WO2014115860A1

WO2014115860A1 - Virucidal agent

Info

Publication number: WO2014115860A1
Application number: PCT/JP2014/051590
Authority: WO
Inventors: 朋子田島; 吉平 ▲高▼畑; 亮門田
Original assignee: 公立大学法人大阪府立大学; 株式会社Tmc
Priority date: 2013-01-25
Filing date: 2014-01-25
Publication date: 2014-07-31
Also published as: JPWO2014115860A1

Abstract

[Problem] To provide an easily handled virucidal agent having a wide virucidal spectrum. [Solution] A virucidal agent having as an active ingredient a water-soluble metasilicate such as sodium metasilicate. Specifically, by a method such as dipping, wiping, or spraying, an aqueous solution of sodium metasilicate containing at least 0.01 mg/g, preferably 0.06 mg/g, is brought into contact with a subject such as a medical instrument, a breeding cage, or animal feces at normal temperature for at least one minute, preferably about 10 minutes.

Description

Viricide

The present invention relates to a virucidal agent.

Dogs and cats are typical companion animals. With the increase of companion animals, nosocomial infections in animal hospitals are becoming a problem. One of the causes of nosocomial infections is inadequate disinfection of bacteria and viruses attached to medical devices, breeding cages, and excreta. For sterilization of bacteria and virus inactivation, surfactants that are quaternary ammonium salts such as ethanol, povidone iodine, sodium hypochlorite, glutaraldehyde, benzalkonium chloride, and glycine surfactants are generally used. Various disinfectants are used.

However, quaternary ammonium salts have almost no effect on viruses, and there is no report that glycine surfactants have demonstrated virucidal action. Ethanol is effective against most bacteria and some viruses, but it causes deterioration of plastic and rubber products. Popidone iodine can be used for plastic products and rubber products, but its effectiveness is weak and it cannot be used for metal products because it is corrosive. Sodium hypochlorite is effective against most bacteria and viruses and inactivates viruses that are resistant to ethanol, such as parvoviruses and noroviruses, but is corrosive, so it is a metal product such as medical devices and cages. Can not be used. Glutaraldehyde also inactivates viruses resistant to ethanol, such as parvovirus and norovirus, but is difficult to handle because it has the property of denaturing proteins.

Thus, at present, there is no virucidal agent that is easy to handle and effective against parvoviruses and noroviruses that are not effective for ethanol targeting the envelope. There is a need for agents.

By the way, sodium metasilicate is also called water glass, and its highly concentrated solution has high viscosity. In addition, it is used for various purposes because it is highly safe for humans and has little environmental load or corrosivity. For example, it is used as an assistant for soap and detergent, an agglomeration aid for water purification, a peptizer for ceramics (dispersing agent), and an inorganic adhesive, and it contains a glycine surfactant. It is also used in cleaning compositions for appliances, dentures, and orthodontic appliances.

Further, a virucidal composition containing sodium metasilicate is disclosed in, for example, Patent Document 1 (Japanese Patent Publication No. 2005-514427), Patent Document 2 (Japanese Patent Publication No. 2005-514427), and Patent Document 3 (Japanese Patent Laid-Open No. 2012-2333058). No.) and Patent Document 4 (Japanese Patent Laid-Open No. 2010-057846).

However, these compositions contain a component having a virucidal activity as an active ingredient in addition to sodium metasilicate, and sodium metasilicate is only used as a so-called builder such as an alkaline agent in the composition.

Patent Document 5 (Japanese Patent Publication No. 2012-522011) and Patent Document 6 (Japanese Patent Publication No. 2012-522012) disclose pharmaceutical compositions for respiratory tract infections caused by bacteria such as influenza virus, rhinovirus, and pneumoniae. Has been. These pharmaceutical compositions include a sodium salt such as sodium metasilicate and a calcium salt such as calcium citrate, and the ratio of sodium ions to calcium ions is adjusted to a specific range. However, there is no suggestion that sodium metasilicate exhibits virucidal activity or bactericidal activity.

Non-Patent Document 1 describes that several types of compositions containing sodium metasilicate (including commercially available products) showed effectiveness against rotavirus. However, these compositions are compositions containing a quaternary ammonium salt, and NPL 1 does not suggest anything that sodium metasilicate exhibits a virucidal action. Non-Patent Document 2 describes that a 5% (w / w) sodium metasilicate solution is ineffective against vesicular stomatitis virus.

