Composition, food, drink, and feed for treating and / or preventing influenza

Abstract

To provide a composition for treating and / or preventing influenza, food and drink, and feed, which are excellent in safety as well as preventive effect and treatment effect.SOLUTION: The present disclosure provides a composition for treating and / or preventing influenza containing a retinoid compound as an active ingredient.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to a composition, food, drink, or feed for treating and / or preventing influenza, which contains a retinoid compound, or a retinoid compound and a lactic acid bacterium as active ingredients. [Background technology]

[0002] Influenza is an abbreviation for influenza virus infection. Influenza viruses are highly contagious, and the incidence of influenza in human populations is high, periodically resulting in large-scale morbidity and mortality. Unlike cold syndromes, influenza is characterized by relatively rapid onset of high fever, blockage, headache, general fatigue, and muscle pain, and is accompanied by upper respiratory tract inflammation symptoms such as sore throat, runny nose, and cough. In addition, complications include bronchitis, pneumonia, and influenza encephalopathy, which may lead to death. In particular, influenza is a threat because it is prone to becoming severe in people with underlying diseases such as chronic heart disease, lung disease, kidney disease, liver disease, and metabolic disease, as well as in the elderly and infants who are not physically strong.

[0003] Influenza vaccines are the main method of preventing influenza. In addition, oseltamivir, zanamivir, peramivir, laninamivir, and the like are used as influenza treatment drugs. However, these vaccines and treatment drugs can cause serious side effects. On the other hand, it has been reported that natural substances, which are highly safe, do not provide therapeutic effects (Non-Patent Document 1). Therefore, there is a demand for the development of new compositions that are not only highly safe but also have sufficient preventive and therapeutic effects.

[0004] Up to now, preventive and therapeutic drugs for influenza infections containing lactic acid bacteria, which have been shown to be highly safe, as active ingredients have been reported. For example, Patent Document 1 discloses a preventive and therapeutic agent for influenza containing Lactobacillus acidophilus CL-92 strain as an active ingredient. In addition, Non-Patent Document 2 discloses that oral intake of lactic acid bacteria suppresses weight loss caused by influenza virus infection. Furthermore, Patent Document 2 discloses that lactic acid bacteria products from Lactobacillus genus lactic acid bacteria suppress the decrease in acquired immune function caused by anti-influenza drugs.

[0005] Incidentally, it is believed that the defense mechanism of lactic acid bacteria against infectious diseases such as influenza virus is due to the regulatory effect of the immune system. For example, Patent Document 3 describes a natural immunity activator containing lactic acid bacteria as an active ingredient. It is believed that orally ingested lactic acid bacteria are taken up into Peyer's patches in the intestinal tract and phagocytosed by macrophages, dendritic cells, etc., thereby activating immune cells and activating natural immunity. In addition, Non-Patent Document 3 describes that lactic acid bacteria are recognized by TLR. It is believed that lactic acid bacteria regulate the functions of macrophages and dendritic cells via TLR signals. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Patent Publication No. 2012-72113 [Patent Document 2] Patent Publication No. 2018-27904 [Patent Document 3] International Publication No. 2018 / 034203 [Non-patent literature]

[0007] [Non-Patent Document 1] Cui, D. et al., High-level dietary vitamin A enhances T-helper type 2 cytokine production and secretory immunoglobulin a response to influenza A virus infection in BALB / c mice, The Journal of nutrition, 2000, 130(5), 1132-1139 [Non-Patent Document 2] Waki, N. et al., Oral administration of lactobacillus brevis KB290 to mice alleviate clinical symptoms following influenza virus infection, Letters in applied microbiology, 2014, 58(1), 87-93 [Non-Patent Document 3] Takagi A. et al., Lipoteichoic acids from lactobacillus strains elicit strong tumor necrosis factor alpha-inducing activities in macrophages through Toll-like receptor 2, Clinical and diagnostic laboratory immunology, 2003, 10(2), 259-266 Summary of the Invention [Problem to be solved by the invention]

[0008] Although compositions containing various active ingredients have been developed to prevent and treat influenza, none have been obtained that are sufficiently safe and have excellent preventive and therapeutic effects. Therefore, there is a demand for compositions that are not only highly safe but also have excellent preventive and therapeutic effects. Therefore, an object of the present invention is to provide a composition, food, drink, or feed for preventing and / or treating influenza that is not only safe but also has excellent preventive and therapeutic effects. [Means for solving the problem]

[0009] As a result of intensive research into the above-mentioned problems, the present inventors have discovered that influenza can be prevented and treated by ingesting a retinoid compound, or a composition containing a retinoid compound and lactic acid bacteria as active ingredients, and have completed the present invention.

[0010] That is, the present invention provides the following [1] to [9]. [1] A composition for preventing and / or treating influenza, comprising a retinoid compound as an active ingredient. This composition can provide a safe and effective composition for preventing and / or treating influenza. [2] A composition for preventing and / or treating influenza described in [1], further characterized in that it contains lactic acid bacteria as an active ingredient. According to this composition, a composition for preventing and / or treating influenza that exhibits excellent safety and efficacy can be provided. [3] The content of the retinoid compound relative to the content of the lactic acid bacteria is 10 10 The composition for preventing and / or treating influenza according to [2], characterized in that the amount is 100 μg to 500 μg per individual. This composition can provide a composition for preventing and / or treating influenza that is more safe and effective. [4] A composition for preventing and / or treating influenza according to any one of [1] to [3], wherein the retinoid compound is vitamin A or a vitamin A derivative. According to this composition, by employing vitamin A and vitamin A derivatives which are easily available, a composition for preventing and / or treating influenza can be easily prepared. [5] The composition for preventing and / or treating influenza according to any one of [1] to [3], wherein the retinoid compound is provitamin A. This composition can provide a composition for preventing and / or treating influenza that is more safe and effective. [6] A composition for preventing and / or treating influenza according to [5], characterized in that the provitamin A is β-carotene. According to this composition, by employing readily available β-carotene, a composition for preventing and / or treating influenza can be easily prepared. [7] A composition for preventing and / or treating influenza described in any of [2] to [6], characterized in that the lactic acid bacterium is Lactobacillus brevis strain KB290 (accession number NITE P-1537). This composition can provide a composition for preventing and / or treating influenza that is more safe and effective. [8] A composition for preventing and / or treating influenza according to any one of [2] to [7], characterized in that the lactic acid bacteria are killed. This composition can provide a composition for preventing and / or treating influenza that exhibits even greater safety and efficacy. [9] A food, drink, and / or feed for preventing and / or treating influenza, comprising the composition according to any one of [1] to [8]. The food, drink, and feed can provide safe and effective food, drink, and feed for preventing and / or treating influenza.

