WO2024010035A1

WO2024010035A1 - Viral infection preventing agent

Info

Publication number: WO2024010035A1
Application number: PCT/JP2023/024948
Authority: WO
Inventors: 夕子中上
Original assignee: 株式会社レゾナック
Priority date: 2022-07-06
Filing date: 2023-07-05
Publication date: 2024-01-11

Abstract

Provided is a viral infection preventing agent containing methyl hesperidin as an active ingredient. Also provided is a method for using the viral infection preventing agent, the method including applying the viral infection preventing agent onto the surface of an article. Also provided is a method for using the viral infection preventing agent, the method including spraying the viral infection preventing agent over the body surface of an animal. Also provided is a method for using the viral infection preventing agent, the method including spraying or blending the viral infection preventing agent over fibers or into the fibers. Also provided is a viral infection preventing composition comprising the viral infection preventing agent and a pharmaceutically acceptable carrier.

Description

Viral infection inhibitor

TECHNICAL FIELD The present invention relates to a virus infection suppressant. The present invention also relates to a method of using the virus infection inhibitor and a composition for preventing virus infection.
This application claims priority based on Japanese Patent Application No. 2022-109090 filed in Japan on July 6, 2022, the contents of which are incorporated herein.

SARS-related coronavirus (hereinafter referred to as "SARS virus") is a coronavirus that causes severe respiratory syndrome (SARS). The virus that causes COVID-19 (SARS-CoV-2), which has become a worldwide infection problem as of 2022, is a type of SARS virus.
The SARS virus infects target cells by fusing its own cell membrane with that of the target cell. The SARS virus has Spike-protein (hereinafter referred to as "S-protein"), which is classified as a class I membrane fusion promoting protein, on its membrane surface. This S-protein is known to play a major role in membrane fusion to target cells.

In order to suppress the infection of viruses including the SARS virus, it is effective to inhibit the binding of viruses to target cells. It has been reported that algae-derived lectins as components that inhibit virus binding are useful for treating or diagnosing viral diseases by inhibiting the binding recognition of viruses to target cells (Patent Document 1).
Hesperidin is a substance classified as a flavonoid that is abundant in citrus fruits such as oranges, oranges, and lemons, and is also known as ``vitamin P.'' Patent Document 2 reports on the virus inhibitory activity of hesperidin, but it is reported that hesperidin does not exhibit virus inhibitory activity even at high concentrations (Patent Document 2).

JP 2021-13375 Publication Special Publication No. 2009-519324

Suppressing the infection of the SARS virus, especially SARS-CoV-2, is an urgent global issue. Therefore, there is a need for a virus infection suppressant that has an infection suppressing effect against SARS-CoV-2 and can be easily used for various daily purposes.
Therefore, the present invention provides a virus infection suppressant, a method for using the same, and a composition for preventing virus infection, which has an infection suppressing effect on SRAS viruses, particularly SARS-CoV-2, and can be easily used for various daily uses. The purpose is to provide.

The present invention includes the following aspects.
[1] A virus infection inhibitor containing methylhesperidin as an active ingredient.
[2] The methylhesperidin is one or more selected from the group consisting of chalcone methylhesperidin represented by the following general formula (1) and flavanone methylhesperidin represented by the following general formula (2). , the virus infection inhibitor according to [1].

[In formula (1), R ¹ to R ⁹ are each independently a methyl group or a hydrogen atom. However, at least one of R ¹ to R ⁹ is a methyl group.
In formula (2), R ¹¹ to R ¹⁸ are each independently a methyl group or a hydrogen atom. However, at least one of R ¹¹ to R ¹⁸ is a methyl group. ]
[3] The methylhesperidin is one or more selected from the group consisting of chalcone methylhesperidin represented by the following general formula (3) and flavanone methylhesperidin represented by the following general formula (4). , the virus infection inhibitor according to [1].

[In formula (3), R ²⁰ to R ²³ are each independently a methyl group or a hydrogen atom.
In formula (4), R ²⁴ to R ²⁵ are each independently a methyl group or a hydrogen atom. ]
[4] The chalcone methylhesperidin represented by the general formula (3) is one or more selected from the group consisting of chalcone-1 to 3 having the combinations of R ²⁰ to R ²³ shown in Table 1 below. The virus infection inhibitor according to [3].

[5] The flavanone methylhesperidin represented by the general formula (4) is one or more selected from the group consisting of flavanones-1 to 4 having the combinations of R ²⁴ to R ²⁵ shown in Table 2 below. The virus infection inhibitor according to [3] or [4].

