Compositions, foods, and feeds 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 or beverage, or animal feed for treating and / or preventing influenza, comprising a retinoid compound, or a retinoid compound and lactic acid bacteria as active ingredients. [Background technology]

[0002] Influenza is an abbreviation for influenza virus infection. The influenza virus is highly contagious, leading to a high incidence of influenza in human populations and regularly causing large-scale morbidity and mortality rates. Unlike the common cold, influenza is characterized by a relatively rapid onset of high fever, nasal congestion, headache, general malaise, and muscle pain, accompanied by upper respiratory tract symptoms such as sore throat, runny nose, and cough. Furthermore, complications such as bronchitis, pneumonia, and influenza encephalopathy can occur, and can even be fatal. In particular, it poses a threat to people with underlying conditions such as chronic heart disease, lung disease, kidney disease, liver disease, and metabolic disorders, as well as to the elderly and infants with weakened immune systems, as they are more likely to develop severe illnesses.

[0003] The primary method of preventing influenza is vaccination. Oseltamivir, zanamivir, peramivir, and laninamivir are used as treatments for influenza. However, these vaccines and treatments can cause serious side effects. On the other hand, it has been reported that highly safe natural substances do not provide therapeutic effects (Non-Patent Literature 1). Therefore, there is a need for the development of new compositions that not only offer high safety but also sufficient preventive and therapeutic effects.

[0004] To date, preventive and therapeutic drugs for influenza infection containing lactic acid bacteria as active ingredients, which have demonstrated high safety, have been reported. For example, Patent Document 1 discloses an influenza preventive and therapeutic agent containing Lactobacillus acidophilus CL-92 strain as an active ingredient. 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 produced by Lactobacillus lactic acid bacteria suppress the decline in acquired immune function caused by anti-influenza drugs.

[0005] Incidentally, the defense mechanism of lactic acid bacteria against infectious diseases such as influenza viruses is thought to involve the regulatory action of the immune system. For example, Patent Document 3 describes an innate immune activator that contains lactic acid bacteria as an active ingredient. It is thought that orally ingested lactic acid bacteria are taken up by Peyer's patches in the intestinal tract and phagocytosed by macrophages and dendritic cells, thereby activating immune cells and stimulating innate immunity. Non-Patent Document 3 also describes that lactic acid bacteria are recognized by TLRs. As a result, it is thought that lactic acid bacteria regulate the function of macrophages and dendritic cells via TLR signaling. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] Japanese Patent Publication No. 2012-72113 [Patent Document 2] Japanese 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 alleviates 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] [Problems to be Solved by the Invention]

[0008] So far, compositions containing various active ingredients have been developed to prevent and treat influenza, but those having sufficient safety, preventive effect, and therapeutic effect have not been obtained. Therefore, a composition with high safety as well as excellent preventive and therapeutic effects is desired. Therefore, an object of the present invention is to provide a composition, a food or drink, and a feed for preventing and / or treating influenza, which are excellent not only in safety but also in preventive and therapeutic effects. [Means for Solving the Problems]

[0009] As a result of intensive studies on the above problems, the present inventors have found that influenza can be prevented and treated by ingesting a composition containing a retinoid compound or a composition containing a retinoid compound and lactic acid bacteria as active ingredients, and thus 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, characterized by containing a retinoid compound as an active ingredient. According to this composition, a composition for preventing and / or treating influenza, which is safe and effective, can be provided. [2] The composition for preventing and / or treating influenza according to [1], further characterized by containing lactic acid bacteria as an active ingredient. According to this composition, a composition for preventing and / or treating influenza, which exhibits excellent safety and effectiveness, can be provided. [3] The composition for preventing and / or treating influenza according to [2], characterized in that the content of the retinoid compound relative to the content of the lactic acid bacteria is 100 μg to 500 μg per 10 10 lactic acid bacteria. According to this composition, a composition for preventing and / or treating influenza, which exhibits even more excellent safety and effectiveness, can be provided. [4] The composition for preventing and / or treating influenza according to any one of [1] to [3], characterized in that the retinoid compound is vitamin A or a vitamin A derivative. This composition allows for the easy preparation of a composition for preventing and / or treating influenza by employing readily available vitamin A and vitamin A derivatives. [5] A composition for preventing and / or treating influenza according to any one of [1] to [3], characterized in that the retinoid compound is provitamin A. This composition provides a composition for preventing and / or treating influenza that exhibits superior safety and efficacy. [6] The composition for preventing and / or treating influenza according to [5], characterized in that the provitamin A is beta-carotene. This composition allows for the easy preparation of a composition for preventing and / or treating influenza by employing readily available beta-carotene. [7] A composition for preventing and / or treating influenza according to any one of [2] to [6], characterized in that the lactic acid bacterium is Lactobacillus brevis strain KB290 (accession number NITE P-1537). This composition provides a composition for preventing and / or treating influenza that exhibits superior safety and efficacy. [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 dead bacteria. This composition provides a composition for preventing and / or treating influenza that exhibits even greater safety and efficacy. Food and / or feed for preventing and / or treating influenza, comprising any of the compositions described in [9][1] to [8]. This food and feed can provide safe and effective food and feed for preventing and / or treating influenza.

