Viral prevention of respiratory infection and therapeutic agent

Abstract

To provide a prophylactic and therapeutic agent for viral respiratory infectious diseases comprising a new component.SOLUTION: The features of the present invention for solving the above problems are as follows. 1. A prophylactic or therapeutic agent for viral respiratory infection, comprising glucosylceramide and / or glucosylceramide as an active ingredient. (2) A prophylactic or therapeutic agent for viral respiratory infections, which comprises β - sitosterol glycoside as an active ingredient. (3) The prophylactic and therapeutic agent for viral respiratory infection according to (1) or (2) above, wherein the viral respiratory infection is influenza.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a preventive and therapeutic agent for viral respiratory infections, which is widely used in medicines, quasi-drugs, health foods, general merchandise, etc. [Background technology]

[0002] There are various epidemic acute respiratory infections caused by viruses, including influenza and coronavirus infections. Influenza is an epidemic acute respiratory infection caused by infection with influenza A or B viruses. Its clinical manifestations are characterized by upper respiratory tract symptoms as well as sudden onset of high fever, general fatigue, headache, muscle pain, and joint pain, resulting in severe illness that can leave patients bedridden for several days. Influenza epidemics occur every winter, affecting millions of people in a short period of time. Furthermore, high-risk groups such as those aged 65 and over, infants, pregnant women, and those with various underlying diseases are at several hundred times higher risk of developing pneumonia, hospitalization, and death when infected with influenza than healthy adults. Therefore, influenza is recognized as a significant disease with significant socioeconomic impacts due to the extent of the damage to patients' health and the scale of its epidemic.

[0003] Influenza vaccines have traditionally been used as a method for preventing influenza. However, the preventive effect is not always satisfactory, as the vaccine strain differs from the prevalent strain, and even if the strain matches, the host defense mechanism is insufficient. Furthermore, vaccine recipients may experience side effects such as redness, swelling, pain, fever, headache, chills, and fatigue. Furthermore, people with egg allergies may experience hives, rash, oral numbness, and anaphylactic shock, and gelatin allergies to gelatin, which is contained in vaccines as a stabilizer, have also been reported. Other reported side effects include Guillain-Barré syndrome, acute encephalopathy, convulsions, and purpura. Therefore, vaccinations are administered under the supervision and guidance of a physician (Patent Document 1). Therefore, there is a need for a preventive and therapeutic agent for influenza that has few side effects and can be ingested as a food.

[0004] [Patent Document 1] Japanese Patent Application Laid-Open No. 2008-297320 DISCLOSURE OF THE INVENTION [Problem to be solved by the invention]

[0005] Against this background, the present inventors discovered that glucosylceramide-containing rice extract and β-sitosterol glucoside, which are known as food ingredients, have preventive and therapeutic effects against viral respiratory infections such as influenza, and completed the present invention. That is, an object of the present invention is to provide a novel agent for preventing and treating viral respiratory infections, which contains a food material with few side effects as an active ingredient. [Means for solving the problem]

[0006] The features of the present invention for solving the above problems are as follows. 1. A preventive and therapeutic agent for viral respiratory infections, containing glucosylceramide and / or glucosylceramide-containing rice extract as an active ingredient. 2. A preventive and therapeutic agent for viral respiratory infections containing β-sitosterol glycoside as an active ingredient. 3. The preventive and therapeutic agent for viral respiratory infections according to 1. or 2. above, wherein the viral respiratory infection is influenza. [Brief explanation of the drawings]

[0007] [Figure 1] 1 is a graph showing the survival rate after virus inoculation in this example. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present invention will be described in detail below. The present invention is characterized in that it contains glucosylceramide and / or a glucosylceramide-containing rice extract as an active ingredient. Glucosylceramide, the active ingredient of the present invention, is a ceramide to which a sugar (mainly galactose) is bound via a glycosyl ether bond.

[0009] The glucosylceramide is not particularly limited, but is preferably one represented by the following chemical formula (1). [ka]

[0010] A large amount of glucosylceramide-containing rice extract can be extracted from rice bran. The rice used as the raw material is not limited to any particular variety, but sweet-tasting Japonica varieties are preferred. Staple rice varieties such as Koshihikari and Sasanishiki are particularly preferred. Typically, the outermost layers of brown rice, the pericarp, seed coat, and aleurone layer, are removed using a rice polisher to produce bran.

[0011] The method for extracting a glucosylceramide-containing rice extract from rice bran is not particularly limited, but the following steps A to C are preferable. A. A process of extracting rice bran with an organic solvent; B. adding an acid to the extract along with water and precipitating gum in the acid solution; C. Extracting the gum with alcohol and purifying sphingolipids from the alcohol extract.