[6] The virus is at least one virus selected from the group consisting of Orthomyxoviridae, Paramyxoviridae, Retroviridae, Filoviridae, and Coronaviridae, [1] to [5] ] The virus infection inhibitor according to any one of the above.
[7] The virus infection inhibitor according to [6], wherein the virus is a SARS-related coronavirus.
[8] A method for using a virus infection inhibitor, which comprises applying the virus infection inhibitor according to any one of [1] to [7] to the surface of an article.
[9] A method for using a virus infection inhibitor, which comprises spraying the virus infection inhibitor according to any one of [1] to [7] onto the body surface of an animal.
[10] A method for using a virus infection inhibitor, which comprises spraying or blending the virus infection inhibitor according to any one of [1] to [7] onto fibers.
[11] A composition for suppressing viral infection, comprising the viral infection suppressing agent according to any one of [1] to [7], and a pharmaceutically acceptable carrier.

According to the present invention, a virus infection inhibitor is provided that can be easily used for various daily uses. Furthermore, a method for using the virus infection inhibitor and a composition for preventing virus infection containing the virus infection inhibitor are provided.

Hereinafter, embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments shown below.

(Viral infection inhibitor)
The virus infection inhibitor of this embodiment contains methylhesperidin as an active ingredient. Methylhesperidin has the effect of suppressing virus binding to target cells. For example, methylhesperidin has the effect of suppressing the binding of the S-protein of SARS viruses, including SARS-CoV-2, to the ACE2 protein on the surface of human cells. Therefore, when methylhesperidin is used as a viral infection inhibitor, it can be expected to have the effect of inhibiting viruses, particularly the SARS virus, from infecting human cells.

<Methylhesperidin>
The virus infection inhibitor of this embodiment is not particularly limited as long as it contains methylhesperidin as an active ingredient. As the methylhesperidin used in the virus infection inhibitor of the present embodiment, it is preferable to methylate hesperidin and solubilize it in water.
Before being methylated, hesperidin is a substance classified as a flavonoid that is abundant in citrus fruits such as oranges, oranges, and lemons. It is well known that hesperidin cooperates with vitamin C to maintain the effects of vitamin C, and has actions such as strengthening capillaries and promoting blood circulation. In addition, it has been reported that hesperidin is involved in various physiological effects such as antioxidant and antiallergic effects. However, hesperidin is poorly soluble in water. Methyl hesperidin is obtained by methyl derivatizing hesperidin. Since methylhesperidin is highly water-soluble, it can be expected to have various effects depending on its characteristics. Methylhesperidin is also considered to be highly effective when used as a virus infection suppressant due to its high water solubility.
Methylhesperidin mainly includes a chalcone type compound (chalcone methylhesperidin) represented by the following general formula (1) and a flavanone type compound (flavanone methylhesperidin) represented by the following general formula (2). It is known.

[In formula (1), R ¹ to R ⁹ are each independently a methyl group or a hydrogen atom. However, at least one of R ¹ to R ⁹ is a methyl group.
In formula (2), R ¹¹ to R ¹⁸ are each independently a methyl group or a hydrogen atom. However, at least one of R ¹¹ to R ¹⁸ is a methyl group. ]

The methylhesperidin used in the virus infection inhibitor of the present embodiment is selected from the group consisting of chalcone methylhesperidin represented by the above general formula (1) and flavanone methylhesperidin represented by the above general formula (2). It is preferable that one or more types are used.

In the general formula (1), R ¹ to R ⁹ are each independently a methyl group or a hydrogen atom, and at least one of R ¹ to R ⁹ is a methyl group. Among R ¹ to R ⁹ , any one to six are preferably methyl groups, and more preferably any two to five are methyl groups.

In the general formula (2), R ¹¹ to R ¹⁸ are each independently a methyl group or a hydrogen atom, and at least one of R ¹¹ to R ¹⁸ is a methyl group. Among R ¹¹ to R ¹⁸ , any one to four are preferably methyl groups, and more preferably any one to three are methyl groups.

Among the compounds represented by the general formula (1), the compound represented by the following general formula (3) is preferable as chalcone methylhesperidin. Among the compounds represented by the general formula (2), the flavanone methylhesperidin is preferably a compound represented by the following general formula (4).

[In formula (3), R ²⁰ to R ²³ are each independently a methyl group or a hydrogen atom.
In formula (4), R ²⁴ to R ²⁵ are each independently a methyl group or a hydrogen atom. ]

In the general formula (3), R ²⁰ to R ²³ are each independently a methyl group or a hydrogen atom. The chalcone methylhesperidin represented by the general formula (3) is one or more selected from the group consisting of chalcone-1 to 3 having the combinations of R ²⁰ to R ²³ shown in Table 3 below. is preferred.

In the general formula (4), R ²⁴ to R ²⁵ are each independently a methyl group or a hydrogen atom. The flavanone methylhesperidin represented by the general formula (4) is one or more selected from the group consisting of flavanones-1 to 4 having combinations of R ²⁴ to R ²⁵ shown in Table 4 below. It is preferable.

