JPH07265064A - Composition for improving enterobacterial flora - Google Patents

Abstract

PURPOSE:To obtain a composition for improving enterobacterial floral, containing a live microbial cell of an enterobacterium as an antiallergic component, useful for treating, etc., allergic diseases such as atopic dermatitis or pollinosis and useful as a food or a medicine. CONSTITUTION:This composition contains a live microbial cell of a nonpathogenic enterobacterium (preferably a bacterium of the genus Lactobacillus, Clostridium, Bifidobacterium, Streptococcus or Enterococcus) as an antiallergic ingredient. Furthermore, the daily intake thereof preferably contains >=100000000000 live strains.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition (food or drug) which achieves an antiallergic effect by improving intestinal bacterial flora and a novel intestinal bacterial flora improving composition.

[0002]

2. Description of the Related Art Allergies such as hay fever and atopic dermatitis, which have been rapidly increasing in recent years, have been considered as diseases of the immune system. However, since the cause and the mechanism of its onset are not known yet, a fundamental and effective treatment method has not been established, and the reality is that the treatment is symptomatic.

In recent years, the relationship between the immune system and other organs has been gradually clarified, and it has come to be considered that the immune system functions while being closely related to other organs.

[0004] For example, it is known that the immune system is affected by the cranial nervous system via hormones, and that the activity of natural killer (NK) cells is reduced under conditions of high mental stress (Matsuda). , Hirano, Brain and Immunity (Metabolism, Vol. 26, Special Issue, pages 45 to 61), R. Ader
Et al. Interactions betw-ee
n the brain and the immune system) Annual Report of Pharmacology and Toxicology (Annu. Rev. Phama-col. Toxicol.) 1990.30, 561.
602).

The Peyer's patches present in the intestine are important tissues that control the immune system by the antigen components present in the digestive organs, and it is considered that the intestinal tract and immune functions are deeply involved.

[0006]

With respect to allergies, it is necessary to comprehensively consider the relationship with other organs and the relationship with the living environment. We have
From this point of view, when the patient with allergic disease was treated for many years, the following findings were obtained.

Noget Ear Examination (Blu Dior Translated by Yukio Nagasu Ear Needle Method-P. Nozier's Theory and Clinical (Published by Enterprise Co., Ltd. in 1985), Nozier Ikuhiro Fukuda Translation of Auricles, 1989 Issued by Taniguchi Shoten , Min Han Cho, Theory and Clinic of Ear Needle Method in Europe and America in 1979.
Although many have no subjective symptoms, they have abnormalities in the digestive organs, mainly the intestinal tract, and at the same time are in a state of being mentally stressed by an interview. That is, there are many people who have constipation or its tendency, or conversely, are likely to have diarrhea, and excrete stool or gas having an abnormally strong odor.

In summary, it has been found that many patients with allergic diseases actually have abnormalities in the intestinal flora and intestinal damage. The intestinal flora and gastrointestinal epithelium easily become abnormal due to mental stress and are easily damaged.

The present inventor has treated various diseases caused by autonomic imbalance by means of electrification from the ear and cured extremely excellent treatment results. However, in allergic diseases, electrification and anti-inflammatory to skin. Although a therapeutic effect was obtained by applying the agent and its combined use, it often took a long treatment period.

In recent years, it has been often pointed out that pesticides, antibiotics, synthetic antibacterial agents and the like, which are frequently used in the fields of agriculture, fisheries and livestock, remain in foods, and these are also enteric. It is a factor that damages the flora. Modern people are in a situation where intestinal bacteria are prone to abnormalities. It is considered that abnormalities of intestinal microbiota and damage to the mucosal epithelium adversely affect the immune system, nervous system, and metabolic system, causing various damages of allergic diseases in a cyclical manner.

Conventionally, there are intestinal regulating agents containing intestinal bacterial flora, but these are insufficient in improving the intestinal bacterial flora due to a small number of bacterial species or a small number of bacteria. Therefore, no consideration is given to allergic diseases.

[0012] Therefore, it is an object of the present invention to provide a composition capable of treating and preventing allergic diseases by improving the intestinal microflora. Another object of the present invention is to provide a novel intestinal microflora-improving composition.

[0013]

Means for Solving the Problems The present inventor has found that in allergic diseases such as atopic dermatitis and hay fever that are not known in origin, stress from the cranial nerves and abnormal intestinal bacterial flora present in the digestive tract are Obtaining the finding that they are often related to each other, we found that ingestion of intestinal bacteria has a great effect on the treatment or prevention of allergies, and completed the present invention that achieves the above object. It is a thing.