[0011] Furthermore, since changes in cytokine production are observed when using retinoid compounds, or when using retinoid compounds and lactic acid bacteria, the present invention also addresses the challenge of providing a composition containing a retinoid compound that regulates cytokine production, or a composition containing a retinoid compound and lactic acid bacteria. Furthermore, it was found that retinoid compounds, or compositions containing retinoid compounds and lactic acid bacteria, can suppress the production of TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5.

[0012] In other words, the present invention provides the following

[10] to

[19] .

[10] A composition for regulating cytokine production due to viral infection, characterized by containing a retinoid compound as an active ingredient. This composition provides a safe and effective way to regulate cytokine production in response to viral infection.

[11] The composition for regulating cytokine production due to viral infection according to

[10] , further characterized by containing lactic acid bacteria as an active ingredient. This composition provides a composition for regulating cytokine production caused by viral infection, exhibiting excellent safety and efficacy.

[12] The composition for regulating cytokine production due to viral infection according to

[10] or

[11] , characterized in that the regulation of cytokine production due to viral infection is the suppression of the 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 provides a safe and effective composition for suppressing the production of cytokines induced by viral infection, including TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5.

[13] The content of the retinoid compound relative to the content of the lactic acid bacteria is such that 10 10A composition for regulating cytokine production due to viral infection as described in

[11] or

[12] , characterized in that the amount per unit is 100 μg to 500 μg. This composition provides a composition for regulating cytokine production induced by viral infection, exhibiting 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. This composition allows for the easy preparation of a composition for regulating cytokine production caused by viral infection by employing readily available vitamin A and vitamin A derivatives.

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

[10] to

[13] , characterized in that the retinoid compound is provitamin A. This composition provides a composition for preventing and / or treating influenza that exhibits superior safety and efficacy.

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

[15] , characterized in that the provitamin A is beta-carotene. This composition allows for the easy preparation of a composition for preventing and / or treating influenza by employing readily available beta-carotene.

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

[11] to

[16] , characterized in that the lactic acid bacterium is Lactobacillus brevis strain KB290 (accession number NITE P-1537). This composition provides a composition for regulating cytokine production induced by viral infection, exhibiting superior safety and efficacy.

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

[11] to

[17] , characterized in that the lactic acid bacteria are dead bacteria. This composition provides a composition for regulating cytokine production induced by viral infection that exhibits even greater safety and efficacy. Foods and / or feeds for regulating cytokine production due to viral infection, comprising any of the compositions described in

[19] ,

[10] , to

[18] . This food and feed can provide safe and effective food and feed for regulating cytokine production caused by viral infection. [Effects of the Invention]

[0013] According to the present invention, it is possible to provide novel compositions for the safe and effective prevention and / or treatment of influenza. [Brief explanation of the drawing]