In addition, sodium metasilicate is approved by the FDA as a food disinfectant and is effective against Escherichia coli O157, Salmonella and Listeria in meat (Non-Patent Documents 3 to 5) However, so far, there has been no report yet that sodium metasilicate exhibits virucidal activity.

JP 2005-514427 A JP 2005-514427 A JP 2012-233058 A JP 2010-057846 A Special table 2012-522011 gazette Special table 2012-522012 gazette

In light of the above-mentioned background art, the inventors of the present invention have made extensive studies and found that sodium metasilicate, which is widely used as an alkaline agent, is not only a herpes virus or coronavirus having an envelope, but also a parvo having no envelope. It has been found that viruses and caliciviruses also have a virucidal action, and the present invention has been completed.

That is, an object of the present invention is to provide a virucidal agent having a broad virucidal spectrum and easy to handle.

The virucidal agent according to the present invention contains a metasilicate such as sodium metasilicate as an active ingredient.

The present invention provides a drug that exhibits a virucidal action not only on herpesviruses and coronaviruses having an envelope, but also on parvoviruses and caliciviruses that do not have an envelope, and is easy to handle.

The virucidal agent according to the present invention contains water-soluble metasilicate as an active ingredient. The metasilicate is, for example, a sodium salt and may be a potassium salt, but sodium metasilicate is preferred.

The object to which the virusicide according to the present invention is applied is not particularly limited as long as it is a place or object in which the presence of a virus is suspected. For example, it may be a dental instrument such as a dental reamer or a denture, a medical instrument such as a scalpel, tweezers or scissors, and may be a cage for various animals including pets. Also, it is the floors and walls of various facilities such as hospitals, general households, companies, etc., door knobs, window handles, sofas, beds, desks, switches such as lighting switches, It can be a toy. In addition, feces and vomits of animals including humans can be objects. Furthermore, perishable foods such as beef, chicken, and vegetables, processed foods, and animals including humans themselves can be objects. Since metasilicate is not corrosive, it can be applied to objects in the anti-range including metal products. The virucidal agent according to the present invention acts on viruses present in such objects. Moreover, since the sodium metasilicate should just contact a virus, not only when a virus exists on the surface of a target object, but also when it exists in a target object like feces and a vomit, a virucidal effect is exhibited.

The virusicide according to the present invention is mainly used as an aqueous solution of metasilicate. The concentration of the aqueous solution of metasilicate varies depending on the object to be used and the contact time, but the minimum concentration of sodium metasilicate is, for example, 0.01 mg / g, 0.05 mg / g, 0 It can be 1 mg / g. The preferred concentration is 0.06 mg / g or more, desirably 0.30 mg / g or more, more desirably 0.6 mg / g or more. Moreover, the upper limit concentration is not particularly limited, but for example, sodium metasilicate is 200 mg / g, 100 mg / g, 50 mg / g, or 10 mg / g.

The virucidal agent according to the present invention is effective by bringing an aqueous solution of sodium metasilicate into contact with the object. The method of contacting is not particularly limited. For example, an object is immersed in an aqueous solution of metasilicate, an aqueous solution of metasilicate is sprayed on the object, an aqueous solution of metasilicate is applied to the object, an aqueous solution of metasilicate The method may include wiping with a wiper that contains metasilicate in the object. Of course, sodium metasilicate may be sprinkled on the object.

Although the contact time varies depending on the concentration of metasilicate and the contact temperature, it is preferably at least 1 minute at room temperature, preferably 5 minutes or more, and more preferably 10 minutes or more at a preferred concentration of 1.2 mg / g solution. Yes, for a 0.6 mg / g solution, it is preferably 5 minutes or longer. The contact temperature may be room temperature or a room temperature of about 5 to 35 ° C., and no special heating or cooling is required.

The virucidal composition according to the present invention comprises a metasilicate and a base necessary for contacting with an object. Such a base is, for example, water for making an aqueous solution, a support for making a wiper, and an excipient for making tablets or granules. The support is, for example, a woven fabric and may be a non-woven fabric. A composition formulated with an excipient is dissolved in water to contain a predetermined concentration of metasilicate at the time of use.

