JP5232404B2 - Anti-cold virus or anti-influenza virus composition containing sporic lactic acid bacteria - Google Patents

Description

  The present invention relates to a composition for anti-cold virus or anti-influenza virus containing spore-forming lactic acid bacteria.

  Administering lactic acid bacteria for the purpose of improving constipation and loose stool is a widely used technique, and this intestinal effect is basically expressed in any lactic acid bacteria species and strains.

  As functions other than the above, lactic acid bacteria species or strains having specific pharmacological actions such as blood pressure lowering, blood glucose lowering, antiallergic, immune enhancement, anticancer, anti-Helicobacter pylori, antivirus, etc. are being discovered, Has been put to practical use.

Moreover, the following reports have been reported so far regarding the anti-cold virus or anti-influenza virus action of lactic acid bacteria.
(1) It has been clarified by an in vitro test that antibacterial virus action is exerted on the cell components of Felicalis, Acidophilus, and Bifidobacteria (Patent Document 1).
(2) It has been reported that an inhibitory component of plaque formation expressed by herpes simplex virus contained in the cells of genus Streptococcus lactic acid bacteria has a protective action against adenovirus infection (Patent Document 2).
(3) It has been found that oral ingestion of L. casei leads to an increase in the proliferation ability of T cells and CD3 + against influenza virus antigens (Patent Document 3).

  On the other hand, in 1949, Nakayama et al. Isolated lactic acid bacteria (hereinafter referred to as sporic lactic acid bacteria) that form spores as a kind of Bacillus coagulans from green malt (Non-patent Document 1). Although sporic lactic acid bacteria are literally spore-shaped lactic acid bacteria, they are sometimes called Lactobacillus sporogenes, but are taxonomically renamed Bacillus coagulans (for example, Non-Patent Document 2). That is, it is a very unique bacterium having a face of lactic acid bacteria under anaerobic conditions and a face of the genus Bacillus under aerobic conditions (Non-patent Document 3).

  Furthermore, spore-forming lactic acid bacteria preparations are resistant to gastric acid and bile because they have the characteristics of heat resistance, aerobicity and acid resistance. Widely blended in food products. In addition to the above-mentioned effects, sporic lactic acid bacteria are effective for diseases mainly having diarrhea such as enteritis and colitis, diarrhea caused by antibiotic administration (for example, Non-Patent Document 4), and skin quality improving effects (for example, Non-patent document 3), and is reported to be effective for hyperlipidemia and aphthous stomatitis (for example, non-patent document 5). Furthermore, it has been reported that the survival rate is improved when cytomegalovirus is infected to mice orally administered with spore-forming lactic acid bacteria (Non-patent Document 6).

However, it has not been known so far whether or not an anti-cold virus action or an anti-influenza action is exhibited by oral administration of spore-forming lactic acid bacteria.
Japanese Patent Laid-Open No. 50-76295 JP-A-53-52611 Special Table 2003-534284 Japanese Journal of Agricultural Chemistry Vol.23 No.12 (1949) p.513-517 Nutrition Research Vol.22 (2002) p.71-84 Biotechnology Vol.80 No.12 (2002) p.581-583 Sankyo Bibliography (1965) p.7-11 Altern Med Rev, Vol.7 No.4 (2002) p.340-342 Probiotic Symposium '05 Preliminary Lectures (2005) p.6

  The present inventors have intensively studied to discover new uses of spore-forming lactic acid bacteria for oral administration. As a result, the present inventors have found that sporic lactic acid bacteria exhibit an excellent antiviral effect against cold-related viruses and influenza-related viruses, and have completed the present invention.

The present invention is (1) a composition for preventing or treating a cold virus or influenza virus infectious disease, comprising a spore-forming lactic acid bacterium,
(2) a composition for preventing or treating various symptoms associated with cold virus or influenza virus infection, comprising a spore-forming lactic acid bacterium;
(3) The composition according to (1) or (2), wherein the sporic lactic acid bacterium is Bacillus coagulans,
(4) The composition according to (1) or (2), wherein the sporic lactic acid bacterium is Bacillus coagulans SANK70258,
(5) The composition according to any one of (1) to (4), wherein the cold virus is a coronavirus, rhinovirus or adenovirus,
(6) The composition according to any one of (1) to (4) selected from (1) to (4), and (7) (1) to (1) to be used as a food or medicine The composition according to any one of (6).

