JP2022130161A - Anti-coronavirus agent - Google Patents

Abstract

To provide an anti-coronavirus agent: particularly an anti-coronavirus agent against SARS-CoV-2.SOLUTION: An anti-coronavirus agent contains at least one selected from the group consisting of alkyl gallate, gallic acid, and derivatives thereof as an active ingredient.SELECTED DRAWING: None

Description

The present invention relates to anti-coronavirus agents and the like.

Coronaviruses, a species of virus belonging to the subfamily Coronavirinae of the family Coronavirus, are positive-stranded RNA viruses that infect humans and animals and cause respiratory, gastrointestinal, vascular, or neurological disease. Coronaviruses can spread from any infected host animal and cause large-scale epidemics in humans. Coronaviruses include, in recent years, for example, SARS coronavirus 1 (SARS-CoV-1), which prevalent in 2002 and 2003, and Middle East respiratory syndrome coronavirus (MERS-CoV). Recently, the SARS coronavirus 2 (SARS-CoV-2) has become a global epidemic, and since no treatment technology has been established, it continues to cause enormous damage in many fields, including medical care and economics.

An object of the present invention is to provide an anti-coronavirus agent, particularly an anti-coronavirus agent against SARS-CoV-2.

As a result of intensive research in view of the above problems, the present inventors have found that an anti-coronavirus agent containing at least one selected from the group consisting of alkyl gallate and derivatives thereof as an active ingredient solves the above problems. I found a solution. The present inventor has completed the present invention as a result of further research based on this finding. That is, the present invention includes the following aspects.

Section 1. An anti-coronavirus agent containing, as an active ingredient, at least one selected from the group consisting of alkyl gallate, gallic acid, and derivatives thereof.

Section 2. Item 2. The anti-coronavirus agent according to Item 1, wherein the coronavirus is SARS-CoV-2.

Item 3. Item 3. The anti-coronavirus agent according to Item 1 or 2, comprising the alkyl gallate as an active ingredient.

Section 4. The alkyl gallate and/or gallic acid has the general formula (1):

[In the formula, R ¹ represents a hydrogen atom or an alkyl group. ]
Item 4. The anti-coronavirus agent according to any one of items 1 to 3, which is a compound represented by

Item 5. Item 5. The anti-coronavirus agent according to Item 4, wherein said R ¹ is a hydrogen atom or an alkyl group having 1 to 7 carbon atoms.

Item 6. Item 6. The anti-coronavirus agent according to Item 4 or 5, wherein said R ¹ is an alkyl group having 1 to 4 carbon atoms.

Item 7. Item 7. The anti-coronavirus agent according to any one of Items 1 to 6, which is used for application to a living body.

Item 8. Item 8. The anti-coronavirus agent according to Item 7, which is a pharmaceutical, cosmetic, food composition, food additive, disinfectant or cleaning agent.

Item 9. Item 9. The anti-coronavirus agent according to Item 7 or 8, which is for prevention or amelioration of COVID-19.

Item 10. Item 7. The anti-coronavirus agent according to any one of Items 1 to 6, which is used for application to an article.

Item 11. Item 11. The anti-coronavirus agent according to Item 10, which is a disinfectant or detergent.

According to the present invention, it is possible to provide an anti-coronavirus agent, particularly an anti-coronavirus agent against SARS-CoV-2.

The outline of the test method (Mix protocol) of Test Example 1 is shown. The results of Test Example 1 are shown. The vertical axis indicates the cell death inhibition rate. The test substance is shown above each graph. The horizontal axis indicates the concentration of the test substance. The outline of the test method (Post-infection protocol) of Test Example 2 is shown. The results of Test Example 2 are shown. The vertical axis indicates the cell death inhibition rate. The test substance is shown above each graph. The horizontal axis indicates the concentration of the test substance.

As used herein, the expressions "contain" and "include" include the concepts "contain", "include", "consist essentially of" and "consist only of".

In one aspect of the present invention, an anti-coronavirus agent containing as an active ingredient at least one selected from the group consisting of alkyl gallate, gallic acid, and derivatives thereof (herein referred to as "the It may be indicated as "agent".) This will be explained below.

1. Active Ingredient The active ingredient is at least one selected from the group consisting of alkyl gallate, gallic acid, and derivatives thereof.

