JP7838819B2 - Antiviral drugs - Google Patents

Description

This invention relates to antiviral agents, etc.

Coronaviruses are a species of virus belonging to the Coronavirinae subfamily of the Coronaviridae family. They are positive-strand RNA viruses that infect humans and animals, causing respiratory, gastrointestinal, or neurological diseases. Coronaviruses emerge from carrier animals and can cause large-scale epidemics in humans. Recent examples of coronaviruses include SARS-CoV-1, which caused outbreaks in 2002 and 2003, and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). More recently, a global pandemic of SARS-CoV-2 has occurred, and because no treatment technology has been established, it continues to cause immense damage in various fields, including healthcare and the economy.

The object of this invention is to provide an antiviral agent, particularly an antiviral agent against coronaviruses such as SARS-CoV-2.

In view of the above problems, the inventors diligently conducted research and found that the above problems can be solved with an antiviral agent containing at least one selected from the group consisting of (Component A) resveratrol, resveratrol derivatives, resveratrol polymers, derivatives of resveratrol polymers, or sirtuin activators, (Component B) plant extract of the genus Orychophragmus, (Component C) valligenol or its glycoside, (Component D) apigenin or its glycoside, (Component E) plant extract of the genus Bellis, (Component F) plant extract of the genus Ilex, (Component G) ursolic acid or its glycoside, (Component H) coumaric acid or its glycoside, (Component I) 2-phenylethanol or its glycoside, (Component J) tamarixetine or its glycoside, and (Component K) salmenthol or its glycoside. Based on this finding, the inventors furthered their research and completed the present invention. That is, the present invention encompasses the following embodiments.

Item 1. (Component A) Resveratrol, resveratrol derivatives, resveratrol polymers, derivatives of resveratrol polymers, or sirtuin activators.
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component D) Apigenin or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(H component) Coumaric acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
An antiviral agent containing (component J) tamarixetine or its glycosides, and (component K) salmenthol or its glycosides, selected from the group.

Item 2. The antiviral agent described in Item 1, wherein the virus is a coronavirus.

Item 3. The antiviral agent according to item 1 or 2, wherein the virus is SARS-CoV-2.

Item 4. (Component A) Resveratrol tetramer,
(Component B) Catalan extract,
(Component C) A1-Valigenol,
(Component D) Apigenin,
(Component E) Daisy flower extract,
(F component) Yerba mate extract,
(Component G) Matesaponin 3,
(H component) p-coumaric acid,
(Component I) 2-phenylethyl-β-D-glucopyranoside,
(Component J) Tamalixetine 7-O-β-D-glucopyranoside, and (Component K) Sesmoside E2 or Sesmoside E3
An antiviral agent according to any one of claims 1 to 3, comprising at least one selected from the group consisting of the following.

Item 5. An antiviral agent described in any of items 1 to 4, used for application to living organisms.

Item 6. An antiviral agent as described in Item 5, which is a pharmaceutical, cosmetic, disinfectant, or cleaning agent.

Item 7. An antiviral agent as described in item 5 or 6, which is used for the prevention or treatment of COVID-19.

Item 8. An antiviral agent as described in any of items 1 to 4, used for application to an article.

Item 9. An antiviral agent as described in Item 8, which is a disinfectant or cleaning agent.

The present invention also encompasses the following aspects.

Item 1. (Component A) Resveratrol polymer, derivative of resveratrol polymer, or sirtuin activator,
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
An antiviral agent containing (component J) tamarixetine or its glycosides, and (component K) salmenthol or its glycosides, selected from the group.

Item 2. The antiviral agent described in Item 1, wherein the virus is a coronavirus.

Item 3. The antiviral agent according to item 1 or 2, wherein the virus is SARS-CoV-2.

Item 4. (Component A) Resveratrol tetramer,
(Component B) Catalan extract,
(Component C) A1-Valigenol,
(Component E) Daisy flower extract,
(F component) Yerba mate extract,
(Component G) Matesaponin 3,
(Component I) 2-phenylethyl-β-D-glucopyranoside,
(Component J) Tamalixetine 7-O-β-D-glucopyranoside, and (Component K) Sesmoside E2 or Sesmoside E3
An antiviral agent according to any one of claims 1 to 3, comprising at least one selected from the group consisting of the following.

Item 5. An antiviral agent according to any one of items 1 to 4, comprising at least one selected from the group consisting of Hopeaphenol, Isohopeaphenol, and derivatives thereof.

Item 6. An antiviral agent described in any of items 1 to 5, used for application to living organisms.

Item 7. An antiviral agent as described in Item 6, which is a food composition, food additive, pharmaceutical, cosmetic, disinfectant, or cleaning agent.