[0014] [Figure 1] This figure shows the changes in body weight of mice. (A) Shows the change in body weight over time due to influenza virus infection. (B) Shows the change in body weight over time due to influenza virus infection when retinoic acid is ingested. (C) Shows the change in body weight over time due to influenza virus infection when Lactobacillus brevis KB290 strain is ingested. (D) Shows the change in body weight over time due to influenza virus infection when retinoic acid and Lactobacillus brevis KB290 strain are ingested. (E) Shows the change in body weight over time of mice that were not treated with influenza virus infection. (F) A figure overlaying (A) to (E) above. [Figure 2] This figure shows the viral titer resulting from influenza virus infection. [Figure 3]This figure shows the amount of viral RNA produced by influenza virus infection. (A) Shows the time course of hemagglutinin RNA expression. (B) Shows the time course of neuraminidase RNA expression. (C) Shows the time course of RNA polymerase α subunit RNA expression. (D) Shows the time course of M1 protein RNA expression. (E) Shows the time course of M2 protein RNA expression. (F) Shows the time course of NS1 protein RNA expression. (G) Shows the time course of NS2 protein RNA expression. [Figure 4] This figure shows the effect of retinoic acid on cytokine production induced by influenza virus infection. (A) Time course of TNF-α concentration. (B) Time course of IL-1β concentration. (C) Time course of IL-6 concentration. (D) Time course of IL-10 concentration. (E) Time course of IL-33 concentration. (F) Time course of CXCL1 concentration. (G) Time course of CXCL10 concentration. (H) Time course of CCL2 concentration. (I) Time course of CCL5 concentration. [Figure 5] This figure shows the effect of Lactobacillus brevis strain KB290 on cytokine production induced by influenza virus infection. (A) Time course of IFN-α concentration. (B) Time course of IFN-β concentration. (C) Time course of IL-1β concentration. (D) Time course of IL-6 concentration. (E) Time course of IL-10 concentration. (F) Time course of CCL5 concentration. [Figure 6] This figure shows the effects of retinoic acid and Lactobacillus brevis strain KB290 on cytokine production induced by influenza virus infection. (A) Time course of TNF-α concentration. (B) Time course of IL-1β concentration. (C) Time course of IL-6 concentration. (D) Time course of IL-10 concentration. (E) Time course of IL-33 concentration. (F) Time course of CXCL1 concentration. (G) Time course of CXCL10 concentration. (H) Time course of CCL2 concentration. (I) Time course of CCL5 concentration. [Modes for carrying out the invention]

[0015] The following describes the compositions, foods, and feeds for preventing and / or treating influenza according to the present invention.

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

[0017] (Retinoid compounds) Retinoid compounds are a group of compounds that have a chemical structure and physiological function similar to vitamin A. Vitamin A also includes retinol (vitamin A1) and 3-dehydroretinol (vitamin A2). Compounds with a chemical structure similar to vitamin A are not particularly limited, but include, for example, those having the structure represented by formula (I).

[0018] TIFF0007837371000001.tif1233 formula: ...(I)

[0019] Vitamin A derivatives that have a similar basic structure to vitamin A and exhibit similar physiological activity include retinoic acid, retinal, retinoic acid ester, retinoic acid amide, 3-dehydroretinal, 3-dehydroretinoic acid, 3-dehydroretinoic acid ester, and 3-dehydroretinoic acid amide.

[0020] The retinoid compounds used in the present invention are not particularly limited, and examples include vitamin A, retinol, retinoic acid, retinal, and vitamin A derivatives such as retinoic acid esters. The retinoid compound may be derived from a natural product or chemically synthesized. Furthermore, the retinoid compound may be any geometric isomer, such as an all-trans isomer or a cis-trans isomer, or it may be in the form of provitamin A, which is converted into vitamin A or a vitamin A derivative in the body.

[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, and 3,4-didehydro-13-cis-retinol.

[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-cisretinal, 11-cisretinal, 13-cisretinal, 3,4-dehydroretinal, 13-ethylretinal, 9-dm-retinal, 3-hydroxyretinal, 4-hydroxyretinal, naphthylretinal, 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, for example, 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 heptadecanate, retinyl stearate, retinyl isostearate, retinyl nonadecanate, retinyl arachidonic acid, retinyl behenate, retinyl linoleate, and retinyl oleate.

[0025] Specific examples of naturally derived provitamin A include alpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin, and echinenone. From the standpoint of availability and safety, the retinoid compounds are preferably vitamin A, retinoic acid, all-trans retinal, and beta-carotene. Particularly preferred are vitamin A and retinoic acid. Furthermore, these retinoid compounds may be formulated individually 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 effects of the present invention, which provide a composition for preventing and / or treating influenza with excellent preventive and therapeutic effects against influenza, are further enhanced.

[0027] (lactic acid bacteria) The lactic acid bacteria used in this invention are not particularly limited as long as they are lactic acid bacteria strains that exhibit preventive and therapeutic effects against influenza. Specific examples of lactic acid bacteria include, for example, Lactobacillus brevis strain KB290, Lactobacillus brevis strain NC-4, Lactobacillus brevis strain SAM2447, Lactobacillus brevis strain IFO-12005, Lactobacillus brevis strain CD2, Lactobacillus brevis strain DNBL1871, Lactobacillus brevis strain ATCC14869, Lactobacillus brevis strain JCM1559, Lactobacillus brevis strain NBRC12005, Lactobacillus brevis strain 1059T, Lactobacillus brevis strain 1170, Lactobacillus brevis strain NTM003, Lactobacillus brevis strain 1059T, and Lactobacillus brevis strain 1170. From the standpoint of safety and efficacy, Lactobacillus brevis strain KB290 is preferred as the lactic acid bacterium. Lactobacillus brevis strain KB290 was deposited on February 13, 2013 (accession number NITE P-1537) at the Patent Microorganism Depository Center of the National Institute of Technology and Evaluation (2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture). Furthermore, these lactic acid bacteria may be blended individually or in combination of two or more types.