Alkyl gallate is an alkyl ester of gallic acid, and is not particularly limited in this respect. Examples of alkyl gallate include methyl gallate, ethyl gallate, propyl gallate, butyl gallate, pentyl gallate, hexyl gallate, heptyl gallate, isopropyl gallate, isobutyl gallate, sec-butyl gallate, tert-butyl gallate, isoamyl gallate and the like. Among these, methyl gallate, ethyl gallate, propyl gallate and butyl gallate are preferred, and methyl gallate, ethyl gallate and propyl gallate are more preferred. It is recognized as a food additive, and propyl gallate is particularly preferred from the viewpoint of safety.

Alkyl gallate, gallic acid, typically has the general formula (1):

[In the formula, R ¹ represents a hydrogen atom or an alkyl group. ]
It is a compound represented by

The alkyl group represented by R ¹ includes any of straight-chain, branched-chain and cyclic ones. From the viewpoint of antiviral action and the like, the alkyl group is preferably a chain alkyl group, and particularly preferably a linear alkyl group. However, it may also be a cyclic alkyl group. The number of carbon atoms in the alkyl group (in the case of linear or branched chain) is not particularly limited, and is, for example, 1-12. In one embodiment of the present invention, the number of carbon atoms is preferably 1-9, more preferably 1-7, even more preferably 1-5, still more preferably 1-4, and most preferably 1-3. The number of carbon atoms in the alkyl group (in the case of cyclic) is not particularly limited, and is, for example, 3-7, preferably 4-6. Specific examples of the alkyl group include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, sec-butyl group, n-pentyl group, neopentyl group and isopentyl. group, n-hexyl group, 3-methylpentyl group, n-heptyl group, n-octyl group and the like.

R ¹ is preferably an alkyl group.

Alkyl gallate and derivatives of gallic acid are not particularly limited as long as they can exhibit antiviral effects as they are or through reactions such as decomposition.

Derivatives of alkyl gallate and gallic acid include, for example, in general formula (1), one to three hydroxyl groups are —OR (R is an optionally substituted hydrocarbon group or an optionally substituted heterocyclic ring ) can be mentioned.

Examples of derivatives of alkyl gallate and gallic acid include glycosides of alkyl gallate and gallic acid. The glycoside of alkyl gallate or gallic acid is not particularly limited as long as it is a compound in which a sugar is linked to an alkyl gallate or gallic acid as an aglycone via a bond (usually a glycosidic bond). Sugars include, for example, glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, gentiobiose, digitoxose, galactose, xylose, and sugars formed by linking a plurality of these. The number of monosaccharide residues constituting sugar is, for example, 1-8, 1-6.

Derivatives of alkyl gallates and gallic acid also include, for example, prodrugs of alkyl gallates or gallic acid. A prodrug refers to a compound that is converted into the compound of the present invention by a reaction with an enzyme, gastric acid, or the like in vivo. Prodrugs of alkyl gallate or gallic acid include, for example, compounds in which the hydroxyl group is acylated, alkylated, phosphorylated, or borated (for example, the hydroxyl group of alkyl gallate is acetylated, palmitoylated, propanoylated, pivaloylated, succinylated, compound, fumarylated, alanylated, dialkyl (e.g. methyl) aminoalkyl (e.g. methyl) carbonylated compounds, etc.); (e.g. ethyl) esterification, aryl (e.g. phenyl) esterification, carboxyalkyl (e.g. methyl) esterification, dialkyl (e.g. methyl) aminoalkyl (e.g. methyl) esterification, pivaloyloxyalkyl (e.g. methyl) esterification, 1-{(alkoxy (e.g. ethoxy)carbonyl)oxy}alkyl (e.g. ethyl) esterification, phthalidyl esterification, (5-alkyl (e.g. methyl)-2-oxo-1,3-dioxolen-4-yl)alkyl (eg, methyl)esterified, 1-{[(cycloalkyl (eg, cyclohexyl)oxy)carbonyl]oxy}alkyl (eg, ethyl)esterified, alkyl (eg, methyl)amidated compounds), and the like. is mentioned. These compounds can be produced by methods known per se. In addition, the prodrug of the compound of the present invention changes to the compound of the present invention under physiological conditions as described in Hirokawa Shoten 1990, "Drug Development", Vol. 7, "Molecular Design", pp. 163-198. can be anything.

Among active ingredients, alkyl gallate is preferred.

The active ingredient may be used singly or in combination of two or more.

As the alkyl gallate and its derivatives, commercially available ones can be used, and those synthesized according to or according to known methods can also be used.

