Coronavirus cell entry inhibitor

Abstract

To provide agents that inhibit coronavirus invasion into cells by effectively inhibiting an infection factor on a human (host) side and are useful for preventing or suppressing coronavirus infection.SOLUTION: The invention provides an inhibitor of the invasion of coronavirus into cells, where the inhibitor contains at least one ingredient selected from tetradecene sulfonate, lauryl sulfonate, lauroylmethyl taurine salt, aluminum lactate, potassium nitrate, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, coconut oil fatty acid amidopropyl betaine, lauroyl sarcosine salt, tranexamic acid, epsilon-aminocaproic acid and copper gluconate.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an inhibitor of coronavirus cell entry.

Background Art

[0002] Since the oral cavity can be an infection route for upper respiratory tract infections including coronavirus disease 2019 (COVID-19), daily oral care behaviors such as brushing teeth may help prevent infection. So far, it has been reported that components contained in oral care products such as dentifrices and mouthwashes act on the novel coronavirus and reduce its infectivity by destroying the virus's function (Non-Patent Documents 1 and 2).

[0003] On the other hand, in the oral cavity, core factors involved in the intracellular infection of the novel coronavirus on the human (host) side, such as angiotensin-converting enzyme 2 (ACE2) as a receptor and transmembrane serine protease 2 (TMPRSS2), are expressed (Non-Patent Document 3), and the oral cavity is considered to be an infection route for the novel coronavirus (promoting invasion into the living body and infection of the upper respiratory tract due to an increase in the number of viruses).

[0004] As an antiviral agent containing components included in oral care products such as dentifrices and mouthwashes, Patent Document 1 describes that tranexamic acid has anti-influenza virus activity.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Non-Patent Documents

[0006]

Non-Patent Document 1

[0007] Although research has been conducted focusing on the effects of ingredients in oral care products such as toothpaste and mouthwash on the virus itself, the effects of oral care ingredients on human (host) infectious agents, which are important for the sustained prevention of coronavirus infection, have not been clarified.

[0008] The present invention aims to provide a viral cell entry inhibitor that can exert effective inhibitory and deterrent effects against infectious agents on the human (host) side, and is useful for inhibiting, preventing, or suppressing coronavirus infection. [Means for solving the problem]

[0009] The present invention provides the following [1] to

[13] . [1] A coronavirus cell entry inhibitor comprising at least one selected from tetradecenesulfonate, lauryl sulfate, lauroyl methyl taurate, aluminum lactate, potassium nitrate, 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, coconut oil fatty acid amidopropyl betaine, lauroyl sarcosine, tranexamic acid, epsilon-aminocaproic acid, and copper gluconate. [2] The agent described in [1], which inhibits the formation of a binding between the coronavirus spike protein and a receptor on the cell surface. [3] The agent according to [2], comprising at least one selected from 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, coconut oil fatty acid amidopropyl betaine, tetradecene sulfonate, lauryl sulfate, lauroyl methyl taurate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine. [4] The agent described in [1], which inhibits the activation of spike proteins bound to cell surface receptors by transmembrane serine protease 2 (TMPRSS2). [5] The agent described in [4], wherein inhibition of spike protein activation is inactivation of transmembrane serine protease 2 (TMPRSS2). [6] The agent according to [4] or [5], comprising at least one selected from lauroyl methyl taurate, tetradecene sulfonate, tranexamic acid, epsilon-aminocaproic acid, lauryl sulfate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine salt. [7] The agent described in any one of the items [2] to [6], wherein the receptor is angiotensin-converting enzyme 2. [8] The agent described in [1] that inhibits the expression of transmembrane serine protease 2 (TMPRSS2). [9] The agent described in [8], comprising lauryl sulfate.

[10] An agent described in any one of items [1] to [9] for the prevention of coronavirus infection, or for the suppression or alleviation of its symptoms.

[11] The agent according to any one of items [1] to

[10] , wherein the coronavirus is SARS-CoV-1, CoV-NL63, or SARS-CoV-2.

[12] A preparation for external use on mucous membranes, as described in any one of items [1] to

[11] .

[13] An oral, nasal, or ophthalmic preparation as described in any one of items [1] to

[12] . [Effects of the Invention]

[0010] According to the present invention, a cell entry inhibitor is provided that can exert an effective inhibitory effect on the core factors involved in infection on the human (host) side, and is useful for preventing or suppressing coronavirus infection. [Modes for carrying out the invention]

[0011] [1. Active ingredients] The agent of the present invention comprises at least one selected from tetradecenesulfonate, lauryl sulfate, lauroyl methyl taurate, aluminum lactate, potassium nitrate, 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, coconut oil fatty acid amidopropyl betaine, lauroyl sarcosine, tranexamic acid, epsilon-aminocaproic acid, and copper gluconate.

[0012] Tetradecenesulfonate, lauryl sulfate, lauroylmethyltaurate, and lauroyl sarcosine salt may be any pharmacologically acceptable salt of their respective type. Examples of pharmacologically acceptable salts include base addition salts and amino acid salts. Specific examples include inorganic base salts such as sodium salt, potassium salt, calcium salt, magnesium salt, and ammonium salt; organic base salts such as triethylammonium salt, triethanolammonium salt, pyridinium salt, and diisopropylammonium salt; and basic amino acid salts such as arginine salt. Among these, inorganic base salts are preferred, with alkali metal salts (e.g., sodium salt, potassium salt) or ammonium salts being preferred, alkali metal salts being more preferred, and sodium salts being even more preferred.

[0013] The agent of the present invention may contain one or more of the above-mentioned active ingredients, and preferably contains any of the following A to E. A) At least one selected from 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, coconut oil fatty acid amidopropyl betaine, tetradecenesulfonate, lauryl sulfate, lauroyl methyl taurate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine; B) At least one selected from lauroyl methyl taurate, tetradecene sulfonate, tranexamic acid, epsilon-aminocaproic acid, lauryl sulfate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine salt; C) At least one selected from tetradecenesulfonate, lauryl sulfate, lauroyl methyltaurate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine; D) A combination of at least one selected from 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, coconut oil fatty acid amidopropyl betaine, tetradecene sulfonate, lauryl sulfate, lauroyl methyl taurate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine salt, and at least one selected from tranexamic acid and epsilon-aminocaproic acid; E) A combination of at least one selected from 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine and coconut oil fatty acid amidopropyl betaine, and at least one selected from lauroyl methyl taurate, tetradecene sulfonate, tranexamic acid, epsilon-aminocaproic acid, lauryl sulfate, aluminum lactate, potassium nitrate, copper gluconate, and lauroyl sarcosine salt; and F) Lauryl sulfate.

