JP4355592B2 - Antiviral agent - Google Patents

Description

  The present invention relates to an antiviral agent characterized by containing lactoperoxidase as an active ingredient, and an antiviral agent characterized by containing lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients. . Furthermore, another invention of the present invention relates to a food and drink and a feed containing an antiviral agent characterized by containing lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients.

  There are various microorganisms in nature that cause infections in humans and animals. These microorganisms are classified into bacteria, mycoplasma, rickettsia, chlamydia, fungi, protozoa and viruses. The genetic material of the virus is deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). Bacteria and other microorganisms have an energy metabolism system and proliferate in heterotropy, whereas viruses are parasitic on host cells. Can be propagated only by (for example, Non-Patent Document 1).

  Influenza virus infections are prevalent in winter, but patients with malignant tumors, heart diseases, cerebrovascular diseases, etc. as basic diseases, elderly people, and infants are high-risk groups of influenza infections, and they can cause serious life-threatening infections. Often connected. RS (Respiratory Syncytial) virus does not normally infect healthy adults, but is a major cause of upper respiratory infections in children.

  Among the infections caused by microorganisms, effective antibiotics have been developed particularly for bacterial infections, and effective treatments have been carried out, but antibiotics do not act effectively on viral infections. Compared to the rapid progress of chemotherapy with antibiotics, the development of antiviral agents as therapeutic agents for viral infections is still not progressing so far. Antiviral agents developed so far include nucleic acid analogues acyclovir, azidothymidine, ribavirin, neuraminidase inhibitor zanamivir (4-guanidinoneuraminic acid), a protease inhibitor indinavir, etc. There is. Although these antiviral agents, which are pharmaceuticals, have a strong medicinal effect, side effects due to administration have a great clinical problem. Under such circumstances, search for an active ingredient having an antiviral action that is safe, has few side effects, and can be ingested over a long period of time has been continued.

  Lactoperoxidase is an oxidoreductase contained in milk of mammals such as cow milk and secretory fluids such as saliva, tears and airway mucus (for example, Non-Patent Document 2). Lactoperoxidase is a protein having a molecular weight of about 80,000, and contains one heme in the molecule as a coenzyme (for example, Non-Patent Document 3). Lactoperoxidase has been reported to have various biological functions such as antibacterial activity, antioxidant activity, anticancer activity, and immunoregulatory activity, and has been shown to be an important protein involved in biological defense. (For example, Non-Patent Document 4 or Non-Patent Document 5).

  Lactoperoxidase generates hypothiocyanic acid (OSCN-), which is an antibacterial substance, by oxidizing thiocyanic acid (SCN-) widely contained in animal tissues and body fluids in the presence of hydrogen peroxide (for example, Non-patent document 6). A reaction system composed of lactoperoxidase, thiocyanic acid, and hydrogen peroxide (or a compound capable of donating hydrogen peroxide: in the present invention, sometimes referred to as a hydrogen peroxide donor) has various effects. The use of lactoperoxidase, peroxide donors and thiocyanates to produce a medicament for the treatment of Helicobacter pylori infection (for example, Patent Document 1), application of peroxidase to prevention and treatment (for example, Patent Document) 2) Method for producing lactic acid bacteria fermented food containing peroxidase-thiocyanate ion and / or halogenoion-hydrogen peroxide system (for example, Patent Document 3), two-enzyme tooth brushing (for example, Patent Document 4), bactericidal composition (For example, Patent Document 5), lactic acid bacteria starter containing peroxidase, fermented milk And their preparation (e.g., Patent Document 6), such as, for industrial applications, is disclosed application to already variety of applications. Further, the present inventors have disclosed a technique relating to a urease inactivating composition and a food and drink (for example, Patent Document 7).

  In addition, it has already been clearly shown that the combination of the three components of lactoperoxidase, thiocyanate, and hydrogen peroxide exhibits antiviral activity against type 1 herpes simplex virus, RS virus, echovirus, human immunodeficiency virus, and the like. (For example, Non-Patent Document 7 or Non-Patent Document 8).