[0028] The form of lactic acid bacteria is not particularly limited, as long as it is a strain of lactic acid bacteria that exhibits preventive and therapeutic effects against influenza. Examples include bacterial cells, bacterial cultures, powdered bacterial cells, crushed bacterial cells, and bacterial cell extracts. Furthermore, lactic acid bacteria may be live bacteria, dead bacteria, or a mixture of live and dead bacteria. Methods for preparing dead bacteria include, for example, heat treatment, ultra-high temperature heat treatment, pressurized treatment, high-pressure steam treatment, and dry heat treatment. From a quality control standpoint, the form of lactic acid bacteria should preferably be dead.

[0029] The lactic acid bacteria content in the composition is not particularly limited; for example, 0.001 × 10⁶ per gram of composition. 10 1000×10 pieces or more10 It is less than the following number. As the lower limit value, preferably 0.01×10 per 1 g of the composition 10 is more than the following number, more preferably 0.1×10 per 1 g of the composition 10 is more than the following number, still more preferably 0.5×10 per 1 g of the composition 10 is more than the following number, particularly preferably 1×10 per 1 g of the composition 10 is more than the following number. On the other hand, as the upper limit value, preferably 500×10 per 1 g of the composition 10 is less than the following number, more preferably 100×10 per 1 g of the composition 10 is less than the following number, still more preferably 50×10 per 1 g of the composition 10 is less than the following number, particularly preferably 10×10 per 1 g of the composition 10 is less than the following number. 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, which has an excellent preventive effect and therapeutic effect against influenza, is more exerted.

[0030] The ratio of the content of the retinoid compound to the content of lactic acid bacteria in the composition is not particularly limited. For example, per 10 10 lactic acid bacteria, it is 1 μg or more and 100 g or less. As the lower limit value, preferably per 10 10 lactic acid bacteria, it is 10 μg or more, more preferably per 10 10 lactic acid bacteria, it is 50 μg or more, still more preferably per 10 10 lactic acid bacteria, it is 100 μg or more, particularly preferably per 10 10 lactic acid bacteria, it is 200 μg or more. On the other hand, as the upper limit value, preferably per 10 10 lactic acid bacteria, it is 1 g or less, more preferably per 10 10 lactic acid bacteria, it is 10 mg or less, still more preferably per 10 10 lactic acid bacteria, it is 1 mg or less, particularly preferably per 10 10 lactic acid bacteria, it is 500 μg or less, 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 effects of the present invention, which provides a composition for preventing and / or treating influenza with excellent preventive and therapeutic effects against influenza, are further enhanced.

[0031] Next, the components that may be contained in the composition of the present invention will be described in detail. The present invention's composition for preventing and / or treating influenza may contain any other active ingredients, additives, etc., as long as the effects of retinoid compounds and lactic acid bacteria on influenza are not impaired.

[0032] (Other active ingredients) The active ingredients other than retinoid compounds and lactic acid bacteria contained in the composition of the present invention are not particularly limited. Other specific examples of 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 type B; Lactobacillus rhamnosus, Lactobacillus sakei, Lactobacillus casei, Lactobacillus delbrueckii subspecies bulgaricus, Bifidobacterium longum, Bifidobacterium breve, and Propionibacterium freudenereich. Examples include fungi such as *Freudenreichii*, as well as symptomatic treatments such as antipyretics, analgesics, anti-inflammatory drugs, anti-allergic drugs, cough suppressants, and expectorants. Furthermore, these active ingredients may be formulated individually or in combination of two or more.

[0033] (Additives) The additives contained in the composition of the present invention are not particularly limited, and include, for example, pharmaceutically acceptable bases, carriers, excipients, binders, disintegrants, lubricants, colorants, pH adjusters, buffers, stabilizers, and preservatives. Furthermore, these additives may be blended individually or in combination of two or more types.

[0034] Pharmaceutically acceptable bases are not particularly limited and include, for example, water, polar solvents such as ethanol, and oily bases.