[0014] Since the active ingredient is the above A, it is possible to inhibit the formation of the binding between the spike protein (S protein) of the coronavirus and the receptor ACE2. Since it is B, it is possible to inhibit the TMPRSS2 activity that contributes to the activation of the S protein bound to the receptor ACE2. By any of the combinations of C to E, it is possible to inhibit the formation of the binding between the S protein of the coronavirus and the receptor ACE2 and to inhibit the TMPRSS2 activity that contributes to the activation of the S protein bound to the receptor ACE2. Since it is F, it is possible to suppress the expression of TMPRSS2.

[0015] [2. Content of active ingredient] The agent of the present invention may be in the form of a so-called composition containing other optional components as necessary. The optional components will be described later. The content of the active ingredient (amount relative to 100% by mass of the agent) when the agent of the present invention contains optional components varies depending on the type of the active ingredient. For example, in an oral agent, when a human 15 years old or older uses about 1 g or more (toothpaste) or about 10 mL or more (for example, liquid toothpaste) once a day, it is as follows.

[0016] - Content of tetradecene sulfonate - The amount of tetradecene sulfonate is preferably 0.007% by mass or more, or 0.01% by mass or more, more preferably 0.02% by mass or more, still more preferably 0.03% by mass or more, or 0.1% by mass or more. The upper limit is preferably 25% by mass or less, more preferably 10% by mass or less, still more preferably 5% by mass. From the aspect of inhibiting the formation of the binding between the S protein and the receptor, it is preferably 0.007% by mass to 25% by mass, more preferably 0.02% by mass to 10% by mass, still more preferably 0.1% by mass to 5% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.01% by mass to 25% by mass, more preferably 0.02% by mass to 10% by mass, still more preferably 0.03% by mass to 5% by mass.

[0017] - Content of lauryl sulfate - The amount of lauryl sulfate is preferably 0.01% by mass or more, or 0.02% by mass or more, more preferably 0.05% by mass or more, and even more preferably 0.5% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 2% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.02% by mass to 10% by mass, more preferably 0.05% by mass to 5% by mass, and even more preferably 0.5% by mass to 2% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.01% by mass to 10% by mass, more preferably 0.02% by mass to 5% by mass, and even more preferably 0.05% by mass to 2% by mass.

[0018] - Lauroyl methyl taurate content - The amount of lauroyl methyl taurate salt is preferably 0.01% by mass or more, or 0.03% by mass or more, more preferably 0.05% by mass or more, and even more preferably 0.08% by mass or more, or 0.1% by mass or more. The upper limit is preferably 25% by mass or less, more preferably 10% by mass or less, and even more preferably 5% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.01% by mass to 25% by mass, more preferably 0.05% by mass to 10% by mass, and even more preferably 0.1% by mass to 5% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.03% by mass to 25% by mass, more preferably 0.05% by mass to 10% by mass, and even more preferably 0.08% by mass to 5% by mass.

[0019] -Aluminum lactate content- The amount of aluminum lactate is preferably 0.02% by mass or more, or 0.05% by mass or more, preferably 0.1% by mass or more, or 0.2% by mass or more, and more preferably 0.3% by mass or more, or 0.5% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, or 3% by mass or less, and even more preferably 2.5% by mass or less, or 2% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.2% by mass to 5% by mass, more preferably 0.3% by mass to 3% by mass, and even more preferably 0.5% by mass to 2% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.02% by mass to 10% by mass, more preferably 0.05% by mass to 5% by mass, and even more preferably 0.1% by mass to 2.5% by mass.

[0020] -Potassium nitrate content- The amount of potassium nitrate is preferably 0.01% by mass or more, or 0.02% by mass or more, more preferably 0.1% by mass or more, or 0.5% by mass or more, and even more preferably 1% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 2% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.01% by mass to 10% by mass, more preferably 0.5% by mass to 5% by mass, and even more preferably 1% by mass to 2% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.02% by mass to 10% by mass, more preferably 0.1% by mass to 10% by mass, and even more preferably 0.5% by mass to 5% by mass.

[0021] -2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine content- The amount of 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine is preferably 0.05% by mass or more, more preferably 0.09% by mass or more, and even more preferably 0.15% by mass or more. The upper limit is preferably 6% by mass or less, more preferably 3% by mass or less, and even more preferably 1.2% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, the amount is preferably 0.05% to 6% by mass, more preferably 0.09% to 3% by mass, and even more preferably 0.15% to 1.2% by mass.

[0022] -Coconut oil fatty acid amidopropyl betaine content- The amount of coconut oil fatty acid amidopropyl betaine is preferably 0.025% by mass or more, more preferably 0.08% by mass or more, and even more preferably 0.4% by mass or more. The upper limit is preferably 8% by mass or less, more preferably 4% by mass or less, and even more preferably 1.6% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, the amount is preferably 0.025% by mass to 8% by mass, more preferably 0.08% by mass to 4% by mass, and even more preferably 0.4% by mass to 1.6% by mass.

[0023] - Lauroyl sarcosine salt content - The amount of lauroyl sarcosine salt is preferably 0.01% by mass or more, or 0.03% by mass or more, more preferably 0.05% by mass or more, and even more preferably 0.07% by mass or more, or 0.12% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 2% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.03% by mass to 10% by mass, more preferably 0.05% by mass to 5% by mass, and even more preferably 0.07% by mass to 2% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.01% by mass to 10% by mass, more preferably 0.03% by mass to 5% by mass, and even more preferably 0.12% by mass to 2% by mass.

[0024] -Tranexamic acid content- The amount of tranexamic acid is preferably 0.005% by mass or more, more preferably 0.01% by mass or more, and even more preferably 0.015% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 1% by mass or less. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.005% by mass to 10% by mass, more preferably 0.01% by mass to 5% by mass, and even more preferably 0.015% by mass to 1% by mass.

[0025] -Epsilon-aminocaproic acid content- The amount of epsilon-aminocaproic acid is preferably 0.005% by mass or more, more preferably 0.01% by mass or more, and even more preferably 0.015% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 5% by mass or less, and even more preferably 1% by mass or less. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.005% by mass to 10% by mass, more preferably 0.01% by mass to 5% by mass, and even more preferably 0.015% by mass to 1% by mass.