[0011] Furthermore, since changes in the amount of cytokines produced have been observed when a retinoid compound, or a retinoid compound and lactic acid bacteria, the present invention also has an objective of providing a composition containing a retinoid compound, or a retinoid compound and lactic acid bacteria, that regulates the production of cytokines. The present inventors have further discovered that a retinoid compound, or a composition containing a retinoid compound and lactic acid bacteria, can suppress the production of TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5.

[0012] That is, the present invention provides the following

[10] to

[19] .

[10] A composition for regulating cytokine production caused by viral infection, comprising a retinoid compound as an active ingredient. This composition can provide a safe and effective composition for regulating cytokine production due to viral infection.

[11] A composition for regulating cytokine production caused by viral infection described in

[10] , further characterized by containing lactic acid bacteria as an active ingredient. This composition can provide a composition for regulating cytokine production due to viral infection, which exhibits excellent safety and efficacy.

[12] A composition for regulating cytokine production due to viral infection described in

[10] or

[11] , characterized in that the regulation of cytokine production due to viral infection is suppression of production of at least one cytokine selected from the group consisting of TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2 and CCL5. This composition can provide a safe and effective composition for suppressing the production of cytokines, such as TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5, due to viral infection.

[13] The content of the retinoid compound relative to the content of the lactic acid bacteria is 10 10The composition for regulating cytokine production due to viral infection according to

[11] or

[12] , characterized in that the amount is 100 μg to 500 μg per individual. This composition can provide a composition for regulating cytokine production due to viral infection that exhibits superior safety and efficacy.

[14] A composition for regulating cytokine production due to viral infection according to any one of

[10] to

[13] , characterized in that the retinoid compound is vitamin A or a vitamin A derivative. According to this composition, by employing vitamin A and vitamin A derivatives which are easily available, a composition for regulating cytokine production due to viral infection can be easily prepared.

[15] The composition for preventing and / or treating influenza according to any one of

[10] to

[13] , wherein the retinoid compound is provitamin A. This composition can provide a composition for preventing and / or treating influenza that is more safe and effective.

[16] A composition for preventing and / or treating influenza according to

[15] , characterized in that the provitamin A is β-carotene. According to this composition, by employing readily available β-carotene, a composition for preventing and / or treating influenza can be easily prepared.

[17] A composition for regulating cytokine production caused by viral infection described in any of

[11] to

[16] , characterized in that the lactic acid bacteria is Lactobacillus brevis strain KB290 (accession number NITE P-1537). This composition can provide a composition for regulating cytokine production due to viral infection that exhibits superior safety and efficacy.

[18] A composition for regulating cytokine production caused by viral infection described in any one of

[11] to

[17] , characterized in that the lactic acid bacteria are killed bacteria. This composition can provide a composition for regulating cytokine production due to viral infection that exhibits even greater safety and efficacy.

[19] A food, drink, and / or feed for regulating cytokine production caused by viral infection, comprising the composition according to any one of

[10] to

[18] . The food, drink, and feed can provide safe and effective food, drink, and feed for regulating cytokine production caused by viral infection. Effect of the Invention

[0013] According to the present invention, a novel composition for preventing and / or treating influenza that is safe and effective can be provided. [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 shows changes in mouse body weight. (A) Changes in body weight over time due to influenza virus infection. (B) Changes in body weight over time due to influenza virus infection when retinoic acid is taken. (C) Changes in body weight over time due to influenza virus infection when Lactobacillus brevis strain KB290 is taken. (D) Changes in body weight over time due to influenza virus infection when retinoic acid and Lactobacillus brevis strain KB290 are taken. (E) Changes in body weight over time in mice that were not infected with influenza virus. (F) A superposition of (A) to (E) above. [Diagram 2] FIG. 1 shows the viral titer due to influenza virus infection. [Diagram 3]FIG. 1 shows the amount of viral RNA due to influenza virus infection. (A) Changes in hemagglutinin RNA expression levels over time. (B) Changes in neuraminidase RNA expression levels over time. (C) Changes in RNA polymerase α subunit RNA expression levels over time. (D) Changes in M1 protein RNA expression levels over time. (E) Changes in M2 protein RNA expression levels over time. (F) Changes in NS1 protein RNA expression levels over time. (G) Changes in NS2 protein RNA expression levels over time. [Figure 4] FIG. 1 shows the effect of retinoic acid on cytokine production induced by influenza virus infection. (A) Changes in TNF-α concentration over time. (B) Changes in IL-1β concentration over time. (C) Changes in IL-6 concentration over time. (D) Changes in IL-10 concentration over time. (E) Changes in IL-33 concentration over time. (F) Changes in CXCL1 concentration over time. (G) Changes in CXCL10 concentration over time. (H) Changes in CCL2 concentration over time. (I) Changes in CCL5 concentration over time. [Diagram 5] FIG. 1 shows the effect of Lactobacillus brevis strain KB290 on cytokine production induced by influenza virus infection. (A) Changes in IFN-α concentration over time. (B) Changes in IFN-β concentration over time. (C) Changes in IL-1β concentration over time. (D) Changes in IL-6 concentration over time. (E) Changes in IL-10 concentration over time. (F) Changes in CCL5 concentration over time. [Figure 6] FIG. 1 shows the effect of retinoic acid and Lactobacillus brevis strain KB290 on cytokine production induced by influenza virus infection. (A) Shows changes in TNF-α concentration over time. (B) Shows changes in IL-1β concentration over time. (C) Shows changes in IL-6 concentration over time. (D) Shows changes in IL-10 concentration over time. (E) Shows changes in IL-33 concentration over time. (F) Shows changes in CXCL1 concentration over time. (G) Shows changes in CXCL10 concentration over time. (H) Shows changes in CCL2 concentration over time. (I) Shows changes in CCL5 concentration over time. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The composition, food and drink, and feed for preventing and / or treating influenza of the present invention will be described below.