2. Use The target coronavirus of the agent of the present invention is not particularly limited as long as it belongs to the subfamily Orthocoronavirus. Examples of coronaviruses include alphacoronavirus, betacoronavirus, gammacoronavirus, deltacoronavirus, etc. Among these, betacoronavirus is preferred. The betacoronavirus genus includes SARS-associated coronavirus (SARSr-CoV), coronavirus HKU1, MERS coronavirus, etc. Among these, SARSr-CoV is preferred. SARSr-CoV includes SARS-CoV-2, SARS-CoV-1, etc. Among these, SARS-CoV-2 is preferred. The agent of the present invention can be preferably used especially against SARS-CoV-2.

The agent of the present invention can exert an antiviral effect against coronavirus. Specifically, the antiviral action includes viral infection inhibitory action, viral cell infection titer-lowering action, viral cell death inhibitory action, viral inactivation action, viral multiplication inhibitory action, and viral budding inhibitory action. , viral resistance-inducing action, and the like. In addition, due to this action, it can also be used as a prophylactic or ameliorating agent (or prophylactic or therapeutic agent) for coronavirus infections (in particular, COVID-19).

The agent of the present invention can be widely used in various fields requiring antiviral properties. The agent of the present invention can be used in various fields such as industrial, cleaning, medical, food, and daily necessities. Applications of the agent of the present invention are mainly divided into applications applied to living bodies and applications applied to articles described later.

In one aspect of the present invention, the agent of the present invention may contain less alkyl gallate having 8 or more or 5 or more carbon atoms in the alkyl group. In the agent of the present invention, the content of alkyl gallate having 8 or more carbon atoms in the alkyl group or 5 or more carbon atoms in the alkyl group is 100% by mass (or mol %), for example, 0 to 50% by mass (or mol%), 0 to 20% by mass (or mol%), 0 to 10% by mass (or mol%), 0 to 5% by mass (or mol%) , 0 to 2% by mass (or mol%), 0 to 1% by mass (or mol%), 0 to 0.1% by mass (or mol%), 0 to 0.01% by mass (or mol%), or 0% by mass ( or mol %).

In one aspect of the present invention, the agent of the present invention can contain less antiviral components other than alkyl gallate. In the agent of the present invention, the content of antiviral components other than alkyl gallate is 100% by mass (or mol%) of the content of alkyl gallate (for example, alkyl gallate having 7 or less or 4 or less carbon atoms in the alkyl group). For example, 0 to 50% by mass (or mol%), 0 to 20% by mass (or mol%), 0 to 10% by mass (or mol%), 0 to 5% by mass (or mol%), 0 to 2% by mass (or mol%), 0 to 1% by mass (or mol%), 0 to 0.1% by mass (or mol%), 0 to 0.01% by mass (or mol%), or 0% by mass (or mol%) ).

2-1. Uses applied to living bodies Examples of uses applied to living bodies include pharmaceuticals, cosmetics, food compositions (including health foods, health promoting agents, dietary supplements (supplements, etc.)), food additives, disinfectants, and detergents. etc. The subject of application in this case is not particularly limited, and examples thereof include various mammals such as humans, monkeys, mice, rats, dogs, cats, rabbits, pigs, horses, cows, sheep, goats, and deer.

By applying the agent of the present invention to a living body, an antiviral effect can be exerted at the site where the active ingredient comes into contact.

The form of the agent of the present invention is not particularly limited, and depending on the use of the agent of the present invention, it can take a form commonly used in each application.

As for the form, when the application is a medicine, for example, injection, drip, gargle, inhalant, patch (plasters, tapes such as plasters (reservoir type, matrix type, etc.), poultices, patches microneedles, etc.), ointments, external liquids (liniments, lotions, etc.), sprays (external aerosols, pump sprays, etc.), creams, gels, eye drops, ophthalmic ointments, nasal drops, Formulations suitable for parenteral intake, such as suppositories, rectal semisolids, and enema tablets; ), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including drinks, suspensions, syrups), jellies, etc. suitable for oral intake Formulations (oral formulations), tube feeding agents, enteral feeding agents, nasal catheter agents, esophageal catheter agents, gastrostomy catheter agents, and the like can be mentioned.

Examples of forms include liquids, gels, creams, ointments, sprays, sticks and the like when the application is cosmetics.

As for the form, when the application is a food composition, liquid, gel or solid foods such as juice, soft drinks, tea, soup, beverages such as soy milk, salad oil, dressing, yogurt, jelly, pudding, sprinkle, Powdered milk for infants, cake mixes, dairy products (eg, powder, liquid, gel, solid, etc.), bread, sweets (eg, cookies, etc.), and the like. In addition, when the application is a food additive, health promoting agent, dietary supplement (supplement, etc.), for example, tablets (orally disintegrating tablets, chewable tablets, effervescent tablets, lozenges, jelly drops, etc.) ), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including suspensions and syrups), and jellies.

