JP7778996B2 - virus inactivating agents - Google Patents

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This international application claims priority to Japanese Patent Application No. 2020-208804, filed with the Japan Patent Office on December 16, 2020, the entire contents of which are incorporated herein by reference.

The present disclosure relates to a virus inactivating agent that inactivates viruses.

Recently, a new coronavirus known as SARS-CoV-2 has become widespread. To prevent viral infection, people are washing their hands with water and soap, and using alcohol-based disinfectants, as described in Non-Patent Document 1.

The inventors have conducted extensive research into substances that can inactivate viruses even more effectively.

The National Institute of Technology and Evaluation (NITE) “Final Report on Efficacy Assessment of Disinfecting Substances Alternative to Alcohol for Use Against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)” The Committee on Efficacy Assessment of Disinfecting Substances Alternative to Alcohol for Use Against Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2)June, 2020

One aspect of the present disclosure preferably provides a virus inactivating agent capable of inactivating the novel coronavirus.

Another aspect of the present disclosure is to preferably provide a virus inactivating agent that can inactivate the novel coronavirus and is also gentle on human skin.

A first aspect of the present disclosure provides:
i) Alkaline ion electrolyzed water containing purified water and at least sodium, silicon, and chlorine as components, and containing more than 99.5% by weight of the purified water; or
ii) a diluted solution obtained by diluting the alkaline ionized water with purified water in an amount of not more than twice the volume of the alkaline ionized water;
It is a virus inactivating agent capable of inactivating viruses.

A second aspect of the present disclosure is a virus inactivation agent capable of inactivating viruses, comprising alkaline ionized water,
The alkaline ionized water is a virus inactivating agent containing at least sodium, silicon, and chlorine.

1 is a photograph showing the state in Example 1 where a subject sample of a test sample is measured by a plaque measurement method. 1 is a photograph showing a state in which a test sample is measured by a plaque measurement method in Example 1.

An exemplary embodiment of the present disclosure (hereinafter referred to as "this embodiment") is described below.

The virus inactivating agent of this embodiment inactivates viruses, such as SARS-CoV-2 (novel coronavirus) which causes the disease COVID-19 (novel coronavirus infection). "Inactivation" refers to killing a virus or eliminating its infectivity and toxicity.

The virus inactivating agent of this embodiment is either i) alkaline ionized water containing purified water and at least sodium, silicon, and chlorine as ingredients, with the purified water content being greater than 99.5% by weight, or ii) a diluted solution obtained by diluting alkaline ionized water with purified water in an amount equal to or less than twice the volume of the alkaline ionized water. It is preferable that the purified water content of the alkaline ionized water be 99.7% by weight or more.

The virus inactivating agent of this embodiment contains alkaline ionized water, which contains at least sodium, silicon, and chlorine as ingredients.

Alkaline ionized water is generally said to have a beneficial effect on the living body. For example, alkaline ionized water is said to have the effect of improving gastrointestinal symptoms, and has been used for medicinal purposes. While independently developing alkaline ionized water that is beneficial to the living body, the inventor discovered that a certain type of alkaline ionized water has the ability to inactivate viruses. This was a major departure from the inventor's expectations that alkaline ionized water would improve the functioning of the living body. This led the inventor to come up with the idea of using alkaline ionized water as a virus inactivating agent.

The virus inactivating agent of this embodiment has virus inactivation performance comparable to or superior to that of detergents, disinfectants, and ethanol. The virus inactivating agent of this embodiment exhibits excellent virus inactivation effects even without containing known virus infection inhibitors such as detergents, disinfectants, and ethanol.

The alkaline ionized water of this embodiment is characterized by containing at least sodium, silicon, and chlorine as ingredients. This characteristic allows the alkaline ionized water of this embodiment to exhibit superior virus inactivation effects, unlike other alkaline ionized waters.

Alkaline ionized water refers to alkaline water that contains, for example, cations and ^OH- ions. Alkaline ionized water is also called reducing ionized water, and can be said to be water with a high electron content. This type of alkaline ionized water has a high electron content and is said to promote normal functioning of the living body.

Alkaline ionized water may be any alkaline water, and is preferably alkaline ionized water produced by electrolysis. Alkaline ionized water is obtained by electrolyzing base water containing electrolytes, and contains cations. Alkaline ionized water contains more cations than anions, and exhibits alkaline properties.

The alkaline ionized water of this embodiment contains at least sodium, silicon, and chlorine as ingredients. In this embodiment, "ingredients" refers to solutes contained in the alkaline ionized water. When the water (H ₂ O) in the alkaline ionized water is used as a solvent, the ingredients are contained in the alkaline ionized water as solutes. Most of the ingredients are inorganic and dissolved in water. By containing these ingredients, alkaline ionized water has a higher virus inactivation effect than water that does not contain these ingredients.