[0016] [Composition for preventing and / or treating influenza] The composition for preventing and / or treating influenza of the present invention is characterized by comprising a retinoid compound, or a retinoid compound and lactic acid bacteria, as active ingredients. First, the active ingredient used in the present invention will be described in detail.

[0017] (Retinoid Compounds) Retinoid compounds are a group of compounds that have similar chemical structures and physiological functions to vitamin A. Vitamin A also includes retinol (vitamin A1) and 3-dehydroretinol (vitamin A2). The compound having a chemical structure similar to that of vitamin A is not particularly limited, and examples thereof include those having a structure represented by formula (I).

[0018] formula: JPEG0007632995000001.jpg61153 (I)

[0019] Vitamin A derivatives that have the same basic structure as vitamin A and exhibit similar physiological activities include retinoic acid, retinal, retinoic acid esters, retinoic acid amides, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid esters, and 3-dehydroretinoic acid amides.

[0020] The retinoid compound used in the present invention is not particularly limited, and examples thereof include vitamin A and vitamin A derivatives such as retinol, retinoic acid, retinal, and retinoic acid esters. The retinoid compound may be naturally derived or chemically synthesized. The retinoid compound may be any geometric isomer such as an all-trans isomer or a cis-trans isomer, or may be in the form of provitamin A that is converted in vivo to vitamin A or a vitamin A derivative.

[0021] Specific examples of retinol include all-trans-retinol, 9-cis-retinol, 11-cis-retinol, 13-cis-retinol, 3,4-didehydro-retinol, 3,4-didehydro-9-cis-retinol, 3,4-didehydro-11-cis-retinol, 3,4-didehydro-13-cis-retinol, and the like.

[0022] Specific examples of retinoic acid include all-trans-retinoic acid, 9-cis retinoic acid, 11-cis retinoic acid, 13-cis retinoic acid, and 3-dehydroretinoic acid.

[0023] Specific examples of retinal include all-trans-retinal, 9-cis retinal, 11-cis retinal, 13-cis retinal, 3,4-dehydroretinal, 13-ethyl retinal, 9-dm-retinal, 3-hydroxyretinal, 4-hydroxyretinal, naphthyl retinal, 3,7,11-trimethyl-dodeca-2,4,6,8,10-pentaenal, 3,7 dimethyl-deca-2,4,6,8-tetraenal, and 3,7-dimethyl-octa-2,4,6-trienal.

[0024] Specific examples of retinoic acid esters include retinyl palmitate, retinyl formate, retinyl acetate, retinyl propionate, retinyl butyrate, retinyl valerate, retinyl isovalerate, retinyl hexanoate, retinyl heptanoate, retinyl octanoate, retinyl nonanoate, retinyl decanoate, retinyl undecanoate, retinyl laurate, retinyl tridecanoate, retinyl myristate, retinyl pentadecanoate, retinyl heptadecanoate, retinyl stearate, retinyl isostearate, retinyl nonadecanoate, retinyl arachidonate, retinyl behenate, retinyl linoleate, and retinyl oleate.

[0025] Specific examples of provitamin A derived from natural products include α-carotene, β-carotene, γ-carotene, β-cryptoxanthin, and echinenone. From the viewpoints of availability and safety, the retinoid compound is preferably vitamin A, retinoic acid, all-trans-retinal, or β-carotene, and more preferably vitamin A or retinoic acid. These retinoid compounds may be used alone or in combination of two or more.

[0026] The content of the retinoid compound in the composition is not particularly limited, and is, for example, 0.01% by mass or more and 20% by mass or less. The lower limit is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, even more preferably 1.0% by mass or more, and particularly preferably 3.0% by mass or more. On the other hand, the upper limit is preferably 115.0% by mass or less, more preferably 10.0% by mass or less, even more preferably 7.5% by mass or less, and particularly preferably 5.0% by mass or less. By setting the content of the retinoid compound in the composition within the above range, the effect of the present invention of providing a composition for preventing and / or treating influenza that has excellent preventive and therapeutic effects against influenza can be more effectively achieved.

[0027] (lactic acid bacteria) The lactic acid bacteria used in the present invention are not particularly limited as long as they are lactic acid bacteria strains that have preventive and therapeutic effects against influenza. Specific examples of lactic acid bacteria include Lactobacillus brevis KB290 strain, Lactobacillus brevis NC-4 strain, Lactobacillus brevis SAM2447 strain, Lactobacillus brevis IFO-12005 strain, Lactobacillus brevis CD2 strain, Lactobacillus brevis DNBL1871 strain, Lactobacillus brevis ATCC14869 strain, Lactobacillus brevis JCM1559 strain, Lactobacillus brevis NBRC12005 strain, Lactobacillus brevis 1059T strain, Lactobacillus brevis 1170 strain, Lactobacillus brevis NTM003 strain, Lactobacillus brevis 1059T strain, Lactobacillus brevis 1170 strain, and the like. From the viewpoints of safety and effectiveness, the lactic acid bacteria is preferably Lactobacillus brevis KB290 strain, which was deposited at the National Institute of Technology and Evaluation, Patent Microorganisms Deposit Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) on February 13, 2013 (accession number NITE P-1537). These lactic acid bacteria may be used alone or in combination of two or more kinds.

[0028] The form of the lactic acid bacteria is not particularly limited as long as it is a lactic acid bacteria strain that has a preventive and therapeutic effect against influenza, and examples thereof include bacterial cells, cultures of bacterial cells, powdered bacterial cells, crushed bacterial cells, and extracts of bacterial cells. The lactic acid bacteria may be live bacteria, dead bacteria, or a mixture of live bacteria and dead bacteria. Methods for preparing dead bacteria include, for example, heat treatment, ultra-high temperature heat treatment, pressure treatment, high pressure steam treatment, dry heat treatment, etc. From the viewpoint of quality control, the lactic acid bacteria are preferably in the form of killed bacteria.