[0026] -Copper gluconate content- The amount of copper gluconate is preferably 0.004% by mass or more, or 0.008% by mass or more, more preferably 0.01% by mass or more, or 0.02% by mass or more, and even more preferably 0.05% by mass or more, or 0.09% by mass or more. The upper limit is preferably 10% by mass or less, more preferably 2% by mass or less, and even more preferably 1% by mass or less. From the viewpoint of inhibiting the formation of binding between the S protein and the receptor, it is preferably 0.008% by mass to 10% by mass, more preferably 0.02% by mass to 2% by mass, and even more preferably 0.09% by mass to 1% by mass. From the viewpoint of inhibiting the activation of the S protein bound to the receptor, it is preferably 0.004% by mass to 10% by mass, more preferably 0.01% by mass to 2% by mass, and even more preferably 0.05% by mass to 1% by mass.

[0027] [3. Cell invasion inhibitory effect] The agent of the present invention can suppress the entry of coronavirus into cells. In this specification, suppressing cell entry means inhibiting, removing, or reducing the activity of at least one factor involved in the entry of the virus into cells. The general process of coronavirus cell entry is as follows: The S protein, a membrane protein of the coronavirus, binds to a receptor on the surface of human cells (on the cell membrane) to form an S protein-receptor binding. Subsequently, the S protein is activated by TMPRSS2 on the human cell membrane (a portion of the S protein is cleaved, causing the fusion site to protrude to the human cell membrane), and membrane fusion is induced. The agent of the present invention preferably inhibits at least one factor involved in the above process, and preferably inhibits the formation of the S protein-receptor binding, inhibits the activation of the S protein bound to the receptor (preferably inactivates TMPRSS2), inhibits TMPRSS2 expression, or a combination of two or more selected from these, i.e., it preferably is an S protein-receptor binding inhibitor, an S protein activation inhibitor (preferably a TMPRSS2 inactivator), and a TMPRSS2 expression inhibitor. As a result, the entry of the virus into cells can be suppressed, making it possible to prevent or suppress coronavirus infection.

[0028] Examples of receptor proteins on the cell membrane include angiotensin-converting enzyme 2 (ACE2), aminopeptidase N (CD13), dipeptidyl peptidase 4 (DPPIV, CD26), and O-acetylated sialic acid, with ACE2 being preferred.

[0029] Examples of coronaviruses include coronaviruses that use ACE2 as a receptor, such as SARS-CoV-1, CoV-NL63, and SARS-CoV-2; coronaviruses that use CD13 as a receptor, such as CoV-OC43 and CoV-HKU1; and coronaviruses that use DPPIV as a receptor, such as MERS-CoV. Coronaviruses that use ACE2 as a receptor are preferred, and SARS-CoV-2 is more preferred.

[0030] [4. Dosage Form] Examples of dosage forms include liquid, syrup, cream, paste, tablets, capsules, powder (granules, fine granules), soft capsules (soft capsules such as gelatin-based capsules), and hard capsules. The appropriate dosage form should be selected according to the administration method.

[0031] [5. Method of administration and target population] Methods of administering the drug include, for example, oral administration (e.g., oral administration, sublingual administration) and parenteral administration (e.g., transdermal administration, intravenous administration, intramuscular administration, subcutaneous administration, nasal administration, transpulmonary administration). Among these, less invasive administration methods are preferred, with transdermal administration (topical application) and oral administration (oral administration) being more preferred, and mucosal administration (topical application to the mucosa) being even more preferred.

[0032] The target of administration can be any animal, including humans, but is usually humans. The target of administration may be healthy individuals, but may also be individuals infected with or suspected of being infected with coronavirus infection (preferably infection caused by SARS-CoV-1, CoV-NL63, or SARS-CoV-2). Examples of animals other than humans include mammals such as mice, rats, hamsters, dogs, cats, sheep, goats, cattle, pigs, and monkeys.

[0033] [6.Optional ingredients] When the agent of the present invention is in the form of a so-called composition containing other optional components, examples of other components include, but are not limited to, pharmaceutically active ingredients, buffering agents, solubilizers, isotonic agents, stabilizers, chelating agents, pH adjusters, preservatives, oily components, excipients, disintegrants, binders, lubricants, coating agents, colorants, color fixatives, flavoring agents (acidulants, fragrances, sweeteners), antioxidants, strengthening agents, leavening agents, thickeners, surfactants, abrasives, wetting agents, moisturizing agents, cooling agents, abrasives, binders, astringents, plant extracts, ultraviolet absorbers, aqueous solvents, preservatives, seasonings, and food ingredients (including food additives). The type and content of optional components may be selected according to the intended use of the pharmaceutical, quasi-drug, food composition, and cosmetic, as well as the dosage form, administration method, etc., and may be one type or a combination of two or more types.

