JP2022157082A - Viral respiratory infection preventing or treating agent - Google Patents

Abstract

To provide a viral respiratory infectious disease preventing or treating agent of a novel component.SOLUTION: The present invention is characterized by the followings. 1. Glucosylceramide and/or viral respiratory infectious disease preventing or treating agent having glucosylceramide as an active ingredient. 2.Viral respiratory infectious disease preventing or treating agent having β-sitosterol glycoside as an active ingredient. 3. Viral respiratory infectious disease preventing or treating agent described in 1 or 2 above in which the viral respiratory infectious disease is influenza.SELECTED DRAWING: None

Description

TECHNICAL FIELD The present invention relates to agents for preventing and treating viral respiratory infections. These are widely used in pharmaceuticals, quasi-drugs, health foods, general merchandise, and the like.

There are various types of epidemic acute respiratory infections caused by viral infections, such as influenza and coronavirus infections. Influenza is a prevalent acute respiratory infection caused by infection with influenza A or B viruses. The clinical picture is characterized by systemic symptoms such as sudden high fever, general malaise, headache, muscle pain, and joint pain, in addition to symptoms of upper respiratory tract inflammation, accompanied by a severe feeling of bed rest for several days. Influenza epidemics repeat every winter season, and many patients occur on the scale of millions in a short period of time. Furthermore, when high-risk people aged 65 and older, infants, pregnant women, and other high-risk groups with various underlying diseases contract influenza, the risks of complication of pneumonia, hospitalization, and death are several hundred times higher than those of healthy adults. Therefore, influenza is recognized as an important disease that exerts a great impact on society and economy because of the magnitude of the health damage to patients and the scale of its epidemic.

Influenza vaccines have been conventionally used as methods for preventing influenza. However, the vaccine strain and epidemic strain are different, and even if the strain matches, the ability to induce biological defense is insufficient, and the preventive effect is not always satisfactory. In addition, vaccinees may experience side effects such as redness, swelling, pain, fever, headache, chills, and malaise. In addition, people with egg allergy may experience hives, rashes, numbness in the mouth, anaphylactic shock, etc. Gelatin allergy to gelatin, which is included as a stabilizer in vaccines, has also been reported. There are also reports of side effects such as Guillain-Barre syndrome, acute encephalopathy, convulsions, and purpura. Therefore, vaccination is performed under the supervision and guidance of a doctor (Patent Document 1).
Therefore, there is a demand for a prophylactic and therapeutic agent for influenza that is less likely to cause side effects and that can be ingested as food.

Japanese Patent Application Laid-Open No. 2008-297320

Against this background, the present inventor discovered that glucosylceramide-containing rice extract and β-sitosterol glucoside, which are known as food materials, have preventive and therapeutic effects on viral respiratory infections such as influenza. and completed the present invention.
That is, it is an object of the present invention to provide a novel preventive and therapeutic agent for viral respiratory infection, which contains food materials with few side effects as active ingredients.

The features of the present invention for solving the above problems are as follows.
1. A prophylactic or therapeutic agent for viral respiratory infections containing glucosylceramide and/or a 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 method of claim 1, wherein the viral respiratory infection is influenza. or the above 2. 3. The agent for preventing and treating viral respiratory infections according to .

It is a graph which shows the survival rate after virus inoculation of a present Example.

The present invention will be described in detail below.
The present invention is characterized by using glucosylceramide and/or a glucosylceramide-containing rice extract as an active ingredient.
Glucosylceramide, which is the active ingredient of the present invention, is obtained by glycosylether-bonding sugar (mainly galactose) to ceramide.

Although the glucosylceramide is not particularly limited, it is preferably represented by the following chemical formula (1).

A large amount of the glucosylceramide-containing rice extract can be extracted from rice bran. Although the variety of rice used as the raw material is not limited, japonica, which exhibits sweetness, is suitable. In particular, staple rice varieties such as Koshihikari and Sasanishiki are preferred. Usually, the pericarp, seed coat and aleurone layer, which are the outermost layers of brown rice, are removed by a rice polishing machine to form bran.

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

The steps A and B can utilize the production process of rice bran oil. In general, 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, glucosylceramide can be efficiently extracted by applying the above step C to the gum produced in the production process of such rice bran oil.

