows, doorknobs, walls, drains, pipes, and garbage collection areas; hard surfaces of special vehicles such as garbage trucks and sanitation vehicles such as garbage inlets, work surfaces, and switch surfaces; hard surfaces of transport vehicles such as railway cars and aircraft bodies such as floors, handrails, and door surfaces; hard surfaces of various appliances, tools, and miscellaneous goods such as kitchenware, furniture, telephones, and toys; and soft surfaces of textile products (carpets, area rugs, curtains, seats, fabric furniture, clothing, masks, etc.). Examples of waste include general waste (household waste such as food scraps, tissues, and masks) and industrial waste (sludge, human waste, medical waste, etc.). If the waste is contained within a bag, the bag surface may be targeted. Other examples of living spaces include the entrance hall, dining kitchen, bedroom, children's room, bathroom, toilet, etc. within ordinary homes; the interiors of facilities such as shops, restaurants, inns, hospitals, workshops, factories, garbage collection points, quarantine stations, livestock farms, and markets; the interiors of vehicles such as cars, trains, and airplanes; and semi-enclosed spaces (lockers, storage rooms, closets, etc.; storage boxes (for toys, karaoke microphones, dishes, condiments, writing instruments, stationery)).

[0028] In the present invention, the virus inactivator, maleic acrylate copolymer or a salt thereof, or a composition containing the same, is applied to objects where viral contamination is a concern, but the embodiment is not particularly limited, and the maleic acrylate copolymer or a salt thereof may be brought into contact with or reacted with the virus in a liquid or gas phase. Methods for contacting the virus with maleic acrylate copolymer or its salt in a liquid phase include directly applying maleic acrylate copolymer or its salt, or a composition containing the same, to the object to be treated; diffusing maleic acrylate copolymer or its salt, or a composition containing the same, and sprinkling it onto the object to be treated; or wiping the surface of the object with a sheet, gauze, towel, wet wipe, tissue, or wet tissue impregnated with maleic acrylate copolymer or its salt, or a composition containing the same. Another method involves filling a known spray container, such as a trigger spray container (direct pressure or pressurized type), a dispenser-type pump spray container, or an aerosol spray container equipped with a pressure-resistant vessel, with maleic acrylic acid copolymer or a salt thereof, or a composition containing the same, and spraying it onto the target object while appropriately adjusting the spray volume. Another method involves filling an atomizing device such as a pressurized air atomizing sprayer, nebulizer, or diffuser, or a diffusion device such as a washer nozzle or misting machine, and spraying it into a space where the virus is present. By dispersing it in a mist form into a space where the virus is present, the volatilization rate can be increased, and the virus inactivation effect can be rapidly exerted.

[0029] Methods for contacting or reacting acrylate-maleic acid copolymer or its salt with viruses in the gas phase include, for example, allowing the acrylate-maleic acid copolymer or its salt to volatilize naturally or by forced volatilization. This allows for the inactivation of viruses present in living spaces, enabling simple virus removal (deviralization) of the air. When the virus inactivator in the present invention is used for natural volatilization, conventionally known methods can be applied, such as impregnating a core rod, filter paper, etc., with maleic acrylate copolymer or its salt, or a composition containing the same, and allowing it to volatilize, or using a permeable membrane for volatilization. Alternatively, maleic acrylate copolymer or its salt, or a composition containing the same, can be kneaded into a resin and used. Examples of resins that can be kneaded include natural, petroleum-based, and synthetic waxes, rosin-based resins, ethylene-vinyl acetate copolymers, ethylene-vinyl alcohol copolymers, polyesters, polyolefins, and acrylic resins. The above kneaded material can be used as is, or it can be supported on a porous carrier, formed into a sheet, or used as a laminate of the sheet. Examples of porous carriers include natural polymers such as cellulose and chitosan, the above-mentioned synthetic resins, and inorganic porous materials such as calcium silicate, all in any shape such as granules or sheets. The above-mentioned mixtures and laminates can be used, for example, by installing them in air conditioning equipment, toilets, bathrooms, living rooms, hospital rooms, waiting rooms, dustbins, etc., allowing the maleic acrylate copolymer or its salts to gradually volatilize. Alternatively, the maleic acrylate copolymer or its salts, or compositions containing them, can be supported on products made of paper, nonwoven fabrics, etc. (such as air purifier filters) and used. When using acrylic acid maleic acid copolymer or its salt by forced volatilization, such means include, for example, volatilization using a fan, heating volatilization using a heater, and volatilization using ultrasound. When performing airborne virus removal treatment, the amount of maleic acrylate copolymer or its salt, or a composition containing it, used can be appropriately adjusted depending on the treatment method, the ambient environment such as temperature and humidity, the vapor pressure of the compound, etc. It is also possible to use a concentration above the saturation concentration of the compound in the space. For example, the concentration of maleic acrylate copolymer or its salt in the target space is 0.1% by mass or more, preferably 1% by mass or more, more preferably 5% by mass or more, even more preferably 10% by mass or more, preferably 100% by mass or less, preferably 50% by mass or less, more preferably 25% by mass or less, and even more preferably 20% by mass or less, when it volatilizes. The concentration of acrylic acid maleic acid copolymer or its salt in a target space can be detected by measuring the concentration of the compound in a gas sample taken from the space. Methods for determining the concentration include using a volatile organic compound (VOC) concentration meter (VOC meter, odor sensor, etc.), or using gas chromatography or gas chromatography / mass spectrometry in combination with a gas collection tube.