[0034] -Medicinal ingredients- The active ingredients include, for example, enzymes such as dextranase, mutanase, amylase, protease, and Litec enzyme; fluorides such as sodium fluoride, sodium monofluorophosphate, and tin fluoride; anti-inflammatory agents such as allantoin, glycyrrhizinate (e.g., dipotassium glycyrrhizinate), glycyrrhetinic acid, glycyrrhetinic acid derivatives (e.g., stearyl glycyrrhetinate), allantoin chlorohydroxyaluminum, azulene, and dihydrocholesterol; metal salts such as zinc salts, copper salts, and tin salts; condensed phosphates, ethane hydroxydiphosphate Examples of effective ingredients include: tartar preventatives such as ionate; blood flow promoters such as vitamin E (e.g., tocopherol acetate); hypersensitivity inhibitors such as strontium chloride; coating agents such as hydroxyethylcellulose dimethyldiallylammonium chloride; astringents such as vitamin C (e.g., ascorbic acid or its salts), lysozyme chloride, and sodium chloride; water-soluble copper compounds such as copper chlorophyll; tartar preventatives; amino acids such as alanine, glycine, and proline; plant extracts such as thyme, scutellaria baicalensis, clove, and witch hazel; caropeptides; and polyvinylpyrrolidone. Other examples include decongestants, anti-inflammatory agents, astringents, antihistamines, vitamins, amino acids, disinfectants, local anesthetics, components other than the active ingredients in the present invention that have an inhibitory effect on coronavirus cell entry, and combinations of two or more of these. Examples of decongestants include naphazoline hydrochloride, tetrahydrozoline hydrochloride, phenylephrine hydrochloride, epinephrine, epinephrine hydrochloride, ephedrine hydrochloride, dl-methylephedrine hydrochloride, tetrahydrozoline nitrate, and naphazoline nitrate. Examples of anti-inflammatory and astringent agents include neostigmine methyl sulfate, allantoin, berberine chloride, berberine sulfate, zinc sulfate, zinc lactate, lysozyme chloride, bromelain, dipotassium glycyrrhizinate, ammonium glycyrrhizinate, glycyrrhetinic acid, methyl salicylate, sodium azulene sulfonate, chamomile, and sodium cromoglycate. Examples of antihistamines include iproheptin hydrochloride, diphenhydramine hydrochloride, diphenhydramine, isotipendyl hydrochloride, and chlorpheniramine maleate.Examples of vitamins include flavin adenine dinucleotide sodium, pyridoxine hydrochloride, cyanocobalamin, vitamin A compounds (e.g., retinol acetate, retinyl palmitate), and vitamin E compounds (e.g., tocopherol acetate (e.g., d-α-tocopherol acetate)). Examples of amino acids include potassium L-aspartate, magnesium L-aspartate, aminoethylsulfonic acid, and sodium chondroitin sulfate. Examples of disinfectants include cetylpyridinium chloride and chlorhexyl gluconate. Examples of pharmacokinetics include din, decalinium chloride, benzalkonium chloride, benzethorium chloride, iodine, potassium iodide, sulfamethoxazole, sulfamethoxazole sodium, sulfisoxazole, sulfisomidine sodium, isopropylmethylphenol, and hinokitiol. Examples of local anesthetics include lidocaine, lidocaine hydrochloride, dibucaine hydrochloride, and chlorobutanol. Each active ingredient may be used individually or in combination of two or more. The effective amount of each active ingredient can be appropriately determined according to conventional methods.

[0035] - Surfactants - The surfactant may be anionic, nonionic, or amphoteric.

[0036] Examples of anionic surfactants include alkyl sulfates, acyl amino acid salts, acyl taurine salts, α-olefin sulfonates, hydrogenated coconut fatty acid monoglyceride monosulfates, and lauryl sulfoacetate. The alkyl and acyl groups may be linear or branched, saturated or unsaturated, and typically have 10 to 20 carbon atoms, preferably 12 to 18, and more preferably 12 to 14. The salt can be selected from pharmacokinetically acceptable salts. Examples of pharmacokinetically acceptable salts include base addition salts and amino acid salts. Specific examples include inorganic base salts such as sodium salts, potassium salts, calcium salts, magnesium salts, and ammonium salts; organic base salts such as triethylammonium salts, triethanolammonium salts, pyridinium salts, and diisopropylammonium salts; and basic amino acid salts such as arginine salts. Among these, inorganic base salts are preferred, alkali metal salts (e.g., sodium salts, potassium salts) or ammonium salts are more preferred, and sodium salts are even more preferred. Examples of alkyl sulfates include myristyl sulfate. Examples of acyl amino acid salts include acyl glutamates such as lauroyl glutamate, myristoyl glutamate, and palmitoyl glutamate; acyl glycine salts such as N-lauroyl-N-methylglycine salt and cocoyl glycine salt; acyl alanine salts such as N-lauroyl-β-alanine salt, N-myristyl-β-alanine salt, N-cocoyl-β-alanine salt, N-lauroyl-N-methyl-β-alanine salt, N-myristoyl-N-methyl-β-alanine salt, and N-methyl-N-acyl alanine salt; and acyl aspartates such as lauroyl aspartate. Examples of acyl taurine salts include N-methyl-N-acyl taurine salt and N-cocoyl methyl taurine salt. Examples of anionic surfactants include, for instance, hydrogenated coconut fatty acid monoglyceride sodium monosulfate and sodium lauryl sulfoacetate.

[0037] Examples of nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene hydrogenated castor oil, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monostearate), alkylolamides, polyoxyethylene fatty acid esters, polyoxyethylene alkenyl ethers, polyglycerin fatty acid esters, sucrose fatty acid esters (e.g., maltose fatty acid esters), sugar alcohol fatty acid esters (e.g., maltitol fatty acid esters, lactitol fatty acid esters), fatty acid diethanolamides (e.g., lauric acid mono or diethanolamide), polyoxyethylene polyoxypropylene copolymers, and polyoxyethylene polyoxypropylene fatty acid esters. The number of carbon atoms in the alkyl chain of polyoxyethylene alkyl ethers is usually 14 to 18, and the average number of moles of ethylene oxide added is usually 5 to 30 moles. The average number of moles of ethylene oxide added to polyoxyethylene hydrogenated castor oil is usually 20 to 100 moles, preferably 20 to 60 moles. The number of carbon atoms in the fatty acids of sorbitan fatty acid esters is typically 12 to 18. The number of carbon atoms in the fatty acids of polyoxyethylene sorbitan fatty acid esters is typically 16 to 18, and the average number of moles of ethylene oxide added is typically 10 to 40 moles. The number of carbon atoms in the alkyl chain of alkylolamides is typically 12 to 14.

[0038] Examples of amphoteric surfactants include betaine-type amphoteric surfactants such as alkyldimethylaminoacetic acid betaine (e.g., lauryldimethylaminoacetic acid betaine) and fatty acid amidopropyldimethylaminoacetic acid betaine; imidazoline-type amphoteric surfactants such as N-fatty acid acyl-N-carboxymethyl-N-hydroxyethylethylenediamine salts and coconut oil fatty acid imidazolinium betaine; and alkylbetaines such as lauryldimethylaminoacetic acid betaine.

[0039] When surfactants are included, the content of anionic, nonionic, and amphoteric surfactants is usually 0.01 to 10% by mass of the total agent, preferably 0.1 to 5% by mass, and more preferably 0.2 to 3% by mass.