  In the present invention, the “spore-forming lactic acid bacterium” generally refers to a lactic acid bacterium that forms a spore, and is preferably a dried spore or viable cell of Bacillus coagulans. Usually, a lactic acid bacteria bulk powder produced by mixing spore-forming lactic acid bacteria with a suitable excipient such as lactose, sucrose, dextrin, starch, or a mixture thereof is used.

  In the present invention, the “cold virus” generally refers to a virus that causes the common cold, and is preferably a coronavirus, rhinovirus, adenovirus, coxsackie virus, or the like, and more preferably a coronavirus, rhinovirus. Virus or adenovirus.

  In the present invention, “influenza virus” generally indicates a virus that causes influenza, preferably influenza virus A, influenza virus B, or influenza virus C, and more preferably influenza virus A or influenza virus. Virus B.

  In the present invention, a “cold virus or influenza virus infectious disease” is a disease that generally develops as a result of infection with a cold virus or influenza virus.

  In the present invention, the “symptoms associated with the cold virus or influenza virus infection” are not particularly limited as long as they are various symptoms associated with the cold virus or influenza virus infection. Pain, cough, rash, sneeze, chills, fever, headache, joint pain or muscle pain.

  In the present invention, “treatment” means to cure, improve or alleviate a disease or symptom, or to suppress a symptom, and “prevention” means to prevent the onset of the disease or symptom. To do.

  Oral administration of spore-forming lactic acid bacteria is useful because it is effective in preventing or treating symptoms caused by infection with a cold-related virus or influenza-related virus.

  Spore-forming lactic acid bacteria are listed in Japanese Pharmacopoeia Standards for Drugs 2002.

  The spore-forming lactic acid bacteria of the present invention can be administered in the form of an oral preparation or food.

The spore-forming lactic acid bacteria bulk powder usually contains 5 × 10 ^{9 to} 1 × 10 ¹² spore-forming lactic acid bacteria in 1 g.

The daily dose of spore-forming lactic acid bacteria varies depending on the indication or purpose, but is usually 1 × 10 ⁵ to 1 × 10 ¹³ , preferably 1 × 10 ⁶ to 1 × 10 ¹² This is administered in 1 to 3 divided doses per day.

  When the composition of the present invention is a solid preparation, the content of the spore-forming lactic acid bacterium bulk powder is usually 10 mg to 1000 mg, preferably 20 mg to 500 mg.

  In the present invention, in addition to the above active ingredients, anti-influenza drugs, cold medicines, traditional Chinese medicines, herbal medicines or herbs, vitamins, and the like can be contained as long as they do not impair the effects of the present invention.

  Specific examples of these forms include tablets, chewing tablets, fine granules (including granules and powders), capsules, etc. for pharmaceuticals, jelly agents, cookies, gums, chocolates, candy, drops, marmalades, Mayonnaise, dressing, bread, pudding, tableted confectionery, powdered juice, sprinkles or toppings can be raised, and additives and base materials suitable for each dosage form are used as appropriate. According to the method of, it can manufacture.

  In the above dosage forms, various commonly used additives can be used depending on the dosage form. For example, excipients, stabilizers, coating agents, lubricants, adsorbents, binders, disintegrants, surfactants, colorants, pH adjusters, fragrances, sugar, starch syrup, cacao butter, butter , Margarine, edible fats and oils, salt, condensed milk, flour, other food materials and food additives, and the like can be blended.

  Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, but the scope of the present invention is not limited thereto.

Example 1 Tablet (1) Ingredient (Table 1)
9 tablets (mg)
---------------------
Sporulated lactic acid bacteria powder 45
White sugar 140
Talc 30
Magnesium stearate 10
Hydroxypropyl cellulose 5
Crystalline cellulose 150
Lactose Appropriate amount of fragrance Trace amount ---------------------.
(2) Manufacturing method Take the above ingredients and the amount, and manufacture tablets according to the section of “General Tablets” “Tablet”.