The metasilicate alone exhibits a virucidal action, and the virucidal composition according to the present invention does not require other virucidal or bactericidal agents. In the present invention, the virucidal action means that the virus cannot be propagated. Moreover, a virucidal agent means the compound which has such a virucidal action. However, the composition according to the present invention may contain other virucidal agents and other bactericides. However, since the virucidal composition according to the present invention is used not only in medical instruments and medical facilities but also in homes, it is preferable to use other virucidal agents and sterilizing agents, particularly in handling. Contains no difficult glutaraldehyde, corrosive iodine and hypochlorite. Moreover, since the composition which concerns on this invention may produce precipitation of metasilicate when it contains bactericidal alcohols, such as ethanol and isopropyl alcohol, Preferably it does not contain the alcohol concerned. On the other hand, metasilicate has bactericidal effect on bacteria such as Escherichia coli and Salmonella. Therefore, when other fungicides are used in combination, it is preferable to use an alkyl polyaminoethyl glycine salt and / or an alkyl diaminoethyl glycine salt having a moderate effect.

The composition for virucidal use according to the present invention contains only the necessary base for bringing the metasilicate into contact with the object and the above-mentioned bactericidal agent added as necessary. It may contain optional ingredients that do not affect the virucidal action, such as agents, fragrances, and chelating agents. In addition, since the aqueous solution of sodium metasilicate exhibits alkalinity, precipitation occurs in an aqueous solution around pH 7, and the virucidal action is lost, so there is no need for neutralization when used as an aqueous solution. Therefore, the composition according to the present invention does not contain acids such as hydrochloric acid, phosphoric acid and sulfuric acid necessary for neutralization.

The virucidal agent or virucidal composition according to the present invention acts on various viruses with or without an envelope. The virus of interest can be, for example, a herpes virus, a coronavirus, a parvovirus, a calicivirus, a feline infectious peritonitis virus, or an influenza virus.

Next, the present invention will be further described based on the following examples, but it goes without saying that the present invention is not limited to the following examples.

[Virucidal action of sodium metasilicate]
Feline parvovirus PLI-IV strain, feline calicivirus F9 strain, feline herpesvirus 80-1 strain, feline infectious peritonitis virus 79-1146 strain, Aujeszky's disease virus Yamagata S81 strain were used. Feline parvovirus, feline calicivirus, feline herpes virus and Aujeszky's disease virus were propagated in feline kidney-derived cell line CRFK, and feline infectious peritonitis virus was propagated in fetal fetal cell line fcwf-4.

To 0.5 ml of the virus solution, 0.5 ml of sodium metasilicate having a concentration of 1.2 mg / g was added and reacted at room temperature for 5 minutes. As a control, a virus solution containing sterilized distilled water instead of sodium metasilicate was reacted in the same manner. Dilute the treated specimen 10-fold stepwise, inoculate the cells, and for feline calicivirus, feline herpesvirus, feline infectious peritonitis virus, after culturing for 2 days, determine the presence or absence of viral infection by the cytopathic effect of the virus 50% tissue culture infection concentration (TCID ₅₀ / ml) was determined. For feline parvovirus, after culturing for 4 days, the cells were detached with trypsin and washed with PBS. After re-floating in PBS, it was smeared on a 12-hole highly water-repellent printing slide glass and fixed with acetone. Thereafter, the presence or absence of infection was determined by the indirect fluorescent antibody method using feline serum vaccinated with FPV as the primary serum and FITC-labeled anti-cat IgG goat serum as the secondary serum, and TCID ₅₀ / ml was determined. The results are shown in Table 1.

As shown in Table 1, feline parvovirus, feline calicivirus, feline herpes virus, Aujeszky's disease virus is reacted with an equivalent amount of 1.2 mg / g sodium metasilicate aqueous solution at room temperature for 5 minutes, Growth was completely prevented. The feline infectious peritonitis virus was less effective than the above four types of viruses, but the infection concentration was reduced to 1 / 10,000.

[Changes in virucidal action by concentration of sodium metasilicate aqueous solution and reaction time]
Feline calicivirus and feline herpesvirus were used to examine the virucidal action by the concentration of sodium metasilicate. Prepare sodium metasilicate in aqueous solution of 1.2mg / g, 0.6mg / g, 0.12mg / g, add 0.5ml of sodium metasilicate aqueous solution to 0.5ml of virus solution and at room temperature for 1 minute or Reacted for 5 minutes. CRFK cells were inoculated after 10-fold serial dilution. After culturing for 2 days, the presence or absence of viral infection was determined by cytopathic effect, and TCID ₅₀ / ml was determined. The results are shown in Table 2.

