JP7840658B2 - Virus inactivator - Google Patents

Description

This invention relates to a virus inactivator that inactivates viruses present in a space.

Viral infections, ranging from common cold symptoms to serious conditions such as pneumonia, hepatitis, and encephalitis, remain a perpetual threat to humanity. In recent years, influenza viruses have ravaged the world, sometimes leading to pandemics due to the emergence of novel influenza strains with altered antigenicity. Furthermore, in 2019, SARS-CoV-2 emerged, causing a pandemic that impacted not only life and health but also economic activity and social functions.

To address situations like this, vaccines and antiviral drugs are being developed, but the development of vaccines and treatments takes time and is not always successful.
Viruses spread when they are brought into living spaces by infected individuals, either directly from the patient or through the environment, including 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.

Traditionally, ethanol, sodium hypochlorite, chlorine dioxide, and glutaraldehyde 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, while spraying chlorine dioxide has been considered as a method to chemically inactivate viruses in the air, its effectiveness is not yet confirmed.

Some alcohols, aldehydes, and carboxylic acids are known to have virus inactivation effects in liquid phase evaluation. For example, hexanol is known to inactivate vesicular stomatitis virus, and phenylethyl alcohol is known to inactivate enveloped viruses at -20°C (Patent Document 1, Non-Patent Document 1). Furthermore, octanal and dodecanal have been reported to inactivate herpes simplex virus type 1, and hexanoic acid has been reported to inactivate hepatitis C virus (Non-Patent Documents 2, 3).

However, while these compounds exhibit activity in the liquid phase, they exert a virus-inactivating effect when mixed with viruses in a liquid. It remains unclear whether, or to what extent, they exert a virus-inactivating effect when diffused into the air as volatile compounds.

U.S. Patent No. 4,909,940

Roihel et al., Acta Virol. 1969;13(2):139-141 Hayashi K, et al., Planta Med. 1995;61(3):237-241 Pfaender et al., J Infect Dis. 2013;208(12):1943-1952

This invention relates to providing a virus inactivator that enables the inactivation of viruses present in a space.

The inventors have discovered that certain volatile compounds have the effect of inactivating influenza viruses in the gas phase and are useful as virus inactivators.

In other words, the present invention relates to the following 1) to 2).
1) A virus inactivator that inactivates viruses in the gas phase, comprising one or more compounds selected from benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal as an active ingredient.
2) A method for inactivating a virus, comprising contacting or reacting a virus in the gas phase with one or more compounds selected from benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal, or a composition containing the same.

The virus inactivating agent of the present invention can inactivate viruses attached to hard and soft surfaces in the living environment, viruses dispersed in living spaces as droplets, and viruses floating in the air as aerosols, thereby preventing or reducing the spread of infection caused by these viruses.

Influenza virus inactivation effect (gas phase). Influenza virus inactivation effect (liquid phase).

The benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal (hereinafter also referred to as "the compounds of the present invention") are all compounds used as fragrances and are commercially available as follows.
- Benzyl alcohol (also known as phenylmethanol): Tokyo Chemical Industries Co., Ltd. - Phenyleethyl alcohol (also known as 2-phenylethanol): Sigma-Aldrich GmbH, Tokyo Chemical Industries Co., Ltd. - Isoamyl alcohol (also known as 3-methyl-1-butanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich GmbH - Amyl alcohol (also known as 1-pentanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich GmbH
Octanal (also known as caprylaldehyde): Sigma-Aldrich, Tokyo Chemical Industries Co., Ltd. Hexanol (also known as 1-hexanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich Hexanoic acid (also known as caproic acid): Tokyo Chemical Industries Co., Ltd., Fujifilm Wako Pure Chemical Industries, Ltd. Heptanol (also known as 1-heptanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich Octanol (also known as 1-octanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich Nonanol (also known as 1-nonanol): Tokyo Chemical Industries Co., Ltd., Sigma-Aldrich Decanol (also known as 1-decanol): Sigma-Aldrich, Tokyo Chemical Industries Co., Ltd. - 1-Dodecanol (also known as lauryl alcohol): Tokyo Chemical Industries, Ltd., Sigma-Aldrich GmbH - Dodecanal (also known as lauryl aldehyde): Sigma-Aldrich GmbH, Fujifilm Wako Pure Chemical Industries Ltd.