That is, the present invention is characterized by being a composition containing live bacterial cells of enterobacteria as an antiallergic component.

The enterobacteria of the present invention refer to various non-pathogenic bacteria existing in the human intestinal tract, such as Streptococcus, Enterococcus, Bifidobacterium,
The bacterium is preferably included in at least one genus of Lactobacillus and Clostridium. More preferably, the anti-allergic composition of the present invention comprises bacteria of all these genera. However, even a bacterium contained in one of these genera achieves a superior effect (anti-allergic effect) as compared with conventional intestinal regulating agents according to the examples described below.

[0016] By ingesting or taking these intestinal bacteria, the intestinal bacterial flora is improved and it becomes possible to treat or prevent allergic diseases. In this application, the anti-allergic action includes not only the treatment of allergies but also the prevention of allergies (maintaining the state where allergies are treated).

The antiallergic action of the composition of the present invention is
It provides good therapeutic and preventive effects against various allergic diseases such as atopic dermatitis, hay fever, respiratory diseases such as allergic rhinitis, and allergic diseases caused by drugs, or symptoms that may cause them.

In this application, the improvement of intestinal flora means normalization of intestinal bacteria (number of intestinal bacteria, balance of intestinal bacterial species, etc.) as well as normal intestinal bacterial flora. It also includes actively maintaining the state of being, or supplementing intestinal bacteria. The improvement of the intestinal microflora itself leads to the antiallergic effect as described above, and also leads to a wide range of effects such as intestinal regulating action, nourishing and tonic action, physical strength promotion / maintenance, and beauty and health.

The present invention also provides a novel composition for improving intestinal flora. That is, the present invention is
An intestinal microflora-improving composition containing intestinal bacteria belonging to all of Streptococcus, Enterococcus, Bifidobacterium, Lactobacillus, and Clostridium as live cells. Further, the present invention is a composition for improving intestinal flora, which comprises 100 billion or more viable bacterial cells of intestinal bacteria in daily intake. As described above, the intestinal microflora is improved and the antiallergic effect is exhibited by these novel compositions, and in addition, the intestinal regulating action, the nutritional tonic action and the like are achieved.

The above-mentioned Streptococcus, Enterococcus, Bifidobacterium, Lactobacillus,
Bacterial species belonging to any one of the genus Clostridium (bacterial species belonging to the present invention) include Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus ebium, Streptococcus salivarius, Streptococcus.
Bovis, Streptococcus ikunus, Lactobacillus acidophilus, Lactobacillus gasserie, Lactobacillus salivarius, Lactobacillus
Fermentum, Lactobacillus reuteri, Lactobacillus arabinosus, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium adrensetis, Bifidobacterium infantes, Bifidobacterium bifidum, etc. Can be illustrated.

Further, the composition of the present invention may contain other intestinal bacteria such as known yogurt bacteria. Examples of such yogurt bacteria include at least one of Streptococcus thermophilus and Lactobacillus bulgaricus.

Examples of specific strains of the above-mentioned strains include Enterococcus faecalis ATCC 19433 and Enterococcus faecium ATC.
C 19434, Enterococcus durans IID 677, Streptococcus salivarius IID 5223, Enterococcus hirae ATCC 8043, Enterococcus hirae ATCC 9790, Lactobacillus acidophilus ATCC
4356, Lactobacillus arabinosas ATCC 8014, Lactobacillus fermentum IAM 1148, Lactobacillus casei ATCC 393, Bifidobacterium adrensetis ATCC 15705, Bifidobacterium longum JCM 1217, Bifidobacterium bifidum
JCM 1254, Bifidobacterium breve JCM 119
2. Enterococcus hirae ATCC 8043 and Clostridium butyricum ATCC 6015. All of these strains are easily available. These strains are managed and stored in the following microorganism preservation institutions. These institutions distribute the lots on request. The English letters IAM for strains indicate the Institute of Applied Microbiology, University of Tokyo (currently Institute for Molecular and Cellular Biology), IID indicates the Institute of Medical Science, University of Tokyo, and ATCC indicates The American Type.
Shows Culture Collection, JCM is Japan Collection
Indicates of Microorganisms. The numbers after these English letters are numbers for identifying the strain. In short, regardless of whether they are Japanese citizens, these strains can be easily obtained because they are offered for distribution to the institutions shown here or are commercially available.

These preservation organizations are the Japanese Federation of Preservation Machines (JFCC).
Since I am a member of the Ollection of Microorganisms),
JFCC Catalog of Cu
ltuers, 1966 and 1968), you can find out in which conservation institution the desired fungus is stored. The contact information (address) of each of the above institutions is as follows.