As the organic solvent, besides hexane, heptane, petroleum ether, ether, chloroform, acetone, and the like can be used. As the acid for treating the organic solvent extract, organic acids such as acetic acid, oxalic acid, citric acid and formic acid can be used, and inorganic acids such as phosphoric acid, sulfuric acid, hydrochloric acid and nitric acid can be used. Desirably, phosphoric acid is used. This is because phosphoric acid has the following advantages.
a. The manufacturing process of rice bran oil can be used as it is.
b. Since it is not a strong acid, tanks and pipes are less likely to corrode, improving the durability of manufacturing equipment.
c. No irritating odor like acetic acid and formic acid.
d. Since the phospholipids contained in the organic solvent extract can be easily removed, the concentration of glucosylceramide contained in the sediment (gum) increases.

As the alcohol, ethanol, methanol, isopropyl alcohol, butanol, hydrous alcohols thereof, and the like can be used. When using glucosylceramide for food and drink, it is good to use ethanol. As means for purifying glucosylceramide from the alcoholic extract of gum in step C, column chromatography or other suitable purification means can be used.

Moreover, the glucosylceramide-containing rice extract may be a commercially available product, for example, Oryza Ceramide (registered trademark) manufactured by Oryza Oil & Chemicals Co., Ltd. may be used.

The present invention is characterized by using 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, β-sitosterol sambubioside and the like, with β-sitosterol glucoside being particularly preferred. β-sitosterol glucoside is a compound represented by the following chemical formula (2).

The method for obtaining β-sitosterol glucoside is not particularly limited, and commercially available products may be used, or they may be extracted from plants. As a commercially available product, "Sitosterol-3-O-glucoside, β- (AHP Verified)" manufactured by FUJIFILM Wako Pure Chemical Industries, Ltd. can be used.

Plant raw materials for obtaining β-sitosterol glucoside are not particularly limited, and examples thereof include rice, wheat, beet, konjac, corn, and the like. Of these, rice is preferred.
The site used to obtain β-sitosterol glucoside from rice is not particularly limited, but it is particularly preferable to use rice bran and rice germ.

The method of obtaining β-sitosterol glucoside from rice bran or rice germ is not particularly limited. and then partitioning this fermented extract. Alternatively, it can be extracted from “Oryza Ceramide” (registered trademark) manufactured by Oryza Oil & Chemical Co., Ltd.

The agent for preventing viral respiratory infections of the present invention can be used as a material for various foods and drinks. Examples of food and drink include confectionery (gum, candy, caramel, chocolate, cookies, snacks, jelly, gummies, tablets, etc.), noodles (soba, udon, ramen, etc.), dairy products (milk, ice cream, yogurt, etc.). ), seasonings (miso, soy sauce, etc.), soups, beverages (juice, coffee, black tea, tea, carbonated drinks, sports drinks, etc.), general foods, health foods (tablets, capsules, etc.), nutrition Supplementary foods (energy drinks, etc.) can be mentioned. The preventive agent for viral respiratory infections of the present invention may be appropriately added to these foods and drinks.

Various ingredients can be blended into these foods and drinks depending on their type, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, corn syrup, lactose, citric acid, tartaric acid, malic acid, and succinate. acid, lactic acid, L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, gum arabic, Food materials such as carrageenan, casein, gelatin, pectin, agar, B vitamins, nicotinamide, calcium pantothenate, amino acids, calcium salts, pigments, flavors and preservatives can be used.

As a specific production method, the agent of the present invention is spray-dried or freeze-dried together with powdered cellulose, and this is made into powder, granules, tablets, or a solution, so that it can be easily incorporated in foods and drinks (instant foods, etc.). can. Also, the agent of the present invention can be dissolved in, for example, fat, ethanol, glycerin, or a mixture thereof to form a liquid, which can be added to beverages or solid foods. If necessary, it can be mixed with a binder such as gum arabic or dextrin to form powder or granules, which can be added to beverages or solid foods.

When the agent for preventing viral respiratory infections of the present invention is applied to food and drink, the main purpose is to prevent disease and maintain health. It is preferably 20 wt% or less.

The agent for preventing viral respiratory infections of the present invention may be used as a material for drugs (including pharmaceuticals and quasi-drugs). The agent of the present invention can be appropriately blended with raw materials for drug formulations for production. Pharmaceutical raw materials that can be incorporated into the agent of the present invention include, for example, excipients (glucose, lactose, sucrose, sodium chloride, starch, calcium carbonate, kaolin, crystalline cellulose, cacao butter, hydrogenated vegetable oil, kaolin, talc, etc.), Binders (distilled water, physiological saline, ethanol water, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, potassium phosphate, polyvinylpyrrolidone, etc.), disintegrants (sodium alginate, agar, sodium bicarbonate, carbonate) Calcium, sodium lauryl sulfate, monoglyceride stearate, 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, stearate, polyethylene glycol, etc.).