(Example 2) Granule (1) Ingredient (Table 2)
1 to 3 packs (mg)
---------------------
Sporulated lactic acid bacteria powder 45
Sucrose 110
Mannitol 900
Tartaric acid 2
Hydroxypropyl cellulose 50
Lactose Appropriate amount perfume Trace amount --------------------.
(2) Production method Take the above ingredients and quantities, and produce a fine granule according to the section of the Japanese Pharmacopoeia General Rules “Granule”.

Example 3 Capsule (1) Ingredient (Table 3)
1 to 2 capsules (mg)
---------------------
Sporulated lactic acid bacteria powder 45
Hydroxypropyl cellulose 20
Magnesium stearate 10
Lactose appropriate amount ---------------------.
(2) Manufacturing method After taking the above components and quantities and preparing a fine granule according to the section of the Japanese Pharmacopoeia General Rules “Granules”, it is filled into capsules to produce hard capsules.

(Example 4) White chocolate (1) component (Table 4)
Of 100g per sheet (g)
---------------------
Sporulated lactic acid bacteria raw powder 1
Bitter chocolate 18
Cocoa butter 22
Whole milk powder 20
Lecithin 0.3
Vanillin trace sugar remainder ---------------------.
(2) Manufacturing method The said component and quantity are taken, and it manufactures according to a normal chocolate manufacturing method. Add spores of lactic acid bacteria bulk powder, knead again and mold.

(Example 5) Candy (1) component (Table 5)
Of 100g per sheet (g)
---------------------
Sporulated lactic acid bacteria raw powder 1
Mizuame 55
Citric acid Appropriate amount of fragrance Trace sugar The remainder ---------------------.
(2) Manufacturing method The said component and quantity are taken, and it manufactures according to a normal candy manufacturing method. When the temperature reaches 80 ° C. or lower in the final step, the spore-forming lactic acid bacteria bulk powder is added, and again kneaded and molded.

(Example 6) Bread (1) ingredients (Table 6)
Of 100g per sheet (g)
---------------------
Sporulated lactic acid bacteria raw powder 1
Flour 85
Salt Suitable amount Shortening Suitable amount Yeast 3
Water balance ---------------------.
(2) Manufacturing method The above components and quantity are taken and manufactured according to a normal bread manufacturing method.

(Test example) Anticoronavirus and anti-influenza virus effect test of spore-forming lactic acid bacteria (1) Test substance The powder of spore-forming lactic acid bacteria (Bacillus coagulans SANK70258) manufactured by Sankyo Lifetech Co., Ltd. was used.

The test substance was prepared to be 20 mg / 0.2 mL / animal using pharmacopoeia water for injection on the test day. The dose of sporic lactic acid bacteria was 1.6 × 10 ⁸ cells / mouse, and the same amount of water for injection was administered to the control group.
(2) Animals BALB / c female mice (made by Charles River Japan) 3 or 5 weeks old, purchased at a P3 facility controlled at a temperature of 23 ± ° C., a humidity of 55 ± 10%, and a lighting time of 8:00 to 20:00 The animals were bred by placing them in polycarbonate breeding cages sterilized by autoclaving. The feed was placed in a water bottle made of polycarbonate sterilized by autoclaving special solid feed for experimental animals (CRF-1, manufactured by Oriental Yeast Co., Ltd.) and tap water passed through a water filter, and each was allowed to ingest freely.

After the acclimatization, the group was divided into 15 groups per group based on the latest body weight, and the following three groups were used. Group 1 is negative control group (no virus intake and spore lactic acid bacteria non-administration), Group 2 is positive control group (virus intake and spore lactic acid bacteria non-administration), group 3 is test substance administration group (virus intake and spore lactic acid bacteria administration) ).
(3) virus types used used virus, the route of administration and dosage level: shows a (50% Tissue Culture Infectious Dose TCID 50 / 10μL) in Table 7. All viruses were purchased from the American Type Culture Collection (ATCC).