JP 2000-509367 Japanese Patent No. 3478821 JP 62-228224 A Japanese Patent Publication No. 4-25924 Japanese Unexamined Patent Publication No. 1-61427 Japanese Patent No. 3007686 JP 2002-238554 A

Medical Virology, Revised 2nd edition, edited by Toshiro Osato, published by Nanedo, 2000, p. 15-24 American Journal of Respiratory and Critical Care Medicine, USA, 166, 2002, p. S57-S61 British Journal of Nutrition, UK, Vol. 84, 2000, p. S19-S25 Life Sciences, UK, 47, 1990, p. 703-709 Journal of Dairy Research, UK, Vol. 63, 1996, p. 257-267 American Journal of Respiratory Cell and Molecular Biology, USA, Vol. 20, 2000, p. 642-644 Antiviral Research, Netherlands, Vol. 26, 1995, p. 161-171 Journal de biologie buccale, France, Vol. 18, 1990, p. 251-253

  In Non-Patent Document 7 or Non-Patent Document 8, a combination of three components of lactoperoxidase, thiocyanic acid, and hydrogen peroxide is used for type 1 herpes simplex virus, RS virus, echo virus, human immunodeficiency virus, etc. However, these three components have not yet been fully examined for prevention and treatment of influenza virus infection. Therefore, the present inventors consider that influenza virus infection is prevalent in winter and patients, elderly people, and infants who have malignant tumors, heart disease, cerebrovascular disease, etc. as basic diseases are high-risk groups for influenza infection. As a result of diligent investigation of antiviral action on a system in which thiocyanic acid and hydrogen peroxide were added as active ingredients for lactoperoxidase, a milk protein that is a highly versatile material with few side effects, As a result, the present invention was completed.

  The object of the present invention is excellent in safety, can be taken and ingested for a long period of time on a daily basis, and is antiviral against pathogenic viruses that cause infectious diseases in animals including humans, particularly influenza viruses. It is to provide an antiviral agent having an action.

  Another object of the present invention is to provide a food or drink or feed containing the antiviral agent.

  A first invention of the present invention for solving the above-mentioned problems is an antiviral agent characterized by containing lactoperoxidase as an active ingredient.

  A second invention of the present invention for solving the above-mentioned problems is an antiviral agent characterized by containing lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients, and is antiviral against influenza virus. Having a function is a preferred embodiment.

  A third invention of the present invention that solves the above-mentioned problems is a food or drink or feed containing the antiviral agent according to the first invention or the second invention.

  The present invention contains a highly safe lactoperoxidase that can be ingested daily as an active ingredient, and contains lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients. Further, the present invention relates to an antiviral agent characterized by the above, and a food and drink or feed containing the antiviral agent, and the effects exhibited by the present invention are as follows.

(1) Inactivate the infectivity of pathogenic viruses that cause infectious diseases in animals including humans.
(2) It is highly safe for humans and can increase the infection protection effect against viruses by daily intake.
(3) An antiviral agent in which thiocyanic acid and hydrogen peroxide are added as active ingredients to lactoperoxidase has a remarkable antiviral action against influenza virus.

  Next, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the following preferred embodiments, and can be freely changed within the scope of the present invention. In the present specification, percentages are expressed by mass unless otherwise specified.

  In the present invention, the antiviral action means an action that inactivates the infectivity of a pathogenic virus that causes human or animal infections. In the present invention, the antiviral agent means a drug having a component having an action of inactivating the infectivity of a pathogenic virus that causes infection in humans and animals, and is highly safe for humans. It also has the characteristic that an infection protection effect against viruses is exhibited by administration.

  The antiviral agent of the present invention preferably contains lactoperoxidase, lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients. In the antiviral agent containing lactoperoxidase, thiocyanic acid, and hydrogen peroxide as active ingredients, the thiocyanic acid and hydrogen peroxide should contain the compound and pharmaceutically acceptable salts of the compound as active ingredients. Foods that originally contain the compound and that can produce the compound (in the present invention, these are collectively referred to as a donor; a thiocyanate donor, a hydrogen peroxide donor) May be substituted for thiocyanic acid and hydrogen peroxide, which are the active ingredients of the antiviral agent of the present invention.