Item 8. An antiviral agent as described in item 6 or 7, which is used for the prevention or treatment of COVID-19.

Item 9. An antiviral agent as described in any of items 1 to 5, used for application to an article.

Item 10. An antiviral agent as described in item 9, which is a disinfectant or cleaning agent.

Item 11. An antiviral food composition containing at least one selected from the group consisting of hopeaphenol, isohopeaphenol, and derivatives thereof.

Item 12. (Component A) Resveratrol polymer, derivative of resveratrol polymer, or sirtuin activator,
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
An antiviral method comprising applying at least one selected from the group consisting of (component J) tamarixetine or its glycosides, and (component K) sarmenthol or its glycosides, to a living organism or article.

Item 13. For use as an antiviral agent,
(Component A) Resveratrol polymer, derivative of resveratrol polymer, or sirtuin activator,
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
A preparation containing (component J) tamarixetine or its glycosides, and (component K) salmenthol or its glycosides, selected from the group.

Item 14. (Component A) Resveratrol polymer, derivative of resveratrol polymer, or sirtuin activator,
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
Use of at least one selected from the group consisting of (component J) tamarixetine or its glycosides, and (component K) sarmenthol or its glycosides, for the manufacture of an antiviral agent.

Item 15. (Component A) Resveratrol polymer, derivative of resveratrol polymer, or sirtuin activator,
(Component B) Extract from plants of the genus *Erythrina*,
(Component C) Valigenol or its glycosides,
(Component E) Daisy plant extract,
(Component F) Extract from plants of the genus Ilex,
(Component G) Ursolic acid or its glycosides,
(Component I) 2-phenylethanol or its glycoside,
Use as an antiviral agent of at least one selected from the group consisting of (component J) tamarixetine or its glycosides, and (component K) salmenthol or its glycosides.

According to the present invention, it is possible to provide an antiviral agent, particularly an antiviral agent against coronaviruses such as SARS-CoV-2.

The results of Test Example 2, using hopeaphenol and isohopeaphenol as samples, are shown. The vertical axis shows the virus inhibition rate based on CPE. The horizontal axis shows the sample name and the virus concentration of the sample in the solution. The results of Test Example 2, using methanol extract from daisy flowers as a sample, are shown. The vertical axis shows the virus inhibition rate based on CPE. The horizontal axis shows the sample name and the virus concentration of the sample in the solution.

In this specification, the terms “contains” and “includes” include the concepts of “contains,” “includes,” “substantially consist of,” and “consist solely of.”

In one embodiment, the present invention relates to an antiviral agent (sometimes referred to as "the formulation of the present invention" in this specification) containing at least one selected from the group consisting of components A to K (sometimes referred to as "the active ingredient" in this specification). This will be described below.

1. Active ingredients
1-1. Component A
Component A is resveratrol, a resveratrol derivative, a resveratrol polymer, a derivative of a resveratrol polymer, or a sirtuin activator.

The resveratrol derivative is not particularly limited, and known derivatives can be used without restriction. Examples of resveratrol derivatives are not particularly limited, but include cis derivatives, derivatives in which a hydroxyl group on the benzene ring (e.g., 1-2 or 1 hydroxyl group) is substituted with another group (e.g., a hydrogen atom, an alkoxy group, an alkyl group, an amino group, a carboxyl group, etc.), derivatives in which a hydrogen atom on the benzene ring (e.g., 1-2 or 1 hydrogen atom) is substituted (e.g., a hydroxyl group, an alkoxy group, an alkyl group, an amino group, a carboxyl group, etc.), derivatives in which the benzene ring is condensed with another ring, and derivatives obtained by combining one or more (e.g., 1-3, 1-2, or 1) modifications of these derivatives.

Resveratrol polymers are compounds having multiple resveratrol or derivative skeletons, and are not particularly limited in that respect. Examples of resveratrol polymers include resveratrol dimers (e.g., Amurensin A and H, Ampelopsin A, B, D, and F, Balanocarpol, Cyphostemmin A and B, δ-viniferin, ε-viniferin, Gnetin A and C, Leachinol F, Malibatol A, Pallidol, Parthenocissin A, Quadrangularin A, Restrytisol A, B, and C, Vitisinol D), resveratrol trimers (α-viniferin, Ampelopsin C, Amurensin C, D, and G, Hopeanolin, Malaysianol A, Miyabenol C, Resviniferin A and B, trans-diptoindonesin B), and resveratrol tetramers (Ampelopsin H, Amurensin K, Caraphenol B, Carasinol B, Flexuosol Examples include A, Hopeaphenol, Isohopeaphenol, Kobophenol A, Miyabenol A, Vaticanol B and C, Vaticaphenol A, Vitisin A, B and C, β-viniferin (cyclic resveratrol tetramer), resveratrol pentamer (Amurensin E and F), resveratrol hexamer (Chunganenol), γ-viniferin, Valeriaphenol A, etc.