[0029] The content of lactic acid bacteria in the composition is not particularly limited, and may be, for example, 0.001 × 10 per 1 g of the composition. 10 pcs or more 1000×1010 The lower limit is preferably 0.01 × 10 per 1 g of the composition. 10 More preferably, the number is 0.1×10 per gram of the composition. 10 or more, more preferably 0.5×10 10 or more, and particularly preferably 1×10 10 On the other hand, the upper limit is preferably 500×10 per 1 g of the composition. 10 More preferably, the number of 10 More preferably, the number of 10 10×10 or less per gram of the composition. 10 1 or less. By setting the content of lactic acid bacteria in the composition within the above range, the effect of the present invention of providing a composition for preventing and / or treating influenza that has excellent preventive and therapeutic effects against influenza can be more effectively achieved.

[0030] The ratio of the content of the retinoid compound to the content of the lactic acid bacteria in the composition is not particularly limited. For example, 10 The lower limit is preferably 1 μg or more and 100 g or less per lactic acid bacteria. 10 More preferably, the amount of lactic acid bacteria is 10 μg or more per unit. 10 More preferably, the amount of lactic acid bacteria is 10 10 More preferably, the amount of lactic acid bacteria is 10 10 On the other hand, the upper limit is preferably 10 10 Each piece should contain 1 g or less of lactic acid bacteria, and more preferably 10 10 More preferably, the amount of lactic acid bacteria is 10 mg or less per unit. 10 The amount of lactic acid bacteria is preferably 10 10 The amount is 500 μg or less per piece, and most preferably 400 μg or less. By setting the ratio of the retinoid compound content to the lactic acid bacteria content in the composition within the above range, the effect of the present invention of providing a composition for preventing and / or treating influenza that has excellent preventive and therapeutic effects against influenza is further exerted.

[0031] Next, the components that may be contained in the composition of the present invention will be described in detail. The composition for preventing and / or treating influenza of the present invention may contain any ingredients such as other active ingredients and additives, so long as the action of the retinoid compound and lactic acid bacteria against influenza is not impaired.

[0032] (Other active ingredients) The active ingredients contained in the composition of the present invention, other than the retinoid compound and the lactic acid bacteria, are not particularly limited. Specific examples of other active ingredients include, for example, anti-influenza drugs such as oseltamivir, zanamivir, peramivir, laninamivir, and baloxavir marboxil, influenza virus antigens such as A / H1N1, A / H3N2, A / H5N1, and B, Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus casei, Lactobacillus delbrueckii subspecies bulgaricus, Bifidobacterium longum, Bifidobacterium breve, Propionibacterium freudenreichii, and the like. freudenreichii), and symptomatic treatments such as antipyretics, analgesics, anti-inflammatory agents, antiallergic agents, cough suppressants, and expectorants. Furthermore, these active ingredients may be used alone or in combination of two or more.

[0033] (Additives) The additives contained in the composition of the present invention are not particularly limited, and examples thereof include pharma- ceutically acceptable bases, carriers, excipients, binders, disintegrants, lubricants, colorants, pH regulators, buffers, stabilizers, and preservatives. These additives may be used alone or in combination of two or more.

[0034] The pharma- ceutically acceptable base is not particularly limited, and examples thereof include water, polar solvents such as ethanol, and oily bases.

[0035] The carrier and excipient are not particularly limited, and examples thereof include lactose, glucose, sucrose, mannitol, potato starch, corn starch, calcium carbonate, calcium phosphate, calcium sulfate, and crystalline cellulose.

[0036] The binder is not particularly limited, and examples thereof include starch, gelatin, syrup, tragacanth gum, polyvinyl alcohol, polyvinyl ether, polyvinylpyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, and carboxymethyl cellulose.

[0037] The disintegrant is not particularly limited, and examples thereof include starch, agar, gelatin powder, crystalline cellulose, calcium carbonate, sodium bicarbonate, sodium alginate, sodium carboxymethylcellulose, and calcium carboxymethylcellulose.

[0038] The lubricant is not particularly limited, and examples thereof include magnesium stearate, hydrogenated vegetable oil, talc, macrogol, and the like.

[0039] The coloring agent is not particularly limited, and examples thereof include cochineal, carmine, curcumin, riboflavin, annatto, titanium oxide, iron oxide, talc, calcined silica, and magnesium carbonate.

[0040] The pH adjuster is not particularly limited, and examples thereof include citric acid, gluconic acid, succinic acid, potassium carbonate, and lactic acid.

[0041] The buffering agent is not particularly limited, and examples thereof include phosphate, arginine, and histidine.

[0042] The stabilizer is not particularly limited, and examples thereof include arginine, polysorbate 80, macrogol 4000, and the like.

[0043] The preservative is not particularly limited, and examples thereof include benzoic acid, phenoxyethanol, and thimerosal.

[0044] Other additives include solubilizing agents, surfactants, emulsifiers, antioxidants, gloss agents, foaming agents, moisture-proofing agents, preservatives, sweetening agents, flavoring agents, refreshing agents, flavoring agents, fragrances, aromatics, disintegration aids, etc. These additives may be used alone or in combination of two or more.

[0045] Due to the above-mentioned characteristics, the composition for preventing and / or treating influenza of the present invention can safely and effectively prevent and treat influenza.

[0046] [Use and form of composition] The form and use of the composition of the present invention for preventing and / or treating influenza will now be described in detail. The use of the composition of the present invention is not particularly limited, and it can be incorporated, for example, into medicines, quasi-drugs, cosmetics, foods and beverages, feeds, and the like. Furthermore, pharmaceuticals, quasi-drugs, cosmetics, foods, beverages, feeds, etc., containing the composition of the present invention are used for humans or non-human animals. Non-human animals are not particularly limited, but include, for example, mammals, birds, reptiles, amphibians, fish, etc., and preferably chickens, cows, pigs, dogs, cats, etc.