The methylhesperidin used in the virus infection suppressant of this embodiment may be used alone or in a mixture of two or more. Methylhesperidin is a chalcone methylhesperidin represented by the general formula (1) or a chalcone methylhesperidin represented by the general formula (3), and a flavanone methylhesperidin represented by the general formula (2) or It may contain both or only one of the flavanone methylhesperidin represented by the general formula (4). Methylhesperidin may include chalcone methylhesperidin represented by the general formula (3) and flavanone methylhesperidin represented by the general formula (4). Furthermore, methylhesperidin may contain one or more of the chalcone bodies-1 to 3, or may contain one or more of the flavanone bodies-1 to 4.
Furthermore, the virus infection inhibitor of the present embodiment may contain a mixture of chalcone bodies-1 to 3 and flavanone bodies-1 to 4 as methylhesperidin.

Methylhesperidin can be produced by a known method. Methyl hesperidin is produced by dissolving hesperidin, which is produced from citrus peels, in an aqueous sodium hydroxide solution, reacting a corresponding amount of dimethyl sulfate with the alkaline solution, neutralizing the resulting reaction solution with sulfuric acid, and producing n- It can be produced by extracting with butyl alcohol, distilling off the solvent, and recrystallizing with isopropyl alcohol (Sakiyuka, Nippon Kagaku Zasshi, (1958) Vol. 79, pp. 733-736; Japanese Patent No. 6312333). However, the manufacturing method is not limited to this.

Methylhesperidin is a commercially available product (for example, those distributed as pharmaceutical additives, food additives, and cosmetic raw materials, or "Methylhesperidin" (Resonac Co., Ltd.), "Methylhesperidin" (Tokyo Kasei Kogyo Co., Ltd.), "Hesperidin Methyl Chalcone" (Sigma) etc.) can also be purchased and used.

The virus infection inhibitor of this embodiment has the effect of suppressing binding of the virus to target cells. In particular, the viral infection inhibitor of this embodiment has the effect of inhibiting the binding between the SARS virus and human ACE2. Therefore, the virus infection inhibitor of the present embodiment has the effect of suppressing the infection of viruses, particularly the SARS virus, to humans.

By applying the virus infection suppressing agent of the present embodiment to the body surface of an animal (such as human skin) or an article, it is possible to impart antiviral properties to the applied surface. Therefore, by applying the virus infection suppressing agent of this embodiment to the body surface of an animal or the surface of an article, it is possible to suppress virus infection due to contact with the skin or article.
Animals to which the virus infection inhibitor of the present embodiment can be applied are not particularly limited, but include, for example, humans and non-human animals (pets such as dogs, cats, rabbits, and hamsters; cows, pigs, horses, sheep, and goats). livestock, etc.).
Articles to which the virus infection suppressant of this embodiment can be applied are not particularly limited. Examples of the goods include clothing such as clothes; daily necessities such as towels, handkerchiefs, toothbrushes, and tableware; food packages; industrial instruments such as various machine tools, various tools, and various measuring instruments; medical gloves, and medical equipment. Examples include medical supplies such as medical clothing and medical protective equipment; various medical instruments, etc.

Examples of methods for applying the virus infection inhibitor of this embodiment include a method of applying the virus infection inhibitor of this embodiment to the application surface using tissue paper, kitchen paper, or nonwoven fabric impregnated with the virus infection inhibitor of this embodiment; A method in which the virus infection inhibitor of the present embodiment is blended into a fabric softener or the like, and the clothes are dipped in the fabric softener and washed, thereby soaking it into the clothes; A method of immersing the virus in a solution containing the virus and then drying it; A method of spraying a spray containing the virus infection inhibitor of this embodiment on the applied surface; A method of prescribing the virus infection inhibitor of this embodiment in a nasal spray and directly treating the human body ; Examples include a method of preparing the virus infection suppressant of this embodiment as a spray agent and disposing it in a space.
When the article is a textile product such as clothing or a towel, the virus infection inhibitor of this embodiment may be blended with the fiber, and the textile product may be manufactured using the fiber. Alternatively, the textile product may be immersed in a solution containing the virus infection inhibitor of this embodiment and dried. Alternatively, a solution containing the virus infection inhibitor of this embodiment may be sprayed onto textile products.
The virus infection inhibitor of this embodiment may be used by being sprayed on the body surface of an animal. Animals include humans and non-human animals. Further, the virus infection suppressing agent of the present embodiment may be contained in a nasal drop and applied to the nasal mucosa as a nasal drop.

Examples of the method of using the virus infection suppressant of this embodiment include a method that includes applying it to the surface of an article. Thereby, it is possible to impart antiviral properties to the surface of the article and suppress viral infection of humans or animals that come into contact with the article.
Examples of methods for using the virus infection suppressant of this embodiment include methods that include spraying it on the body surface of animals (including humans). Thereby, antiviral properties can be imparted to the body surface of the animal, thereby suppressing viral infection in the animal or in humans or non-human animals that have come into contact with the animal.
Examples of the method of using the virus infection suppressant of this embodiment include a method including spraying or blending it on fibers. Thereby, antiviral properties can be imparted to the fibers, and virus infection of humans or non-human animals that come into contact with the fibers can be suppressed.
The invention also provides the use of methylhesperidin to impart antiviral properties to articles, animal (including human) body surfaces, or textiles.