  The food originally containing thiocyanic acid and hydrogen peroxide and the food capable of producing the compound can be appropriately selected depending on the form of the antiviral agent, and examples thereof include the following. . Examples of thiocyanic acid include thiocyanic acid contained in mammalian milk and thiocyanic acid contained in vegetables selected from one or more of cabbage, broccoli, cauliflower, turnip, etc. Various forms of thiocyanic acid can be used. In addition, when the antiviral agent of the present invention is contained in a food or drink, in consideration of safety, it is preferable that thiocyanic acid is supplied from the vegetable, and the liquid is obtained by grinding the vegetable or grinding it with a mixer. About the vegetable juice prepared by making it, it is possible to contain it as an active ingredient of the antiviral agent of this invention using the powder obtained by drying as it is or by freeze-drying method.

  As for hydrogen peroxide, for example, it has a low redox potential and can generate hydrogen peroxide by polyphenols such as hydroxyhydroquinone, which can reduce oxygen molecules and generate hydrogen peroxide, or oxidase reaction. Examples include glucose oxidase and oxidase / substrate mixtures of glucose, xanthine oxidase, and xanthine in combination, and lactic acid bacteria such as Lactobacillus or Streptococcus that produce hydrogen peroxide. In addition, when the antiviral agent of the present invention is contained in a food or drink, in consideration of safety, hydrogen peroxide is preferably supplied from an oxidase / substrate mixture or lactic acid bacteria. In the following, thiocyanic acid produced from vegetables and hydrogen peroxide produced from lactic acid bacteria are exemplified as Reference Example 1 and Reference Example 2, respectively.

[Reference Example 1]
Vegetable juice was prepared by applying 1 kg of cabbage to a home mixer and filtering the liquid obtained by pulverization with a filter paper. Using the method of Bath et al. [Indian Journal of Physiology and Pharmacology, Vol. 30, pp. 241-247, 1986] As a result, it was 0.0048%.

[Reference Example 2]
10 kg of raw milk having a milk fat content of 3.5% and a non-fat milk solid content of 9.2% was homogenized and heat-sterilized at 90 to 92 ° C. for 10 minutes. After cooling to about 42 ° C., 100 g each of commercially available Streptococcus thermophilus and Lactobacillus delbuleckii subsp. Bulgaricus milk cultures was added as a starter. The mixture was sufficiently stirred in the fermentation tank and allowed to stand at 42 to 45 ° C. for 4 hours for fermentation. The fermented milk was cooled to 5-8 ° C. with stirring, and then homogenized with a homogenizer to obtain homogenized fermented milk. The hydrogen peroxide concentration of this fermented milk was measured by the Gililand method [Journal of Dairy Science, Vol. 52, pp. 321 to 324, 1969]. there were.

  Lactoperoxidase, which is an active ingredient of the antiviral agent of the present invention, is defatted as a colostrum, transitional milk, normal milk, terminal milk or processed product of these milks of mammals such as cows, humans, goats, sheep, horses Lactoperoxidase separated from milk, whey, etc. by a conventional method (for example, ion chromatography) can be used. In addition, a commercially available lactoperoxidase (for example, manufactured by Biopol) can be used more simply. It is also possible to use recombinant lactoperoxidase produced by microorganisms, animal cells, transgenic animals, and the like based on genetic manipulations as long as they have antiviral activity.

  The antiviral activity in the antiviral agent of the present invention can be measured by a conventional method, for example, a plaque suppression method using animal cultured cells. For example, when a culture system in which no virus is added and a culture system in which a virus is added but no antiviral agent is used as a control and the number of plaque formation is measured, the ratio (%) to the control is suppressed to 50% or less. Can be determined to have antiviral activity [for example, [Antiviral Research, The Netherlands, Vol. 22, 1993, p. 175-188], [Journal of General Virology, England, Vol. 71, 1990, p. 2149-2155].

  The amount of lactoperoxidase contained in the preparation of the antiviral agent of the present invention is not particularly limited and may be appropriately selected. For example, it is usually 0.001% by mass or more, preferably 0.01 to 10% by mass in the preparation. It is good to do. The administration method of the preparation of the present invention is not particularly limited, and is determined according to various preparation forms, the patient's age, sex, other conditions, the degree of symptoms of the patient and the like. The dosage of the active ingredient in the preparation of the present invention is appropriately selected depending on the usage, patient age, sex, disease severity, other conditions, and the like. Usually, the amount of lactoperoxidase as an active ingredient should be 0.02-200 mg / kg / day, preferably 0.2-20 mg / kg / day, and once a day. Alternatively, it can be administered in multiple doses.