When the application is a disinfectant or cleaning agent, for example, liquid (solution, milky lotion, suspension, spray, etc.), semi-solid (gel, cream, paste, etc.), solid (tablet, particulate, capsules, films, kneaded products, molten solids, waxy solids, elastic solids, etc.). For example, when applied to the oral cavity, more specifically, dentifrice (toothpaste, liquid dentifrice, liquid dentifrice, toothpaste, etc.), mouthwash, coating agent, patch, mouth freshener, food ( Examples include chewing gum, tablet candy, candy, gummies, films, troches, etc.). In the case of application to the nasal cavity, more specific examples include nasal sprays and the like. For skin application, soaps, body soaps, shampoos, rinses, spray agents and the like can be mentioned.

The agent of the present invention may further contain other ingredients as necessary. Other ingredients are not particularly limited as long as they are ingredients that can be blended in pharmaceuticals, cosmetics, disinfectants, detergents, etc. Examples include bases, carriers, solvents, dispersants, emulsifiers, buffers, Stabilizers, excipients, binders, disintegrants, lubricants, thickeners, humectants, colorants, perfumes, chelating agents and the like.

The buffering agent is not particularly limited, and any acceptable and appropriate one can be adopted depending on the application. Phosphate buffer, acetate buffer and the like are preferred.

The content of the active ingredient in the agent of the present invention depends on the type of active ingredient, application, mode of use, application target, condition of application target, etc., and is not limited. Preferably, it can be 0.01 to 50% by weight.

The amount of application (e.g., administration, ingestion, inoculation, etc.) of the agent of the present invention is not particularly limited as long as it is an effective amount that exhibits the desired effect, and is generally 0.1 to 0.1 per day as the weight of the active ingredient. 1000 mg/kg body weight. The above dosage is preferably administered once a day or in 2 to 3 divided doses, and can be adjusted appropriately according to age, condition and symptoms.

2-2. Uses applied to articles Examples of uses applied to articles include disinfectants and detergents. Applications in this case are not particularly limited, and include industrial products and their raw materials used in various fields.

Specific examples of articles include masks, face masks, gloves, and the like. In addition, endotracheal intubation tubes, bite blocks, laryngoscopes, electrodes, measuring devices, drip devices, oxygen masks, nasal catheters, tubes, catheters, white coats, gowns, caps, protective clothing, protective shields, surgical instruments and other medical instruments, surgery Tables, beds, stretchers, wheelchairs, etc. can be mentioned. In addition, OA equipment, home appliances, air conditioners, vacuum cleaners, desks, chairs, sofas, benches, windows, straps, handles, seats, automatic ticket gates, automatic ticket vending machines, vending machines, doors, fences, handrails, tableware, Cooking utensils, packaging films, packaging bags, bottles, bottles, packaging packs, sinks, toilet bowls, stationery, books, shelves, toothbrushes, mirrors, air conditioning filters, masks, coats, jackets, pants, skirts, dress shirts, knit shirts, blouses, Sweaters, cardigans, nightwear, underwear, underwear, diapers, supporters, socks, tights, stockings, hats, scarves, mufflers, scarves, stoles, gloves, linings, interlinings, batting, work clothes, Clothing such as uniforms and school uniforms, curtains, nets, futon fabrics, futon cotton, futon covers, pillowcases, sheets, mats, carpets, towels, handkerchiefs, wall cloths, band-aids, bandages, and the like.

By applying the agent of the present invention to an article, an antiviral effect can be exerted at the site where the active ingredient comes into contact. The site with which the active ingredient comes into contact is not limited to the site on the article, and includes sites in the living body when the article is applied to the living body.

The dosage form of the agent of the present invention is not particularly limited, and can be appropriately selected according to its use. Examples of dosage forms include liquids such as liquids, emulsions, suspensions, dispersants, sprays, and aerosols; be done. Various articles can be painted or coated.

The agent of the present invention may further contain other ingredients as necessary. Other ingredients are not particularly limited as long as they are ingredients that can be blended in detergents, disinfectants, etc. for articles, for example, bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers. , excipients, binders, disintegrants, lubricants, thickeners, humectants, coloring agents, fragrances, chelating agents and the like.

The buffering agent is not particularly limited, and any acceptable and appropriate one can be adopted depending on the application. Phosphate buffer, acetate buffer and the like are preferred.

The content of the active ingredient in the agent of the present invention depends on the type of active ingredient, application, mode of use, application target, condition of application target, etc., and is not limited. Preferably, it can be 0.001 to 50% by weight.