[0035] The carriers and excipients are not particularly limited and include, for example, 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 includes, for example, starch, gelatin, syrup, tragacanth gum, polyvinyl alcohol, polyvinyl ether, polyvinylpyrrolidone, hydroxypropyl cellulose, methylcellulose, ethylcellulose, and carboxymethylcellulose.

[0037] The disintegrant is not particularly limited and examples 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 may include, for example, magnesium stearate, hydrogenated vegetable oil, talc, and macrogol.

[0039] The coloring agents are not particularly limited and include, for example, cochineal, carmine, curcumin, riboflavin, annat, titanium dioxide, iron oxide, talc, calcined silica, and magnesium carbonate.

[0040] pH adjusters are not particularly limited and include, for example, citric acid, gluconic acid, succinic acid, potassium carbonate, and lactic acid.

[0041] Buffering agents are not particularly limited and include, for example, phosphates, arginine, and histidine.

[0042] The stabilizers are not particularly limited and include, for example, arginine, polysorbate 80, and macrogol 4000.

[0043] The preservatives are not particularly limited and include, for example, benzoic acid, phenoxyethanol, and thimerosal.

[0044] Other additives include solubilizers, surfactants, emulsifiers, antioxidants, glazing agents, foaming agents, moisture-proofing agents, preservatives, sweeteners, flavoring agents, cooling agents, flavoring agents, fragrances, air fresheners, and disintegration aids. Furthermore, these additives may be blended individually or in combination of two or more types.

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

[0046] [Uses and forms of the composition] The form and use of the composition for preventing and / or treating influenza of the present invention will be described in detail. The uses of the composition of the present invention are not particularly limited, and it can be incorporated into pharmaceuticals, quasi-drugs, cosmetics, food and beverages, animal feed, and the like. Furthermore, pharmaceuticals, quasi-drugs, cosmetics, food and beverages, animal feed, etc., containing the composition of the present invention may be used for human or non-human animals. Non-human animals are not particularly limited, but examples include mammals, birds, reptiles, amphibians, and fish, and preferably include chickens, cattle, pigs, dogs, and cats.

[0047] (Pharmaceuticals, quasi-drugs, cosmetics) The forms of pharmaceuticals, quasi-drugs, and cosmetics containing the composition of the present invention are not particularly limited, and examples include, as oral administration forms, tablets such as sugar-coated tablets, buccal tablets, coated tablets, and chewable tablets; lozenges, pills, powders, capsules including soft capsules; granules, suspensions, emulsions, syrups including dry syrups; and as parenteral administration forms, injectable preparations such as intravenous injection, subcutaneous injection, intraperitoneal injection, and intramuscular injection; transdermal absorption tapes for transdermal administration, transnasal administration, transpulmonary administration, enteral administration, oral administration, and transmucosal administration; aerosols; and suppositories. From the standpoint of convenience and versatility, the form of pharmaceuticals, quasi-drugs, and cosmetics should preferably be in the form of oral administration.

[0048] The dosage of pharmaceuticals, quasi-drugs, and cosmetics containing the composition of the present invention is not particularly limited. For example, the retinoid compound may be 1 μg / kg (body weight) or more and 50 mg / kg (body weight) or less per day, and the lactic acid bacteria may be 0.0001 × 10 per day. 10 pieces / kg (weight) or more 1000×10 10 The amount is less than or equal to 10 μg / kg (body weight) per day. The lower limit for retinoid compounds 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 for retinoid compounds 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. The lower limit for lactic acid bacteria is preferably 0.0005 × 10 per day. 10 The amount should be at least one unit / kg (body weight), and more preferably 0.001 × 10⁻¹⁶ per day. 10 The amount is at least one unit / kg (body weight), and more preferably 0.005 × 10 per day. 10 The amount is at least one unit / kg (body weight), and particularly preferably 0.01 × 10⁶ units per day.10 The amount should be 500 x 10¹ / kg (body weight) or more. On the other hand, the upper limit for lactic acid bacteria is preferably 500 x 10¹ per day. 10 The number of particles per kg (body weight) is less than or equal to 100 x 10 per day, more preferably 100 x 10 10 The number of units / kg (body weight) is less than or equal to 50 x 10 per day, and more preferably 50 x 10 10 The number of particles per kg (body weight) is less than or equal to 10 x 10 per day, and is particularly preferable. 10 The number is less than or equal to the number of individuals / kg (body weight).

[0049] The number of times 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, three times a day, etc. The duration of administration of pharmaceuticals, quasi-drugs, and cosmetics 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 a duration of administration. Furthermore, the administration during the administration period is not particularly limited and may be administered daily, every other day, every two days, every three days, etc.