[0047] (Medicines, quasi-drugs, cosmetics) The form of the pharmaceutical, quasi-drug, or cosmetic containing the composition of the present invention is not particularly limited, and examples of the oral administration forms include tablets such as sugar-coated tablets, buccal tablets, coated tablets, and chewable tablets, troches, pills, powders, capsules including soft capsules, granules, suspensions, emulsions, syrups including dry syrups, and liquids such as elixirs, and the like; and parenteral administration forms include injections such as intravenous injection, subcutaneous injection, intraperitoneal injection, and intramuscular injection, transdermal absorption tapes, aerosols, and suppositories for transdermal, nasal, pulmonary, enteral, oral, and transmucosal administration. From the viewpoint of convenience and versatility, the pharmaceutical, quasi-drug, and cosmetic are preferably administered orally.

[0048] The dosage of the drug, quasi-drug, or cosmetic containing the composition of the present invention is not particularly limited. For example, the dosage of the retinoid compound is 1 μg / kg (body weight) or more and 50 mg / kg (body weight) or less per day, and the dosage of the lactic acid bacteria is 0.0001 × 10 10 pieces / kg (weight) or more 1000×10 10 The lower limit of the retinoid compound is preferably 10 μg / kg (body weight) or more per day, more preferably 50 μg / kg (body weight) or more per day, even more preferably 100 μg / kg (body weight) or more per day, and particularly preferably 500 μg / kg (body weight) or more per day. On the other hand, the upper limit of the retinoid compound is preferably 25 mg / kg (body weight) or less per day, more preferably 10 mg / kg (body weight) or less per day, even more preferably 5 mg / kg (body weight) or less per day, and particularly preferably 1 mg / kg (body weight) or less per day. In addition, the lower limit of the lactic acid bacteria is preferably 0.0005×10 10 10 / kg (body weight) or more, more preferably 0.001 x 10 10 10 / kg (body weight) or more, and more preferably 0.005×10 10 10 / kg (body weight) or more, and particularly preferably 0.01×1010 The upper limit of the number of lactic acid bacteria is preferably 500 × 10 10 per kg of body weight or less, and more preferably 100×10 10 per kg of body weight or less, and more preferably 50×10 10 per kg (body weight) or less, and particularly preferably 10×10 10 per kg (body weight) or less.

[0049] The number of times that a pharmaceutical, quasi-drug, or cosmetic containing the composition of the present invention is administered is not particularly limited, and may be, for example, once a day, twice a day, or three times a day. The administration period of the pharmaceutical, quasi-drug, or cosmetic containing the composition of the present invention is not particularly limited, and may be, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, or 10 days. In particular, when the composition of the present invention is used for the purpose of preventing influenza, it may be used continuously without specifying the administration period. In addition, the administration during the administration period is not particularly limited, and may be, for example, daily administration, or every other day, every third day, or every third day.

[0050] (Food and beverages, feed) The form of the food, drink, or feed containing the composition of the present invention is not particularly limited, and examples include processed foods, health foods (nutritional supplements, nutritional functional foods, foods for the sick, foods for specified health uses, foods with functional claims, etc.), supplements, foods for the sick (hospital food, food for sick people, nursing care food, etc.), confectionery, oils and fats, dairy products, retort foods, microwave foods, frozen foods, seasonings, dietary supplements, beverages, and nutritional drinks.

[0051] The shape and properties of the food, drink, and feed containing the composition of the present invention are not particularly limited, and examples thereof include solid, semisolid, gel, liquid, powder, etc. In particular, when used as a health food or supplement, it is preferable to make it in the form of granules, capsules, tablets, chewable agents, beverage powders, drinks, smoothies, jellies, gummies, etc. so that it can be continuously and easily ingested.

[0052] Food, drink, and feed containing the composition of the present invention may be labeled with a label indicating that the composition may have a beneficial effect against influenza. Such labeling can be attached to the container packaging means by a known method, and this clearly indicates that the food, drink, and feed containing the composition of the present invention are used to improve influenza, making it clear to distinguish them from ordinary food, drink, and feed.

[0053] The intake amount of food, drink, or feed containing the composition of the present invention is not particularly limited. For example, the retinoid compound is 1 μg / kg (body weight) or more and 50 mg / kg (body weight) or less per day, and the lactic acid bacteria is 0.0001 × 10 10 pieces / kg (weight) or more 1000×10 10 The lower limit of the retinoid compound is preferably 10 μg / kg (body weight) or more per day, more preferably 50 μg / kg (body weight) or more per day, even more preferably 100 μg / kg (body weight) or more per day, and particularly preferably 500 μg / kg (body weight) or more per day. On the other hand, the upper limit of the retinoid compound is preferably 25 mg / kg (body weight) or less per day, more preferably 10 mg / kg (body weight) or less per day, even more preferably 5 mg / kg (body weight) or less per day, and particularly preferably 1 mg / kg (body weight) or less per day. In addition, the lower limit of the lactic acid bacteria is preferably 0.0005×10 10 10 / kg (body weight) or more, more preferably 0.001 x 10 10 10 / kg (body weight) or more, and more preferably 0.005×10 10 10 per kg (body weight) or more, and particularly preferably 0.01 × 10 10 The upper limit of the number of lactic acid bacteria is preferably 500 × 10 10 per kg of body weight or less, and more preferably 100×10 10 per kg of body weight or less, and more preferably 50×1010 per kg (body weight) or less, and particularly preferably 10×10 10 per kg (body weight) or less. The amount of intake can be changed depending on the type of non-human animal.

[0054] The number of times that the food, drink, or feed containing the composition of the present invention is ingested is not particularly limited, and may be, for example, once a day, twice a day, or three times a day. The intake period of the food, drink, or feed containing the composition of the present invention is not particularly limited, and may be, for example, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, etc. In particular, when the composition of the present invention is used for the purpose of preventing influenza, it may be used continuously without specifying the intake period. In addition, the administration during the intake period is not particularly limited, and may be, for example, daily intake, intake every other day, intake every third day, intake every third day, etc. The intake period and intervals can be changed depending on the type of non-human animal.