The alkaline ionized water of this embodiment may further contain at least one of phosphorus and potassium as an ingredient. The alkaline ionized water of this embodiment may also contain at least one of calcium and magnesium as an ingredient. Here, the ingredient may be contained in the virus inactivating agent in the form of a cation, anion, or hydroxide, or in the form of a salt. The ingredient may be produced by electrolysis of an electrolyte or by adding a salt to purified water.

The virus inactivating agent of this embodiment can inactivate viruses.

The virus inactivating agent of this embodiment has been confirmed to have low cytotoxicity, i.e., to have little effect on human skin.

The pH of the virus inactivating agent of this embodiment is preferably 10-13, and more preferably 11.5-12.5 (12±0.5). For reasons unknown, this effectively inactivates viruses while reducing the impact on human skin when the virus inactivating agent comes into contact with the skin.

The alkaline ionized water contained in the virus inactivation agent should have a pH of 12±0.5. Of the alkaline ionized water, it is preferable that the alkaline ionized water be adjusted to a pH of 12.0±0.5. The reason for this is unclear, but it is believed that this stabilizes the pH of the diluted solution formed by diluting the alkaline ionized water.

Virus inactivating agents include purified water. Purified water is water whose purity has been increased through purification methods such as distillation, ion exchange, and filtration. Examples of purified water include distilled water, ion-exchanged water, filtered water, and pure water.

The purified water content of the virus inactivator may be 98% by weight or more, preferably greater than 99.5% by weight, and more preferably 99.7% by weight or more. Such a virus inactivator can effectively inactivate viruses.

The virus inactivating agent of this embodiment contains alkaline ionized water, such as alkaline ionized water. When the virus inactivating agent of this embodiment is taken as 100% by weight, the total content of ingredients contained in the virus inactivating agent is preferably 0.001% by weight or more, more preferably 0.01% by weight or more, and particularly preferably 0.1% by weight or more. The total content of ingredients contained in the virus inactivating agent may be 2% by weight or less, and more preferably 0.5% by weight or less. In this case, viruses can be effectively inactivated.

If the total content of ingredients is too high, it may have an effect on skin cells when a person comes into contact with the virus inactivating agent. "Total content of ingredients" refers to the total content of all ingredients contained in the virus inactivating agent of this embodiment, and specifically refers to the total content of all ingredients contained in the virus inactivating agent, such as sodium, silicon, chlorine, phosphorus, potassium, calcium, and magnesium.

When the virus inactivating agent is taken as 100% by weight, the sodium content may be 0.0001-1% by weight, or may be 0.0001-0.3% by weight, and more preferably 0.001-0.2% by weight, with 0.01-0.2% by weight being preferred. When the total of the ingredients in the virus inactivating agent is taken as 100% by weight, the sodium content is preferably 10-60% by weight, and more preferably 20-50% by weight. "Total of ingredients" refers to the sum of all ingredients contained in the virus inactivating agent. If the sodium content is too low, virus inactivation may be reduced, while if the sodium content is too high, it may have an effect on skin cells.

When the virus inactivator is taken as 100% by weight, the silicon content can be 0.00001-0.01% by weight, or alternatively, 0.00001-0.001% by weight, preferably 0.00005-0.005% by weight, and even more preferably 0.0001-0.005% by weight. When the total contents of the virus inactivator is taken as 100% by weight, the silicon content is preferably 0.2-5% by weight, and even more preferably 0.5-0.9% by weight. In this case, virus inactivation by the virus inactivator is further enhanced. If the silicon content is too low, there is a risk of reduced virus inactivation, while if the silicon content is too high, there is a risk of adverse effects on skin cells.

When the virus inactivating agent is taken as 100% by weight, the chlorine content may be 0.0001-0.1% by weight, preferably 0.001-0.05% by weight, and more preferably 0.005-0.05% by weight. When the total content of the virus inactivating agent is taken as 100% by weight, the chlorine content should be 5-15% by weight, and more preferably 7-12% by weight. If the chlorine content is too low, there is a risk of reduced virus inactivation, and if the chlorine content is too high, there is a risk of it affecting skin cells.

When the virus inactivating agent is taken as 100% by weight, the potassium content may be 0-1% by weight, or may be 0-0.2% by weight, or preferably 0.003-0.15% by weight, and preferably 0.01-0.15% by weight. When the total contents of the virus inactivating agent is taken as 100% by weight, the potassium content is preferably 0-50% by weight, and more preferably 20-40% by weight. If the potassium content is too low, there is a risk of reduced virus inactivation, and if the potassium content is too high, there is a risk of it affecting skin cells.