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

[0051] The shape and properties of foods, beverages, and animal feeds containing the composition of the present invention are not particularly limited, and examples include solid, semi-solid, gel, liquid, and powder forms. In particular, when used as a health food or supplement, it is preferable to use it in the form of granules, capsules, tablets, chewable tablets, beverage powders, drinks, smoothies, jellies, gummies, etc., to allow for continuous and easy intake.

[0052] Foods, beverages, and animal feeds containing the composition of the present invention may be labeled as potentially having beneficial effects against influenza. These labels can be attached to containers and packaging in known ways, thereby clearly indicating that the foods, beverages, and animal feeds containing the composition of the present invention are intended for use in improving influenza, thus clearly distinguishing them from ordinary foods, beverages, and animal feeds.

[0053] The intake of food, beverages, and feed containing the composition of the present invention is not particularly limited. For example, the intake of retinoid compounds may be 1 μg / kg (body weight) or more and 50 mg / kg (body weight) or less per day, and lactic acid bacteria may be 0.0001 × 10 per day. 10 pieces / kg (weight) or more 1000×10 10 The amount is less than or equal to 10 μg / kg (body weight) per day. The lower limit for retinoid compounds 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 for retinoid compounds 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. The lower limit for lactic acid bacteria is preferably 0.0005 × 10 per day. 10 The amount should be at least one unit / kg (body weight), and more preferably 0.001 × 10⁻¹⁶ per day. 10 The amount is at least one unit / kg (body weight), and more preferably 0.005 × 10 per day. 10 The amount is at least one unit / kg (body weight), and particularly preferably 0.01 × 10⁶ units per day. 10 The amount should be 500 x 10¹ / kg (body weight) or more. On the other hand, the upper limit for lactic acid bacteria is preferably 500 x 10¹ per day. 10 The number of particles per kg (body weight) is less than or equal to 100 x 10 per day, more preferably 100 x 10 10 The number of units / kg (body weight) is less than or equal to 50 x 10 per day, and more preferably 50 x 1010 The number of particles per kg (body weight) is less than or equal to 10 x 10 per day, and is particularly preferable. 10 The number is less than or equal to the number of individuals / kg (body weight). Furthermore, the intake amount can be adjusted depending on the type of non-human animal.

[0054] The frequency of consumption of food, beverages, or feed containing the composition of the present invention is not particularly limited and may be, for example, once a day, twice a day, three times a day, etc. The period of intake of food, beverages, 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 an intake period. Furthermore, the administration during the intake period is not particularly limited and may be, for example, daily intake, intake every other day, intake every two days, intake every three days, etc. Furthermore, the duration and interval of intake can be adjusted depending on the type of non-human animal.

[0055] Due to the above 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 caused by viral infection] Compositions for regulating cytokine production due to viral infection can have the same composition, uses, and forms as compositions for preventing and / or treating influenza.

[0057] The cytokines that regulate 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 may involve, for example, promoting or suppressing cytokine production at any point in time after viral infection.

[0059] The viruses that cause infection are not particularly limited and include, for example, 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 characteristics, compositions for regulating cytokine production can safely and effectively regulate cytokine production, thus contributing to the prevention and treatment of viral infections. [Examples]

[0061] [Example 1: The effect of the present invention on influenza] (raising mice) Seven-eight-week-old female BALB / c mice (AgResearch Ruakura Small Animal Facility, Palmerston North, New Zealand) fed with standard solid feed (Prolab RMH 1800, LabDiet, USA) and sterile water were used. The mice were housed in plastic cages with 3-4 mice per cage, and maintained under a 12-hour light / dark cycle, room temperature of 21°C, and humidity of 50%. The mice were divided into five groups of 30 mice each: an untreated influenza virus infection group (hereinafter referred to as the "untreated group"), a control group treated with influenza virus infection (hereinafter referred to as the "control group"), a group administered retinoic acid before influenza virus infection (hereinafter referred to as the "RA-administered group"), a group administered Lactobacillus brevis KB290 strain before influenza virus infection (hereinafter referred to as the "KB-administered group"), and a group administered retinoic acid and Lactobacillus brevis KB290 strain before influenza virus infection (hereinafter referred to as the "RA+KB-administered group").

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

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

[0064] (Methods of influenza virus transmission) The influenza virus used for treating mice was influenza A / PR / 8 / 34(H1N1). The influenza virus solution used for intranasal infection was prepared in PBS to a dose of 0.5 × LD50. The influenza virus infection method involved administering the test food to mice for 14 days, then anesthetizing the mice the following day, and finally administering an influenza virus solution into the nasal cavity.