Derivatives of resveratrol polymers are treated similarly to resveratrol derivatives.

The sirtuin activator is not particularly limited, but for example, the above components can be used. In addition to these, other examples of sirtuin activators include Resveratrol, Quercetin, Butein, SRT1720, SRT1460, SRT2183, STAC-5, STAC-9, STAC-10, Piceatannol, SRT2104, 1,4-DHP derivative, UBCS039, SRT2379, SRT3025, β-nicotinamide mononucleotide (NMN), etc.

As component A, plant extracts containing the above-mentioned components can also be used.

Component A is preferably a resveratrol polymer, more preferably a resveratrol tetramer, and particularly preferably Hopeaphenol and Isohopeaphenol.

Resveratrol and its polymers can be obtained by methods known to produce them, or by using commercially available products. Resveratrol and its polymers can be obtained, for example, by extraction and purification from various plants (e.g., grapes, Shorea roxburghii). Specifically, they can be obtained by methods following or similar to those previously reported (Bioorganic & Medicinal Chemistry 20 (2012) 832-840).

Component A may be a single type or a combination of two or more types.

If the formulation of the present invention contains component A, the amount thereof is not particularly limited, as long as it is an amount capable of exerting an antiviral effect against the virus. The amount thereof may be such that the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.01 μM or more, preferably 0.1 μM or more, more preferably 1 μM or more (the upper limit is not particularly limited, but for example 100 mM, 10 mM, 1 mM, 100 μM).

1-2． B component
Component B is an extract from plants of the genus *Erythrina*.

The plant extracts of the genus *Mallotus* are not particularly limited, as long as they are obtained by extraction from plants of the genus *Mallotus*.

Examples of plants belonging to the genus Orychophragmus include Orychophragmus violaceu, Orychophragmus limprichtianus, and Orychophragmus ziguiensis. Among these, Orychophragmus violaceu is particularly preferred.

It is preferable to cut the plants of the genus *Erythrina* used for extraction as needed.

The extraction solvent is not particularly limited. Examples of extraction solvents include water; alcohols such as methanol, ethanol, propanol, and butanol; alkyl acetates such as methyl acetate, ethyl acetate, propyl acetate, and butyl acetate; and supercritical carbon dioxide. Among these, alcohols (especially methanol) are preferred. The extraction solvent may be a single solvent or a combination of two or more solvents.

The temperature of the solvent during extraction is not particularly limited, but is preferably 50 to 100°C, and more preferably 60 to 100°C. The extraction time varies depending on the extraction method, solvent, temperature, etc., and is not particularly limited.

Component B may be a single type or a combination of two or more types.

If the formulation of the present invention contains component B, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content (calculated on a dry weight basis of the extract) may be such that the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.1 mg/mL or more, preferably 1 mg/mL or more, more preferably 5 mg/mL or more (the upper limit is not particularly limited, but may be, for example, 1 g/mL, 100 mg/mL, 10 mg/mL).

1-3． C component
Component C is barigenol or its glycoside.

Examples of barrigenol include A1-barrigenol and R1-barrigenol.

The barigenol glycoside is not particularly limited as long as it is a compound in which barigenol is used as the aglycone and a sugar is linked to it by a sugar bond (usually a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6.

As component C, plant extracts containing the above components can also be used.

The C component is preferably valigenol, and particularly preferably A1-valigenol.

As barigenol and its glycosides, those obtained according to known manufacturing methods may be used, or commercially available products may be used. Barigenol and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., tea flowers). Specifically, they can be obtained, for example, by methods following or similar to those previously reported (Chem. Pharm. Bull. 60(5) 674-680 (2012)).

Component C may be a single element or a combination of two or more elements.

If the formulation of the present invention contains component C, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-4. D component
Component D is apigenin or its glycoside.

Apigenin glycosides are not particularly limited as long as they are compounds formed by linking a sugar (usually via glycosidic bonds) to apigenin as the aglycone. Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, and xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6.

As component D, plant extracts containing the above-mentioned components can also be used.

As component D, apigenin is particularly preferred.

Apigenin and its glycosides can be obtained by methods known to produce them, or by using commercially available products. Apigenin and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., daisy flowers). Specifically, they can be obtained by methods following or similar to those previously reported (Chem. Pharm. Bull. 56(4) 559-568 (2008)).

Component D may be a single type or a combination of two or more types.