When the virus inactivator is taken as 100% by weight, the phosphorus content may be 0-0.1% by weight, preferably 0.002-0.1% by weight, and even more preferably 0.01-0.1% by weight. When the total contents of the virus inactivator is taken as 100% by weight, the phosphorus content is preferably 0-40% by weight, and even more preferably 10-30% by weight. In this case, virus inactivation by the virus inactivator is further enhanced. If the phosphorus content is too low, there is a risk of reduced virus inactivation, while if the phosphorus content is too high, there is a risk of it affecting skin cells.

When the virus inactivating agent is taken as 100% by weight, the calcium content may be 0-0.002% by weight, and preferably 0-0.0002% by weight. When the total content of the virus inactivating agent is taken as 100% by weight, the calcium content is preferably 0-20% by weight, and more preferably 0-2% by weight. In this case, the virus inactivation by the virus inactivating agent is improved.

When the virus inactivating agent is taken as 100% by weight, the magnesium content may be 0-0.001% by weight, and preferably 0-0.0001% by weight. When the total content of the virus inactivating agent is taken as 100% by weight, the magnesium content is preferably 0-10% by weight, and more preferably 0-1% by weight. In this case, the virus inactivation by the virus inactivating agent is improved.

The composition of the virus inactivating agent of this embodiment preferably includes any of the following combinations, when the virus inactivating agent is taken as 100% by weight:

i) 0.0001-1 wt% sodium, 0.00001-0.01 wt% silicon, 0.0001-0.1 wt% chlorine, 0-1 wt% potassium, and 0-0.1 wt% phosphorus;
ii) 0.0001-0.3 wt% sodium, 0.00001-0.001 wt% silicon, 0.0001-0.1 wt% chlorine, 0-0.2 wt% potassium, and 0-0.1 wt% phosphorus;
iii) 0.001-0.2 wt.% sodium, 0.00005-0.005 wt.% silicon, 0.001-0.05 wt.% chlorine, 0.003-0.15 wt.% potassium, and 0.002-0.1 wt.% phosphorus; or iv) 0.01-0.2 wt.% sodium, 0.0001-0.005 wt.% silicon, 0.005-0.05 wt.% chlorine, 0.01-0.15 wt.% potassium, and 0.01-0.1 wt.% phosphorus.

The composition of the virus inactivating agent of this embodiment is as follows, when the total amount of the components contained in the virus inactivating agent is 100% by weight:
v) 10-60% by weight sodium, 0.2-5% by weight silicon, 5-15% by weight chlorine, 0-50% by weight potassium, and 0-40% by weight phosphorus; or vi) 20-50% by weight sodium, 0.5-0.9% by weight silicon, 7-12% by weight chlorine, 20-40% by weight potassium, and 10-30% by weight phosphorus.
It is good to include:

In the virus inactivating agent of this embodiment, the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon ^{, and chlorine is preferably 1-3, and more preferably 1.5-2.5. Phosphorus, silicon, and chlorine are elements that, optionally together with oxygen, mainly form anions (e.g., PO4 3- , SiO4 4-} _, ^Cl _- ^, ClO ^- ), and sodium and potassium are elements that form cations (Na ⁺ , K ⁺ ). When the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon, and chlorine is greater than 1, the virus inactivating agent can exhibit alkalinity suitable for inactivating viruses. When the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon, and chlorine is excessively large, the alkalinity is too strong, and the virus inactivating agent may affect the skin upon contact with the skin.

In the virus inactivation agent of this embodiment, to enhance virus inactivation, the weight ratio of phosphorus to silicon is preferably 10-50, and more preferably 20-40.

The components and contents of the ingredients contained in the virus inactivating agent of this embodiment can be determined, for example, by atomic absorption spectrometry, ICP atomic emission spectrometry, or combustion-coulometric titration.

The virus inactivating agent of this embodiment is not particularly limited as long as it has the above-mentioned composition. For example, iW-100 and S-100 manufactured by AI System Products Co., Ltd. can be used. iW-100 is alkaline ionized water with a pH of 11.5-12.5, containing sodium, silicon, chlorine, phosphorus, and potassium, with a total content of these elements of 0.5% by weight or less, and a water content of 99.5% by weight or more. S-100 is alkaline ionized water with a pH of 11.5-12.5, containing silicon, sodium, chlorine, calcium, potassium, phosphorus, and magnesium, with a total content of these elements of 1.2% by weight, and a water content of 99.7% by weight. The commercially available S-100 was able to inactivate the SARS-CoV-2 virus and influenza virus. Furthermore, iW-100 was able to inactivate the SARS-CoV-2 virus, as shown in the experimental examples described below.