EXAMPLES The present invention will be described in detail below based on examples, but the present invention is not limited by these examples.

Test example 1. Antiviral activity evaluation test (Mix protocol)
The outline of this test is shown in FIG. Methyl gallate (General formula (1): R ¹ = methyl group), Propyl gallate (General formula (1): R ¹ = n-propyl group), or Butyl gallate (General formula (1): R ¹ = n-butyl base) was dissolved in filtered sterilized water to 100 mM to obtain a sample solution.

The sample solution was diluted in two stages with filtered sterilized water. 200 μl of SF DMEM (MS) or 200 μl of SARS-CoV-2 (5 X 10^5 TCID50/200 μl) was added to 2 μl of each concentration sample solution and allowed to stand at room temperature for 1 minute. VeroE6/TMPRSS2 cells were seeded at 5×10^4/100 μl/well the day before and cultured for 24 hours on a 96-well plate. After adsorption for 1 hour, the virus solution was removed, MS was added at 100 μl/well, and cultured for 24 hours. Cell viability was quantified by the following method.

Media was removed from the plate and washed with PBS. 20% Cell Count Reagent SF (Nacalai) was mixed with phenol red-free medium, 50 μl/well was added to the cells, and cultured for 60 minutes. As a blank, wells were also prepared in which only the reagent mixture was added to wells in which no cells were cultured. After completion of the reaction, 20 µl/well of 10% SDS was added to inactivate viruses remaining in the wells. Absorbance at a wavelength of 450 nm was measured using a plate reader. Measurements were made in tetraplicates. %Inhibition of cell death to the following formula: %Inhibition of cell death = (OD((Virus + Alkyl gallate)-OD(Virus only))/(OD(Alkyl gallate only)-OD(Virus only)) x 100) calculated based on

<Results>
The results are shown in FIG. Addition of Methyl gallate, Propyl gallate, and Butyl gallate to SARS-CoV-2 all dose-dependently inhibited viral infection-induced cell death.

Test example 2. Antiviral activity evaluation test (Post-infection protocol)
The outline of this test is shown in FIG. Methyl gallate (general formula (1): R ¹ = methyl group) or propyl gallate (general formula (1): R ¹ = n-propyl group) was dissolved in filtered sterilized water to 250 mM. A sample liquid was obtained by treating for 5 minutes in a constant temperature bath at 78°C.

The sample solution was diluted in two stages with 0.5% FBS DMEM (MS). VeroE6/TMPRSS2 cells were seeded in a 96-well plate at 5 X 10^4/100 μl/well and cultured for 15 hours. After removing the supernatant from the cells, SARS-CoV-2 was added at MOI 5 and allowed to adsorb for 1 hour. 100 μl/well of sample solution or MS of each concentration was added except the virus solution, and cultured for 24 hours. Quantification of cell viability and calculation of %Inhibition of cell death were performed in the same manner as in Test Example 1 above.

<Results>
The results are shown in FIG. Both methyl gallate and propyl gallate inhibited cell death due to virus infection in a dose-dependent manner when added to SARS-CoV-2-adsorbed cells after washing.

Claims (11)

An anti-coronavirus agent containing, as an active ingredient, at least one selected from the group consisting of alkyl gallate, gallic acid, and derivatives thereof. 2. The anti-coronaviral agent of claim 1, wherein said coronavirus is SARS-CoV-2. 3. The anti-coronavirus agent according to claim 1, comprising said alkyl gallate as an active ingredient. The alkyl gallate and/or gallic acid has the general formula (1):

[In the formula, R ¹ represents a hydrogen atom or an alkyl group. ]
The anti-coronavirus agent according to any one of claims 1 to 3, which is a compound represented by 5. The anti-coronavirus agent according to claim 4, wherein said R ¹ is a hydrogen atom or an alkyl group having 1 to 7 carbon atoms. 6. The anti-coronavirus agent according to claim 4 or 5, wherein said R ¹ is an alkyl group having 1 to 4 carbon atoms. 7. The anti-coronavirus agent according to any one of claims 1 to 6, which is used for application to living organisms. 8. The anti-coronavirus agent according to claim 7, which is a pharmaceutical, cosmetic, food composition, food additive, disinfectant or cleaning agent. 9. The anti-coronavirus agent according to claim 7 or 8, which is for prevention or amelioration of COVID-19. The anti-coronavirus agent according to any one of claims 1 to 6, which is used for application to articles. 11. The anti-coronaviral agent according to claim 10, which is a disinfectant or cleaning agent.

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