  The amount of lactoperoxidase, thiocyanic acid, and hydrogen peroxide contained in the preparation of the antiviral agent of the present invention is not particularly limited and may be appropriately selected. For example, the content is usually as shown below in the preparation. Is preferred. That is, the content of lactoperoxidase is preferably at least 0.0005% by mass, particularly preferably 0.001-1% by mass. The thiocyanic acid content is preferably at least 0.0005% by mass, particularly preferably 0.005 to 0.05% by mass. Furthermore, the content of hydrogen peroxide is preferably at least 0.0001 to 0.2% by mass, particularly preferably 0.001 to 0.01% by mass. The administration method of the preparation of the present invention is not particularly limited, and is determined according to various preparation forms, the patient's age, sex, other conditions, the degree of symptoms of the patient and the like. The dosage of the active ingredient in the preparation of the present invention is appropriately selected depending on the usage, patient age, sex, disease severity, other conditions, and the like. Usually, as the active ingredient, the amount of lactoperoxidase is 0.01 to 20 mg / kg / day, preferably 0.02 to 2 mg / kg / day, and the amount of thiocyanic acid is 0.01 to 1 mg / day. kg / day, preferably in the range of 0.1-1 mg / kg / day, the amount of hydrogen peroxide is 0.002-0.4 mg / kg / day, preferably 0.02-0.2 mg / kg / day The amount within the above range should be taken as a guideline, and can be administered once a day or divided into a plurality of times. In addition, if the content of each component of the antiviral agent containing lactoperoxidase, thiocyanic acid, and hydrogen peroxide as the active ingredient is within the above range, it exhibits a remarkable antiviral action against influenza virus, and the cause of the virus It has the effect of improving symptoms caused by infection.

  In formulating, additives such as excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents, diluents, and solvents for injections can be used. Examples of the excipient include sugar derivatives such as lactose, sucrose, glucose, mannitol and sorbit; starch derivatives such as corn starch, potato starch, α-starch, dextrin and carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Cellulose derivatives such as hydroxypropylmethylcellulose, carboxymethylcellulose, carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, magnesium magnesium magnesium silicate; phosphate derivatives such as calcium phosphate; Examples thereof include carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate, etc. Examples of the binder include gelatin, polyvinyl pyrrolidone; Examples of disintegrants include, in addition to the above excipients, chemically modified starch or cellulose derivatives such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone. Examples of the agent include talc; stearic acid; stearic acid metal salts such as calcium stearate and magnesium stearate; colloidal silica; waxes such as bee gum and gallow; boric acid; glycol; carboxylic acids such as fumaric acid and adipic acid; Carboxylic acid sodium salts such as sodium sulfate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as anhydrous silicic acid and silicic acid hydrate; starch derivatives and the like As a stabilizer, Examples include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; sorbic acid and the like. , Sweeteners, acidulants, fragrances, and the like. Examples of the solvent for injection include water, ethanol, glycerin, and the like.

  Examples of the administration route of the antiviral agent of the present invention include parenteral administration such as oral administration and enteral administration. Examples of the dosage form include sprays, capsules, tablets, granules, syrups, emulsions, suppositories, injections, ointments, tapes and the like. Moreover, the antiviral agent of this invention can also be mix | blended and administered to food-drinks, feed, etc.

  When the antiviral agent of the present invention is contained in a food or drink, for example, the active ingredient of the antiviral agent of the present invention includes saccharides such as dextrin and starch; proteins such as gelatin, soy protein and corn protein; alanine, glutamine, Foods and drinks can be produced by blending amino acids such as isoleucine; polysaccharides such as cellulose and gum arabic; and fats and oils such as soybean oil and medium chain fatty acid triglycerides. Moreover, as a form of the food / beverage products containing the antiviral agent of this invention, drinks, such as a soft drink, a carbonated drink, a nutritive drink, a fruit drink, and a lactic acid drink (including the concentrated undiluted solution and adjustment powder of these drinks); Ice cream, ice sherbet, shaved ice and other frozen desserts; noodles such as buckwheat, udon, harusame, gyoza skin, sukimai skin, chinese noodles, instant noodles; rice cakes, chewing gum, candy, gum, chocolate, tablet confectionery, snacks, biscuits , Jelly, jam, cream, baked confectionery, etc .; fish and fishery processed foods such as kamaboko, ham, sausage; dairy products such as processed milk and fermented milk; salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream , Dressings and other fats and oils and processed foods; seasonings such as sauces and sauces; soups, stews, salads Prepared foods, pickles, bread; include functional foods and the like; enteral nutrition.