[0040] -Abrasive- The abrasive can be either an inorganic abrasive or an organic abrasive. Examples of inorganic abrasives include abrasive silica such as precipitated silica, aluminosilicate, zirconosilicate, crystalline zirconium silicate, and titanium-bonded silica; calcium phosphate compounds such as dicalcium phosphate dihydrate or anhydrous, monocalcium phosphate, tricalcium phosphate, and calcium pyrophosphate; calcium carbonate abrasives such as calcium carbonate; calcium abrasives other than carbonate / phosphate, such as calcium hydroxide and calcium sulfate; aluminum-based materials such as aluminum oxide, aluminum hydroxide, and alumina; silicate-based materials such as anhydrous silicic acid, zeolite, and zirconium silicate; magnesium-based materials such as magnesium carbonate and trimagnesium phosphate; apatite-based materials such as hydroxyapatite, fluoroapatite, and calcium-deficient apatite; titanium-based materials such as titanium dioxide, titanium mica, and titanium oxide; and minerals such as bentonite. Examples of organic abrasives include polymethyl methacrylate and synthetic resin-based abrasives. Of these, abrasive silica and calcium phosphate compounds are preferred, and anhydrous silicic acid is more preferred. The amount of abrasive is preferably 50% by mass or less of the total amount of the agent, and more preferably 8 to 50% by mass.

[0041] -Wetting agent- Examples of humectants include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of sugar alcohols include sorbitol, erythritol, maltitol, lactitol, and xylitol. Examples of polyhydric alcohols other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and polyethylene glycol (PEG); and reduced starch saccharifies. As for polyethylene glycol, polyethylene glycol with an average molecular weight of 150 to 6000 is preferred, and polyethylene glycol with an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600) is preferred. The average molecular weight is the average molecular weight described in the 2006 Standards for Raw Materials of Quasi-Drugs. The content of the humectant is usually 40% by mass or less of the total amount of the agent, and preferably 1 to 30% by mass.

[0042] - Binding agent - Examples of binders include conventionally known and suitable organic binders, such as polysaccharides, cellulosic binders (e.g., carboxymethylcellulose (CMC), hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, cationized cellulose, etc.), other polysaccharide thickeners (e.g., xanthan gum, guar gum, gellan gum, tragacanth gum, karaya gum, arabic gum, locust bean gum, carrageenan, sodium alginate), and synthetic water-soluble polymers (e.g., sodium polyacrylate, carboxyvinyl polymer, polyvinylpyrrolidone, polyvinyl alcohol, propylene glycol alginate). Furthermore, inorganic binders such as thickening silica and aluminum silicate can also be included. The content of organic binders is preferably 0 to 3% by mass, more preferably 0.1 to 2% by mass, relative to the total amount of the agent. The content of inorganic binders is preferably 0 to 10% by mass, more preferably 1 to 8% by mass.

[0043] - Buffering agent - Examples of buffering agents include citric acid or its salts (e.g., sodium citrate), phosphoric acid or its salts (e.g., sodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium phosphate, potassium dihydrogen phosphate), tartaric acid or its salts (e.g., sodium tartrate), gluconic acid or its salts (e.g., sodium gluconate), acetic acid or its salts (e.g., sodium acetate), carbonic acid or its salts (e.g., sodium bicarbonate), trometamol, amino acids (e.g., potassium aspartate, aminoethylsulfonic acid, glutamic acid, sodium glutamate), and combinations of two or more of these.

[0044] -Solubilizer- Examples of solubilizers include propylene glycol, polyethylene glycol, and combinations of two or more of these.

[0045] -Tonicity agent- Examples of isotonic agents include sodium chloride, potassium chloride, glycerin, and combinations of two or more of these.

[0046] -Stabilizer- Examples of stabilizers include sodium edetate, cyclodextrin, sulfites, citric acid or its salts, dibutylhydroxytoluene, ascorbic acid, and combinations of two or more of these.

[0047] -Chelating agent- Examples of chelating agents include sodium edetate, sodium citrate, and combinations thereof.

[0048] -Moisturizer- Examples of humectants include glycerin, concentrated glycerin, sugar alcohols (e.g., sorbitol, xylitol, maltitol, mannitol, reduced starch syrup, reduced palatinose, erythritol, lactitol, isomalt), and combinations of two or more of these.

[0049] - Flavoring agent - Flavoring agents include, for example, sweeteners (e.g., sodium saccharin, dipotassium glycyrrhizinate, aspartame, stevia, stevioside, paramethoxycinnamaldehyde, neohesperidin dihydrochalcone, perillartin, aspartylphenylalanine methyl ester, thaumatin, acesulfame potassium, sucralose, maltitol, sorbitol, mannitol, reduced starch syrup, reduced palatinose, xylitol, erythritol, lactitol and other artificial sweeteners), and flavorings. (Examples: Natural essential oils such as anise oil, cassia oil, wintergreen oil, mastic oil, neroli oil (orange blossom oil), lemongrass oil, jasmine oil, rose oil, iris oil, clove oil, sage oil, cardamom oil, rosemary oil, laurel oil, chamomile oil, basil oil, marjoram oil, lemon oil, orange oil, lime oil, yuzu oil, nutmeg oil, lavender oil, paraclete oil, vanilla oil, cinnamon oil, pimento oil, cinnamon leaf oil, perilla oil, wintergreen oil, peppermint oil, lychee oil, etc.; menthol, carvone, cinnamic acid) Fragrance components contained in the above natural essential oils, such as sulfehyde, anethole, methyl salicylate, eugenol, linalool, limonene, menthone, menthyl acetate, citral, decanal, camphor, borneol, pinene, spiranthol, n-decyl alcohol, citronellol, α-terpineol, citronellyl acetate, ethyl linalool, vanillin, etc.; ethyl acetate, ethyl butyrate, isoamyl acetate, hexanal, hexenal, methyl anthranilate, ethyl methyl Fragrance components such as ethylphenylglycidate, benzaldehyde, vanillin, ethyl vanillin, furaneol, N-ethyl-p-menthane-3-carboxamide, menthyl lactate, and ethylene glycol-l-menthyl carbonate; and various blended flavors such as mint, fruit, and herb (e.g., Peppermint Micron X-8277-T, Drycoat Matcha #421) formed by combining several fragrance components and natural essential oils, as well as acidulants (e.g., citric acid, tartaric acid, malic acid) and green tea powder.