[0012] The rice bran oil production process can be used for steps A and B. Generally, when producing rice bran oil, the raw material rice bran is extracted with hexane, and then treated with phosphoric acid to separate the oil and obtain a gum. Therefore, by applying step C to the gum produced in this rice bran oil production process, glucosylceramide can be efficiently extracted.

[0013] The organic solvent may be hexane, heptane, petroleum ether, ether, chloroform, acetone, or the like. The acid used to treat the organic solvent extract may be organic, such as acetic acid, oxalic acid, citric acid, or formic acid, or inorganic, such as phosphoric acid, sulfuric acid, hydrochloric acid, or nitric acid. Phosphoric acid is preferably used because it has the following advantages: a. The manufacturing process for rice bran oil can be used as is. b. Because it is not a strong acid, it is less likely to corrode tanks, pipes, etc., improving the durability of manufacturing equipment. c. No irritating odor like acetic acid or formic acid. d. Because the phospholipids contained in organic solvent extracts are easily removed, the concentration of glucosylceramide in the precipitate (gum) increases.

[0014] The alcohol may be ethanol, methanol, isopropyl alcohol, butanol, or any of these hydroalcohols. When glucosylceramide is used in food or beverage products, ethanol is preferably used. In step C, glucosylceramide can be purified from the alcohol extract of the gummy substance by column chromatography or other suitable purification methods.

[0015] Furthermore, the glucosylceramide-containing rice extract may be a commercially available product, for example, Oryza Ceramide (registered trademark) manufactured by Oryza Oil & Fat Chemical Co., Ltd.

[0016] The present invention is characterized in that it contains β-sitosterol glycoside as an active ingredient. The β-sitosterol glycoside is not particularly limited and examples thereof include β-sitosterol glucoside, β-sitosterol rutinoid, and β-sitosterol sambubioside, with β-sitosterol glucoside being particularly preferred. β-Sitosterol glucoside is a compound represented by the following chemical formula (2):

[0017] [ka]

[0018] The method for obtaining β-sitosterol glucoside is not particularly limited, and a commercially available product may be used, or it may be extracted from a plant. An example of a commercially available product that can be used is "Sitosterol-3-O-glucoside, β- (AHP Verified)" manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.

[0019] The plant raw material from which β-sitosterol glucoside is obtained is not particularly limited, but examples thereof include rice, wheat, beet, konjac, corn, etc. Among these, rice is preferred. There are no particular limitations on the parts of rice that can be used to obtain β-sitosterol glucoside, but it is particularly preferable to use rice bran and rice germ.

[0020] The method for obtaining β-sitosterol glucoside from rice bran or rice germ is not particularly limited, but for example, it can be obtained by fermenting rice bran or rice germ to obtain a fermented product, extracting the fermented product with a nonpolar solvent to obtain a fermented product extract, and then distributing the fermented product extract. It can also be extracted from "Oryza Ceramide" (registered trademark) manufactured by Oryza Oil & Fat Chemical Co., Ltd.

[0021] The viral respiratory infection preventive agent of the present invention can be used as an ingredient in various foods and beverages. Examples of foods and beverages include general foods such as confectioneries (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummy candy, tablet candy, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, green tea, carbonated drinks, sports drinks, etc.), as well as health foods (tablets, capsules, etc.), and nutritional supplements (nutrition drinks, etc.). The viral respiratory infection preventive agent of the present invention can be appropriately incorporated into these foods and beverages.

[0022] These foods and beverages can contain various ingredients depending on their type, such as glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, succinic acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid esters, polyglycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, gum arabic, carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, colorants, flavorings, and preservatives.

[0023] Specifically, the agent of the present invention can be spray-dried or freeze-dried together with powdered cellulose, and then formed into a powder, granules, tablets, or solution, which can be easily incorporated into foods and beverages (e.g., instant foods). The agent of the present invention can also be dissolved in, for example, fats and oils, ethanol, glycerin, or a mixture thereof to form a liquid, which can then be added to beverages or solid foods. If necessary, the agent can also be mixed with a binder such as gum arabic or dextrin to form a powder or granules, which can then be added to beverages or solid foods.

[0024] When the viral respiratory infection preventive agent of the present invention is applied to food and drink, the amount of active ingredient added is preferably 1 to 20 wt% in total relative to the food and drink, since the main purpose is disease prevention and health maintenance.

[0025] The preventive agent for viral respiratory infections of the present invention may be used as a material for medicines (including pharmaceuticals and quasi-drugs). Pharmaceutical preparations can be produced by appropriately blending the agent of the present invention with raw materials for pharmaceutical preparations. Examples of pharmaceutical raw materials that can be incorporated into the agent of the present invention include excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cocoa butter, hardened vegetable oil, kaolin, talc, etc.), binders (distilled water, saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, calcium carbonate, sodium lauryl sulfate, stearate monoglyceride, starch, lactose, gum arabic powder, gelatin, ethanol, etc.), disintegration inhibitors (sucrose, stearin, cocoa butter, hydrogenated oil, etc.), absorption enhancers (quaternary ammonium base, sodium lauryl sulfate, etc.), adsorbents (glycerin, starch, lactose, kaolin, bentonite, silicic acid, etc.), lubricants (purified talc, stearates, polyethylene glycol, etc.).