When feline calicivirus was treated with 0.6 mg / g sodium metasilicate aqueous solution for 1 minute, the infection concentration decreased to 1 / 10,000 or less, and the growth was completely prevented by treatment for 5 minutes. In addition, no effect was observed at 0.12 mg / g. On the other hand, feline herpesvirus was not effective at both 0.6 mg / g and 0.12 mg / g. In addition, cytotoxicity to CRFK cells was observed in an aqueous solution having a concentration of 15 mg / g, and the action on these viruses could not be observed. However, the antiviral action of the aqueous sodium metasilicate solution was not lost even at the concentration.

[Influence of protein on virucidal action]
In order to examine the effect of protein on the virucidal action of sodium metasilicate, fetal calicivirus solution with fetal calf serum added at 1v / v%, 5v / v%, 10v / v%, 20v / v% concentration 0 0.5 ml and 0.5 ml of a 1.2 mg / g sodium metasilicate aqueous solution were reacted at room temperature for 5 minutes. As a control, instead of sodium metasilicate aqueous solution, Pulux (trade name: containing 6 w / v% sodium hypochlorite), which is widely used as a disinfectant, was diluted to contain 1% sodium hypochlorite. The added solution was reacted in the same manner. After 10-fold serial dilution, CRFK cells were inoculated and cultured for 2 days, and then the presence or absence of virus infection was determined by cytopathic effect, and TCID ₅₀ / ml was determined. The results are shown in Table 3.

The feline calicivirus was completely inhibited from growing with a 1.2 mg / g sodium metasilicate aqueous solution even in a state in which fetal calf serum was mixed up to 20%. On the other hand, Pulux diluted to contain 1% sodium hypochlorite inhibited the virucidal action only by adding fetal calf serum to a concentration of 1%.

From the above results, a virucidal action was observed against feline parvovirus, feline calicivirus and feline herpesvirus by contact with about 0.3 mg / g sodium metasilicate aqueous solution for about 1 minute or longer. Moreover, although it was weak against the feline infectious peritonitis virus compared with these viruses, the virucidal action was recognized. And it was shown that the virucidal action of the sodium metasilicate aqueous solution is hardly affected by the protein.

[Virucidal action of a virucidal agent containing a bactericidal agent]
Next, the virucidal effect of a virucidal agent having the following composition (containing 9.0 mg / g as sodium metasilicate) was examined. Viralicide and virus solution were mixed at a volume ratio shown in Table 4 and contacted at room temperature for 1 minute or 5 minutes. As a control, distilled water and virus solution were mixed at a volume ratio of 50% and contacted in the same manner. In addition, a diluted solution of Pulux diluted to contain 1% sodium hypochlorite was also brought into contact in the same manner, and the virus titer was determined in the same manner as in Example 1. The results are shown in Tables 4 and 5. As shown in Table 4, the approximately 2-fold diluted solution (4.5 mg / g sodium metasilicate aqueous solution) of Example 2 showed a virucidal action upon contact for 1 minute. When the bactericidal agent was used in combination, the virucidal effect of sodium metasilicate tended to decrease, but even when the bactericidal agent was used in combination, the virucidal effect of sodium metasilicate was not lost.

(Viricide with fungicide)
0.92 g of anhydrous sodium metasilicate
Polyoctyl polyaminoethylglycine 0.06g
Polyoxyethylene alkyl ether 0.04g
100.90 g of water

[Effects on viruses in feces]
Purax solution diluted to contain 0.9 ml of 6.0 mg / g sodium metasilicate aqueous solution or 1% sodium hypochlorite after adding feline parvovirus 10 ⁶ TCID ₅₀ to 0.1 g of dog feces and mixing. 0.9 ml was added and reacted at room temperature for 5 minutes. As a control, sterilized distilled water was reacted in the same manner. After 10-fold serial dilution, CRFK cells were inoculated, cultured for 4 days, the presence or absence of viral infection was determined by the indirect fluorescent antibody method, and TCID ₅₀ / ml was determined. The results are shown in Table 6.