  When the antiviral agent of the present invention is contained in the feed, for example, the active ingredient of the antiviral agent of the present invention includes cereals such as corn, wheat, barley, rye, and milo; soybean oil meal, rapeseed oil meal, coconut oil meal, linseed Vegetable oils such as oil lees; bran, wheat straw, rice bran, defatted rice lees, etc .; production corn such as corn gluten meal, corn jam meal; animal nature such as fish meal, skim milk powder, whey, yellow grease, tallow Feeds can be produced by blending yeasts such as tolal yeast and beer yeast; mineral feeds such as tricalcium phosphate and calcium carbonate; fats and oils; simple amino acids; sugars and the like. Examples of the form of feed containing the antiviral agent of the present invention include pet food, livestock feed, and fish feed.

  In addition, the food / beverage products or feed of this invention can be made into the food / beverage products or feed which displayed the effect shown below. That is, efficacy as an antiviral action or the like can be displayed. Here, “display” means an act of informing the consumer of the effect, for example, an act of attaching the effect to a food / drink or feed product or packaging / advertisement of the product of the present invention, There are acts such as assigning, handing over, and exhibiting the attached items, but especially food for specified health use (12th of the Ordinance for Enforcement of Health Promotion Act (April 30, 2003, Ministry of Health, Labor and Welfare Ordinance No. 86)) (See Article 1, Paragraph 1, Item 5]].

  The antiviral agent of the present invention can be used alone, but may be used in combination with other antiviral agents. By using it together, the antiviral action can be enhanced. The antiviral agent to be used in combination may be contained as an active ingredient in the composition of the present invention, or may not be contained in the antiviral agent of the present invention but may be combined and commercialized as a separate drug.

Next, the present invention will be described in detail with reference to test examples.
[Test Example 1]
This test was conducted in order to examine the antiviral action of lactoperoxidase, which is an active ingredient of the antiviral agent of the present invention, against pathogenic viruses.

(1) Preparation of sample Lactoperoxidase (manufactured by Sigma) was dissolved in purified water at a concentration of 10 mg / ml, and sterilized by filtration with a filter having a pore size of 0.2 µm to prepare test sample 1. As a test virus, RS virus A2 strain belonging to Paramyxoviridae (a single-stranded RNA virus having an envelope) was used. In addition, an influenza virus A / PR / 8/34 (H1N1) strain belonging to the Orthomyxoviridae family (a single-stranded RNA virus having an envelope) was used as a control virus. As cultured animal cells, Hep2 cells are used for RS virus, MDCK cells are used for influenza virus, and cultured in a 60 mm diameter dish until monolayer using serum-added Eagle MEM medium (manufactured by Nissui Pharmaceutical). did.

(2) Test method RS virus was added in 0.2 ml of phosphate buffered saline so as to form 100 plaque forming units (hereinafter abbreviated as PFU). The influenza virus was added to 100 PFU in 0.2 ml of phosphate buffered saline containing 1% bovine serum albumin. To these solutions, 2 μl of test sample 1 (final concentration: 100 μg / ml) was added and incubated at 37 ° C. for 1 hour. Nothing was added to the control, and the mixture was kept at 37 ° C. for 1 hour. The whole amount of these virus treatment solutions was added to the animal cells cultured in a monolayer with a dish having a diameter of 60 mm, and kept at 37 ° C. for 1 hour.