The method for producing the virus inactivating agent of this embodiment includes a step of producing alkaline ionized water containing at least sodium, silicon, and chlorine, and optionally further containing at least one of potassium, phosphorus, calcium, and magnesium.

Furthermore, the method for producing the virus inactivating agent of this embodiment preferably includes a step of obtaining alkaline ionized water by electrolyzing raw water containing at least sodium, silicon, and chlorine, and optionally further containing at least one of potassium, phosphorus, calcium, and magnesium, in an electrolytic cell. The alkaline ionized water is preferably recovered from near the cathode after electrolysis in an electrolytic cell equipped with a cathode and an anode.

Original water is produced by adding ingredients to purified water. The ingredients may be electrolytes, or components derived from seawater, or/and components dissolved from soil or rocks.

An electrolyte is a substance that can be ionized into cations and anions by electrolysis, and examples of such substances include salts. Salts that can serve as electrolytes preferably include at least sodium chloride, and may also include potassium chloride, sodium carbonate, and potassium carbonate, but are not limited to these. Various other salts, such as silicates such as sodium silicate and phosphates such as sodium phosphate, may also be used as electrolytes.

Seawater is primarily composed of water, with approximately 3.5% by weight of salt and trace metals. Seawater contains salts such as sodium chloride, magnesium chloride, magnesium sulfate, calcium sulfate, and potassium chloride. Soil and rock contain a lot of silicon. Soil and rock vary depending on the type, but some contain calcium, potassium, sodium, magnesium, and phosphorus in addition to silicon. The type of soil and rock to be used should be selected so that it matches the composition of the alkaline ionized water you are trying to produce.

To electrolyze the raw water, it is preferable to apply electrical energy of 100 to 300 W to the raw water. Electrolysis is preferably carried out in a sealed electrolytic cell. The electrolytic cell should have an inert atmosphere such as nitrogen or carbon dioxide. Alternatively, an oxygen remover may be added to the electrolytic cell. The oxygen remover may be any material that has the properties to contribute to removing oxygen from the electrolytic cell, such as Ageless (registered trademark, Mitsubishi Gas Chemical Company, Inc.).

Alkaline ionized water may be used as a virus inactivation agent as a stock solution, or it may be diluted before use. In this specification, alkaline ionized water itself is also referred to as the stock solution. Alkaline ionized water can be diluted by adding purified water. When the dilution factor of alkaline ionized water is the ratio of the weight of the virus inactivation agent to the weight of the alkaline ionized water, the dilution factor of alkaline ionized water is preferably 10 times or less, more preferably 5 times or less, and desirably 3 times or less. A virus inactivation agent whose dilution factor with alkaline ionized water is within this range can exhibit high virus inactivation performance.

The virus inactivating agent of this embodiment may further contain additives. Examples of additives include dyes, fragrances, stabilizers, moisturizers, preservatives, surfactants, and ethanol.

Another aspect of this embodiment is a virus inactivation agent having alkaline ionized water containing at least sodium, silicon, and chlorine as ingredients.

Another aspect of this embodiment is the use of a solution containing alkaline ionized water for virus inactivation, wherein the alkaline ionized water contains at least sodium, silicon, and chlorine as components.

Another aspect of this embodiment is a method for inactivating viruses using a solution containing alkaline ionized water, wherein the alkaline ionized water contains at least sodium, silicon, and chlorine as components.

Another aspect of this embodiment is the use of alkaline ionized water to produce a virus inactivating agent, wherein the alkaline ionized water contains at least sodium, silicon, and chlorine as components.

The virus inactivating agent of this embodiment can be used to inactivate viruses. For example, the virus inactivating agent of this embodiment can be suitably used to inactivate SARS-CoV-2 (novel coronavirus), which causes the disease known as COVID-19 (novel coronavirus infection).

The virus inactivating agent of this embodiment can be used to inactivate not only SARS-CoV-2 (novel coronavirus), but also influenza viruses and most viruses.

The virus inactivating agent of this embodiment may be a virus inactivation liquid. The virus inactivating agent of this embodiment may be, for example, a liquid like a virus inactivation liquid, a semi-liquid such as a gel, or a solid. A gel-like virus inactivating agent may further contain a thickener, and a solid virus inactivating agent may further contain a gelling agent. The thickener may be any thickener that turns the virus inactivating agent into a semi-liquid, and for example, a polysaccharide may be used. Examples of polysaccharides that can be used as thickeners include carboxymethylcellulose, dextrin, pectin, guar gum, xanthan gum, tamarind gum, carrageenan, and propylene glycol. The gelling agent may be any gel that turns the virus inactivating agent into a solid, and examples include agar, silica gel, and gelatin.