If the formulation of the present invention contains component D, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-5. E component
Component E is an extract from plants of the daisy genus.

The daisy plant extract is not particularly limited, as long as it is obtained by extracting from a daisy plant.

Examples of plants belonging to the genus Bellis include Bellis perennis (daisy flower), Bellis annua, Bellis bernardii, Bellis caerulescens, Bellis cordifolia, Bellis dubia, Bellis hyrcanica, Bellis longifolia, Bellis microcephala, Bellis pappulosa, Bellis rotundifolia, and Bellis sylvestris. Among these, the daisy flower is particularly preferred.

It is preferable to cut the daisy plants used for extraction as needed.

The extraction solvent is not particularly limited. Examples of extraction solvents include water; alcohols such as methanol, ethanol, propanol, and butanol; alkyl acetates such as methyl acetate, ethyl acetate, propyl acetate, and butyl acetate; and supercritical carbon dioxide. Among these, alcohols (especially methanol) are preferred. The extraction solvent may be a single solvent or a combination of two or more solvents.

The extraction method is not particularly limited, but for example, the method previously reported (Chem.Pharm.Bull. 56(4) 559-568 (2008)) can be used.

Component E may be a single type or a combination of two or more types.

If the formulation of the present invention contains component E, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content (calculated on a dry weight basis of the extract) may be such that the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.1 mg/mL or more, preferably 1 mg/mL or more, more preferably 5 mg/mL or more (the upper limit is not particularly limited, but may be, for example, 1 g/mL, 100 mg/mL, 10 mg/mL).

1-6. Component F
Component F is an extract from plants of the genus Ilex.

The plant extracts of the genus Ilex are not particularly limited, as long as they are obtained by extraction from plants of the genus Ilex.

Examples of plants belonging to the genus Ilex include Ilex paraguariensis (Yerba Mate), Ilex abscondita, Ilex ambigua, Ilex amelanchier, Ilex amygdalina, Ilex anonoides, Ilex aquifolium, Ilex arisanensis, Ilex arnhemensis, and Ilex asprella. Among these, Yerba Mate is particularly preferred.

It is preferable to cut the Ilex genus plants used for extraction as needed.

The extraction solvent is not particularly limited. Examples of extraction solvents include water; alcohols such as methanol, ethanol, propanol, and butanol; alkyl acetates such as methyl acetate, ethyl acetate, propyl acetate, and butyl acetate; and supercritical carbon dioxide. Among these, alcohols (especially butanol) are preferred. The extraction solvent may be a single solvent or a combination of two or more solvents.

The extraction method is not particularly limited, but for example, the method previously reported (Chem.Pharm.Bull. 57(3) 257-261 (2009)) can be used.

The F component may be a single element or a combination of two or more elements.

If the formulation of the present invention contains component F, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content (calculated on a dry weight basis of the extract) may be such that the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 mg/mL or more, preferably 0.5 mg/mL or more, more preferably 2 mg/mL or more (the upper limit is not particularly limited, but may be, for example, 1 g/mL, 100 mg/mL, 10 mg/mL).

1-7. G component
Component G is ursolic acid or its glycoside.

Ursolic acid glycosides are not particularly limited as long as they are compounds formed by linking ursolic acid as the aglycone to a sugar (usually via a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, and xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6. A specific example of an ursolic acid glycoside is matesaponin 3.

As component G, plant extracts containing the above components can also be used.

The G component is preferably an ursolic acid glycoside, and particularly preferably matesaponin 3.

Ursolic acid and its glycosides can be obtained by methods known to produce them, or by using commercially available products. Ursolic acid and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., Yerba mate). Specifically, they can be obtained by methods following or similar to those previously reported (Chem. Pharm. Bull. 57(3) 257-261 (2009)).

The G component may be a single type or a combination of two or more types.

If the formulation of the present invention contains component G, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-8. H component
The H component is coumaric acid or its glycoside.

Examples of coumaric acid include p-coumaric acid, m-coumaric acid, and o-coumaric acid.

Coumaric acid glycosides are not particularly limited as long as they are compounds formed by linking coumaric acid as the aglycone with a sugar (usually via a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, and xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6.

As the H component, plant extracts containing the above components can also be used.

As the H component, coumaric acid is preferred, and p-coumaric acid is particularly preferred.

Coumaric acid and its glycosides can be obtained by methods known to produce them, or by using commercially available products. Coumaric acid and its glycosides can be obtained, for example, by extraction and purification from various plants.

The H component may be a single type or a combination of two or more types.

If the formulation of the present invention contains component H, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-9. Component I
Component I is 2-phenylethanol or its glycoside.