[0050] -Oily components- Examples of oily components include fatty acid esters (e.g., glycerin fatty acid esters), hydrocarbons (e.g., paraffin, liquid paraffin, ceresin, squalane, petrolatum, microcrystalline wax), higher fatty acids (e.g., fatty acids with 8 to 22 carbon atoms such as lauric acid, myristic acid, oleic acid, and isostearic acid), higher alcohols (e.g., alcohols with 8 to 22 carbon atoms such as lauryl alcohol, cetyl alcohol, cetostearyl alcohol, oleyl alcohol, and isostearyl alcohol), vegetable oils (e.g., vegetable oils such as olive oil, castor oil, and coconut oil; fatty acid esters such as isopropyl myristate), beeswax, and combinations of two or more of these.

[0051] - Preservatives - Examples of preservatives include para-hydroxybenzoic acid esters such as methylparaben, ethylparaben, propylparaben, and butylparaben; alcohol derivatives such as phenylethyl alcohol, benzyl alcohol, phenol, and acrinol; sorbic acid and its salts (such as potassium sorbate); benzalkonium chloride; benzethonium chloride; chlorhexidine gluconate; cetylpyridinium chloride; alkyl polyaminoethylglycine; and combinations of two or more of these.

[0052] -Wetting agent- Examples of wetting agents include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of sugar alcohols include sorbitol, erythritol, maltitol, lactitol, and xylitol. Examples of polyhydric alcohols other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and polyethylene glycol (PEG); and reduced starch saccharifies. As for polyethylene glycol, polyethylene glycol with an average molecular weight of 150 to 6000 is preferred, and polyethylene glycol with an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600) is preferred. The average molecular weight is the average molecular weight described in the 2006 Standards for Raw Materials of Quasi-Drugs. The amount of wetting agent is usually 40% by mass or less of the total amount of the agent, and preferably 1 to 35% by mass.

[0053] -Colorants- Examples of coloring agents include natural pigments such as safflower red pigment, gardenia yellow pigment, gardenia blue pigment, perilla pigment, red yeast rice pigment, red cabbage pigment, carrot pigment, hibiscus pigment, cocoa pigment, spirulina blue pigment, and tamarind pigment, as well as legally approved coloring agents such as Red No. 2, Red No. 3, Red No. 104, Red No. 105, Red No. 106, Red No. 227, Yellow No. 4, Yellow No. 5, Green No. 3, and Blue No. 1, and riboflavin, copper chlorophyllin sodium, and titanium dioxide. When coloring agents are included, their content is preferably 0.00001 to 3% by mass of the total agent.

[0054] - pH adjuster - Examples of pH adjusting agents include organic acids such as phthalic acid, citric acid, succinic acid, acetic acid, fumaric acid, malic acid, and lactic acid, or their salts (sodium citrate); inorganic acids such as phosphoric acid (orthophosphoric acid), or their salts (e.g., potassium salts, sodium salts, and ammonium salts); and hydroxides such as sodium hydroxide and potassium hydroxide. Examples of inorganic salts include disodium hydrogen phosphate and sodium dihydrogen phosphate. The amount of pH adjusting agent can usually be such that the pH of the agent after addition is 5 to 9, preferably 6 to 8.5. In this specification, the pH value usually refers to the value obtained 3 minutes after the start of measurement at 25°C. The pH value can be measured, for example, using a pH meter (model number Hm-30S) manufactured by Toa Denpa Kogyo Co., Ltd.

[0055] -solvent- Examples of solvents include water (purified water) and ethanol, with water being preferred. The solvent may be used alone or in combination of two or more.

[0056] - Excipients - Excipients include, for example, cellulose such as hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, crystalline cellulose, ethylcellulose, methylethylcellulose, and low-substituted hydroxypropyl cellulose, and their pharmacologically acceptable derivatives; synthetic polymers such as polyvinylpyrrolidone and partially saponified polyvinyl alcohol; polysaccharides such as gelatin, acacia powder, pullulan, agar, alginic acid, sodium alginate, and xanthan gum; corn starch, potato starch, pregelatinized starch, and hydroxypropyl starch. Starches such as thiosulfate and their pharmacologically acceptable derivatives; lactose, lactose granules, fructose, glucose, sucrose, granulated sugar, hydrated glucose, trehalose, palatinose, mannitol, sorbitol, erythritol, xylitol, maltotetraose, lactitol, isomalt, reduced palatinose, reduced starch syrup, powdered reduced maltose syrup, maltitol; inorganic excipients such as magnesium carbonate, calcium carbonate, light anhydrous silicic acid, silicon dioxide (also known as anhydrous silicic acid, fine-grained silicon dioxide), titanium dioxide, and aluminum hydroxide gel; and combinations of two or more of these.

[0057] -Disintegrant- Examples of disintegrants include crospovidone, carmellose calcium, croscarmellose sodium, low-substituted hydroxypropylcellulose, carboxymethylcellulose, carboxymethyl starch sodium, croscarmellose sodium, cross-linked insoluble polyvinylpyrrolidone, hydroxypropyl starch, partially pregelatinized starch, corn starch, and combinations of two or more of these.

[0058] -Binder- Examples of binders include hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, ethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, dextrin, starch, pregelatinized starch, and combinations of two or more of these.

[0059] -lubricant- Examples of lubricants include calcium stearate, magnesium stearate, sucrose fatty acid esters, light anhydrous silicic acid, sodium stearyl fumarate, polyethylene glycol, talc, stearic acid, and combinations of two or more of these.

[0060] -Other optional components- Examples of optional components other than those mentioned above include polyisobutylene, polybutadiene, urethane, silicone, and natural rubber. The content of these other optional components can be appropriately set within a range that does not hinder the effects of the present invention.

[0061] [Dosage form, application site] The agents of the present invention can be used as pharmaceuticals, quasi-drugs, cosmetics, and foods. Dosage forms include, but are not particularly limited to, liquids, sprays, solids, semi-solids, powders, and granules. In the case of pharmaceuticals and quasi-drugs, application sites include, for example, the oral cavity, nasal cavity, and eyes. For oral use, examples include toothpaste (e.g., toothpaste, gel toothpaste, moistened toothpaste, liquid toothpaste), mouthwash, tongue cleaner, oral spray, oral tablet, gum, mouth freshener, gargle tablets, oral paste, gel, and ointment. For nasal use, examples include nasal drops and nasal lavages. For ophthalmal use, examples include eye drops (e.g., general eye drops, eye drops for contact lens users), eye washes (general eye washes, eye washes for contact lens users), and contact lens preparations (e.g., contact lens preservatives, contact lens cleaners).