The feline parvovirus added to the stool was completely prevented from growing by contact with a 6.0 mg / g sodium metasilicate aqueous solution for 5 minutes. A similar inhibition was seen with 1% Pulux.

[Effects against viruses attached to cloth]
It was investigated whether sodium metasilicate exerted a virucidal action on the virus attached to the cloth. A 2 cm × 2 cm sterilized towel was soaked in 1 ml of the feline calicivirus virus solution, so that the virus solution was firmly immersed. Then, it was immersed in 5 ml of 1.2 mg / g sodium metasilicate aqueous solution or culture solution for 30 minutes. After 10-fold serial dilution, CRFK cells were inoculated, cultured for 2 days, and the presence or absence of virus infection was determined by cytopathic effect, and TCID ₅₀ / ml was determined. As a control, only the virus solution was prepared by reacting 1 ml of the virus solution with 5 ml of a 1.2 mg / g sodium metasilicate aqueous solution. The results are shown in Table 7. The virus adhering to the towel was completely prevented from growing by being immersed in an aqueous sodium metasilicate solution for 30 minutes.

[Virucidal effect against influenza virus]
Influenza A virus A / panama / 2007/99 (H3N2) used as a vaccine was used. The reaction was carried out with an equivalent amount of 1.2 mg / g sodium metasilicate aqueous solution for 5 minutes at room temperature. After 10-fold serial dilution, MDCK cells were inoculated, cultured for 4 days, and the presence or absence of virus infection was determined by cytopathic effect, and TCID ₅₀ / ml was determined. As a control, a virus solution and an equivalent amount of sterile distilled water were reacted. The results are shown in Table 8. The influenza virus was also prevented from growing.

The feline parvovirus used above is a virus belonging to the genus Parvoviridae, and does not have an envelope. Feline calicivirus is a virus belonging to the genus Besivirus of the Caliciviridae family and does not have an envelope. Feline herpesvirus (feline viral rhinotracheitis virus) and Aujeszky's disease virus are viruses belonging to the genus Baricellovirus of the herpesviridae family and have an envelope. The feline infectious peritonitis virus is a virus belonging to the genus Alphacoronavirus of the Coronaviridae family and has an envelope.

Since viruses belonging to the same family and the same genus have similar physicochemical properties, they are considered to have the same susceptibility to virucidal agents. Therefore, canine parvoviruses belonging to the genus Parvoviridae, porcine parvoviruses, B19 viruses belonging to the genus Parvoviridae erythrovirus (infecting humans), noroviruses belonging to the genus Besiviridae (infecting humans), herpes It can be concluded that sodium metasilicate is also effective for varicella / zoster virus (infecting humans) belonging to the genus Baricellovirus genus, and for porcine infectious gastroenteritis virus belonging to the coronavirus family Alpha coronavirus genus. However, it is considered that sodium metasilicate has a virucidal action on various viruses such as influenza virus regardless of the presence or absence of an envelope.

According to the present invention, a virucidal agent having a broad virucidal spectrum and easy to handle is provided.

Claims

Viricide containing water-soluble metasilicate as an active ingredient.
A composition for virucidal activity that exhibits a virucidal action by a method of coating, spraying, dipping, wiping, and impregnation, and comprising metasilicate as an active ingredient.
The composition for virucidal use according to claim 2, which does not contain a compound having a virucidal action other than metasilicate and a compound having a bactericidal action as an active ingredient.
The composition for virus killing according to claim 2 or 3, comprising an alkyl polyaminoethylglycine salt and / or an alkyldiaminoethylglycine salt.
A method of killing a virus present in an object by bringing the metasilicate into contact with the object. However, the object does not include humans.
The contacting method includes immersing an object in an aqueous solution of metasilicate, spraying an aqueous solution of metasilicate onto the object, applying an aqueous solution of metasilicate to the object, and an aqueous solution of metasilicate. 6. The method according to claim 5, wherein the surface of the object is wiped with a wiper or metasilicate is included in the object. However, the object does not include humans.
The method according to claim 5 or 6, wherein the aqueous solution of metasilicate contains 0.01 mg / g or more of metasilicate as sodium metasilicate.
The method according to any one of claims 5 to 7, wherein the aqueous solution of metasilicate contains an alkyl polyaminoethyl glycine salt and / or an alkyl diaminoethyl glycine salt.