Institute of Applied Microbiology, University of Tokyo: 1-1-1 Yayoi, Bunkyo-ku, Tokyo ATCC: 12301 Parklawn Drive, Rockville, Md. 20852 ,
USA JCM: 2-1 Hirosawa, Wako City, Saitama Life Science
Bio-biological department As other preservation organizations that preserve the above strains,
Fermentation Research Institute (Address: Jusannishinomachi, Higashiyodogawa-ku, Osaka-shi, Osaka), The Northern Regional Research Laborato
ry, Peoria, Illinois, USA, Centraalbureau voor sc
himmelcultur-es, Baar, Netherlands.

The ATCC strains described here are the strains issued in 1989.
ATCC catalog (17th edition), JCM strains in JCM catalog issued in 1992, other strains in 1979 or
It is described in the JFCC catalog issued in 1992.

Known culture techniques can be applied to the culture of these strains. The following are all examples of culture methods.

Culturing of the genus Enterococcus, the genus Streptomyces, and the genus Lactobacillus is carried out aerobically using a Rogosa medium. Cultivation of the genus Bifidobacterium and the genus Clostridium is anaerobic, and is performed using, for example, GAM medium (manufactured or sold by Nippon Pharmaceutical Co., Ltd.).

In a preferred embodiment of the composition of the present invention, live cells are separated from this culture solution, and the live cells are frozen with the culture solution and excipients such as skim milk, glucose, lactose and starch. It consists of dried bacterial cells. These strains are preferably stored in a cool dark place (preferably several degrees Celsius or less).

The composition of the present invention is prepared by a known formulation technique, if necessary, together with excipients, etc., into powders, tablets,
It can be provided as a liquid preparation, granules or fine granules. The viable cell count in the obtained composition is preferably 10 ^{9 to} 10 ¹² / g.

The effective amount of the composition of the present invention is tentatively based on the amount which can be ingested from 100 billion to 10 trillion in the daily intake of viable cell count. However, even if ingesting less than or more than this amount of viable bacteria, the effect is observed in most cases. No acute toxicity was observed with the composition of the present invention.

The composition of the present invention may contain a culture obtained by culturing the intestinal bacterium in an edible medium. Thereby, the nutrition of the edible medium can be simultaneously ingested together with the ingestion of the composition of the present invention. In addition, it is preferable that the number of bacteria in the composition is increased by culturing in an edible medium, and then it can be ingested. Examples of such an edible medium include milk and soy milk.

The bacterium is preferably cultured in an edible medium at the optimum growth temperature, and in the case of the strain, the temperature is preferably 40 ± 4 ° C. (36 to 44 ° C.). In the aggregate of all the above-mentioned strains, particularly preferably 37 to 40 ° C. When the culturing temperature is within this range, the enterobacteria can be grown, proliferated, and cultivated while preventing the growth of various bacteria other than the enterobacteria of the present invention.

In the present invention, the duration of heating for culturing is preferably the time until fermentation (growth of microorganisms) reaches almost all regions of the nutrient medium. The culturing time at this time is preferably about 8 to 15 hours. Adjusting the culturing time means that the growth of bacteria (may be referred to as growth and growth), the fermentation of an edible medium (the growth of bacteria in an edible medium) are allowed to proceed to a desired range, and It can be said that it is desirable from the viewpoint of setting the properties (mouthfeel, fluidity, taste, etc.) of the edible medium (nutrient medium) within a desired range or preventing the growth of various bacteria.

The amount of bacteria inoculated into the edible medium is not particularly limited and is selected from the most suitable range depending on the type of bacteria and / or the components of the medium or the amount thereof. Effectively, the composition of the present invention contains a component that improves the growth of bacteria in the composition. This improves the growth of the bacteria on the edible medium and helps to evenly grow on the edible medium. Oligosaccharides are preferred as this component. This is because this oligosaccharide selectively grows intestinal bacteria. Even when ingested by humans, this oligosaccharide is not digested by digestive enzymes and reaches the large intestine as it is to be used for the growth of intestinal bacteria. Such oligosaccharides include lactosucrose (milk fruit oligosaccharide), isomaltooligosaccharide, galactooligosaccharide (raffinose,
Soybean oligosaccharides) are included.

By adding this oligosaccharide to the composition of the present invention, the intestinal bacteria ingested can grow better even in the human intestinal tract. The content of oligosaccharides added to the composition is not particularly limited, but usually, it may be added to the composition so that the daily intake of humans is 10 mg to 100 g of oligosaccharides. Other similar ingredients include skim milk. In addition, components such as oligos may be directly added to the edible medium (nutrient medium). The composition of the present invention has use as a health food, a functional food, or a drug.