[0055] Due to the above-mentioned characteristics, the composition for preventing and / or treating influenza of the present invention can safely and effectively prevent and treat influenza.

[0056] [Composition for regulating cytokine production due to viral infection] A composition for regulating cytokine production due to viral infection may have the same compositional components, uses and forms as a composition for preventing and / or treating influenza.

[0057] The cytokines that regulate the production are not particularly limited. Specific examples of cytokines include TNF-α, IFN-α, IFN-β, IFN-γ, TGFβ, G-CSF, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-11, IL-12, IL-13, IL-18, IL-33, CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL10, CCL1, CCL2, CCL3, CCL4, CCL5, and CCL8.

[0058] The regulation of cytokine production is not particularly limited, and includes, for example, promoting or suppressing the amount of cytokine production at any time point after viral infection.

[0059] The infective virus is not particularly limited, and examples thereof include RNA viruses such as influenza virus, norovirus, rotavirus, and human immunodeficiency virus, and DNA viruses such as hepatitis B virus, adenovirus, herpes simplex virus, and cytomegalovirus.

[0060] Due to the above-mentioned characteristics, the composition for regulating cytokine production can safely and effectively regulate the amount of cytokine produced, and is therefore useful for the prevention and treatment of viral infections. EXAMPLES

[0061] [Example 1: Effect of the present invention on influenza] (Mouse rearing) We used 7-8 week-old female BALB / c mice (AgResearch Ruakura Small Animal Facility, Palmerston North, New Zealand) that were fed normal solid food (Prolab RMH 1800, LabDiet, USA) and sterilized water. The mice were housed in plastic cages with 3-4 mice per cage, and the environment was controlled with a 12-hour light / dark cycle, room temperature of 21°C, and humidity of 50%. The mice were divided into five groups, each with 30 mice: an untreated group infected with influenza virus (hereafter referred to as the "untreated group"), a control group infected with influenza virus (hereafter referred to as the "control group"), a group administered retinoic acid before influenza virus infection (hereafter referred to as the "RA-administered group"), a group administered Lactobacillus brevis KB290 strain before influenza virus infection (hereafter referred to as the "KB-administered group"), and a group administered retinoic acid and Lactobacillus brevis KB290 strain before influenza virus infection (hereafter referred to as the "RA+KB-administered group").

[0062] (Method of administering test food) The test food for the untreated group was a mixture of 185 μL phosphate buffered saline (PBS), 15 μL canola oil. The test food for the RA treatment group was a mixture of 185 μL PBS, 15 μL canola oil containing 300 μg retinoic acid. The test food for the KB administration group was 10 10 A mixture of 185 μL of PBS and 15 μL of canola oil containing Lactobacillus brevis strain KB290 was prepared. The test food for the RA+KB group was 10 10 A mixture of 185 μL of PBS containing Lactobacillus brevis strain KB290 and 15 μL of canola oil containing 300 μg of retinoic acid was prepared. The retinoid compound used in the test food was retinoic acid (Sigma-Aldrich Co. LLC., USA), Lactobacillus brevis KB290 strain was a freeze-dried bacterial liquid produced at the Komaki Factory of Kagome Co., Ltd., and canola oil (Palm's canola oil, Countdown, New Zealand).

[0063] Retinoic acid and Lactobacillus brevis KB290 strain were administered by force feeding 200 μL of the test food once daily using a 16-gauge polyurethane feeding tube (Instech Laboratories Inc., USA) for 14 days prior to influenza virus infection. The test food was administered by force once a day, at 200 μL per day, using a 16-gauge polyurethane feeding tube (Instech Laboratories Inc., USA), for 14 days prior to influenza virus infection.

[0064] (How influenza viruses are transmitted) The influenza virus used for the infection of mice was influenza virus A / PR / 8 / 34 (H1N1). The influenza virus solution used for intranasal infection was prepared in PBS to a dose of 0.5×LD50. Influenza virus infection was performed by anesthetizing the mice on the day after administration of the test food for 14 days and then administering an influenza virus solution intranasally.

[0065] (Weight measurement) The body weight of the mice was measured using an electronic scale (product number 6971956, Sartorius AG, Germany) once every other day during the 14 days during which the test food was administered, once on the day of influenza virus infection, and once daily for 14 days after influenza virus infection.

[0066] Figure 1 (A)-(E) show the predicted curves based on the actual body weights and the average values ​​of each group over time, with the influenza virus infection treatment date being set as day 0, the test food administration period being from -1 to -14, and the period after infection treatment being from 1 to 14 days. Figure 1 (F) shows the calculation of the 84% parametric bootstrap confidence interval for the predicted curves of each group. The width of the 84% confidence interval was set so that there would be a 5% false positive rate when there was no difference between the groups.

[0067] (Measurement of viral titer) For the measurement of virus titer, 10 mice from each group were euthanized by cervical dislocation on the third day after influenza virus infection, and the lungs were promptly excised and homogenized using a stirring and grinding device (TissueLyser II, Qiagen NV, Germany) in the presence of a proteolytic inhibitor (Sigma-Aldrich Co. LLC., USA), 100 U / mL penicillin, and 100 μg / mL streptomycin (Life Technologies New Zealand Ltd., Auckland New Zealand). The supernatant was centrifuged at 4°C and 15,000 × g for 30 minutes. The influenza virus titer in the supernatant fraction of the lung tissue homogenate was determined by a chicken hemagglutination test using MDCK cells and determined as the 50% tissue culture cell infectivity (TCID 50 ) was calculated as

[0068] FIG. 2 shows the log 100% virus titer in lung tissue 3 days after influenza virus infection. 10 (TCID 50 The values ​​are shown as mean ± standard error, and a P value of less than 0.05 was determined to be statistically significant and is marked with an asterisk (*).