The virus inactivating agent of this embodiment may be used alone or mixed with a product. Examples of products that can be mixed with the virus inactivating agent include laundry detergent, disinfectant, kitchen detergent, ethanol, cosmetics, nasal drops, nasal wash, and mouthwash.

When using the virus inactivating agent of this embodiment, it is advisable to bring at least a part of a human body (e.g., hands or fingers) that is at risk of viral infection into contact with the virus inactivating agent. The virus inactivating agent may also be applied or sprayed onto the surface of an object that may come into contact with the human body, or may be sprayed in a human living space.

When applying the virus inactivating agent of this embodiment, for example, the virus inactivating liquid, the application amount is preferably 0.005 to 5.0 ml/1 ^cm2 , more preferably 0.01 to 1.0 ml/1 ^cm2 , and preferably about 0.1 ml/1 ^cm2 . When spraying the virus inactivating liquid of this embodiment, the application amount is preferably 15 to 6000 ml/16 ^m3 , more preferably 30 to 3000 ml/16 ^m3 , and preferably about 300 ml/16 ^m3 .

The time of contact with the virus inactivating agent is preferably 2 seconds or more, and more preferably 10 seconds or more. The time of contact with the virus inactivating agent may be 2 minutes or less.

Examples of objects that may come into contact with the human body include textile products such as clothing, hats, towels, masks, bedding, sofas, wet towels, and cushions; decorative items such as rings, necklaces, earrings, piercings, and hair clips; residential components such as floors, walls, windows, and doors; household products such as kitchen counters, toilet seats, doorknobs, bathtubs, faucets, washbasins, home appliances, and kitchen utensils; furniture such as desks, chairs, shelves, and counters; electronic devices such as multifunction printers, personal computers, and keyboards; public equipment such as ticket machines, vending machines, automatic teller machines, automatic ticket gates, straps, equipment control panels, shopping carts, and handrails, as well as general nursing care equipment and general medical equipment.

Example 1
1. Virus inactivation liquid In the manufacturing process of the virus inactivation liquid of this example, purified water is passed through a filter layer that appropriately contains ingredients such as sodium, silicon, chlorine, phosphorus, and potassium to produce raw water containing these ingredients. The raw water is then electrolyzed with a target pH of 12 to produce a stock solution consisting of alkaline ionized water.

The filter layer can be made of any material (such as soil or rocks, as long as it can be used to form a filter layer) as long as the alkaline ionized water after filtration and electrolysis contains at least 99.5% purified water by weight and no more than 0.5% contaminants by weight. The filter layer is made by agitating the main components and adjusting the size so that the filtered raw water contains at least sodium, silicon, and chlorine.

The ingredients used are rocks containing phosphorus and silicon, and seawater containing sodium, chlorine, and potassium. This makes the virus inactivation liquid a gentle liquid that has little impact on the human body or the natural environment.

The ratio of the above-mentioned substances (sodium, silicon, chlorine, phosphorus, and potassium) to purified water varies depending on the filtration conditions, as does the ratio of the substances to the original water. The pH also varies depending on the amount and ratio of the substances to the purified water.

Based on the inventor's decades of experience, he has now discovered, by applying it himself and his collaborators, that a concentrate consisting of alkaline ionized water produced by electrolyzing raw water containing more than 99.5% purified water by weight and at least sodium, silicon, and chlorine as ingredients, with a target pH of 12.0, is natural and gentle on the skin.

The following experiment has now discovered that the above-mentioned concentrate has antiviral activity, particularly against the SARS-CoV-2 virus (a new use).

Based on this discovery, the following experiment was conducted to prove that the above stock solution can be used as an inactivation solution for the SARS-CoV-2 virus. The following experiment was conducted using a general method for verifying performance such as virus inactivation. The method for producing the stock solution and the experimental method are described below.
<Production method of stock solution>
Purified water was passed through a filter layer containing appropriate amounts of sodium, silicon, chlorine, phosphorus, potassium, and other ingredients to produce raw water containing these ingredients. The filter layer was a tank (60 cm inner diameter) filled to a height of 10 cm with rock crushed to an average diameter of 0.1 mm. The rock type was igneous rock. 80 L of raw water and 80 L of seawater were introduced into a filter tank (tank) equipped with a filter layer filled with rock and stored for 24 hours with appropriate stirring. The composition of the introduced seawater was 30 wt% sodium, 55 wt% chlorine, 1 wt% potassium, and the remainder trace elements, assuming the total solutes contained in the seawater to be 100 wt%.