[0030] With regard to the embodiments described above, the present invention further discloses the following aspects.

[0031] <1> (A) A virus inactivator containing maleic acrylate copolymer or a salt thereof as an active ingredient. <2> (A) A method for inactivating a virus, comprising applying a maleic acrylate copolymer or a salt thereof, or a composition containing the same, to an object suspected of being contaminated by a virus. <3> (A) A virus inactivation composition comprising maleic acrylate copolymer or a salt thereof as an active ingredient, A virus inactivation composition for application to objects suspected of viral contamination, which is diluted with water to form a diluted solution containing component (A) in an amount of 10 ppm to 15,000 ppm. <4> (A) A virus inactivation composition comprising maleic acrylate copolymer or a salt thereof as an active ingredient, A virus inactivating composition in which the concentration of component (A) in the composition is 10 ppm or more and 15,000 ppm or less.

[0032] <5> The molar ratio (acrylic acid:maleic acid) of acrylic acid-derived constituent units to maleic acid-derived constituent units constituting the acrylic acid-maleic acid copolymer or its salt is preferably 0.20:0.80 to 0.80:0.20. <1> ~ <4> A virus inactivating agent, a virus inactivation method, or a virus inactivation composition as described in any of the above. <6> The virus is preferably an enveloped RNA virus, an envelopeless RNA virus, or an envelopeless DNA virus, and more preferably an influenza virus, norovirus, adenovirus, or enterovirus. <1> ~ <5> A virus inactivating agent, a virus inactivation method, or a virus inactivation composition as described in any of the above. <7> Preferably used in a liquid phase state. <1> ~ <6> A virus inactivating agent, a virus inactivation method, or a virus inactivation composition as described in any of the above. <8> The pH of the composition is preferably 4 to 11, more preferably 8 to 11, and even more preferably 10 to 11. <2> ~ <7> A method for inactivating a virus or a virus inactivation composition as described in any of the above. [Examples]

[0033] Test Example 1: Evaluation against Feline Calicivirus (FCV) The polymers were suspended in each pH standard solution (Fujifilm Wako Pure Chemicals) to achieve the concentrations and pH values ​​listed in Tables 1-4. A phthalate pH standard solution was used to prepare the pH 4 test solution, while neutral phosphate pH standard solution, borate pH standard solution, and carbonate pH standard solution were used to prepare the pH 5.5-8, pH 9, and pH 10-11 test solutions, respectively. pH adjustment was performed using 2M hydrochloric acid solution and 1M potassium hydroxide solution. The pH standard solutions adjusted to the pH values ​​listed in the table were used as controls. Units in the table are "mass%". The pH and pfu / mL after mixing the test solution and feline calicivirus solution are shown in the table. The FCV inactivation effect of the test solution was evaluated using the method described below.