Such compounds may be used individually or in mixtures of two or more.

Among the compounds of the present invention, preferred candidates include benzyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, phenylethyl alcohol, hexanoic acid, and heptanol from the viewpoint of virus inactivation effect; benzyl alcohol and phenylethyl alcohol from the viewpoint of fragrance intensity, quality, and volatility; and benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, and octanal from the viewpoint of overall suitability.
In the present invention, such compounds are used in a gaseous state.

The viruses targeted by the virus inactivator of the present 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.

Of these, viruses having an envelope are preferred, viruses having an envelope and RNA as their nucleic acid are more preferred, and influenza virus, human coronavirus, SARS coronavirus, and SARS coronavirus-2 are even more preferred.
SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is a SARS-related coronavirus that causes acute respiratory illness (COVID-19).

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 influenza virus, for example, canine kidney cells (MDCK), African green monkey kidney epithelial cells (Vero), or duck embryonic stem cell-derived cell lines (EB66) can be used. For human coronavirus, for example, human ileocecal adenocarcinoma cells (HCT-8), African green monkey kidney epithelial cells (VeroE6), or human liver cancer-derived cell lines (Huh7) can be used.

As shown in the examples described later, when the compound of the present invention, soaked into a cotton ball, is placed in a glass bottle and filled to capacity, and then dried influenza virus is added, and the compound of the present invention and the virus are brought into contact in the gas phase at room temperature, the viral infectivity titer decreases by 13% or more.
Therefore, the compounds of the present invention can serve as virus inactivators that inactivate viruses in the gas phase. Alternatively, the compounds of the present invention can be used to produce virus inactivators that inactivate viruses in the gas phase.
Furthermore, the compounds of the present invention can be used to inactivate viruses in the gas phase.

The virus inactivating agent of the present invention may be used in the form of the compound of the present invention alone, or in the form of a composition containing it. That is, the virus inactivating agent of the present invention may be an antiviral composition that exhibits a virus inactivation effect in the gas phase, more specifically a composition for deviralizing spaces, or a material or formulation for incorporating such a composition.
Here, examples of spaces include living spaces such as dining kitchens, bedrooms, children's rooms, bathrooms, and toilets in ordinary homes; facilities such as shops, restaurants, inns, hospitals, workshops, and factories; vehicles such as cars, trains, and airplanes; and semi-enclosed spaces (lockers, storage rooms, closets, etc.).

The above antiviral composition may take the form of a liquid or a gel, but a liquid form is preferred. The composition can be prepared by blending the compound of the present invention with a base material and various additives (polyols (dipropylene glycol, propylene glycol, etc.), surfactants, ultraviolet absorbers, antioxidants, preservatives, deodorants, natural extracts, silicones, thickeners, dyes, pigments, colorants, oils, fragrances, etc.). Here, the base material can be conventionally known, whether oily or aqueous, such as water, methanol, ethanol, propanol, isopropanol, butanol, isobutanol, dimethyl ether, liquid propane, petrolatum, lanolin, castor oil, paraffinic hydrocarbons (e.g., liquid paraffin, etc.), and can be used alone or in combination of two or more.
When preparing a gel-like formulation, natural or synthetic gelling agents, such as water-soluble gelling agents like carrageenan or gellan gum, or oil-soluble gelling agents like metal soaps or aluminum octylate, may be added as appropriate according to conventionally known methods.

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 the compound of the present invention relative to the total amount of the composition is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and even more preferably 0.1% by mass or more. Furthermore, it is preferably 99.999% by mass or less, more preferably 50% by mass or less, and even more preferably 10% by mass or less. Also, it is preferably 0.001 to 99.999% by mass, more preferably 0.01 to 50% by mass, and even more preferably 0.1 to 10% by mass.