The raw water was removed from the filtration layer and subjected to electrolysis. To perform the electrolysis, a cathode and an anode were placed in a cathode chamber and an anode chamber, respectively, separated by a diaphragm. The raw water was introduced into the electrolysis cell. The electrolysis conditions were a voltage of 200 V and an electric energy of 200 W. After electrolysis, a stock solution consisting of alkaline ionized water present near the cathode was recovered. The pH of the stock solution consisting of alkaline ionized water was 12.0.
<Test Overview>
1: Test location: Kobe Testing Center, Japan Textile Products Quality Technology Center, General Incorporated Foundation 2: Test virus: NIID isolate provided by the National Institute of Infectious Diseases: JPN/TY/WK-521 (SARS-CoV-2 virus)
3: Host cell: VeroE6/TMPRSS2
4: Cell culture medium: EMEM (Minimum Essential Medium Eagle)
DMEM (Dulbecco's modified Eagle's medium) (low-glucise)
5: Fetal Bovine Serum: FBS (Fetal Bovine Serum)
6: Control sample (negative control): PBS (phosphate buffered saline)
7: Test sample (this example): 1) Virus inactivation solution (undiluted solution)
2) Virus inactivation solution (2x diluted solution)
: 3) Virus inactivation solution (3-fold diluted solution)
*The stock solution is iW-100 (alkaline ionized water), which is diluted with purified water. The dilution ratio is the weight ratio of the virus inactivation solution to the stock solution.
8: Drug inactivating agent: SCDLP (Soybean-Casein Digest Broth with Lecithin & Polysorbate 80) was diluted 10-fold with DMEM containing 2% by weight of FBS to serve as a drug inactivating agent.
9: Infectivity titer measurement method: plaque measurement method (JIS L 1922)
10: Virus suspension: EMEM was added to host cells infected with SARS-CoV-2 virus, and the cells were cultured at 37°C for a predetermined time. The cells were then centrifuged at 1000 xg for 15 minutes at 4°C to obtain a supernatant, which was used as the virus suspension for testing. The virus concentration in the virus suspension was 2.5 x ¹⁰ PFU/ml.
<Test content and results>
[Test 1: Cytotoxicity Confirmation Test]
<Test Contents> (Test to confirm whether the test sample is toxic or not)
1. 0.1 mL of EMEM and 1.9 mL of the test sample were thoroughly mixed to prepare a toxicity test solution.

2. 0.1 mL of toxicity test solution was added to 0.9 mL of drug inactivator and stirred thoroughly.

A dilution series of 10 ^0- fold, 10 ^1- fold, 10 ^2- fold, 10 ^3- fold, and 10 ^4- fold dilutions was prepared using DMEM containing 3.2% by weight of FBS.

4. The presence or absence of cytotoxicity in each dilution series was confirmed using the plaque assay.
<Test Results> Cytotoxicity confirmation tests are essential safety tests for medical devices that come into direct contact with body tissues and blood. Table 1 shows the results of the cytotoxicity confirmation test. In Table 1, if no toxicity was found in the cytotoxicity confirmation test, the result is marked as "None." The test samples of this example were not toxic, even in their undiluted form.

[Test 2: Test to confirm cell susceptibility to viruses] (Test to confirm the presence or absence of the reaction-stopping function of the drug inactivator)
<<Exam Contents>>
1. 0.1 mL of EMEM and 1.9 mL of the test sample were thoroughly mixed to prepare a test solution.

2. 0.5 mL of test solution was added to 4.5 mL of drug inactivator and stirred thoroughly.

Using DMEM containing 3.2% by weight of FBS, a dilution series of 10 ^0- fold, 10 ^1- fold, 10 ^2- fold, 10 ^3- fold, and 10 ^4- fold was prepared.

4. The above-mentioned virus suspension, adjusted to 4.3 x 10 ⁴ PFU/ml using EMEM, was added to each dilution series in a volume of 1/100 of the dilution series.

5. Each dilution series from step 4 above was left to stand at room temperature for 10 minutes.

6. The virus infectivity per mL of each dilution series was measured by plaque assay to confirm the susceptibility of the cells to the virus.
Test Results
Table 1 shows the results of a test to confirm the susceptibility of cells to viruses. The presence or absence of susceptibility was determined by calculating the common logarithm mean value of the virus infectivity titer, and based on this common logarithm mean value, determining whether or not the condition of the calculation formula: Ig (virus infectivity titer of PBS (PFU/mL)) - Ig (virus infectivity titer of test sample (PFU/mL)) ≦ 0.5 was met. Meeting the conditions of this calculation formula indicates that virus growth has stopped, and not meeting the conditions indicates that the virus has been inactivated.