[0069] (Measurement of viral RNA quantity) For the measurement of viral RNA quantity, 10 mice from each group were euthanized by cervical dislocation on days 3, 7, and 14 after influenza virus infection, and the lungs were promptly excised and treated with a stirring and grinding device (TissueLyser II, Qiagen NV, Germany) in the presence of a proteolytic inhibitor (Sigma-Aldrich Co. LLC., USA), 100 U / mL penicillin, and 100 μg / mL streptomycin (Life Technologies New Zealand Ltd., Auckland New Zealand) to obtain homogenates. RNA was prepared from the homogenates using an RNA preparation kit (RNeasy, Qiagen NV, Germany).

[0070] The RNA samples were fluorescently labeled with the seven mouse reference genes in Table 1 and the influenza A virus (IAV) genes in Table 2 using a hybridization kit (PlexSet Reagent, NanoString Technologies Inc., USA). The fluorescently labeled genes were quantified as single molecule numbers using a fluorometer (nCounter Analysis System Technologies Inc., USA).

[0071] [Table 1]

[0072] [Table 2]

[0073] Figure 3 shows the expression of seven viral genes in lung tissues on days 3 (white), 7 (gray), and 14 (black) after influenza virus infection, as expressed as RNA counts. Values ​​are shown as mean ± standard error, and a P value of less than 0.05 compared to the untreated group was determined to be statistically significant and is marked with an *.

[0074] (Result 1: Effect of test food on body weight) As shown in Figure 1, body weight loss occurred over the 10th day after influenza virus infection (Figure 1(A)). On the other hand, weight loss due to influenza virus infection was also observed in the RA and KB groups, but was improved compared to the control group (Figure 1(B) and Figure 1(C)). Furthermore, in the RA+KB group, weight loss was improved to the same level as in the untreated group (Figure 1(D) and (E)).

[0075] Therefore, it was found that the intake of retinoic acid and Lactobacillus brevis KB290 strain improved weight loss caused by influenza virus infection. In addition, it was shown that the improving effect on weight loss caused by influenza virus infection was strongest in the following order: intake of Lactobacillus brevis strain KB290, intake of retinoic acid, and intake of retinoic acid and Lactobacillus brevis strain KB290.

[0076] (Result 2: Effect of test foods on viral titers) As shown in Figure 2, influenza virus infection significantly increased the viral titer in lung tissue. On the other hand, the viral titer in the RA, KB, and RA+KB groups was significantly decreased compared to the control group. In particular, the viral titer in the RA+KB group was significantly decreased compared to the control group.

[0077] Therefore, it was shown that the intake of retinoic acid and Lactobacillus brevis KB290 strain suppressed the increase in viral titer due to influenza virus infection. In addition, it was found that the inhibitory effect on the viral titer caused by influenza virus infection was significantly exhibited by the combined use of retinoic acid and Lactobacillus brevis KB290 strain.

[0078] (Result 3: Effect of test foods on viral RNA levels) As shown in Figures 3(A)-(G), influenza virus infection increased the RNA levels of HA, NA, PA, M1, M2, NS1, and NS2 from days 3 to 7 after infection, and then decreased on day 14 after infection. The increase in all of these influenza virus RNA levels was significantly suppressed in the RA+KB group compared to the control group, RA group, and KB group.

[0079] Therefore, it was shown that the intake of retinoic acid and Lactobacillus brevis KB290 strain significantly suppressed the amount of influenza virus RNA in the lungs.

[0080] As shown in the above results 1 to 3, the composition of the present invention improves weight loss caused by influenza virus infection, suppresses the increase in viral titer, and, when used in combination with retinoic acid and Lactobacillus brevis KB290 strain, significantly suppresses the proliferation of influenza viruses in the lungs. Therefore, a composition containing a retinoid compound and lactic acid bacteria is recognized to be effective in the prevention and treatment of influenza.

[0081] [Example 2: Effect of the present invention on cytokine production induced by virus infection] The mice were bred and the test food was administered in the same manner as in Example 1.

[0082] (Cytokine measurement) For the measurement of cytokines, 10 mice per group were euthanized by cervical dislocation on days 3, 7, and 14 after influenza virus infection, and the lungs were promptly removed and homogenized using a stirring and grinding device (TissueLyser II, Qiagen NV, Germany) in the presence of a proteolytic inhibitor (Sigma-Aldrich Co. LLC., USA), 100 U / mL penicillin, and 100 μg / mL streptomycin (Life Technologies New Zealand Ltd., Auckland New Zealand). The supernatant was then centrifuged at 4°C and 15,000 × g for 30 minutes. TNF-α, IFN-α, IFN-β, IL-1β, IL-6, IL-10, IL-33, CXCL1, CXCL10, CCL2, and CCL5 in the supernatant fraction of lung tissue homogenates were quantified by flow cytometry using a bead assay kit (LEGENDplex, BioLegend Inc., USA). Cytokine concentrations were calculated as the cytokine mass relative to the total protein amount of lung tissue.

[0083] 4 to 6 show the cytokine concentrations in lung tissues on days 3 (white), 7 (gray), and 14 (black) after influenza virus infection in the RA, KB, and RA+KB groups, respectively. Values ​​are expressed as mean ± standard error.

[0084] (Result 4: Effect of retinoic acid on cytokine production) As shown in Figures 4(A), (C), (D), (F), (G), (H), and (I), comparing the control group with the RA-treated group, retinoic acid suppressed the increased production of TNF-α, IL-6, CXCL1, CCL2, and CCL5 on day 3 after influenza virus infection, and TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5 on day 7.

[0085] Therefore, it was revealed that ingestion of retinoic acid suppressed the production of TNF-α, IL-6, CXCL1, CCL2, and CCL5 up to day 3 after influenza virus infection, and TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5 up to day 7. Since cytokine production was suppressed by ingestion of retinoic acid, it is speculated that retinoic acid may suppress the invasion of viruses into the lungs, which causes increased cytokine production.

[0086] (Result 5: Effect of Lactobacillus brevis KB290 on cytokine production) As shown in Figure 5(D), when comparing the control group with the KB-administered group, Lactobacillus brevis KB290 strain suppressed the increase in IL-6 production on days 3 and 7 after influenza virus infection. On the other hand, as shown in Figures 5(A), (B), (C), and (E), when comparing the control group and the KB-administered group, Lactobacillus brevis KB290 strain promoted the production of IFN-α, IFN-β, IL-1β, and IL-10 on the 3rd day after influenza virus infection.