[0034] <Evaluation Method> The tests were conducted with reference to the "FY2015 Survey Report on Norovirus Inactivation Conditions" (National Institute of Health Sciences) and ASTM-E1052, which are generally known as standard methods for evaluating viral inactivation. First, feline calicivirus solutions (F-9 strain, ATCC VR-782) with log pfu / mL values ​​of approximately 6 and 6.3, respectively, were prepared. The former was used in the tests shown in Table 2, and the latter was used in tests other than those shown in Table 2. In a 1.5 mL sample tube, 90 μL of each test solution shown in the table and 10 μL of the above feline calicivirus solution (F-9 strain, ATCC VR-782) were added and mixed thoroughly. After reacting at 30°C for the time indicated in the table, the reaction was stopped by diluting 10-fold with 900 μL of quench solution (SCDLP solution). After stopping the reaction, the solution was serially diluted 10-fold, 100-fold, and 1000-fold in DMEM medium. 0.5 mL of each of these dilutions was then used to infect CRFK cells (feline kidney-derived cell line) (ATCC CCL-94) that had been washed with PBS (phosphate-buffered saline). After 1 hour, the medium was replaced with methylcellulose-containing medium. After culturing at 37°C for 2 days, the cells were stained with crystal violet and the number of plaques was counted.

[0035] [Table 1]

[0036] [Table 2]

[0037] [Table 3]

[0038] [Table 4]

[0039] Test Example 2: Evaluation against Human Norovirus (HNV) Polymers were prepared by suspending them in carbonate pH standard solutions (Fujifilm Wako Pure Chemicals) to the concentrations and pH levels listed in Table 5. pH adjustment was performed using 2M hydrochloric acid solution and 1M potassium hydroxide solution. The carbonate pH standard solutions adjusted to the pH levels listed in the table were used as controls. The units in the table are in "mass percent". The pH values ​​listed in the table are the pH after mixing the test solution and virus solution. The HNV (HuNoV) inactivation effect was evaluated using the method described below.

[0040] <Evaluation Method> (1) Culture of human intestinal epithelial organoids (hIEOs) hIEOs were embedded in Matrigel (Corning) on ​​24-well plates and then cultured in three dimensions under conditions of 37°C and 5% CO2. IntestiCult Organoid Growth Medium (Human) (STEMCELL Technologies) was used as the culture medium. The entire volume of medium (500 μL / 2 days) was changed every 2-3 days, and subculturing was performed every 5-7 days. After 3D culture, the hIEOs were suspended in TrypLE Express (Thermo Fisher Scientific) along with the Matrigel. The hIEOs were dispersed by enzymatic treatment at 37°C for 10 minutes, and to stop the reaction, a basal medium containing Advanced DMEM / F-12 (Gibco) supplemented with 100 U / mL Penicillin-Streptomycin (Thermo Fisher Scientific), 10 mM HEPES (Thermo Fisher Scientific), and 1×GlutaMAX (Thermo Fisher Scientific) was added. After settling the cells by centrifugation (3 minutes, 300×g, 4°C), the above medium was removed. Cells were suspended in differentiation medium supplemented with 1×B27 supplement (Thermo Fisher Scientific), 10nM gastrin I (Sigma-Aldrich), 1mM N-acetylcysteine ​​(Fujifilm Wako), 50ng / mL mouse EGF recombinant protein (Thermo Fisher Scientific), 100ng / mL mouse Noggin (Peprotech), 500μg / mL human R-spondin-1 (R&D Systems), 500nM A83-01 (Tocris), and 10μM Y-27632 (Fujifilm Wako) in basal medium. Furthermore, for monolayer culture, 96-well flat-bottom plates coated with Cell Matrix (registered trademark) type IC (Nitta Gelatin) were prepared. To prepare the plates, 100 μL of Cell Matrix type IC (Nitta Gelatin), diluted 10-fold with 0.1 M acetic acid (Nacalai tesque), was added to each well, and then incubated at 37°C for 90 minutes. After that, the solution was removed and washed three times with 200 μL of D-PBS(-) (Nacalai tesque). 5 × 10⁶ cells were added to the 96-well flat-bottom plates that had undergone the above treatment. 4 Cells were seeded at a concentration of 100 μL / well. Differentiation induction was carried out for a total of 6 days, with the entire differentiation medium being replaced every 2 days, to obtain monolayered hIEOs.