Viruses to which the virus infection inhibitor of the present embodiment can be applied are not particularly limited, but are selected from the group consisting of Orthomyxoviridae, Paramyxoviridae, Retroviridae, Filoviridae, and Coronaviridae, for example. At least one type of virus is mentioned. The virus infection suppressing agent of this embodiment can be suitably applied to viruses of the Orthomyxoviridae family or the Coronaviridae family, among others. Examples of viruses of the Orthomyxoviridae family include influenza viruses (influenza virus genus A, influenza B virus, influenza C virus, etc.), Isavirus genus, Quaranjavirus genus, Togotovirus genus, etc. It will be done. Viruses of the Coronaviridae family include, for example, Alphacoronavirus, Betacoronavirus, Deltacoronavirus, and Gammacoronavirus. The Betacoronavirus genus includes, for example, Enbecovirus subgenus, Hibecovirus subgenus, Merbecovirus subgenus, Nobecovirus subgenus, and Sarbecovirus subgenus. Viruses of the sarbecovirus subgenus include SARS viruses (SARS-related coronaviruses) such as SARS-CoV and SARS-CoV-2.

(Composition for suppressing viral infection)
The composition for suppressing viral infection of this embodiment includes the viral infection suppressing agent of the above embodiment and a pharmaceutically acceptable carrier.

The viral infection inhibitor of the above embodiment may be prepared as a composition for suppressing viral infection by adding a pharmaceutically acceptable carrier and the like as appropriate. Viruses to which the composition for suppressing viral infection can be applied include those mentioned above.

The composition for suppressing viral infection of this embodiment is formulated by mixing methylhesperidin, a pharmaceutically acceptable carrier, and other ingredients as the case may be, according to a conventional method (for example, the method described in the Japanese Pharmacopoeia). It can be manufactured by

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier that does not inhibit the physiological activity of the active ingredient and does not exhibit substantial toxicity to the subject to which it is administered.
"Substantially non-toxic" means that the component exhibits no toxicity to the recipient at doses commonly used.
Pharmaceutically acceptable carriers include, but are not particularly limited to, excipients, binders, disintegrants, lubricants, stabilizers, diluents, solvents for injections, humectants, feel improvers, and surfactants. , polymers/thickeners/gelling agents, solvents, propellants, antioxidants, reducing agents, oxidizing agents, chelating agents, acids, alkalis, powders, inorganic salts, water, metal-containing compounds, unsaturated monomers , polyhydric alcohols, polymeric additives, wetting agents, thickeners, tackifiers, oily raw materials, liquid matrices, fat-soluble substances, polymeric carboxylates, and the like.
One type of pharmaceutically acceptable carrier may be used alone, or two or more types may be used in combination.

Other ingredients include, but are not limited to, preservatives, antibacterial agents, ultraviolet absorbers, skin whitening agents, vitamins and their derivatives, anti-inflammatory agents, anti-inflammatory agents, hair growth agents, blood circulation promoters, stimulants, and hormones. anti-wrinkle agents, anti-aging agents, tightening agents, cooling agents, warming agents, wound healing promoters, irritation relievers, analgesics, cell activators, plant/animal/microbial extracts, seed oils, antipruritics, Exfoliating and dissolving agents, antiperspirants, refreshing agents, astringents, enzymes, nucleic acids, fragrances, dyes, coloring agents, dyes, pigments, anti-inflammatory and analgesic agents, antifungal agents, antihistamines, hypnotic sedatives, tranquilizers, antiseptics Hypertensive agents, antihypertensive diuretics, antibiotics, anesthetics, antibacterial substances, antiepileptic agents, coronary vasodilators, herbal medicines, antipruritic agents, keratin softening exfoliants, ultraviolet blocking agents, bactericidal agents, antioxidants, pH adjustment agents, additives, metal soaps, etc. Specific examples of plant/animal/microbial extracts include Lapusana communis flowers/leaves/stem, tea leaves, and the like. A specific example of the seed oil includes Moringa seed oil. A specific example of the fragrance includes perylaldehyde.
One type of other components may be used alone, or two or more types may be used in combination.

Specific examples of the composition for suppressing viral infection of this embodiment include, for example, a spray composition, an aerosol composition, a skin spray composition, a nasal spray composition, and the like.