2-phenylethanol glycosides are not particularly limited as long as they are compounds formed by linking 2-phenylethanol as the aglycone to a sugar (usually via a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, and xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6. Specifically, an example of a 2-phenylethanol glycoside is 2-phenylethyl-β-D-glucopyranoside.

As component I, plant extracts containing the above components can also be used.

Component I is preferably a 2-phenylethanol glycoside, and particularly preferably 2-phenylethyl-β-D-glucopyranoside.

2-phenylethanol and its glycosides can be obtained by methods known to produce them, or by using commercially available products. 2-phenylethanol and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., Sedum sarmentosum). Specifically, they can be obtained by methods following or similar to those previously reported (HETEROCYCLES, Vol. 71, No. 7, 2007, pp1565-1576).

Component I may be a single type or a combination of two or more types.

If the formulation of the present invention contains component I, its content is not particularly limited as long as it is an amount capable of exerting an antiviral effect against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-10． J component
Component J is tamarixetine or its glycoside.
Tamalixetine glycosides are not particularly limited as long as they are compounds formed by linking tamarixetine as the aglycone with a sugar (usually via a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, and xylose, and sugars formed by linking multiple units of these sugars. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6. Specifically, an example of a tamarixetine glycoside is tamarixetine 7-O-β-D-glucopyranoside.

As component J, plant extracts containing the above components can also be used.

Component J is preferably a tamalixetine glycoside, and particularly preferably tamalixetine 7-O-β-D-glucopyranoside.

Tamalixetine and its glycosides can be obtained by methods known to produce them, or by using commercially available products. Tamalixetine and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., Sedum sarmentosum). Specifically, they can be obtained by methods following or similar to those previously reported (HETEROCYCLES, Vol. 71, No. 7, 2007, pp1565-1576).

Component J may be a single type or a combination of two or more types.

If the formulation of the present invention contains component J, its content is not particularly limited as long as it is an amount capable of exerting antiviral activity against the virus. The content may be such that, for example, the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example, 100 mM, 10 mM, 1 mM, 100 μM).

1-11. K component
Component K is sarmenthol or its glycosides. Sarmenthol glycosides are not particularly limited as long as they are compounds in which sarmenthol is used as the aglycone and a sugar is linked to it by a sugar bond (usually a glycosidic bond). Examples of sugars include glucose, arabinose, pyranose, raffinose, rutinose, apiose, rhamnose, rutinose, primeverose, genthiobiose, digitoxose, galactose, xylose, etc., and sugars formed by linking multiple of these together are also included. The number of monosaccharide residues constituting the sugar is, for example, 1 to 8 or 1 to 6. Specific examples of sarmenthol glycosides include sesumoside E2 and sesumoside E3.

As the K component, plant extracts containing the above components can also be used.

Preferably, the K component is a salmenthol glycoside, and particularly preferably sesumoside E2 and sesumoside E3.

Sarmenthol and its glycosides can be obtained by methods known to produce them, or by using commercially available products. Sarmenthol and its glycosides can be obtained, for example, by extraction and purification from various plants (e.g., Sedum sarmentosum). Specifically, they can be obtained by methods following or similar to those previously reported (Chem. Pharm. Bull. 55(3) 435-441 (2007)).

The K component may be a single type or a combination of two or more types.

If the formulation of the present invention contains component K, the amount is not particularly limited as long as it is sufficient to exert an antiviral effect against the virus. The amount may be such that the concentration at the time of use (blood concentration or tissue concentration after ingestion or administration) is, for example, 0.05 μM or more, preferably 0.5 μM or more, more preferably 5 μM or more (the upper limit is not particularly limited, but for example 100 mM, 10 mM, 1 mM, 100 μM).

2. Uses The target viruses of the formulation of the present invention are not particularly limited. Examples of viruses include enveloped viruses (viruses having an envelope) such as influenza virus (e.g., type A, type B, etc.), rubella virus, Ebola virus, coronavirus, measles virus, varicella-zoster virus, mumps virus, arbovirus, RSV, SARS virus, hepatitis virus (e.g., hepatitis B virus, hepatitis C virus, etc.), yellow fever virus, HIV, rabies virus, hantavirus, dengue virus, nipah virus, lyssavirus, etc.; and non-enveloped viruses (viruses without an envelope) such as adenovirus, norovirus, rotavirus, human papillomavirus, poliovirus, enterovirus, coxsackievirus, human parvovirus, encephalomyocarditis virus, poliovirus, rhinovirus, etc. Among these, coronavirus is particularly preferred.