[0041] (2) Preparation of a 10% emulsion of feces containing HNV (HuNoV) A 10% fecal emulsion was prepared from the feces of patients with HNV type GII.4. One tablet of the protease inhibitor cOmplete protease inhibitor cocktail tablets (Sigma-Aldrich, 11697498001) was suspended in 50 mL of D-PBS(-). 1 g of feces was suspended in 10 mL of cOmplete-containing D-PBS(-) and thoroughly mixed using a test tube mixer. After standing at 4°C for 20 minutes, the mixture was centrifuged at 2,000 × g at 4°C for 10 minutes. The supernatant was collected in a new tube and stored at -80°C until use in infection experiments.

[0042] (3) Inactivation treatment of HNV and infection of differentiated hIEOs A 10% fecal emulsion containing HNV was diluted 10-fold with differentiation medium and filtered using a 1 mL syringe and a Millex HV Filter unit (Millipore, SLHVR04NL). 5 μL of the filtered fecal solution (2.8 × 10⁶) was collected in a PA microcentrifuge tube (Beckman coulter). 6A 45 μL test solution containing HNV genome copy equivalent and polymer at the specified concentrations shown in Table 5 was mixed and reacted at the specified temperature and time shown in Table 5. Next, 0.95 mL of Fetal Bovine Serum (Capricorn Scientific) solution was added to this drug-treated fecal solution. The centrifuge tube was set on a fixed-angle rotor TLA-55 (Beckman coulter) and ultracentrifuged for 1.5 hours at Rmax with a centrifugal force of 186047 × g (equivalent to 55,000 rpm) using an Optima MAX-TL (Beckman coulter), and the supernatant was removed. The pellet was suspended in 100 μL of differentiation medium to prepare the infection solution for hIEOs. After removing the existing medium from the wells (96-well plate), the infection solution prepared in the manner described above was applied to the hIEOs after 6 days of differentiation induction. Incubation was performed at 37°C for 1 hour. After washing three times with 300 μL of basal medium, 250 μL of differentiation medium was added, and the cells were cultured at 37°C under 5% CO2 conditions until sampling. 10 μL of supernatant was collected immediately after the start of culture and again after 6 days of culture. The collected supernatant was centrifuged at 15,000 rpm for 5 minutes to remove cell debris, and then frozen and stored at -80°C until HuNoV genome RNA quantification was performed.

[0043] (4) RT-qPCR The number of HNV genome copies in the recovered supernatant was quantified using the Norovirus Detection Kit G1 / G2 (Toyobo, FIK-273). The procedure followed the protocol. PCR amplification and data measurement were performed using LightCycler480II (Roche).

[0044] [Table 5]

[0045] Test Example 3: Evaluation against adenovirus The polymer was suspended in a carbonate pH standard solution (FUJIFILM Wako Pure Chemical Industries, Ltd.) to achieve the concentrations and pH values shown in Table 6. Further, 2M hydrochloric acid solution and 1M potassium hydroxide solution were used for pH adjustment. Note that the carbonate pH standard solution adjusted to the pH shown in Table 6 was used as a control. The unit in the table is “mass %”. The pH after mixing the test solution and the adenovirus solution and log 10 TCID 50 / mL are shown in Table 6. The adenovirus inactivation effect of the test solution was evaluated by the method shown below.

[0046] <Evaluation method> Each test solution shown in Table 6 and log 10 TCID 50 / mL of an adenovirus solution (Human adeno virus5; Strain: adenoid75 ATCC VR-5) adjusted to 7.8 were mixed at a ratio of 9:1 (v:v) and thoroughly mixed. After reacting at room temperature for the time shown in Table 6, heat-inactivated fetal bovine serum solution was added and diluted 10-fold to stop the reaction. After preparing a 10-fold dilution series with a medium (heat-inactivated fetal bovine serum: Ham’s F-12K medium = 2:98 = v:v), these diluted solutions were each infected with A549 cells (cells derived from human lung epithelial cells, ATCC CCL-185) cultured in a 96-well plate and cultured at 37°C for 6 days. After culturing, the presence or absence of cytopathic effect was confirmed using an inverted phase contrast microscope, and log 10 TCID 50 / mL was calculated.