[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 for 14 days during which the test food was administered, once on the day of influenza virus infection treatment, and once daily for 14 days after influenza virus infection treatment.

[0066] Figures 1(A) to 1(E) show the measured actual body weight and the predictive curves based on the mean values ​​for each group over time, with the influenza virus infection treatment day set as day 0, the test food administration period from -1 to -14 days, and the post-infection treatment period from day 1 to 14 days. Figure 1(F) shows the 84% parametric bootstrap confidence intervals calculated for each group's predictive curve. The width of the 84% confidence interval was set so that there is a 5% false positive rate when there is no difference between the groups.

[0067] (Measurement of viral titer) To measure viral titers, 10 mice from each group were euthanized by cervical dislocation three days after influenza virus infection. The lungs were promptly removed and homogenized using a 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 obtained by centrifugation 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 the 50% tissue culture cell infection rate (TCID) was measured. 50 ) was calculated as follows.

[0068] Figure 2 shows the viral titer in lung tissue 3 days after influenza virus infection treatment (log). 10 (TCID 50 The values ​​are shown as mean ± standard error, and a p-value less than 0.05 indicates a statistically significant difference, and is marked with an asterisk (*).

[0069] (Measurement of viral RNA levels) To measure viral RNA levels, 10 mice from each group were euthanized by cervical dislocation on days 3, 7, and 14 after influenza virus infection. The lungs were promptly removed and homogenated using a 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). RNA was prepared from the homogenates using an RNA preparation kit (RNeasy, Qiagen NV, Germany).

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

[0071] [Table 1]

[0072] [Table 2]

[0073] Figure 3 shows the RNA counts of seven viral genes in lung tissue 3 days (white), 7 days (gray), and 14 days (black) after influenza virus infection treatment. The values ​​are expressed as mean ± standard error, and a statistically significant difference is indicated by an asterisk (*) if the P-value is less than 0.05 compared to the untreated group.

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

[0075] Therefore, the intake of retinoic acid and Lactobacillus brevis strain KB290 was found to improve weight loss caused by influenza virus infection. Furthermore, the effect of improving weight loss caused by influenza virus infection was shown to be enhanced in the following order: intake of Lactobacillus brevis KB290 strain, intake of retinoic acid, and intake of retinoic acid and Lactobacillus brevis KB290 strain.

[0076] (Result 2: Effect of test food on viral titer) As shown in Figure 2, influenza virus infection significantly increased viral titer in lung tissue. On the other hand, viral titer decreased significantly in the RA-administered group, KB-administered group, and RA+KB-administered group compared to the control group. In particular, the RA+KB-administered group showed a statistically significant decrease compared to the control group.

[0077] Therefore, it has been shown that intake of retinoic acid and Lactobacillus brevis strain KB290 suppresses the increase in viral titer caused by influenza virus infection. Furthermore, it was found that the inhibitory effect on viral titer caused by influenza virus infection was significantly demonstrated by using retinoic acid and Lactobacillus brevis strain KB290 in combination.

[0078] (Result 3: Effect of test food on viral RNA levels) As shown in Figures 3(A) to (G), influenza virus infection increased the levels of HA, NA, PA, M1, M2, NS1, and NS2 RNA from day 3 to day 7 post-infection, and decreased on day 14 post-infection. In all of these increases in influenza virus RNA levels, the RA+KB treatment group showed significantly suppressed levels compared to the control group, the RA-only group, and the KB-only group.

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

[0080] As shown in results 1-3 above, the compositions of the present invention improve weight loss caused by influenza virus infection, suppress the increase in viral titer, and significantly suppress the proliferation of influenza virus in the lungs when used in combination with retinoic acid and Lactobacillus brevis strain KB290. Therefore, compositions containing retinoid compounds and lactic acid bacteria are recognized as effective in the prevention and treatment of influenza.

[0081] [Example 2: The effect of the present invention on cytokine production induced by viral infection] The method of raising the mice and administering the test food was the same as in Example 1.

[0082] (Measurement of cytokines) For cytokine measurement, 10 mice from each group were euthanized by cervical dislocation on days 3, 7, and 14 after influenza virus infection treatment. The lungs were then promptly removed and homogenized using a 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 obtained by centrifugation 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 homogenate were quantified by flow cytometry using a bead assay kit (LEGENDplex, BioLegend Inc., USA). Cytokine concentrations were calculated as cytokine mass relative to the total protein content of lung tissue.