Coronaviruses are viruses belonging to the subfamily Orthocoronavirinae. Examples of coronaviruses include the genera Alphacoronavirus, Betacoronavirus, Gammacoronavirus, and Deltacoronavirus, with Betacoronavirus being preferred among these. Examples of Betacoronavirus are SARS-related coronavirus (SARSr-CoV), broad coronavirus HKU1, and MERS coronavirus, with SARSr-CoV being preferred among these. Examples of SARSr-CoV are SARS-CoV-2 and SARS-CoV-1, with SARS-CoV-2 being preferred among these. The formulation of the present invention can be suitably used against SARS-CoV-2 in particular.

The formulation of the present invention can exert antiviral activity against viruses. Specifically, antiviral activity includes suppression of viral infection, suppression of cell death caused by viruses, viral inactivation, and induction of viral resistance. Furthermore, due to this activity, it can also be used as a preventive or therapeutic agent for viral infections (preferably coronavirus infections, particularly COVID-19).

The formulation of the present invention can be widely used in various fields requiring antiviral properties. For example, it can be used in various fields such as industry, cleaning, medicine, food, and daily necessities. The uses of the formulation of the present invention can be mainly divided into applications applied to living organisms and applications applied to the articles described below.

2-1. Applications to living organisms Examples of applications to living organisms include pharmaceuticals, cosmetics, food compositions (including health foods, health enhancers, and nutritional supplements), food additives, disinfectants, and cleaning agents. In this case, the target organisms are not particularly limited and include various mammals such as humans, monkeys, mice, rats, dogs, cats, rabbits, pigs, horses, cattle, sheep, goats, and deer.

By applying the formulation of the present invention to a living organism, the active ingredient can exert an antiviral effect at the site of contact.

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

In terms of form, when the intended use is pharmaceutical, examples include formulations suitable for parenteral administration such as injections, intravenous infusions, gargles, transdermal patches (plasters, ointments, etc., tapes (reservoir type, matrix type, etc.), poultices, patches, microneedles, etc.), ointments, topical solutions (liniments, lotions, etc.), sprays (topical aerosols, pump sprays, etc.), creams, gels, eye drops, eye ointments, nasal drops, suppositories, semi-solid rectal preparations, enema preparations, etc.); and formulations suitable for oral administration (oral formulations) such as tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, lozenges, jelly-like drops, etc.), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including drinks, suspensions, syrups), and jellies.

In terms of form, when the intended use is as a cosmetic, examples include liquids, gels, creams, ointments, sprays, sticks, etc.

In terms of form, when used in food compositions, examples include liquid, gel, or solid foods such as juices, soft drinks, teas, soups, soy milk, salad oils, dressings, yogurts, jellies, puddings, seasonings, infant formula, cake mixes, dairy products (e.g., powders, liquids, gels, solids, etc.), bread, and confectionery (e.g., cookies, etc.). When used as food additives, health enhancers, or nutritional supplements, examples include tablets (including orally disintegrating tablets, chewable tablets, effervescent tablets, lozenges, and jelly drops), pills, granules, fine granules, powders, hard capsules, soft capsules, dry syrups, liquids (including suspensions and syrups), and jellies.

In terms of form, when used as a disinfectant or cleaning agent, it can take any form, such as liquid (solution, emulsion, suspension, spray, etc.), semi-solid (gel, cream, paste, etc.), or solid (tablet, particulate, capsule, film, compound, molten solid, waxy solid, elastic solid, etc.). For example, when applied to the oral cavity, more specifically, examples include toothpaste (paste, liquid toothpaste, powder toothpaste, etc.), mouthwash, topical agents, patches, mouth fresheners, and foods (e.g., chewing gum, tablets, candy, gummies, films, lozenges, etc.). When applied to the nasal cavity, more specifically, examples include nasal sprays. When applied to the skin, examples include soap, body wash, shampoo, conditioner, and sprays.

The formulation of the present invention may further contain other components as needed. These other components are not particularly limited, as long as they are components that can be incorporated into pharmaceuticals, cosmetics, disinfectants, detergents, etc., but examples include bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, thickeners, humectants, colorants, fragrances, and chelating agents.

The buffering agent is not particularly limited, and any appropriate and acceptable one can be used depending on the application. Preferably, it is a phosphate buffer, an acetate buffer, or the like.

The content of the active ingredient in the formulation of the present invention depends on the type of active ingredient, its use, the manner of use, the target of application, the condition of the target of application, etc., and is not limited thereto, but for example it can be 0.000001 to 100% by weight, preferably 0.01 to 50% by weight.

The dosage of the formulation of the present invention (e.g., administration, ingestion, etc.) is not particularly limited as long as it is an effective amount that produces the desired effect, and is generally 0.1 to 1000 mg/kg body weight per day as the weight of the active ingredient. The above dosage is preferably administered once a day or divided into two to three doses, and can be increased or decreased as appropriate depending on age, condition, and symptoms.