In this example, the virus infectivity titers of the undiluted solution, the 2-fold diluted solution, and the 3-fold diluted solution all had a difference of 0.5 or less from the virus infectivity titer of PBS, confirming that the drug inactivating agent stopped the proliferation of the SARS-CoV-2 virus.
[Test 3: Main Test] (Test to confirm whether or not the virus is inactivated)
<<Exam Contents>>
1. 0.1 mL of the virus suspension was mixed with 1.9 mL of the control sample or test sample, and the mixture was left to stand at 25°C for 20 seconds, 1 minute, and 5 minutes to prepare an infection test liquid.

2. 0.1 mL of the infection test solution was added to 0.9 mL of the drug inactivator and thoroughly stirred to stop the growth of the virus in the infection test solution. This was used as the reaction stop solution.

3. Using the reaction stop solution from 2. above as ¹⁰⁰ , a dilution series of 100 ^- fold, ^101- fold, ^102- fold, ^103- fold, and ^104- fold was prepared with DMEM containing 2 wt% FBS. For each dilution series, the virus infectivity per 0.1 ml of the infection test solution was measured by plaque assay, and the virus infectivity per ml of the infection test solution was calculated.

4. Table 2 shows the results of tests to confirm whether or not the virus was inactivated. In Table 2, n1, n2, and n3 indicate that each subject or test sample was tested in triplicate.

Test Results
As shown in Table 2, for the control sample (PBS), the common logarithm of the virus infectivity titer was almost constant. This indicates that the drug inactivator was functioning effectively. In contrast, for the test samples, the undiluted iW-100 (alkaline ionized water) inactivated more than 99.99% of the virus 20 seconds after mixing with the virus suspension. The 2x diluted solution inactivated more than 99.99% of the virus after 1 minute. Even the 3x diluted solution inactivated nearly 99.99% of the virus after 1 minute. This demonstrates that not only the alkaline ionized water (undiluted solution), but also diluted solutions obtained by diluting alkaline ionized water with purified water at a volume less than twice the volume of alkaline ionized water, have excellent virus inactivation performance.
<Measurement using the plaque measurement method>
Figure 1 is a photograph taken of the state of the test virus in a petri dish in Test 2. Figure 2 is a photograph taken of the state of the test virus in a petri dish in Test 3.

As shown in Figure 1, when the reaction stopping solution was not diluted with PBS (dilution series ¹⁰⁰ ), the number of plaques was too large to count, but when it was diluted 10,000 times (dilution series ¹⁰⁴ ), the number of plaques became countable. Thus, the proliferation of the virus was remarkable in PBS.

On the other hand, with iW-100 (alkaline ionized water) of this example, it was possible to count the number of plaques by the plaque measurement method, not only in the original solution but also in a 3-fold diluted solution without dilution with a reaction stop solution (100 ^- fold), as shown in Figure 2. This indicates that iW-100 of this example inactivates viruses.

As explained above, iW-100 of this example exhibits no cytotoxicity even in its undiluted form, meaning that no effects on the human body were confirmed. Even in a 3-fold diluted solution, nearly 99.99% of the virus was inactivated within 1 minute. This demonstrates that the virus inactivation solution of this example is capable of inactivating SARS-CoV-2 to a level where the antiviral activity value increases by 3 or more in the common logarithm mean of the virus infectivity titer within 1 minute. It was also found that diluting the undiluted iW-100 of this example to a volume less than twice the original volume produces a virus inactivation solution that is capable of inactivating the novel coronavirus and is gentle on human skin.

Therefore, by spraying the iW-100 of this embodiment onto human skin, the novel coronavirus can be inactivated while reducing the impact on human skin.
Example 2
<Composition of virus inactivation solution>
iW-100 (a stock solution consisting of alkaline ionized water) was produced using the same method as the stock solution described in Example 1, "Production Method of Stock Solution." The sodium (Na), phosphorus (P), potassium (K), silicon (Si), and chlorine (Cl) contents of the stock solution were measured. Measurements were outsourced to the Japan Food Analysis Center, a general incorporated foundation. The sodium and potassium contained in iW-100 were measured by atomic absorption spectrometry, phosphorus and silicon by ICP atomic emission spectrometry, and chlorine by combustion-coulometric titration. The weight ratio of phosphorus to silicon in the stock solution was also determined, as was the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon, and chlorine. While trace components may be present in the contents, the total weight of the contents was calculated as the sum of the weights of sodium, phosphorus, potassium, silicon, and chlorine. The number of iW-100 samples (n) was five. The measurement results are shown in Table 3.