[0026] The dosage may vary depending on the administration method, the condition, the age of the patient, etc., but typically, adults can be given 0.5 to 5000 mg of the active ingredient per day, and children can be given 0.5 to 3000 mg. The compounding ratio of the viral respiratory infection preventive agent of the present invention can be changed as appropriate depending on the dosage form, but is usually about 0.3 to 15.0 wt% when administered orally or via mucosal absorption, and about 0.01 to 10 wt% when administered parenterally. Note that the dosage varies depending on various conditions, so in some cases a smaller dosage than the above-mentioned dosage is sufficient, and in other cases it may be necessary to administer an amount exceeding the range. [Example]

[0027] Examples of the present invention will be described below. Note that the examples shown below are provided to confirm the various actions and effects of the agent obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

[0028] Test Example: Evaluation of the influenza preventive and therapeutic effects of glucosylceramide-containing rice extract, glucosylceramide, and β-sitosterol glucoside (hereinafter simply referred to as "BSG") 1. Test substance and preparation method 1-1 Test substance 1 Name: Oryza Ceramide (registered trademark) (rice extract containing 30% glucosylceramide) Obtained from: Oryza Oil & Fat Chemical Co., Ltd. 1-2. Test substance 2 Name: Glucosylceramide isolated from the above Oryza ceramide and represented by the following chemical formula (1) [ka] 1-3 Test substance 3 Name: BSG (β-sitosterol glucoside) 1-4 Solvent Name: 0.5w / v% Methylcellulose 400 Solution 1-5.Preparation method The weighed test substance was mixed with the solvent and prepared using a mortar and pestle. The test substance was prepared in a batch for 3 days and stored in a refrigerator until use.

[0029] 2. Viruses Used and Preparation of Inoculum 2-1.Virus used The virus was purchased from ATCC and then mutated by Hamley to a mouse-sensitive virus. Virus name: Influenza virus Strain name: A / PR / 8 / 34 ATCC No.:VR-1469 BSL:2 Viral titer: 1.58 x 10 8 TCID 50 / mL Storage: Stored in a deep freezer (CLN-51UD1, Nippon Freezer Co., Ltd.) 2.2 Preparation of inoculum virus Thaw the frozen virus solution and use PBS to dilute 2 x 10 cells for general observation. 5 TCID 50 / mL (1 × 10 4 TCID 50 / 50 μL) and 6 × 10 for measuring lung viral load. 4 TCID 50 / mL (3 × 10 3 TCID 50 The volume was adjusted to 50 μL / 50 μL.

[0030] 3.Animals used Animal species: Mouse Strain:BALB / c Available from: Charles River Japan Co., Ltd. Gender: Female Age: 4 weeks (at time of arrival) Number of fish used: 48 (50 when shipped) Reason for selecting animal species: Since mouse-sensitive influenza viruses are used, mice are considered to be an appropriate test system.

[0031] 4. Animal management conditions 4-1. Rearing conditions Room temperature 22.0-25.0% (set at 24 ± 3°C), humidity 43.29-58.18% (set at 50 ± 20%), ventilation (10-25 times / hour), lighting 12 hours (8:00-20:00) 4-2. Feed MF (Oriental Yeast Co., Ltd., Lot 200617) was given ad libitum. 4-3.Drinking water Autoclaved city water was provided ad libitum. 4-4.Cage Autoclave-sterilized heat-resistant polysulfone cages (207W x 365D x 140H mm, TECNIPLAST Ltd) were used, and animals were housed 1-5 per cage. 4-5. Quarantine and acclimatization The animals were observed daily for their general condition from the day of arrival until the day of the end of quarantine and acclimation. Body weights were also measured on the day of arrival and the day of the end of quarantine and acclimation. 4-6. Grouping Using the body weight at the end of quarantine acclimation, the animals were assigned to each group by weight-stratified random sampling.

[0032] 5. Study group composition The test group composition was as shown in Table 1 below. [Table 1]

[0033] 6. Virus Inoculation Under anesthesia with isoflurane (Myline Pharmaceuticals, Lot 196AR0), 50 μL / head of the virus solution was intranasally inoculated.

[0034] 7. Administration of the test substance The test substance was administered orally at 10 mL / kg once a day using a syringe and probe from 7 days before virus inoculation until the day before the end of observation. On the day of virus inoculation, the test substance was administered before virus inoculation.