2-2. Applications to Articles Examples of applications to articles include disinfectants and cleaning agents. In this case, there are no particular restrictions on the scope of application, and it can include industrial products and their raw materials used in various fields.

Specific examples of items include office automation equipment, home appliances, air conditioners, vacuum cleaners, desks, chairs, sofas, benches, windows, handrails, steering wheels, seats, automatic ticket gates, automatic ticket vending machines, vending machines, doors, fences, handrails, tableware, cooking utensils, packaging film, packaging bags, bottles, containers, packaging packs, sinks, toilets, stationery, books, shelves, toothbrushes, mirrors, air conditioning filters, masks, coats, jackets, trousers, skirts, dress shirts, knit shirts, blouses, sweaters, cardigans, nightwear, underwear, diapers, supporters, socks, tights, stockings, hats, scarves, mufflers, neck wraps, stoles, gloves, clothing linings, clothing interlinings, clothing padding, work clothes, uniforms, school uniforms, curtains, screen doors, futon fabric, futon cotton, futon covers, pillowcases, sheets, mats, carpets, towels, handkerchiefs, wall coverings, bandages, and more.

By applying the formulation of the present invention to an article, an antiviral effect can be exerted at the site of contact with the active ingredient. The site of contact with the active ingredient is not limited to a site on the article; if the article is applied to a living organism, it also includes sites within the living organism.

The dosage form of the formulation of the present invention is not particularly limited and can be appropriately selected according to its application. Examples of dosage forms include liquid formulations such as liquids, emulsions, suspensions, dispersants, sprays, and aerosols; and solid or semi-solid formulations such as wettable powders, powders, granules, fine granules, and flowable formulations.

The formulation of the present invention may further contain other components as needed. These other components are not particularly limited, as long as they are components that can be incorporated into, for example, cleaning agents, disinfectants, etc., but examples include bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, thickeners, humectants, colorants, fragrances, and chelating agents.

The buffering agent is not particularly limited, and any appropriate and acceptable one can be used depending on the application. Preferably, it is a phosphate buffer, an acetate buffer, or the like.

The content of the active ingredient in the formulation of the present invention depends on the type of active ingredient, its use, the manner of use, the target of application, the condition of the target of application, etc., and is not limited thereto, but for example it can be 0.000001 to 100% by weight, preferably 0.001 to 50% by weight.

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

Test Example 1. Evaluation Test of Antiviral Efficacy 1
(Component A) hopeaphenol (10 μM) and isohopeaphenol (10 μM), (Component B) Catalan lime methanol extract (10 μg/mL), (Component C) A1-valigenol (10 μM), (Component D) apigenin (10 μM), (Component E) Daisy flower methanol extract (10 μg/mL), (Component F) Yerba mate butanol extract (10 μg/mL), (Component G) Mate saponin 3 (10 μM), (Component H) p-coumaric acid (10 μM), (Component I) 2-phenylethyl-β-D-glucopyranoside (10 μM), (Component J) Tamalixetine The antiviral activity against SARS-CoV-2 infection was evaluated for 7-O-β-D-glucopyranoside (10 μM), and (potassium component) sesumoside E2 (10 μM) and sesumoside E3 (10 μM) (all concentrations are final concentrations).

Specifically, the virus and the test sample were mixed and incubated for a certain period of time. The mixture was then added to cell culture wells, and the cells were cultured. After the culture was complete, live cell staining was performed, and antiviral activity was determined to be present if no CPE (cancerous pulmonary embolism) was observed, similar to the control (no virus added). The virus amount was varied in three stages ( ^10⁻² , ^10⁻³ , or ^10⁴ TCID _50/100 μL) and tested. If antiviral activity was determined to be present in all cases ( ^10⁻² , ^10⁻³ , and ^10⁴ TCID _50/100 μL ₎ , it was evaluated as "+++". If antiviral activity was determined to be present only in the cases of ^10⁻² and ^10⁻³ TCID _50/100 μL, it was evaluated as "++". If antiviral activity was determined to be present only in the case of ^10⁻² TCID 50/100 μL, it was evaluated as "+".