In addition, the inventor measured the pH of each iW-100 and found that all were 12.0.

Three types of virus inactivation solutions, an undiluted solution, a 2-fold diluted solution, and a 3-fold diluted solution, were prepared from the five iW-100 solutions shown in Table 3. The average values of the components contained in these virus inactivation solutions and their total contents are shown in Table 4. All three types of virus inactivation solutions prepared (undiluted solution, 2-fold diluted solution, and 3-fold diluted solution) had a pH of 12.0.

a) From the above results, when the virus inactivation solution is taken as 100% by weight,
0.0001-1 wt % sodium, 0.00001-0.01 wt % silicon, 0.0001-0.1 wt % chlorine, 0-1 wt % potassium, and 0-0.1 wt % phosphorus,
0.0001-0.3 wt % sodium, 0.00001-0.001 wt % silicon, 0.0001-0.1 wt % chlorine, 0-0.2 wt % potassium, and 0-0.1 wt % phosphorus,
0.001-0.2% by weight sodium, 0.00005-0.005% by weight silicon, 0.001-0.05% by weight chlorine, 0.003-0.15% by weight potassium, and 0.002-0.1% by weight phosphorus, particularly
0.01-0.2 wt. % sodium, 0.0001-0.005 wt. % silicon, 0.005-0.05 wt. % chlorine, 0.01-0.15 wt. % potassium, and 0.01-0.1 wt. % phosphorus; or
b) When the total content of the virus inactivation solution is taken as 100% by weight,
It has been found that excellent virus inactivation can be achieved when the composition is 10-60% by weight sodium, 0.2-5% by weight silicon, 5-15% by weight chlorine, 0-50% by weight potassium, and 0-40% by weight phosphorus, or when the composition is 20-50% by weight sodium, 0.5-0.9% by weight silicon, 7-12% by weight chlorine, 20-40% by weight potassium, and 10-30% by weight phosphorus.

Furthermore, when the weight ratio of phosphorus to silicon in the raw solution is 10-50, or even 20-40, or/and
It has been found that even better virus inactivation can be achieved when the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon and chlorine is 1-3, or even 1.5-2.5.

Claims (8)

A virus inactivating agent capable of inactivating viruses,
The virus inactivating agent is
i) Alkaline ionized water containing purified water and at least sodium, silicon , chlorine , potassium, and phosphorus as components, containing more than 99.5% by weight of the purified water and having a pH of 10-13 , or
ii) a diluted solution obtained by diluting the alkaline ionized water with purified water in an amount of not more than twice the volume of the alkaline ionized water;
and
When the virus inactivating agent is taken as 100% by weight, the virus inactivating agent contains 0.01-0.2% by weight of sodium, 0.0001-0.005% by weight of silicon, 0.005-0.05% by weight of chlorine, 0.01-0.15% by weight of potassium, and 0.01-0.1% by weight of phosphorus;
The virus to be inactivated by the virus inactivating agent is SARS-CoV-2.
Virus inactivator. 2. The virus inactivating agent according to claim 1 , wherein the content of the purified water in the alkaline ionized water is 99.7% by weight or more. A virus inactivation agent capable of inactivating viruses, comprising alkaline ionized water having a pH of 10-13 ,
The alkaline ionized water contains at least sodium, silicon , chlorine , potassium, and phosphorus as its components,
When the virus inactivating agent is taken as 100% by weight, the virus inactivating agent contains 0.01-0.2% by weight of sodium, 0.0001-0.005% by weight of silicon, 0.005-0.05% by weight of chlorine, 0.01-0.15% by weight of potassium, and 0.01-0.1% by weight of phosphorus;
The virus to be inactivated by the virus inactivating agent is SARS-CoV-2.
Virus inactivator. The virus inactivating agent according to claim 3 , wherein the alkaline ionized water is alkaline ionized water. 5. The virus inactivating agent according to claim 3 , wherein a total amount of ingredients contained in the virus inactivating agent is 0.001% by weight or more and 2% by weight or less when the virus inactivating agent is taken as 100% by weight. 6. The virus inactivating agent according to claim 1 , wherein the weight ratio of phosphorus to silicon in the virus inactivating agent is 10-50. 7. The virus inactivating agent according to claim 1 , wherein the ratio of the total weight of sodium and potassium to the total weight of phosphorus, silicon and chlorine is 1-3. The virus inactivating agent according to any one of claims 1 to 7 , wherein the virus inactivating agent is a virus inactivating liquid.