In the virus inactivator of the present invention, the compound of the present invention or a composition containing the same is applied to objects where viral contamination is a concern. This can be done by contacting or reacting the compound of the present invention or a composition containing the same with the virus in the gas phase, and the compound of the present invention may be volatilized naturally or by forced volatilization.
If the compound of the present invention is in a form that naturally volatilizes, viruses present in the space can be inactivated simply by leaving it in a living space, allowing for easy removal of viruses from the space (virus elimination).
When the virus inactivator of the present invention is used for natural volatilization, conventionally known methods such as impregnating a core rod, filter paper, etc., with the compound of the present invention or a composition containing it and volatilizing it, or using a permeable membrane for volatilization, can be applied. Alternatively, the compound of the present invention or a composition containing it can be kneaded into a resin and used. Examples of resins that can be kneaded include natural, petroleum, and synthetic waxes, rosin resins, ethylene-vinyl acetate copolymers, ethylene-vinyl alcohol copolymers, polyesters, polyolefins, and acrylic resins. The 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 synthetic resins mentioned above, and inorganic porous materials such as calcium silicate, all in any shape such as granules or sheets. The kneaded material or laminate can be used, for example, by installing it in air conditioning equipment, toilets, bathrooms, living rooms, hospital rooms, hospital waiting rooms, dustbins, etc., allowing the compound of the present invention to volatilize gradually. Furthermore, the compound of the present invention or a composition containing the same can be used by supporting it on a product made of paper, nonwoven fabric, or the like (such as an air purifier filter).

When the compound of the present invention is used by forced volatilization, such means include, for example, volatilization using a fan, heating volatilization using a heater, or volatilization using ultrasound.

Furthermore, the compound of the present invention or a composition containing the same can be used by filling it into a container or device for atomization or diffusion, such as a pressurized liquid spray, pressurized air atomizer, diffuser, or nebulizer, and spraying it in a mist-like manner into a space where viruses are present. This allows for an accelerated volatilization rate and rapid activation of the virus. Similar effects can also be obtained by using the compound of the present invention or a composition containing the same in the form of an aerosol or mist spray.

When performing airborne virus removal treatment, the amount of the compound of the present invention 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 each compound, etc. It is also possible to use an amount greater than or equal to the saturation concentration of each compound in the space. For example, the concentration of the compound of the present invention in the target space is 0.1% or more, preferably 1% or more, more preferably 5% or more, even more preferably 10% or more, and preferably 10% or less of the saturation concentration of the compound in the space, and is used so that it volatilizes at 100% or less, more preferably 50% or less, and more preferably 25% or less.
The concentration of the compound of the present invention in a target space can be detected by measuring the concentration of the compound in a gas sample taken from the space. This can be done using a volatile organic compound concentration meter (VOC meter, odor sensor, etc.), or by using gas chromatography or gas chromatography/mass spectrometry in combination with a gas collection tube.

With regard to the embodiments described above, the present invention further discloses the following embodiments.
<1> A virus inactivator that inactivates viruses in the gas phase, comprising one or more compounds selected from benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal as an active ingredient.
<2> Use of one or more compounds selected from the following for producing a virus inactivator that inactivates viruses in the gas phase.
<3> Use of one or more compounds selected from benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal to inactivate the virus in the gas phase.
<4> A method for inactivating a virus, comprising contacting or reacting a virus in the gas phase with one or more compounds selected from benzyl alcohol, phenylethyl alcohol, isoamyl alcohol, amyl alcohol, octanal, hexanol, hexanoic acid, heptanol, octanol, nonanol, decanol, 1-dodecanol, and dodecanal, or a composition containing the same.
In <5>, <1> to <4>, the virus is preferably an enveloped RNA virus.
In <6>, <1> to <4>, the virus is preferably an influenza virus or a coronavirus.
In <7>, <1> to <6>, the content of the compound in the composition containing the compound 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 0.1% by mass or more, and preferably 99.999% by mass or less, more preferably 50% by mass or less, even more preferably 10% by mass or less, or preferably 0.001 to 99.999% by mass, more preferably 0.01 to 50% by mass, even more preferably 0.1 to 10% by mass.
<8> The method of <4>, wherein the compound or a composition containing the same is used such that the concentration of the compound in the target space is 0.1% or more, preferably 1% or more, more preferably 5% or more, even more preferably 10% or more, preferably 100% or less, more preferably 50% or less, and more preferably 25% or less of the saturation concentration of the compound in the space when it is volatilized.

