JPH07145A - Composition of butyric acid bacterium - Google Patents

Abstract

PURPOSE:To obtain the subject composition, composed of a live microorganism for controlling the intestinal function represented by a butyric acid bacterium and glucomannan, excellent in bioavailability and therapeutic effects and useful for acute gastroenteritis, functional foods, etc. CONSTITUTION:This bacterial composition is obtained by blending (A) 1 pt. wt. live microorganism for controlling the intestinal function represented by a butyric acid bacterium such as Clostridium butyricum Miyairi 588 (FERM P-2789) with (B) preferably 0.2-10 pts.wt. glucomannan. Furthermore, the daily dose of this composition is preferably within the range of 1X10<6> to 1X10<10> cells expressed in terms of cell number.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a butyric acid bacterium composition exhibiting excellent effects from the viewpoint of formulation and bioavailability.

[0002]

BACKGROUND OF THE INVENTION Butyric acid bacteria, especially Clostridium butyricum Miyairi 588, were discovered in 1935 by Mr. Konji Miyairi [Chiba Medical Society 13 2311-2315 (1
935)], and then, many researchers found that this bacterium strongly and stably antagonizes various food poisoning-causing bacteria [Chiba Medical Journal 13 2141-2161].
(1935)], Chiba University Corruption Research Institute Report 127-53
(1948), Chiba University Corruption Research Institute Report 1 54-59
(1948), Chiba University Corruption Research Institute Report 243-50
(1949)]. In addition, it has been clinically shown to be effective against various digestive system diseases such as acute or chronic enteritis, dysentery, plague, dyspepsia [Clinical internal medicine 4
973-980 (1935), New drugs and clinical 25 150
5-1509 (1976)].

[0003]

On the other hand, in recent years, the number of overseas travelers has increased, and so-called traveler's diarrhea has become an important issue among travelers to developing countries. It is infected by various enteric pathogens during travel,
Most are acute gastroenteritis with diarrhea as the main symptom. Among these intestinal pathogens, there are many cases in which infection is caused by toxogenic Escherichia coli, and basic experiments have shown that butyric acid bacteria are effective against this toxogenic Escherichia coli diarrhea. And treatment 146073-6080 (1
986), Pharmacology and Treatment 14 4651-4655 (19)
86), Japanese Journal of Pharmacology (Japan. J. Pharmacol.) 50 495-498.
(1989)].

In recent years, antibiotics having a broad antibacterial spectrum and a strong antibacterial activity have been successively introduced into the clinic, and accordingly, colitis and diarrhea caused by antibiotics have become important problems. This is because the administered antibiotics destroy the intestinal flora that is physiologically closely related to the host, and specific antibiotic-resistant bacteria, such as the minor population Clostridium difficile, are manifested and produced. It is caused by a toxin, which is accompanied by high fever, leukocytosis, hypoproteinemia, and severe colitis symptoms such as abdominal pain, diarrhea, and bloody stool, and sometimes has a serious outcome. It was found that butyric acid bacteria were effective against colitis and diarrhea caused by such drugs as a basic experiment [Japanese Journal of Bacteriology 43 829-835 (198).
8)], and clinical [pediatric clinical 41 2409-24
14 (1988)].

Not limited to these examples, it is already well known that butyric acid bacteria act not only in various digestive diseases but also in promoting bowel movements, intestinal epithelial cell repair, water reabsorption in the large intestine, and the like. .

[0006] On the other hand, glucomannan is a dietary fiber (dietary fiber) prepared from konjak, lily root, etc., contains a large amount of mucilage, is not digested and decomposed by human digestive enzymes, and is therefore not a low calorie. It is known as a polysaccharide. Ingested glucomannan takes in salt, lipid, and other harmful bacterial metabolites together with a large amount of water in the crosslinked polymer, swells, and is excreted outside the body. From these things, glucomannan has a normal intestinal function, promotes bowel movements, lowers blood cholesterol, and has the effect of improving obesity by giving a feeling of fullness to obese persons due to overnutrition. It is said that there is. Furthermore, in recent years, the correlation between the occurrence of colorectal cancer and dietary fiber intake has been clarified epidemiologically, and this type of dietary fiber is considered effective for its prevention, and its importance is being reviewed.

[0007]

[Means for Solving the Problems] The inventors of the present invention have made extensive studies on the composition of glucomannan and butyric acid bacterium having this excellent physiological property from the viewpoints of pharmaceutical and bioavailability. As a result, it became clear that the present composition was extremely useful, and the present invention was completed. That is, the gist of the present invention resides in a butyric acid bacterium composition composed of a live intestinal bacterium representative of butyric acid bacterium and glucomannan.

The butyric acid bacterium which can be used in the present invention is, for example, Clostridium butyricum NIP1006 (Ministry of Industrial Science and Technology No. 11868), Clostridium butyricum NIP10.
15 (Microtechnology Research Institute, No. 11869), Clostridium buty
ricum NIP 1017 (Ministry of Microbiology Research No. 11870), and particularly preferred is Clostridium butyricum Miyairi 588 (Ministry of Microbiology Research No. 2789).

Generally, the butyric acid bacterium paste harvested from the culture solution of butyric acid bacterium by the centrifugal separation method has a grayish brown irregular block-like shape when dried in this state, and even if it is crushed, the grain shape is not uniform. There is also a problem in terms of formulation in terms of flavor and odor. Therefore, it is desirable that the fungal paste is kneaded and homogenized in a liquid or mud state with some type of excipient before being dried, and then dried.