[0083] Figures 4-6 show cytokine concentrations in lung tissue on days 3 (white), 7 (gray), and 14 (black) after influenza virus infection treatment in the RA-administered group, KB-administered group, and RA+KB-administered group, 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), when 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 on day 7.

[0085] Therefore, it was revealed that retinoic acid intake suppresses the production of TNF-α, IL-6, CXCL1, CCL2, and CCL5 up to day 3 after influenza virus infection, and up to day 7. Since cytokine production was suppressed by retinoic acid intake, it is inferred that retinoic acid may be preventing viruses that cause increased cytokine production from entering the lungs.

[0086] (Result 5: Effect of Lactobacillus brevis strain KB290 on cytokine production) As shown in Figure 5(D), when comparing the control group and the KB-treated group, Lactobacillus brevis strain KB290 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 with the KB-treated group, Lactobacillus brevis strain KB290 promoted the production of IFN-α, IFN-β, IL-1β, and IL-10 on day 3 after influenza virus infection.

[0087] Therefore, it was revealed that ingestion of Lactobacillus brevis strain KB290 suppressed IL-6 production up to day 7 post-infection with influenza virus and promoted the production of IFN-α, IFN-β, IL-1β, and IL-10 on day 3 post-infection. Since many of the cytokines suppressed by retinoic acid ingestion were promoted by Lactobacillus brevis strain KB290 ingestion, it is inferred 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 strain KB290 on cytokine production) As shown in Figures 6(A), (C), (F), (G), (H), and (I), when comparing the control group with the RA+KB-treated group, retinoic acid and Lactobacillus brevis KB290 strain suppressed the increased production of TNF-α, IL-6, CXCL1, CXCL10, CCL2, and CCL5 on days 3 and 7 after influenza virus infection. Furthermore, as shown in Figure 6(D), when comparing the control group with the RA+KB-treated group, retinoic acid and Lactobacillus brevis KB290 strain suppressed the increased 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 day 7 after influenza virus infection, and suppressed the production of IL-10 on day 7 after infection. Furthermore, when retinoic acid and Lactobacillus brevis KB290 strain were taken together, several cytokines showed significantly suppressed production on days 3 and 7 compared to when each was taken alone. Therefore, it is inferred that the combined use of retinoic acid and Lactobacillus brevis KB290 strain more strongly suppresses influenza virus infection.

[0090] As shown in results 4-6 above, the compositions of the present invention can adjust the amount of cytokine produced by viral infection. Therefore, compositions containing retinoid compounds and lactic acid bacteria are recognized as effective in preventing and treating infectious diseases caused by viruses such as influenza virus by optimizing the immune mechanism. [Industrial applicability]

[0091] This invention makes it possible to safely prevent and treat influenza. This makes it possible to suppress the large-scale spread of influenza virus infection in populations of humans, livestock, and other animals.

Claims

1. It contains a retinoid compound and lactic acid bacteria as active ingredients, and the content of the retinoid compound is 0.01% by mass or more and 20% by mass or less. The aforementioned lactic acid bacterium is Lactobacillus brevis strain KB290 (accession number NITE P-1537), A composition for preventing and / or treating influenza, 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 The composition for preventing and / or treating influenza according to claim 1, characterized in that the amount is 100 μg to 500 μg per unit.

3. The composition for preventing and / or treating influenza according to claim 1 or 2, characterized in that the lactic acid bacteria are dead bacteria.

4. Food and / or feed for preventing and / or treating influenza, comprising the composition according to any one of claims 1 to 3.

5. It contains a retinoid compound and lactic acid bacteria as active ingredients, and the content of the retinoid compound is 0.01% by mass or more and 20% by mass or less. The aforementioned lactic acid bacterium is Lactobacillus brevis strain KB290 (accession number NITE P-1537), A composition for regulating cytokine production due to viral infection, 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.

6. The composition for regulating cytokine production due to viral infection according to claim 5, characterized in that the regulation of cytokine production due to viral infection is the suppression of the production of at least one cytokine selected from the group consisting of TNF-α, IL-6, IL-10, CXCL1, CXCL10, CCL2, and CCL5.

7. The content of the retinoid compound relative to the content of the lactic acid bacteria is １０ A composition for regulating cytokine production due to viral infection according to claim 5 or 6, characterized in that the amount is 100 μg to 500 μg per unit.

8. A composition for regulating cytokine production due to viral infection according to any one of claims 5 to 7, characterized in that the lactic acid bacteria are dead bacteria.

9. Food and / or feed for regulating cytokine production due to viral infection, comprising the composition according to any one of claims 5 to 8.

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