[0035] 8. Sampling (groups 5-8) After euthanasia by overdose of thiamylal sodium (Nichi-Iko Pharmaceutical Co., Ltd., Lot LU1200, Isozole® Injection 0.5 g), lungs were collected and weighed. For viral load measurement, 50-100 mg of lung was collected, immersed in RNAlater (Thermo Fisher Scientific, Lot 00874638), refrigerated for 1 day, and then stored in a deep freezer until use. The remaining lungs were also stored in a deep freezer.

[0036] 9. Observation and measurement items 9-1. General condition The general condition was observed at least once a day. 9-2.Weight measurement At the time of group allocation (first administration: Day -7), Days 0, 3, 7, 10 and 14

[0037] 10.Statistical analysis The measurement data obtained as test data was calculated as the mean ± standard deviation. The average survival time and survival rate of the mice used for general condition observation were calculated, and then a Log-Rank test was performed against the vehicle group.

[0038] 11.Results The survival rates are shown in Figure 1 and Table 2 below. Table 3 below shows the average survival time and survival rate at the end of observation. The mean survival times for groups 1 (vehicle group), 2 (Oryza ceramide group), 3 (glucosylceramide group), and 4 (β-sitosterol glucoside group) were 7.1 days, 8.5 days, 7.8 days, and 8.9 days, respectively. Statistical analysis showed that groups 2 and 4 had significantly longer survival times than group 1. [Table 2] [Table 3]

[0039] 12. Considerations and Effects of Examples To confirm the effects of Oryza Ceramide and its ingredients on influenza-infected mice, we evaluated the survival rate and mean survival time of mice after virus inoculation. As a result, Oryza Ceramide and BSG significantly extended survival times compared to the vehicle group. Glucosylceramide, the main component, also tended to extend survival times. From the above, it was confirmed that glucosylceramide, rice extract containing it, and BSG are useful as agents for preventing and treating influenza.

[0040] The following are formulation examples of the agent (glucosylceramide-containing rice extract) according to the present invention. Note that the following formulation examples do not limit the present invention. In addition, the following formulation examples show examples of only the glucosylceramide-containing rice extract, but BSG and the above-mentioned glucosylceramide can also be formulated in the same way. Example 1: Chewing gum Sugar 53.0wt% Gum base 20.0 Glucose 10.0 Starch syrup 16.0 Fragrance 0.5 Glucosylceramide-containing rice extract 0.5 100.0wt%

[0041] Mixing example 2: Gummies Reduced starch syrup 40.0wt% Granulated sugar 20.0 Glucose 20.0 Gelatin 4.7 Wednesday 9.68 Yuzu juice 4.0 Yuzu flavor 0.6 Dye 0.02 Glucosylceramide-containing rice extract 1.0 100.0wt%

[0042] Mixing example 3: Candy Sugar 50.0wt% Starch syrup 33.0 Wednesday 14.4 organic acid 2.0 Fragrance 0.2 Glucosylceramide-containing rice extract 0.4 100.0wt%

[0043] Blend Example 4: Yogurt (hard / soft) Milk 41.5wt% Skim milk powder 5.8 Sugar 8.0 Agar 0.15 Gelatin 0.1 Lactic acid bacteria 0.005 Glucosylceramide-containing rice extract 0.4 fragrance trace amount water residue 100.0wt%

[0044] Formulation example 5: Soft drink High fructose corn syrup 30.0wt% Emulsifier 0.5 Glucosylceramide-containing rice extract 0.3 Fragrance (appropriate amount) Purified water remainder 100.0wt%

[0045] Formulation example 6: Tablet confectionery Sugar 76.4wt% Glucose 19.0 Sucrose fatty acid ester 0.2 Glucosylceramide-containing rice extract 0.5 Purified water 3.9 100.0wt%

[0046] Formulation example 7: Soft capsule Brown rice germ oil 47.0wt% Yuzu seed oil 40.0 Emulsifier 12.0 Glucosylceramide-containing rice extract 1.0 100.0wt%

[0047] Formulation example 8: Tablets Lactose 54.0wt% Microcrystalline cellulose 30.0 Starch decomposition product 10.0 Glycerin fatty acid ester 5.0 Glucosylceramide-containing rice extract 1.0 100.0wt% [Industrial Applicability]

[0048] As described above, the present invention can provide a novel agent for preventing and treating viral respiratory infections.

Claims

1. An oral agent for preventing and treating viral respiratory infections, the agent comprising a glucosylceramide-containing rice extract as an active ingredient, The glucosylceramide-containing rice extract is extracted from a gum obtained by extracting rice bran with an organic solvent.

2. 2. The oral preventive and therapeutic agent for viral respiratory infections according to claim 1, wherein the viral respiratory infection is influenza.

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