The dosage may vary depending on the administration method, disease state, age of the patient, etc., but for adults the dosage is usually 0.5-5000 mg of the active ingredient per day, and for children it is usually 0.5-3000 mg.
The compounding ratio of the agent for preventing viral respiratory infections of the present invention can be appropriately changed depending on the dosage form, but is usually about 0.3 to 15.0 wt% when administered orally or by mucosal absorption. In the case of parenteral administration, it should be about 0.01 to 10 wt%. Since the dose varies depending on various conditions, there are cases where a dose smaller than the above dose is sufficient, and there are cases where it is necessary to administer a dose exceeding the above range.

Examples of the present invention will be described below. The examples shown below are for confirmation of various functions and effects of the agents obtained by the present invention, and the scope of the present invention is not limited to these products and manufacturing methods.

１‐３ 被験物質3
名称：BSG（β-シトステロールグルコシド）
１‐４ 溶媒
名称：0.5w/v% メチルセルロース400溶液
１－５．調製方法
秤量済みの被験物質に溶媒を混合し、乳鉢と乳棒を用いて調製した。
なお、被験物質は3日分まとめて調製し、調製後は使用するまで冷蔵で保存した。 Test Example: Evaluation of Influenza Prophylaxis and Treatment of Rice Extract Containing Glucosylceramide, Glucosylceramide and β-Sitosterol Glucoside (hereinafter referred to simply as “BSG”) 1. Test substance and preparation method 1-1 Test substance 1
Name: Oryza Ceramide (registered trademark) (rice extract containing 30% glucosylceramide)
Source: Oryza Petrochemical Co., Ltd. 1-2. Test substance 2
Name: Glucosylceramide represented by the following chemical formula (1) isolated from the above oryzaceramide

1-3 Test substance 3
Name: BSG (β-sitosterol glucoside)
1-4 Solvent name: 0.5 w/v% methyl cellulose 400 solution 1-5. Preparation method A solvent was mixed with the weighed test substance and prepared using a mortar and pestle.
The test substance was prepared for 3 days at a time and stored in a refrigerator after preparation until use.

2. Preparation of virus to be used and virus to be inoculated 2-1. Virus used
A virus that was purchased from ATCC and mutated into a mouse hypersusceptible virus at Hamley Co., Ltd. was used.
Virus name: Influenza virus
Strain name: A/PR/8/34
ATCC No.: VR-1469
BSL: 2
Viral titer: 1.58×10 ⁸ TCID ₅₀ /mL
Storage: Store in a deep freezer (CLN-51UD1, Nihon Freezer Co., Ltd.)
2.2 Preparation of virus inoculum Thaw the cryopreserved virus solution, and use PBS to make 2 × 10 ⁵ TCID ₅₀ /mL (1 × 10 ⁴ TCID ₅₀ /50 μL) for general condition observation, and for lung viral load measurement. was prepared to 6×10 ⁴ TCID ₅₀ /mL (3×10 ³ TCID ₅₀ /50 μL).

3. Animal species used: Mouse strain: BALB/c
Source: Charles River Japan Co., Ltd. Gender: Female Age: 4 weeks (at the time of arrival)
Number of animals used: 48 (50 at the time of arrival)
Reason for selection of animal species: Since mouse-susceptible influenza virus is used, mouse selection is considered appropriate as a test system.

4. Animal care conditions 4-1. Breeding conditions Room temperature 22.0-25.0% (setting 24 ± 3 ℃), humidity 43.29-58.18% (setting 50 ± 20%), ventilation (10-25 times/hour), lighting 12 hours (8: 00-20: 00) )
4-2. feed
Ad libitum intake of MF (Oriental Yeast Co., Ltd., Lot 200617).
4-3. Drinking water Autoclave-sterilized city water was available ad libitum.
4-4. Cage Autoclave-sterilized heat-resistant polysulfone cage (207W×365D×140H mm, TECNIPLAST Ltd) is used. Animals were housed 1-5/cage.
4-5. From the arrival of quarantine and acclimatization animals until the end of quarantine and acclimatization, general conditions were observed daily. Body weights were also measured on the day of arrival and on the end of quarantine and acclimatization.
4-6. Grouping Animals were assigned to each group by weight-stratified random sampling using body weights at the end of quarantine acclimation.