  Next, the dish was washed with phosphate buffered saline. The cultured cells to which the RS virus treatment solution was added were layered with 5 ml of 0.8% methylcellulose-added Eagle MEM medium and cultured for 5 days. The cultured cells to which the influenza virus treatment solution was added were layered with 5 ml of Eagle's MEM medium supplemented with 0.8% agarose and cultured for 3 days. The monolayer cultured animal cells after completion of the culture were fixed with a 5% formalin solution and stained with a 0.03% methylene blue solution. The number of plaque formation detected in monolayer cultured animal cells was measured, and the ratio (%) of the test sample 1 to the control [Antiviral Research, Netherlands, Vol. 22, 1993, p. 175-188], [Journal of General Virology, England, Vol. 71, 1990, p. 2149-2155]. These experimental operations were performed twice and the average value was calculated.

(3) Test results The results of this test are as shown in Table 1. The rate of plaque formation relative to the control of lactoperoxidase was 49.4% for RS virus. Moreover, it was 82.9% with respect to the influenza virus which is a control virus. Accordingly, it was revealed that lactoperoxidase has an antiviral action against RS virus.

[Test Example 2]
This test was conducted to examine the antiviral action against influenza virus for lactoperoxidase, thiocyanic acid, and hydrogen peroxide, which are active ingredients of the antiviral agent of the present invention.

(1) Preparation of sample Lactoperoxidase (manufactured by Sigma) was dissolved in purified water at a concentration of 1 mg / ml. Sodium thiocyanate (manufactured by Nacalai Tesque) was dissolved in purified water at a concentration of 1 mg / ml. Catalase (manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in phosphate buffered saline at a concentration of 0.2 mg / ml. Each of these solutions was sterilized by filtration through a filter having a pore size of 0.2 μm. 30% hydrogen peroxide solution (Wako Pure Chemical Industries, Ltd.) was diluted 100 times with purified water. The influenza virus A / PR / 8/34 (H1N1) strain was used as a test virus (Antiviral Research, Netherlands, Vol. 22, pages 175-188, 1993). MDCK cells were cultured using a serum-added Eagle MEM medium (manufactured by Nissui Pharmaceutical Co., Ltd.) in a dish having a diameter of 60 mm until becoming a monolayer.

(2) Test method The influenza virus was added to 0.5 ml of 2% bovine serum albumin-added phosphate buffered saline so as to give 500 PFU. In test group A, 0.5 μl of virus solution, 8 μl of lactoperoxidase solution (final concentration 0.0008%), 53.58 μl of sodium thiocyanate solution (final concentration 0.005%), phosphate buffered saline 391. 7 μl was added and mixed, and 6.8 μl of hydrogen peroxide solution (final concentration 0.002%) was added to make the total volume 1 ml. In the test group B, 53.58 μl of sodium thiocyanate solution and 399.7 μl of phosphate buffered saline are added to 0.5 ml of virus solution and mixed, and 6.8 μl of hydrogen peroxide solution is added to make a total volume of 1 ml. It was. In test group C, 0.5 μl of virus solution was mixed with 8 μl of lactoperoxidase solution and 445.2 μl of phosphate buffered saline, and 6.8 μl of hydrogen peroxide solution was added to make the total volume 1 ml. . In test group D, 0.5 μl of virus solution was mixed with 8 μl of lactoperoxidase solution, 53.58 μl of sodium thiocyanate solution, and 398.5 μl of phosphate buffered saline to make the total volume 1 ml.

  For control, 0.5 ml of phosphate buffered saline was added to 0.5 ml of the virus solution to make the total volume 1 ml. After incubating these solutions at 37 ° C. for 1 hour, 40 μl of catalase solution was added and mixed. 0.2 ml of this virus treatment solution was added to MDCK cells cultured in a monolayer in a dish having a diameter of 60 mm, and incubated at 37 ° C. for 1 hour. Next, the dish was washed with phosphate buffered saline, and 5 ml of Eagle's MEM medium supplemented with 0.8% agarose was overlaid and cultured for 3 days. The cells after completion of the culture were fixed with a 5% formalin solution and stained with a 0.03% methylene blue solution. The number of plaque formation detected in the monolayer cultured animal cells was measured, and the ratio (%) of each test group to the control was calculated in the same manner as in Test Example 1. These experimental operations were performed twice and the average value was calculated.