Further, it is preferable to apply a known acid-resistant coating (enteric coating, enteric coating) to the above-mentioned dried cells in order to prevent decomposition by gastric acid when ingested. Such coating includes covering dry cells with a fat composed of fatty acids (stearic acid, palmitic acid, etc.) and wax (slack, etc.), or coating dry cells with proteins such as zein. Be done. These coating materials may be applied to or sprayed on the dry cells directly or after being dissolved in a solvent (alcohol such as ethanol).

The amount of bacteria inoculated into the edible medium is not particularly limited and is selected from the most suitable range depending on the type of bacteria and / or the components of the medium or the amount thereof.

[0038]

According to the present invention, in allergic diseases such as atopic dermatitis and hay fever, stress coming from cranial nerves and abnormalities of intestinal microflora existing in the digestive tract are often associated with each other. Based on the above findings, the intestinal bacteria are ingested to improve the intestinal microflora, and it has a great effect on the treatment or prevention of allergy.

Since the present invention is a novel composition for improving intestinal microflora, it has an excellent anti-allergic action, an excellent intestinal regulating action, a nutritional tonic action and the like in comparison with the conventional similar compositions. It exerts an effect based on the improvement of the internal flora.

[0040]

【Example】

(Production of Composition) Cultivation of bacteria contained in the genus Enterococcus, the genus Streptococcus, or the genus Lactobacillus is aerobically carried out at 37 ° C. using Rogosa culture. Cultivation of bacteria belonging to the genus Bifidobacterium or Clostridium is anaerobically carried out at 37 ° C. using a GAM medium (manufactured by Nissui Pharmaceutical Co., Ltd.).

The culture medium was previously autoclaved 121
Apply sterilization for 15 minutes at 0 ° C. Dry cells were obtained by the following method for each strain shown in Table 1.

The medium was inoculated with each strain shown in Table 1 (10 ⁶
/)), After culturing for 15 hours, the culture solution is centrifuged at 12,000 rpm to collect the cells, and the cells are dispersed in 10% skim milk of 1/20 of the culture solution and freeze-dried. The viable cell count of the dried cells was 10 ^{9 to} 10 ¹² / g. The viable cell count of each strain is shown in Table 1.

Rogosa medium: in 1 liter of water Trypticase 10 g Yeast extract 5 g Tryptose 3 g K ₂ HPO ₄ 3 g KH ₂ PO ₄ 2 g Ammonium citrate 2 g Tween 80 1 g Glucose 20 g Cysteine 0.2 g Salt solution 5 ml Salt solution: Purified water 1000 ml MgSO ₄ · 7H ₂ O 11.5 g FeSO ₄ · 7H ₂ O 0.68 g MnSO ₄ · 7H ₂ 0 2.4 g (Anti-allergic test 1) Next, for patients with atopic dermatitis, energization as described above. When only therapeutic treatment and application treatment of anti-inflammatory ointment (Shiunkaku: manufactured or sold by Nagakura Seiyaku Co., Ltd. or Matsuura Seiyaku Co., Ltd.), this treatment and a commercially available rectifying agent (Levenin: Wakamoto Pharmaceutical) are used. , Biosley:
Experiments were carried out on the antiallergic effect when the dry bacterial cells shown in Table 1 were ingested as compared with the case where the drug was taken according to the usage of Toa Pharmaceutical Co., Ltd.). The results are shown in Table 2.

Experimental Example 1: Dried bacterial cells of the strains shown in Table 1 are mixed in equal amounts to give a mixed dried bacterial cell powder. This was packed in a gelatin capsule at a dose of 500 mg and taken 3 times daily.

Experimental Example 2: The dried bacterial cells of Experimental Example 1 were added to 1 liter of commercially available milk that had been refrigerated, placed in a warmer, and kept at about 40 ° C. for 8 to 12 hours. Milk coagulates weakly, but this was refrigerated and taken in an amount of 500 ml daily. At the time of administration, sugar, aspartame and other sweetening amounts were added, or jam and fruit sugar were added according to taste, and they were seasoned appropriately before eating.

Experimental Example 3: Enterococcus hirae A
500 mg of dry cells of TCC 9790 was filled in a gelatin capsule, and was taken 3 times daily.

Experimental Example 4: 500 mg of dry cells of Lactobacillus acidophilus JCM 1217 was packed in a gelatin capsule and taken three times daily.

Example 5. Bifidoacterium longum JCM 1217 500 mg of dried bacterial cells was packed in a gelatin capsule and taken 3 times daily.