5. Test group composition The test group composition was as shown in Table 1 below.

6. Inoculation of Virus Under anesthesia with isoflurane (Lot 196AR0, Myline Pharmaceutical Co., Ltd.), 50 μL/head of the inoculated virus solution was nasally inoculated.

7. Administration of Test Substance From 7 days before virus inoculation to the day before the end of observation, 10 mL/kg of test substance was orally administered once a day using a syringe and sonde. On the day of virus inoculation, the test substance was administered before virus inoculation.

8. Specimen collection (5-8 groups)
After euthanasia by overdose of thiamylal sodium (Nichi-Iko Co., Ltd., Lot LU1200, Isosol® injection 0.5 g), lungs were collected and weighed. 50-100 mg of lungs were harvested for viral load measurement, 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.

9. Observation/measurement items 9-1. General Condition General condition was observed at least once a day.
9-2. At the time of body weight measurement grouping (first administration: Day -7), Days 0, 3, 7, 10 and Day 14

10. Statistical Analysis Weighing data obtained as test data was calculated as mean ± standard deviation.
After calculating the average survival days and survival rate of the mice for general condition observation, the Log-Rank test was performed on the vehicle group.

11. Results Fig. 1 and Table 2 below show survival rates.
Table 3 below shows the average survival days and the survival rate at the end of observation.
The average survival days in Group 1 (vehicle group), Group 2 (oryzaceramide group), Group 3 (glucosylceramide group) and Group 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.

12. Discussion and Effect of Examples For the purpose of confirming the effects of oryzaceramide and its ingredients using an influenza-infected mouse model, the survival rate and average survival days of mice after virus inoculation were evaluated.
As a result, oryzaceramide and BSG significantly extended the survival time compared to the vehicle group. In addition, it was confirmed that the main component of glucosylceramide also tended to prolong survival.
From the above, it was confirmed that glucosylceramide, the rice extract containing it, and BSG are useful as agents for preventing and treating influenza.

Fig. 2 shows a formulation example of the agent (glucosylceramide-containing rice extract) according to the present invention. In addition, the following formulation examples do not limit the present invention. In the formulation examples below, only the glucosylceramide-containing rice extract is used, but BSG and the above-mentioned glucosylceramide can also be blended in the same manner.
Formulation Example 1: Chewing gum
Sugar 53.0 wt%
gum base 20.0
glucose 10.0
Starch syrup 16.0
Perfume 0.5
Glucosylceramide-containing rice extract 0.5
100.0 wt%

Formulation Example 2: Gummy
Reduced starch syrup 40.0 wt%
Granulated sugar 20.0
Glucose 20.0
gelatin 4.7
Water 9.68
Yuzu Juice 4.0
Yuzu flavor 0.6
Pigment 0.02
Glucosylceramide-containing rice extract 1.0
100.0 wt%

Formulation Example 3: Candy
Sugar 50.0 wt%
Starch syrup 33.0
Water 14.4
Organic acid 2.0
Perfume 0.2
Glucosylceramide-containing rice extract 0.4
100.0 wt%

Formulation Example 4: Yogurt (hard/soft)
Milk 41.5 wt%
Skimmed 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
100.0 wt%

Formulation Example 5: Soft drink
Fructose glucose liquid sugar 30.0 wt%
emulsifier 0.5
Glucosylceramide-containing rice extract 0.3
Perfume Appropriate amount
purified water residue
100.0 wt%

Formulation Example 6: Tablet candy
Sugar 76.4 wt%
Glucose 19.0
Sucrose fatty acid ester 0.2
Glucosylceramide-containing rice extract 0.5
Purified water 3.9
100.0 wt%

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

Formulation Example 8: Tablet
Lactose 54.0 wt%
Crystalline cellulose 30.0
Starch hydrolyzate 10.0
Glycerin fatty acid ester 5.0
Glucosylceramide-containing rice extract 1.0
100.0 wt%

As described above, the present invention can provide a novel preventive and therapeutic agent for viral respiratory infections.

Claims (3)

Glucosylceramide and/or an agent for preventing and treating viral respiratory infections containing glucosylceramide as an active ingredient. A preventive and therapeutic agent for viral respiratory infections containing β-sitosterol glycoside as an active ingredient. 3. The agent for preventing or treating a viral respiratory infection according to claim 1 or 2, wherein the viral respiratory infection is influenza.

Images