The details of the test protocol are as follows:
Day -1
VeroE6/TMPRSS2 cells were seeded in 96-well plates at a rate of 5.0 x ^10⁴ cells/100 μL. The culture medium was DMEM medium supplemented with 0.5% fetal bovine serum. Cells were incubated at 37°C under 5% _CO₂ /95% air for 24 hours.
Day 0
120 μL of virus solution ( ^10₂ , ^10₃ , or ^10₄ TCID _50/100 μL) was added to a 96-well plate (with 1.2 μL of sample already added) and allowed to stand at room temperature for 5 minutes. The culture supernatant was discarded from the 96-well plate in which cells had been seeded and culture started the previous day (Day 1), and 100 μL of the sample-virus mixture obtained above was added. The cells were incubated at 37°C under 5% _CO₂ /95% air for 72 hours.
Day 3
100 μL of glutaraldehyde solution (25% W/V) was added to the cells (96-well plate) and allowed to stand at room temperature for 30 minutes. The culture medium and glutaraldehyde mixture was discarded, and the wells were washed with tap water. The plates were allowed to stand at room temperature to dry. 100 μL of 1% crystal violet staining solution was added to the fixed cells (96-well plate) and allowed to stand at room temperature for 30 minutes. The staining solution was discarded, and the wells were washed with tap water. After allowing to stand at room temperature to dry, the plates were photographed to observe whether or not staining occurred.

For component B, the methanol extract of sweet potato was obtained by hot extraction of 1.0 kg of whole sweet potato (Orychophragmus violaceus) with methanol (heating under reflux for 2 hours each, 3 times), followed by removal of the solvent under reduced pressure. For component E, the methanol extract of daisy flower was obtained according to the method previously reported (Chem. Pharm. Bull. 56(4) 559-568 (2008)). For component F, the butanol extract of yerba mate was obtained according to the method previously reported (Chem. Pharm. Bull. 57(3) 257-261 (2009)).

The results are shown in Table 1.

Test Example 2. Evaluation Test of Antiviral Efficacy 2
The antiviral activity against SARS-CoV-2 was evaluated for (Component A) hopeaphenol (10 μM, 1 μM, 0.1 μM) and isohopeaphenol (10 μM, 1 μM, 0.1 μM), and (Component E) daisy flower methanol extract (10 μg/mL, 1 μg/mL, 0.1 μg/mL) (all concentrations are final concentrations).

Specifically, in addition to the same method as in Test Example 1 (Pre-infection protocol: a method of reacting the sample with the virus before infecting the cells), the test was also conducted using a Post-infection protocol: a method of adding the sample after infecting the cells with the virus and culturing them. The test was performed using Quadruplication. A virus infection score was assigned to each of the four wells of the Quadruplication as shown in Table 2. The average score of the four wells was then calculated.

The CPE reduction rate was calculated using the following formula.

Furthermore, as a control, wells containing only the test samples, without virus infection, were prepared using Quadruplication. If cell death was observed in these wells, it was possible that the cytotoxicity was caused by DMSO, the solvent in the samples. Therefore, the test samples at that concentration were judged as "undetermined."

The pre-infection protocol is the same as in Example 1, except that a ^10⁻² TCID _50/100 μL virus solution is used and the incubation time on Day 0 is 96 hours.

The post-infection protocol is the same as in Example 1, except that Day 0 follows the protocol shown below.
Day 0
The culture supernatant was discarded from a 96-well plate in which VeroE6/TMPRSS2 cells had been seeded and cultured the previous day (Day 1), and 100 μL of virus solution ( ^10⁻² TCID _50/100 μL) was added. The cells were incubated at 37°C under 5% _CO₂ /95% air for 30 minutes. 1 μL of the sample was added, and the cells were incubated at 37°C under 5% _CO₂ /95% air for 96 hours.

The results are shown in Figures 1 and 2.

Claims (11)

An anticoronavirus agent containing at least one selected from the group consisting of Hopeaphenol and Isohopeaphenol . The anticoronavirus agent according to claim 1, wherein the coronavirus is SARS-CoV-2. An anticoronavirus agent according to claim 1 or 2, used for application to a living organism. The anticoronavirus agent according to claim 3 , which is a food composition, food additive, pharmaceutical, cosmetic, disinfectant, or cleaning agent. An anticoronavirus agent according to claim 3 or 4 , which is a preventive or therapeutic agent for COVID-19. An anticoronavirus agent according to claim 1 or 2, used for application to an article. The anticoronavirus agent according to claim 6 , which is a disinfectant or cleaning agent. An anti-coronavirus food composition containing at least one selected from the group consisting of Hopeaphenol and Isohopeaphenol. An anticoronavirus method comprising applying at least one substance selected from the group consisting of hopeaphenol and isohopeaphenol to an article. A formulation containing at least one selected from the group consisting of Hopeaphenol and Isohopeaphenol, for use as an anticoronavirus agent. Use of at least one selected from the group consisting of Hopeaphenol and Isohopeaphenol for the manufacture of an anticoronavirus agent.