The energization treatment method and the application treatment of anti-inflammatory ointment (purple cloud), this treatment and the administration of a commercially available rectifying intestinal medicine, and each of the experimental examples were conducted for 3 months. Place the dried cells or a serratin capsule filled with dried cells in a sealed bottle 4
Stored below ℃.

The results are shown in Table 2. With regard to the composition of the present invention, an obvious antiallergic action was observed. Moreover, a high antiallergic action was observed for the composition containing a plurality of strains. In addition, D to H shown in Table 2
As a result of an interview with the patients in the group, an intestinal regulating effect was observed in the constipated person.

[0051]

[Table 1]

[0052]

[Table 2]

(Anti-allergenicity test 2) A patient (12-year-old girl) suffering from allergic purpura and having internal hemorrhage from the end of March 1993 to the beginning of April of the same year, and then, based on Example 2, When the composition was taken, internal bleeding began to disappear about 1 week after the administration, and when the composition was further taken, no bleeding was observed until the end of August 1994.

As described above, it was confirmed that the composition of the present invention has an antiallergic action against various allergic diseases including atopic dermatitis.

Dry cells containing oligosaccharide (composition)
Can provide the dried bacterial cells of each strain shown in Table 2 with the following composition, for example.

Lactose oligosaccharides 1.0 g Crystalline cellulose 1.5 g Lactose 0.5 g Dry cells 0.02 g

[0057]

As described above, the composition of the present invention has an excellent antiallergic action. Moreover, according to this invention, the novel composition for improving intestinal flora is provided.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display part (C12N 1/20 C12R 1: 225) (C12N 1/20 C12R 1: 145) (C12N 1/20 (C12R 1:01) (C12N 1/20 C12R 1:23) (C12N 1/20 C12R 1: 245)

Claims (13)

[Claims] 1. A composition for improving intestinal flora, which contains live bacterial cells of intestinal bacteria as an antiallergic component. 2. The intestinal flora composition according to claim 1, which contains viable cells of a bacterium belonging to at least one of the genus Streptococcus, Enterococcus, Bifidobacterium, Lactobacillus, and Clostridium. 3. A composition according to claim 2 or 3 containing bacteria of all genera according to claim 2. 4. An intestinal flora improving composition containing live bacterial cells of bacteria belonging to the genus Streptococcus, Enterococcus, Bifidobacterium, Lactobacillus, and Clostridium. 5. The intestinal microflora-improving composition according to claim 4, which has an antiallergic effect. 6. The intestinal microflora-improving composition according to any one of claims 1 to 5, which contains a viable cell count of 100 billion or more per day. 7. An intestinal microflora-improving composition containing 100 billion or more viable bacterial cells of intestinal bacteria per daily intake. 8. The intestinal microflora-improving composition according to claim 7, which has an antiallergic effect. 9. The intestinal microflora-improving composition according to claim 1, which comprises a culture obtained by culturing the viable cells in an edible medium. 10. A bacterium belonging to any one of the genus Streptococcus, Enterococcus, Bifidobacterium, Lactobacillus, and Clostridium is Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, Enterococcus ebium. , Streptococcus salivarius, Streptococcus bovis, Streptococcus ikuinus, Lactobacillus acidophilus, Lactobacillus gasserie, Lactobacillus salivarius, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus arabinus burebidobacterium, Bifidobacterium. , Bifidobacterium longum, Bifidobacterium adrensetis, Bifidobacte The composition for improving intestinal flora according to any one of claims 2 to 6, which is Lithium infantes or Bifidobacterium bifidum. 11. A specific strain of the strain is Enterococcus faecalis ATCC 19433, Enterococcus faecium ATCC 19434, Enterococcus durans IID 677, Streptococcus salivarius IID.
5223, Enterococcus hirae ATCC 8043, Enterococcus hirae ATCC 9790, Lactobacillus acidophilus ATCC 4356, Lactobacillus arabinosus ATCC 8014, Lactobacillus fermentum IAM 1
148, Lactobacillus casei ATCC 393, Bifidobacterium adresentis ATCC 15705, Bifidobacterium longum JCM 1217, Bifidobacterium bifidum JCM 1254, Bifidobacterium
Breve JCM 1192, Enterococcus hirae ATCC
8043, Clostridium butyricum ATCC 6015, The composition for improving intestinal flora according to claim 10. 12. The intestinal flora improving composition according to claim 1, which further comprises an oligosaccharide. 13. The intestinal microflora-improving composition according to claim 1, wherein the viable cell count is 10 ^{9 to} 10 ¹² / g.