≪Spray composition≫
A spray composition is a composition that is used by spraying. The spray composition is prepared as a spray liquid to be filled into a spray container, and then used by being filled into the spray container. The spray solution may contain ethanol, an antibacterial agent, and the like. The spray composition is applied to the surface of the article, for example, by spraying. The methylhesperidin contained in the spray composition imparts antiviral properties to the surface of the article coated with the spray composition, thereby reducing the risk of virus infection due to contact with the surface of the article. Applicable articles include those mentioned above for the virus infection inhibitor.

≪Aerosol composition≫
An aerosol composition is a composition that is used by being sprayed by aerosol. The aerosol composition is prepared as an aerosol spray solution to be filled into an aerosol container, and then used by being filled into the aerosol container. The aerosol spray solution may contain anionic sodium salt additives, titanium oxide, and the like. The aerosol composition is applied to the surface of the article by, for example, aerosol spraying. The methylhesperidin contained in the aerosol composition imparts antiviral properties to the surface of the article coated with the aerosol composition, thereby reducing the risk of virus infection due to contact with the surface of the article. Applicable articles include those mentioned above for the virus infection inhibitor.

≪Skin spray composition≫
A skin spray composition is a composition that is used by spraying it onto human skin. A skin spray composition is prepared as a skin spray dispersion liquid to be filled into a spray container, and then used by being filled into a spray container. The skin spray dispersion liquid may contain titanium oxide, ethanol, sodium hyaluronate, and the like. Skin spray compositions are applied to human skin, for example, by spraying. The methylhesperidin contained in the skin spray composition imparts antiviral properties to the skin surface to which the skin spray composition is applied, reducing the risk of viral infection due to contact with the skin. . Application sites include, but are not particularly limited to, hands, arms, face, neck, feet, legs, torso, and the like.

≪Nasal medicated composition≫
A nasal spray composition is a composition used as a nasal spray. A nasal spray composition is prepared as a nasal spray solution to be filled into a nasal spray container, and then used by being filled into a nasal spray container. The nasal medicinal solution may contain antiallergic agents, antibacterial agents, and the like. The nasal spray composition is applied to the nasal mucosa as a nasal spray. Methylhesperidin contained in the nasal spray composition inhibits binding of the virus to target cells, thereby suppressing virus infection via the nasal mucosa.

≪Composition for medical supplies≫
A composition for medical supplies is a composition used by applying it to the surface of a medical product. The composition for medical supplies is used by applying it to medical supplies. The methylhesperidin contained in the medical product composition imparts antiviral properties to the surface of the medical product coated with the medical product composition, reducing the risk of viral infection due to contact with the medical product. do. Examples of medical supplies include medical gloves, medical clothing, medical protective equipment, and the like. When used in medical gloves, the gloves may be dried after being dipped in the medical product composition. Methylhesperidin remains on the surface of the glove after drying, so it exhibits antiviral properties.

<<Pharmaceutical composition>>
The composition for suppressing viral infection of this embodiment may be a pharmaceutical composition for preventing viral infection or viral infectious disease.

In addition to those listed above, carriers commonly used for pharmaceuticals can be used as pharmaceutically acceptable carriers in the pharmaceutical composition. For example, the Japanese Pharmacopoeia, non-Japanese Pharmacopoeial drug standards, Pharmaceutical Excipient Standards 2013 (Yakuji Nippo Sha, 2013), Pharmaceutical Excipient Dictionary 2016 (edited by the Japan Pharmaceutical Excipients Association, Yakuji Nippo Sha, 2016), Handbook of General raw materials described in Pharmaceutical Excipients, 7th edition (Pharmaceutical Press, 2012) and the like can be used. In the pharmaceutical composition, one type of pharmaceutically acceptable carrier may be used alone, or two or more types may be used in combination.

The pharmaceutical composition may contain other components in addition to the viral infection inhibitor and the pharmaceutically acceptable carrier. Other components are not particularly limited, and general pharmaceutical additives can be used. Moreover, active ingredients other than the above-mentioned virus infection inhibitors can also be used as other ingredients. Pharmaceutical additives and active ingredients as other ingredients include those listed above, such as the Japanese Pharmacopoeia, Pharmaceutical Standards outside the Japanese Pharmacopoeia, Pharmaceutical Excipient Standards 2013 (Yakuji Nipposha, 2013), Pharmaceutical Additives Common raw materials listed in Dictionary of Pharmaceutical Excipients 2016 (edited by Japan Pharmaceutical Excipients Association, Yakuji Nipposha, 2016), Handbook of Pharmaceutical Excipients, 7th edition (Pharmaceutical Press, 2012), etc. can be used. In the pharmaceutical composition, one type of other components may be used alone, or two or more types may be used in combination.

The dosage form of the pharmaceutical composition is not particularly limited, and can be any dosage form commonly used for pharmaceutical preparations. For example, orally administered dosage forms such as tablets, coated tablets, pills, powders, granules, capsules, solutions, suspensions, and emulsions; and injections, suppositories, mouthwashes, eye drops, Examples include dosage forms for parenteral administration such as external preparations for skin and nasal drops. Pharmaceutical compositions in these dosage forms can be formulated according to conventional methods (for example, the method described in the Japanese Pharmacopoeia). The pharmaceutical composition is preferably a parenteral preparation, more preferably a skin preparation or a nasal spray.