The present invention will be described in more detail below with reference to examples.
Example 1 Inactivation of influenza virus in the gas phase by fragrance compounds 1. Method Influenza virus type A (A/Puerto Rico/8/1934, H1N1) strain was used as the test virus strain. 75 μL of the compounds or mineral oil shown in Table 1 below was soaked into a cotton ball and attached to the lid of a 15 mL glass bottle (Maruemu Co., Ltd.) with double-sided tape to seal it, and it was filled for 30 minutes. 1.5 μL of influenza virus (8.3 × ^10⁵ FFU) was dried on the lid of a cryovial (Thermo Fisher Scientific) for 30 minutes. The lid of the vial with the virus attached was placed in the glass bottle, and the compound and virus were reacted at room temperature (approximately 23°C) for 30 minutes. During this time, the compound was placed on the lid of the bottle, and the virus was placed at the bottom of the bottle, maintaining a state where the compound and virus did not come into direct contact. After the reaction, the virus was recovered in Hybridoma-SFM medium (Thermo Fisher Scientific) and inoculated into MDCK cells (derived from canine renal tubular epithelial cells) that had been pre-cultured in 12-well plates. After incubation at 37°C under 5% _CO2 conditions for approximately 18 hours, the number of foci formed was measured to determine the viral infectivity titer. The infectivity titer when reacted with the control mineral oil was set to 100%, and the viral inactivation activity of each compound was calculated. The test was performed three times.

2. Results As shown in Figure 1, compared to the control mineral oil, compounds 1 to 13 reduced the detectable viral load to 0.001%, 0.04%, 0.001%, 0.001%, 0.01%, 0.01%, 0.13%, 0.67%, 23.7%, 44.7%, 65.5%, 67.3%, and 86.6%, respectively.

Reference Example: Inactivation of influenza virus in liquid phase with fragrance compounds 1. Method Influenza virus type A (A/Puerto Rico/8/1934, H1N1) strain was used as the test virus strain. The compounds shown in Table 1 were dissolved in Hybridoma-SFM medium (Thermo Fisher Scientific) containing 1% (v/v) dipropylene glycol as a solvent so that the final concentration when reacting with the virus was 0.1% (v/v). 60 μL each of the compound solution or the 1% (v/v) dipropylene glycol solution and the influenza virus solution (8.3 × ^10⁵ FFU) using the medium as a solvent were added to a 96-well plate and reacted at room temperature (approximately 23°C) for 30 minutes. After the reaction, the virus was diluted with culture medium and inoculated into MDCK cells (derived from canine renal tubular epithelial cells) that had been pre-cultured in 48-well plates. After culturing at 37°C under 5% _CO2 conditions for approximately 18 hours, the number of foci formed was measured to determine the viral infectivity titer. The infectivity titer when reacted with a control 1% (v/v) dipropylene glycol solution was set as 100%, and the viral infectivity titer of each compound was calculated. The test was performed three times.

2. Results As shown in Figure 2, compared to the control 1% dipropylene glycol solution, compounds 1 to 9 reduced the detected viral load to 76.2%, 84.1%, 69.9%, 73.0%, 67.0%, 81.5%, 82.0%, 65.3%, 73.4%, 76.9%, 20.6%, 21.1%, and 33.8%, respectively.

Claims (4)

A virus inactivator that uses phenylethyl alcohol as its active ingredient and is used to inactivate viruses in the gas phase. The virus inactivator according to claim 1, wherein the virus is an enveloped RNA virus. The virus inactivator according to claim 1 or 2, wherein the virus is an influenza virus or a coronavirus. A method for inactivating a virus, comprising contacting or reacting a virus with phenylethyl alcohol or a composition containing the same in the gas phase.