[0062] As cosmetics, it can be used in dosage forms such as creams, lotions, packs, gels, aerosols, and sheets. Specifically, examples include skin cosmetics such as lotions, serums, whitening agents, moisturizers, face masks, emulsions, foundations, eyeshadows, mascaras, eyebrow pencils, eyeliners, cheek powders, lipsticks, and lip balms; and hair cosmetics such as hair rinses, hair conditioners, hair treatments, hair lotions, hair tonics, hair packs, hair creams, conditioning mousses, hair mousses, hair sprays, shampoos, leave-in treatments, hair dyes, and hair styling products.

[0063] Examples of food products (food compositions) include food compositions with specific uses such as health foods, functional foods, health supplements, nutritional supplements, foods for specified health uses, foods with nutritional function, medical foods, foods for the sick, infant foods, foods for elderly care, and foods for the elderly.

[0064] [Manufacturing method] The method for producing the agent of the present invention may be determined according to the dosage form, use, and application site. For example, when used as a toothpaste, one method is to prepare the components that dissolve in the solvent, then mix in the other insoluble components, and degas as necessary (e.g., by reducing pressure). Another example is to prepare a composition by dispersing the active ingredient and other components used as needed in an aqueous solvent (e.g., purified water, sterile water, etc.), dissolving them, and filling it into a suitable container (e.g., glass, resin). For oral preparations, a laminate tube is an example of a container, and the material may be, for example, polyethylene, polypropylene, polyethylene terephthalate, nylon, or other resins. In the case of a spray, a container equipped with a spraying mechanism (e.g., trigger type, pump type, aerosol type container) may be selected. The resulting toothpaste can be contained in a container to make a product. The shape and material of the container are not particularly limited, and containers used for ordinary oral compositions can be used.

[0065] [How to use] The method of using the agent of the present invention may be, for example, to administer the agent to the application site. The number of administrations per day is not particularly limited, and may be, for example, 1 to 6 times, or more. In the case of an oral agent, an appropriate amount of the agent may be placed on a toothbrush and the tooth surface may be brushed, and then rinsed with water after use (toothpaste), or an appropriate amount of the agent may be taken into the mouth and gargled and then spit out (mouthwash).

Examples

[0066] Examples 1 to 16 and Comparative Examples 1 to 4 For each evaluation sample shown in Table 1, the inhibition rate of S protein-ACE2 binding and the inhibition rate of TMPRSS2 activity were evaluated.

[0067]

Table 1

[0068] The inhibition rate of S protein-ACE2 binding and the inhibition rate of TMPRSS2 activity were evaluated using the following evaluation systems, respectively (Tables 2 and 3).

[0069] <S protein-ACE2 binding evaluation system> The experiment was carried out according to the protocol of Spike S1(SARS-CoV-2):ACE2 Inhibitor Screening Colorimetric Assay Kit(Cat#.79954,BPS Bioscience)〔https: / / bpsbioscience.com / pub / media / wysiwyg / Other / 79954_2.pdf〕. (All concentrations of the control, blank, and each evaluation sample were carried out in triplicate)

[0070] 1. Add 50 μL / well of Spike S1 solution to coat Spike S1 on a 96-well plate and incubate overnight at 4°C. After removing the reaction solution, wash 3 times with 100 μL / well of 1× Immuno Buffer, add 100 μL / well of Blocking Buffer, and incubate for 1 hour at room temperature with shaking.

[0071] After washing three times with 2.1×Immuno Buffer, 20 μL / well of 1×Immuno Buffer was added, followed by 10 μL / well of the evaluation sample solution. For the control and blank wells, only PBS was added instead of PBS containing the evaluation sample. The samples were then incubated at room temperature for 1 hour with shaking.

[0072] 3. 20 μL / well of ACE2-Biotin (1 ng / μL) was added to each well except the blank. 20 μL / well of 1×Immuno Buffer was added to the blank. The mixtures were incubated at room temperature for 1 hour with shaking.

[0073] 4. After removing the reaction solution, the wells were washed three times with 100 μL / well of 1×Immuno Buffer.

[0074] 5. Add 100 μl of Blocking Buffer per well and incubate at room temperature for 10 minutes.

[0075] 6. Add 100 μL / well of streptoavidin-HRP solution and incubate at room temperature for 1 hour with shaking.

[0076] 7. After the reaction, the samples were washed three times with 100 μL / well of 1×Immuno Buffer.

[0077] 8. Add 100 μl / well of Blocking Buffer and incubate at room temperature for 10 minutes.

[0078] 9. After removing the Blocking Buffer, 100 μL / well of Colorimetric HRP substrate was added and incubated at room temperature until the control well turned blue.

[0079] 10. After the color change, 100 μL / well of 1 N HCl was added to stop the reaction. The absorbance of each well was measured at a wavelength of 450 nm using a plate reader.

[0080] 11. The value obtained by subtracting the absorbance of the blank well from the absorbance of the wells of the control and each evaluation sample was used as the measured value of the control and each evaluation sample.

[0081] The inhibition rate was calculated using the following formula. Inhibition rate (%) = (Measured value of control - Measured value of evaluation sample) / Measured value of control × 100

[0082] <TMPRSS2 evaluation system> Based on the following references, an evaluation method was formulated and implemented. (All concentrations of the control, blank, and each evaluation sample were performed in triplicate) Hoffmann et al.: Version 1. bioRxiv. Preprint. 2020 Aug (https: / / www.biorxiv.org / CONTENT / 10.1101 / 2020.08.05.237651v1) Shrimp et al.: Version 2. bioRxiv. Preprint 2020 Jun 23 (https: / / www.biorxiv.org / content / 10.1101 / 2020.06.23.167544v1.full)

[0083] 1. An assay was performed on a 384-well black plate such that the solution was 10 μL / well.

[0084] 2. First, a 384-well black plate was prepared by adding Recombinant Human TMPRSS2 (N-terminus 6xHis, aa106-492; LifeSpan Biosciences) prepared with buffer (50 mM Tris-HCl pH 8.0, 154 mM NaCl) to a final concentration of 4 μg / mL. Only the buffer was added to the blank wells.

[0085] 3. The evaluation samples, dissolved in phosphate buffer (PBS), were added to the 384-well black plate described in (2.) above, to reach their respective final concentrations. Note that in the control and blank wells, only PBS was added instead of the PBS containing the evaluation samples.