The method of administering the pharmaceutical composition is not particularly limited, and it can be administered by a method commonly used for administering pharmaceuticals. For example, the pharmaceutical composition may be administered orally, intravenously, intraarterially, intramuscularly, intradermally, subcutaneously, intraperitoneally, etc. as an injection or infusion preparation, or rectally as a suppository. It may be administered internally, it may be administered to the eyes as eye drops, it may be used to clean the mouth as a mouthwash, it may be administered to the skin as an external preparation, and it may be administered to the nose as a nasal spray. You can.

The dosage of the pharmaceutical composition can be a therapeutically effective amount. The therapeutically effective amount may be appropriately determined depending on the patient's symptoms, body weight, age, sex, etc., the dosage form of the pharmaceutical composition, the administration method, etc.
For example, in the case of an external preparation for the skin, the dosage of the pharmaceutical composition is 0.01 to 100 μg/cm ² , 0.1 to 50 μg/cm 2 , or 0.1 to 50 μg/cm ² per applied skin area as the total content of methylhesperidin. Examples include 1 to 20 μg/cm ² . For example, in the case of a nasal spray, the dosage of the pharmaceutical composition is 0.01 to 100 μg/cm ² , 0.1 to 50 μg/cm 2 , 0.1 to 50 μg/cm ² , or 1 to 20 μg/cm ² and the like. The pharmaceutical composition may include a therapeutically effective amount of methylhesperidin. A therapeutically effective amount of methylhesperidin in a pharmaceutical composition can be an amount of methylhesperidin effective to prevent viral infection. A therapeutically effective amount of methylhesperidin is, for example, an amount capable of inhibiting binding of a virus to a target cell, an amount capable of inhibiting binding of the SARS virus to human ACE2, an amount capable of reducing viral activity, or an amount capable of inhibiting viral infection. It can be the amount you get.

The administration interval of the pharmaceutical composition is not particularly limited and may be determined as appropriate depending on the administration method and the like. For example, it can be done once a day or about 2 to 3 times a day.

Since the pharmaceutical composition of this embodiment contains methylhesperidin as an active ingredient, it is effective in preventing viral infection or viral infectious disease. For example, by applying the pharmaceutical composition of this embodiment as an external preparation to the skin of the hands or face, it is possible to reduce the virus activity at the application site and suppress viral infection through the application site. .
By applying the pharmaceutical composition of this embodiment as a nasal drop to the nasal mucosa, viral infection from the nasal mucosa can be suppressed.
Therefore, the pharmaceutical composition of this embodiment is effective in preventing viral infection.

(Other embodiments)
In one embodiment, the invention provides a method for preventing viral infection, comprising administering methylhesperidin to a subject. The present invention also provides a method for preventing a viral infection or viral infection, comprising the step of applying methylhesperidin to the skin of a subject. The present invention provides a method for preventing viral infection or viral infection, which includes the step of nasally administering methylhesperidin to a subject.
In one embodiment, the invention provides a method of imparting antiviral properties to a surface of an article, the method comprising applying methylhesperidin to the surface of the article. The present invention also provides a method for imparting antiviral properties to the surface of an article, which includes the step of spraying or aerosolizing methylhesperidin onto the surface of the article. The present invention also provides a method for manufacturing an antiviral article, which includes the steps of adding methylhesperidin to a raw material for the article, and manufacturing the article from the raw material. The present invention also provides a method for imparting antiviral properties to the surface of an article, which includes the steps of immersing the article in an aqueous solution containing methylhesperidin, and drying the article after the immersion.
In one embodiment, the invention provides methylhesperidin for inhibiting viral infection. The present invention also provides methylhesperidin for the prevention of viral infections or viral infections.
In one embodiment, the present invention provides the use of methylhesperidin for the manufacture of a composition for inhibiting viral infection. The invention also provides the use of methylhesperidin for the manufacture of a pharmaceutical composition for preventing viral infections. The invention also provides the use of methylhesperidin for the manufacture of a pharmaceutical composition for preventing viral infections.
In one embodiment, the invention provides the use of methylhesperidin for inhibiting viral infection. The invention also provides the use of methylhesperidin to prevent viral infections. The invention also provides the use of methylhesperidin to prevent viral infections.

Hereinafter, the present invention will be explained in more detail with reference to Examples and Comparative Examples, but the present invention is not limited to the following Examples.

[Methylhesperidin]
Methylhesperidin (product name: Methylhesperidin) sold by Resonac Co., Ltd. was used. In this product, the total content of the chalcone bodies-1 to 3 and the flavanone bodies-1 to 4 is 97.5% by mass or more based on the total amount of the composition.