[0087] Therefore, it was revealed that ingestion of Lactobacillus brevis strain KB290 suppressed the production of IL-6 up to 7 days after influenza virus infection, and promoted the production of IFN-α, IFN-β, IL-1β, and IL-10 on day 3 after infection. Since the production of many of the cytokines suppressed by the ingestion of retinoic acid was promoted by the ingestion of Lactobacillus brevis strain KB290, it is speculated that Lactobacillus brevis strain KB290 contributes to the improvement of influenza through a mechanism different from that of retinoic acid.

[0088] (Result 6: Effects of retinoic acid and Lactobacillus brevis KB290 strain on cytokine production) As shown in Figures 6(A), (C), (F), (G), (H), and (I), when comparing the control group with the RA+KB administration group, retinoic acid and Lactobacillus brevis KB290 strain suppressed the increase in production of TNF-α, IL-6, CXCL1, CXCL10, CCL2, and CCL5 on days 3 and 7 after influenza virus infection. In addition, as shown in Figure 6(D), when comparing the control group with the RA+KB administration group, retinoic acid and Lactobacillus brevis KB290 strain suppressed the increase in production of IL-10 on day 7 after influenza virus infection.

[0089] Therefore, it was revealed that the intake of retinoic acid and Lactobacillus brevis KB290 strain suppressed the production of TNF-α, IL-6, CXCL1, CXCL10, CCL2, and CCL5 up to 7 days after influenza virus infection, and the production of IL-10 on the 7th day after infection. In addition, when retinoic acid and Lactobacillus brevis KB290 strain were used in combination, the production of several cytokines was confirmed to be significantly suppressed on the 3rd and 7th days compared to when either was taken alone. Therefore, it is presumed that the combined use of retinoic acid and Lactobacillus brevis KB290 strain more strongly suppresses influenza virus infection.

[0090] As shown in the above Results 4 to 6, the composition of the present invention can regulate the amount of cytokines produced by viral infection. Therefore, the composition containing a retinoid compound and lactic acid bacteria is recognized to be effective in preventing and treating viral infections such as influenza virus by optimizing the immune mechanism. [Industrial Applicability]

[0091] The present invention makes it possible to safely prevent and treat influenza, thereby making it possible to suppress the large-scale spread of influenza virus infection in populations of humans, livestock, and the like.

Claims

1. Contains retinoid compounds and lactic acid bacteria as active ingredients. The lactic acid bacterium is Lactobacillus brevis KB290 strain (accession number NITE P-1537), A composition for preventing and / or treating influenza (excluding compositions containing lactic acid bacteria, cranberry extract, and propolis extract), characterized in that the retinoid compound is at least one selected from vitamin A, retinol, 3-dehydroretinol, retinoic acid, retinal, retinoic acid ester, retinoic acid amide, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid ester, 3-dehydroretinoic acid amide, α-carotene, β-carotene, γ-carotene, β-cryptoxanthin, and echinenone.

2. The content of the retinoid compound relative to the content of the lactic acid bacteria is 10 １０ The composition for preventing and / or treating influenza according to claim 1, characterized in that the amount is 100 μg to 500 μg per individual.

3. 3. The composition for preventing and / or treating influenza according to claim 1 or 2, wherein the retinoid compound is at least one selected from vitamin A, retinol, 3-dehydroretinol, retinoic acid, retinal, retinoic acid ester, retinoic acid amide, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid ester, and 3-dehydroretinoic acid amide.

4. the retinoid compound is provitamin A; 3. The composition for preventing and / or treating influenza according to claim 1 or 2, characterized in that the provitamin A is at least one selected from alpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin, and echinenone.

5. The composition for preventing and / or treating influenza according to claim 4, wherein the provitamin A is β-carotene.

6. The composition for preventing and / or treating influenza according to any one of claims 1 to 5, wherein the lactic acid bacteria are killed.

7. A food, drink, and / or feed for preventing and / or treating influenza, comprising the composition according to any one of claims 1 to 6.

8. Contains retinoid compounds and lactic acid bacteria as active ingredients. The lactic acid bacterium is Lactobacillus brevis KB290 strain (accession number NITE P-1537), A composition for regulating cytokine production caused by viral infection (excluding compositions containing lactic acid bacteria, cranberry extract, and propolis extract), characterized in that the retinoid compound is at least one selected from vitamin A, retinol, 3-dehydroretinol, retinoic acid, retinal, retinoic acid ester, retinoic acid amide, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid ester, 3-dehydroretinoic acid amide, α-carotene, β-carotene, γ-carotene, β-cryptoxanthin, and echinenone.

9. The composition for regulating cytokine production caused by viral infection according to claim 8, characterized in that the regulation of cytokine production caused by viral infection is suppression of production of at least one cytokine selected from the group consisting of TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2 and CCL5.

10. The content of the retinoid compound relative to the content of the lactic acid bacteria is 10 １０ The composition for regulating cytokine production due to viral infection according to claim 8 or 9, characterized in that the amount is 100 μg to 500 μg per individual.

11. The composition for regulating cytokine production due to viral infection according to any one of claims 8 to 10, characterized in that the retinoid compound is at least one selected from vitamin A, retinol, 3-dehydroretinol, retinoic acid, retinal, retinoic acid ester, retinoic acid amide, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid ester, and 3-dehydroretinoic acid amide.

12. the retinoid compound is provitamin A; The composition for regulating cytokine production due to viral infection according to any one of claims 8 to 10, characterized in that the provitamin A is at least one selected from alpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin, and echinenone.

13. The composition for regulating cytokine production due to viral infection according to claim 12, characterized in that the provitamin A is beta-carotene.

14. The composition for regulating cytokine production due to viral infection according to any one of claims 8 to 13, characterized in that the lactic acid bacteria are killed bacteria.

15. A food, drink, and / or feed for regulating cytokine production caused by viral infection, comprising the composition according to any one of claims 8 to 14.

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