(3) Test results The results of this test are as shown in Table 2. The plaque formation rate relative to the control was test group A: 0.3%, test group B: 107.9%, test group C: 97.8%, test group D: 106.3%. Therefore, it was clarified that the three components of lactoperoxidase, thiocyanic acid, and hydrogen peroxide have an antiviral action against influenza virus. When any two of these three components are added, the antiviral action is not confirmed, and as is clear from the test results of Test Example 1, lactoperoxidase alone (one component) is effective against influenza virus. On the other hand, since no antiviral action has been confirmed, the antiviral agent of the present invention comprising three components of lactoperoxidase, thiocyanic acid, and hydrogen peroxide as an active ingredient has a remarkable antiviral action against influenza virus. It turned out to show.

  The same antiviral effect was confirmed when hydrogen peroxide donors glucose and glucose oxidase were used instead of hydrogen peroxide.

  EXAMPLES Next, although an Example is shown and this invention is demonstrated further in detail, this invention is not limited to a following example.

  Lactoperoxidase (manufactured by Biopol) 50 g, lactulose powder (manufactured by Morinaga Milk Industry Co., Ltd.) 100 g, maltodextrin (manufactured by Matsutani Chemical Industry Co., Ltd.) 735 g, skim milk powder (manufactured by Morinaga Milk Industry Co., Ltd.) 85 g, stevia sweetener Each powder of 1 g of F eye), 5 g of yogurt flavor (manufactured by Saneigen F.F.I.), and 24 g of glycerin fatty acid ester preparation (manufactured by Riken Vitamin Co., Ltd.) was added and mixed uniformly. (Made by Hata Steel Co., Ltd.), 0.5 g per tablet, tableting the mixed powder continuously at a tableting speed of 12 tablets / min and a pressure of 9.8 KPa, and lactoperoxidase 1800 tablets (about 900 g) were produced. In addition, it was confirmed that the manufactured tablet has an antiviral action against RS virus.

  10 kg of raw milk having a milk fat content of 3.5% and a non-fat milk solid content of 9.2% was homogenized and heat-sterilized at 90 to 92 ° C. for 10 minutes. After cooling to about 42 ° C., 100 g each of commercially available Streptococcus thermophilus and Lactobacillus delbrueckii subsp. Bulgaricus milk cultures was added as a starter. The mixture was sufficiently stirred in the fermentation tank and allowed to stand at 42 to 45 ° C. for 4 hours for fermentation. Homogenized fermented milk was prepared by cooling the fermented milk to 5-8 ° C. with stirring and then homogenizing with a homogenizer. Next, 60 g of pectin (manufactured by Saneigen FFI) is added to 5 kg of 17% granulated sugar (manufactured by Toyo Seika Co., Ltd.), sterilized at 90-92 ° C. for 10 minutes, and then cooled to 5-8 ° C. A sugar solution was prepared. Further, 3 kg of cabbage was passed through a household mixer, filtered with a filter paper (manufactured by Whatman), sterilized at 90 to 92 ° C for 10 minutes, and then cooled to 5 to 8 ° C to prepare 900 g of vegetable juice. Furthermore, 0.8 g of lactoperoxidase (manufactured by Biopol) was dissolved in 40 g of water, and sterile filtered through a membrane filter (manufactured by Millipore) to obtain a lactoperoxidase solution. All the homogenized fermented milk, sugar solution, vegetable juice, and lactoperoxidase solution prepared by these methods were thoroughly mixed in a tank, filled in a paper container, and sealed to produce a drink yogurt. In addition, as a result of examining the antiviral action with respect to influenza virus in the system of Test Example 2 about the manufactured drink yogurt, it was confirmed that this drink yogurt has an antiviral action with respect to influenza virus.

The antiviral agent of the present invention is highly safe for humans and can inactivate the infectivity of pathogenic viruses that cause infectious diseases in animals including humans. The effect of protecting against viruses can be increased. In addition, since it exhibits a remarkable antiviral action against influenza virus, it is possible to provide an influenza virus therapeutic agent that replaces the existing antiviral agent.

Claims (2)

  An anti-RS virus agent comprising lactoperoxidase as an active ingredient.   An anti-RS virus agent comprising 0.01% by mass or more of lactoperoxidase as an active ingredient.