(Example 1)
[Inhibitory effect on binding of SARS-CoV-2 to membrane protein]
Using RayBiotech Life, Inc.'s COVID-19 Spike-ACE2 binding Assay Kit, the inhibitory effect of methylhesperidin on the binding of the SARS-Cov-2 virus to human ACE2 was investigated.

A 0.5% (V/V) aqueous solution of methylhesperidin was prepared by dissolving liquid methylhesperidin in purified water. In addition, hesperidin (manufactured by Tokyo Kasei Kogyo Co., Ltd.) was dissolved in 10% (V/V) dimethyl sulfoxide (DMSO) (manufactured by Fuji Film Wako Pure Chemical Industries, Ltd.), and 0.5% (V/V) hesperidin was added. A solution was prepared. According to the instructions provided with the kit, the methylhesperidin aqueous solution or hesperidin solution was mixed with the ACE2 protein solution provided with the kit. The mixture was added to the COVID-19 S-protein adsorption plate included in the kit, and reacted at 4° C. with shaking overnight. A methylhesperidin control was prepared by adding the same amount of water instead of the methylhesperidin aqueous solution. A hesperidin control was prepared by adding the same amount of 10% (V/V) DMSO solution instead of the hesperidin solution. Thereafter, a horseradish-derived peroxidase-conjugated anti-goat IgG antibody, which is a secondary antibody included in the kit, was added to the plate and allowed to react. After washing the plate, a peroxidase coloring substrate 3,3',5,5'-tetramethylbenzimide (attached to the kit) was added to the plate to perform a coloring reaction. After the color reaction, absorbance was measured at a wavelength of 450 nm using a microplate reader (manufactured by TECAN).

The results are shown in Table 5. The virus binding rate in each case was shown as a relative binding rate when the virus binding rate in the control was set as 1.00. From the results shown in Table 5, it was confirmed that methylhesperidin has the effect of inhibiting the binding of SARS-CoV-2 to ACE2. On the other hand, no binding inhibitory effect was observed with hesperidin. These results indicate that methylhesperidin has an inhibitory effect on the binding of SARS-CoV-2 to membrane proteins.

(Example 2)
[Example of application to medical supplies]
Commercially available natural rubber disposable gloves (TouchNTuff69-318, manufactured by Ansell Corporation) were immersed in a 1% (V/V) methylhesperidin aqueous solution or a 1% (V/V) hesperidin solution for 10 minutes. After soaking, the water was drained from the gloves, and the gloves were dried in an oven at about 60° C. for 3 hours. A 5 cm x 5 cm square film was cut from the dried glove, and its antiviral properties against influenza virus (influenza A/Kitakyushu/159/93 (H3N2)) were measured using a plaque method.

The cut sample was placed in a plastic Petri dish, 50 μL of influenza virus suspension was added dropwise thereto, and the mixture was allowed to react at room temperature for 1 hour. Thereafter, 950 μL of SCDLP medium was added, and the virus was washed out and collected by pipetting. Thereafter, each virus washout solution was serially diluted 10-fold with MEM dilution solution until the concentration of each virus was 1/1-1,000,000 to 1/1,000,000. 100 μL of the sample solution was inoculated into Madin-Darby Canine Kidney (MDCK: derived from dog kidney tubular epithelial cells) cells cultured in a petri dish. After allowing the virus to adsorb to the cells by standing for 1 hour, a 0.7% agar medium was overlaid and cultured for 48 hours at 34° C. in a 5% CO ₂ incubator. Thereafter, formalin fixation was performed, and the formed plaques were stained with methylene blue staining, and the number of plaques was counted. Based on the counted number of plaques, the viral infectivity (PFU/0.1 mL, Log10); (PFU: plaque-forming units) was calculated. As a control, a virus solution that had not come into contact with the sample was used instead of the washing solution from the sample cut out from the glove.

The results are shown in Table 6. When contacted with the sample coated with methylhesperidin, the virus infectivity titer was lower than when contacted with the control and the sample coated with hesperidin. From these results, it was confirmed that applying methylhesperidin to medical supplies has a high antiviral effect against influenza viruses.

(Example 3)
[Example of application to nonwoven fabric]
A nonwoven fabric made of polyethylene terephthalate having a basis weight of 80 g/m ² was immersed in a 2% (V/V) methylhesperidin aqueous solution or a 2% (V/V) hesperidin solution so that the pickup rate was 200%. Thereafter, the nonwoven fabric was dried at 110°C to prepare a processed nonwoven fabric. The pickup rate is calculated by the following formula.
Pickup rate (%) =
[(Nonwoven fabric weight after dipping - Nonwoven fabric weight before dipping)/Nonwoven fabric weight before dipping] x 100

The processed nonwoven fabric prepared as described above was subjected to an antiviral test against coronavirus (SARS-CoV-2) in accordance with ISO 18184. The test method is almost the same as in Example 2. As a control, a virus solution that had not come into contact with the processed nonwoven fabric was used instead of the washing solution from the processed nonwoven fabric. Based on the number of plaques formed, the infectious titer (PFU/0.1 mL, Log10) of the virus was calculated.