  Cells purchased from ATCC were cultured, and after confirming the monolayer state under the microscope, the virus was added and the Cytopathic Effect (CPE) was confirmed under the microscope. The culture supernatant was collected and stored at -80 ° C or lower.

(Table 7)
Virus type Administration route Administration concentration At the time of inoculation ―――――――――――――――――――――――――――――――――――――
Influenza virus A (H1N1) Nasal 1 × 10 ⁸ 5-week-old influenza virus B (Hong Kong / 5/72) Nasal 1 × 10 ⁸ 5-week-old coronavirus (MHV) In the brain 1 × 10 ⁸ 7-week-old ―――――――――――――――――――――――――――――――――――.
(4) Method From 2 weeks before the virus inoculation to the 10th day after the inoculation, the test substance was forcibly orally administered once a day for a total of 24 times using a gastric sonde and a syringe.

  On the second week after administration of the test drug, the virus was inoculated intranasally or intracerebrally at a concentration that caused death in the preliminary test. The mice were observed for 10 days after the inoculation, and the survival rate of the mice was calculated by the following formula.

Survival rate of mice after virus infection = 100 × [number of surviving mice / (total: 15)]

(5) Test result The result of the obtained survival rate is shown in FIGS. 1-3 and Tables 8-10. Each value is an average value of 15 animals per group.

  Table 8 shows the results of the survival rate on the 10th day after the coronavirus infection.

(Table 8)
Presence or absence of infection Spore lactic acid bacteria Survival rate (%)
―――――――――――――――――――――――
Non-infection Non-administration 100
Infection Administration 67
Infection Not administered 47
―――――――――――――――――――――――.

  From Table 8, the improvement of the survival rate was recognized by administration of the spore lactic acid bacteria.

FIG. 1 is a graph in which the horizontal axis represents the number of days elapsed since virus infection and the vertical axis represents the change in survival rate (%). From FIG. 1, the survival rate was improved by the pre-infection administration of spore-forming lactic acid bacteria.

Table 9 shows the results of survival rate on the 10th day after infection with influenza virus A.

(Table 9)
Presence or absence of infection Spore lactic acid bacteria Survival rate (%)
―――――――――――――――――――――――
Non-infection Non-administration 100
Infection Administration 100
Infection Not administered 87
―――――――――――――――――――――――.

  From Table 9, death cases were observed in the non-administration group of spore-forming lactic acid bacteria, but not in the administration group.

FIG. 2 is a graph in which the horizontal axis represents the number of days elapsed since virus infection and the vertical axis represents the change in survival rate (%). From FIG. 2, the survival rate was improved by the pre-infection administration of spore-forming lactic acid bacteria.

Table 10 shows the results of survival rate on the 10th day after influenza virus B infection.

(Table 10)
Presence or absence of infection Spore lactic acid bacteria Survival rate (%)
―――――――――――――――――――――――
Non-infection Non-administration 100
Infection Administration 100
Infectious non-administration 87
―――――――――――――――――――――――.

  From Table 10, death cases were observed in the non-administration group of spore-forming lactic acid bacteria, but not in the administration group.

  FIG. 3 is a graph in which the horizontal axis represents the number of days elapsed since virus infection and the vertical axis represents the change in survival rate (%). From FIG. 3, the survival rate was improved by the pre-infection administration of spore-forming lactic acid bacteria.

  Oral administration of spore-forming lactic acid bacteria is useful because it is effective in preventing or treating symptoms caused by infection with a cold-related virus or influenza-related virus.

FIG. 1 is a graph showing the survival rate during coronavirus infection. The horizontal axis represents the number of days that have passed since coronavirus infection, and the vertical axis represents the survival rate (%). FIG. 2 is a graph showing the survival rate during influenza virus A infection. The horizontal axis represents the number of days elapsed from influenza virus A infection, and the vertical axis represents the survival rate (%). FIG. 3 is a graph showing the survival rate during influenza virus B infection. The horizontal axis represents the number of days elapsed since influenza virus B infection, and the vertical axis represents the survival rate (%).

Claims (1)

Containing Bacillus coagulans SANK70258, composition for preventing or treating Koronauiru scan infectious diseases.