[0047]

Table 6

[0048] Test Example 4 Evaluation against Enterovirus The polymer was suspended in a carbonate pH standard solution (FUJIFILM Wako Pure Chemical Industries, Ltd.) to achieve the concentrations and pH values shown in Table 7. Further, 2M hydrochloric acid solution and 1M potassium hydroxide solution were used for pH adjustment. Note that the carbonate pH standard solution adjusted to the pH shown in Table 7 was used as a control. The unit in the table is “mass %”. The pH after mixing the test solution and the enterovirus solution and log 10TCID 50 Table 7 shows the values ​​per mL. The enterovirus inactivation effect of the test solution was evaluated using the method described below.

[0049] <Evaluation Method> Each test solution shown in Table 7 and log 10 TCID 50 Enterovirus solution (Enterovirus A71; Strain:BrCr, ATCC VR-1775) adjusted to 8.8 / mL was added in a 9:1 (v:v) ratio and thoroughly mixed. After reacting at room temperature for the time indicated in Table 7, the reaction was stopped by adding inactivated fetal bovine serum to dilute 10-fold. A 10-fold dilution series was prepared in culture medium (inactivated fetal bovine serum:Eagle's Minimum Essential Medium = 2:98 = v:v), and these dilutions were used to infect VERO cells (African green monkey kidney epithelial cells, ATCC CCL-81) cultured in 96-well plates, respectively, and cultured at 37°C for 6 days. After culturing, the presence or absence of cytopathic effects was confirmed by inverted phase-contrast microscopy, and log 10 TCID 50 The / mL value was calculated.

[0050] [Table 7]

[0051] Test Example 5: Evaluation against influenza virus The polymer was suspended in a carbonate pH standard solution (Fujifilm Wako Pure Chemicals) to the concentrations and pH shown in Table 8. pH adjustment was performed using a 2M hydrochloric acid solution and a 1M potassium hydroxide solution. The carbonate pH standard solution adjusted to the pH shown in Table 8 was used as a control. Units in the table are "mass%". pH and log after mixing the test solution / influenza virus solution. 10 TCID 50 Table 8 shows the values ​​per mL. Note that the units in the table are in "mass percent". The influenza virus inactivation effect of the test solution was evaluated using the method described below.

[0052] <Evaluation Method> Each test solution shown in Table 8 and log 10 TCID 50 Influenza A virus solution (H1N1, A / PR / 8 / 34 ATCC VR-1469) adjusted to 8.9 / mL was added in a 9:1 (v:v) ratio and thoroughly mixed. After reacting at room temperature for the time indicated in Table 8, the reaction was stopped by adding SCDLP medium to dilute 10-fold. A 10-fold dilution series was prepared in Eagle's Minimum Essential Medium, and these dilutions were used to infect MDCK cells (canine kidney-derived cells, ATCC CCL-34) cultured in 96-well plates, respectively, and cultured at 37°C for 4 days. After culturing, the presence or absence of cytopathic effects was confirmed by inverted phase-contrast microscopy, and log 10 TCID 50 The / mL value was calculated.

[0053] [Table 8]

Claims

1. (A) A virus inactivator containing maleic acrylate copolymer or a salt thereof as an active ingredient.

2. The virus inactivator according to claim 1, wherein the virus is an enveloped RNA virus, an envelopeless RNA virus, or an envelopeless DNA virus.

3. The virus inactivator according to claim 1 or 2, wherein the virus is an influenza virus, norovirus, adenovirus, or enterovirus.

4. The virus inactivator according to claim 1 or 2, wherein the concentration of component (A) is 10 ppm or more and 15,000 ppm or less.

5. (A) A method for inactivating a virus, comprising applying a maleic acrylate copolymer or a salt thereof, or a composition containing the same, to an object suspected of being contaminated with a virus.

6. (A) A virus inactivation composition comprising maleic acrylate copolymer or a salt thereof as an active ingredient, A virus inactivation composition for application to objects suspected of viral contamination, which is diluted with water to form a diluted solution containing component (A) in an amount of 10 ppm to 15,000 ppm.

7. (A) A virus inactivation composition comprising maleic acrylate copolymer or a salt thereof as an active ingredient, A virus inactivating composition in which the concentration of component (A) in the composition is 10 ppm or more and 15,000 ppm or less.

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