The results are shown in Table 7. When a processed nonwoven fabric coated with methylhesperidin was used, the viral infection titer of the coronavirus was below the detection limit, demonstrating high antiviral properties. From these results, it was confirmed that applying methylhesperidin to a nonwoven fabric has a high antiviral effect against coronavirus.

(Example 4)
The spray liquid of Formulation Example 1 shown in Table 8 was filled into a spray container. A spray liquid was sprayed from this spray container onto the evaluation surface. Methylhesperidin was detected in an amount of 0.1 to 10 μg/cm ² on the evaluation surface sprayed with the spray solution. This indicates that a sufficient effect of suppressing viral infection can be expected by spraying a spray solution containing 2% by mass of methylhesperidin.

(Example 5)
The aerosol spray liquid of Formulation Example 2 shown in Table 9 was filled into a spray container. A spray liquid was sprayed onto the evaluation surface from this aerosol spray container. Methylhesperidin was detected in an amount of 0.5 to 10 μg/cm ² on the evaluation surface sprayed with the spray solution. This indicates that a sufficient effect of suppressing viral infection can be expected by spraying an aerosol spray solution containing 2% by mass of methylhesperidin.

(Example 6)
A spray container was filled with the skin spray dispersion liquid of Formulation Example 3 shown in Table 10. A spray liquid was sprayed from this spray container onto the evaluation surface. Methylhesperidin was detected in an amount of 1 to 10 μg/cm ² on the evaluation surface sprayed with the spray solution. This indicates that a sufficient effect of suppressing viral infection can be expected by spraying a skin spray containing 1% by mass of methylhesperidin.

(Example 7)
A nasal spray solution of formulation 4 shown in Table 11 was filled into a nasal spray container. After applying this nasal spray, methylhesperidin was detected in an amount of 0.1 to 5 μg/cm ² on the applied surface. This indicates that application of a nasal medicated solution containing 0.5% by mass of methylhesperidin can be expected to have a sufficient effect of suppressing viral infection.

Although preferred embodiments of the present invention have been described above, the present invention is not limited to these embodiments. Additions, omissions, substitutions, and other changes to the configuration are possible without departing from the spirit of the invention. The invention is not limited by the foregoing description, but only by the scope of the appended claims.

Claims

A virus infection inhibitor containing methylhesperidin as an active ingredient.
The methylhesperidin is one or more selected from the group consisting of chalcone methylhesperidin represented by the following general formula (1) and flavanone methylhesperidin represented by the following general formula (2). 1. The virus infection inhibitor according to 1.

[In formula (1), R 1 to R 9 are each independently a methyl group or a hydrogen atom. However, at least one of R 1 to R 9 is a methyl group.
In formula (2), R 11 to R 18 are each independently a methyl group or a hydrogen atom. However, at least one of R 11 to R 18 is a methyl group. ]
The methylhesperidin is one or more selected from the group consisting of chalcone methylhesperidin represented by the following general formula (3) and flavanone methylhesperidin represented by the following general formula (4). 1. The virus infection inhibitor according to 1.

[In formula (3), R 20 to R 23 are each independently a methyl group or a hydrogen atom. In formula (4), R 24 to R 25 are each independently a methyl group or a hydrogen atom. ]
The chalcone methylhesperidin represented by the general formula (3) is one or more selected from the group consisting of chalcone-1 to 3 having a combination of R 20 to R 23 shown in Table 1 below, The virus infection inhibitor according to claim 3.
The flavanone methylhesperidin represented by the general formula (4) is one or more selected from the group consisting of flavanones-1 to 4 having a combination of R 24 to R 25 shown in Table 2 below. The virus infection inhibitor according to claim 3 or 4.
Any one of claims 1 to 4, wherein the virus is at least one virus selected from the group consisting of Orthomyxoviridae, Paramyxoviridae, Retroviridae, Filoviridae, and Coronaviridae. Viral infection inhibitors described in section.
The virus infection inhibitor according to claim 6, wherein the virus is a SARS-related coronavirus.
A method for using a virus infection inhibitor, which comprises applying the virus infection inhibitor according to any one of claims 1 to 4 to the surface of an article.
A method for using a virus infection inhibitor, which comprises spraying the virus infection inhibitor according to any one of claims 1 to 4 on the body surface of an animal.
A method for using a virus infection inhibitor, which comprises spraying or blending the virus infection inhibitor according to any one of claims 1 to 4 onto fibers.
A composition for suppressing viral infection, comprising the viral infection suppressing agent according to any one of claims 1 to 4, and a pharmaceutically acceptable carrier.