[0086] 4. The substrate (Boc-Gln-Ala-Arg-MCA (manufactured by Peptide Research Institute Co., Ltd.)) was added to the 384-well black plate described in (3.) above to a final concentration of 10 μM.

[0087] 5. The reaction was allowed to proceed at room temperature for 60 minutes.

[0088] 6. Fluorescence values ​​were measured using a microplate reader (SpectraMax M5; manufactured by molecular devices) at an excitation wavelength of 380 nm and an emission wavelength of 460 nm.

[0089] 7. The measured values ​​for the control and each evaluation sample were obtained by subtracting the fluorescence value of the blank well from the fluorescence value of the well of the control and each evaluation sample.

[0090] The inhibition rate was calculated using the following formula. Inhibition rate (%) = (Control measurement - Evaluation sample measurement) / Control measurement × 100

[0091] [Table 2]

[0092]

Table 3

[0093] In Comparative Example 1, no inhibitory effect on S protein-ACE2 binding was observed. In contrast, in Examples 1 to 9, a concentration-dependent inhibitory effect was observed (Table 2). Also, in Comparative Example 2, no inhibitory effect on TMPRSS2 activity was observed. In contrast, in Examples 10 to 18, an inhibitory effect was observed, and the higher the added concentration, the greater the inhibition rate was shown (Table 3). Sodium lauroyl methyl taurine, sodium tetradecene sulfonate, sodium lauryl sulfate, aluminum lactate, potassium nitrate, sodium lauroyl sarcosine, and copper gluconate showed an inhibitory effect on both S protein-ACE2 binding and TMPRSS2 activity (Examples 3 to 9, 10, and 11, 14 to 18).

[0094] These results indicate that the agent of the present invention exhibits an effect of suppressing the cell invasion of coronaviruses and is useful as a preventive agent for coronavirus infections or as an agent for suppressing or alleviating symptoms.

[0095] Example 19 The expression of TMPRSS2 was evaluated for the evaluation samples having the composition shown in Table 4.

[0096]

Table 4

[0097] <TMPRSS2 Expression Evaluation System> The expression of TMPRSS2 was evaluated according to the following procedure.

[0098] 1. Saliva was collected from the subjects who brushed their teeth with the evaluation samples using salivette cotton (manufactured by Zalstat) before toothbrushing, 1 hour after toothbrushing, and 3 hours after toothbrushing.

[0099] 2. Saliva protein concentration was measured using the BCA Protein Assay Kit (manufactured by Takara Bio Inc.), and the protein concentrations between samples were adjusted using UPW (ultrapure water).

[0100] 3. Sample buffer (NuPAGE® LDS Sample Buffer, manufactured by Thermo Fisher) and a reducing agent (NuPAGE® Sample Reducing Agent, manufactured by Thermo Fisher) were added to (2) above, and the mixture was heated at 95°C for 5 minutes.

[0101] 4. The samples from (3) above were added to Bolt® Bis-Tris Plus gel (manufactured by Thermo Fisher) and electrophoresis was performed.

[0102] 5. The proteins in the gel described in (4) above were transferred to a membrane (TransBlot Turbo® Transfer Pack PVDF, manufactured by Bio-Rad).

[0103] 6. The membrane described in (5.) above was blocked for 1 hour with 5% skim milk-containing TBS-T (Tris Buffered Saline with Tween® 20), and then reacted with the primary antibody (Anti-TMPRSS2 antibody [EPR3861], Abcam) overnight at 4°C.

[0104] 7.2 The secondary antibody (Anti-rabbit IgG, HRP-linked Antibody, manufactured by Cell Signaling Technologies) was reacted at room temperature for 1 hour.

[0105] 8. The detection reagent (ECL Prime Western Blotting Detection Reagent, manufactured by Cytiva) was added, and the TMPRSS2 protein band was obtained using an imager (Amersham® ImageQuant® 800, manufactured by Cytiva).

[0106] 9. Using the imaging software attached to (8) above, the band signal intensity (expression level) of the TMPRSS2 protein was analyzed.

[0107] 10. Taking the expression level before toothbrushing as 1, the expression levels 1 hour and 3 hours after toothbrushing were calculated (Table 5).

[0108] Comparative Example 3 In Example 19, instead of the subjects who brushed their teeth with the evaluation samples in Table 5, samples were collected and evaluated at the same time points for the subjects who were kept quiet without toothbrushing.

[0109] [Table 5]

[0110] Compared with Comparative Example 3, in Example 19, the expression level of TMPRSS2 decreased after toothbrushing (Table 5).

[0111] Comparative Example 4 When an evaluation sample obtained by diluting a blank preparation obtained by removing sodium lauryl sulfate from the composition of Table 5 15-fold with purified water assuming a dentifrice was evaluated in the <TMPRSS2 evaluation system>, no inhibitory effect on TMPRSS2 activity was observed.

[0112] These results indicate that the agent of the present invention can exert various actions such as inhibition of the formation of the binding between the spike protein of the coronavirus and the receptor on the cell surface, inhibition of the activation of the spike protein bound to the receptor on the cell surface by TMPRSS2, and inhibition of the expression of TMPRSS2, and is useful as a preventive agent for coronavirus infection or an agent for suppressing or alleviating symptoms.

Claims

1. comprising at least one selected from aluminum lactate, potassium nitrate, tranexamic acid, epsilon-aminocaproic acid, and copper gluconate, It inhibits the formation of binding between the coronavirus spike protein and the cell surface receptor, or It inhibits the activation of spike proteins bound to cell surface receptors by transmembrane serine protease 2 (TMPRSS2). It is a cell entry inhibitor for coronavirus, The aforementioned receptor is angiotensin-converting enzyme 2, The aforementioned coronavirus is a coronavirus that uses angiotensin-converting enzyme 2 as its receptor. The aforementioned agent.

2. The agent according to claim 1, comprising at least one selected from aluminum lactate, potassium nitrate, and copper gluconate.

3. The agent according to claim 1 or 2, wherein inhibition of spike protein activation is inactivation of transmembrane serine protease 2 (TMPRSS2).

4. The agent according to any one of claims 1 to 3, for the prevention of coronavirus infection that uses angiotensin-converting enzyme 2 as a receptor.

5. The agent according to any one of claims 1 to 4, wherein the coronavirus that uses angiotensin-converting enzyme 2 as a receptor is SARS-CoV-1, CoV-NL63, or SARS-CoV-2.

Images