Then, various types of excipients and fungal paste were kneaded, and the properties of the composition were examined. As a result, it was found that the composition obtained by kneading glucomannan and butyric acid bacteria paste was milky white and had a very uniform fine grain. In addition, as a dosage form, other pharmaceutical additives,
It became clear that it is excellent in terms of formulation as well, for example, it is well dispersed even if it is granulated by adding a diluent or the like.

On the other hand, the composition of the butyric acid bacterium was examined from the viewpoint of bioavailability. Orally administered butyric acid bacteria are exposed in the stomach to the low pH environment of protein digestive fluid pepsin and gastric acid, but butyric acid bacteria, which are resistant to spores, pass through here without dying and enter the duodenum. To reach. There, the pH rises to near neutrality, the food is digested by various digestive fluids, and a nutrient-rich environment is created. However, there, a large number of indigenous bacteria compete with each other for nutrition and antagonize each other to form so-called intestinal microflora. Under these circumstances, butyric acid bacteria are required to germinate and proliferate and reach a population capable of expressing a pharmacological action. The effect is enhanced when a large amount is administered, but the dose is limited. Therefore, by incorporating a component having selective availability that is not assimilated by other indigenous bacteria but is specifically assimilated by butyric acid bacteria, the population of only butyric acid bacteria is increased. That is, the composition was examined for the purpose of enhancing the pharmacological action.

First, according to the method of Mitsuoka et al. From human feces, Enterobacteriace such as Escherichia, Klebsiella, etc.
ae, Enterococcus, Lactobacillus, Bifidobacterium,
Eubacterium, Bacteroidaceae, Peptococcaceae, Veillo
Intestinal bacteria such as nella, Megasphaera and Clostridium were isolated. In these liquid media in which the intestinal bacteria can grow, media in which glucomannan was added instead of the sugar component were prepared, and the utilization of glucomannan by each intestinal bacterium was examined. As a result, it was revealed that among the various intestinal bacteria, only the butyric acid bacterium Clostridium butyricum favorably assimilates glucomannan.

Further, in order to observe whether butyric acid bacteria selectively grow in human feces, a fecal suspension was prepared, and corn starch or glucomannan was added to this to inoculate and culture the butyric acid bacteria. As a result, the glucomannan additive solution was compared with the control corn starch additive solution,
The pH was markedly decreased and it was observed that butyric acid bacteria were growing.

Furthermore, when the butyric acid bacterium was administered to a mouse which was allowed to freely ingest glucomannan for 3 days, and the number of butyric acid bacteria in the intestinal contents of the mouse was examined, it was found that the mice ingested glucomannan were better than those not ingested. It was also confirmed that the number of butyric acid bacteria in the intestine was high. The results of these experiments indicate that glucomannan is not assimilated by resident intestinal bacteria in the digestive tract,
It is shown that the composition that is selectively assimilated only to the butyric acid bacterium used in combination and that contains glucomannan and butyric acid bacterium can increase the population.

As described above, the present composition has excellent characteristics from the viewpoint of pharmaceutical or bioavailability.

The optimum blending ratio (weight) of this composition is 0.1 to 1000 parts by weight of glucomannan, particularly 0.2 to 10 parts by weight, based on 1 part by weight of the bacterial paste. However, the amount of glucomannan can be appropriately increased or decreased in order to prepare it according to the desired bacterial concentration.

The dosage form of this composition for use as a medicine can be used as it is, but it can be formulated into powders, granules, fine granules, tablets, sugar-coated tablets, capsules, enteric coating agents and the like. Good. Diluents include
Excipients, binders, disintegrants and the like used in general pharmaceutical preparations are used, and in addition thereto, coloring agents, corrigents, stabilizers, preservatives, lubricants and the like may be added.

Although it can be used as it is for functional foods, it is suitable for food by adding other dietary fibers, oligosaccharides, grains, vitamins and the like, and further adding flavors, coloring agents, flavoring agents and the like. It is also possible to form and process the shape.

Although it can be used as it is for animals, various excipients and additives are added in the same manner as for humans.
It is also used as a similar dosage form. Corn flour, soybean meal, barley flour, naked wheat flour, soybean flour, rice bran, defatted rice bran,
It is also possible to mix and use it in livestock feed such as chaff, potato powder, taro powder, tofu dregs, starch, yeast and fish meal.

The dosage is appropriately in the range of 1 × 10 ⁶ to 1 × 10 ¹⁰ cells per day, but depending on the age of the patient, the symptoms of the disease, and for functional foods and animals. The usage can be increased or decreased depending on the purpose. As for the administration method, it is preferable to take the medicine once a day or three times after meals, but it is also possible to take it appropriately. For functional foods and animals, they can be taken freely once to several times a day.

[0021]

【Example】

Example 1 A composition of butyric acid bacterium and glucomannan was prepared as follows. 100 g of glucomannan was added to 100 g of a butyric acid bacterium (Clostridium butyricum Miyairi 588) bacterial paste cultivated in a known CS medium (Japanese Patent Laid-Open No. 59-187784) and kneaded with a kneader until uniform. Next, vacuum drying (10 mmHg) was performed at 50 ° C. for 5 hours using a shelf vacuum dryer. The obtained dried product was pulverized with a pulverizer to obtain the composition. The composition was milky white, uniform fine granules, and had almost no taste and odor. The viable cell count in this composition was 1.8 × 10 ¹⁰ cells / g.

Example 2 From the human feces, Enterobacteriaceae showed DHL agar, TS aga
r medium, Enterococcus is TATAC agar, TSagar medium,
Lactobacillus was prepared from LBS agar, BL agar medium and Bifidoba
For cterium, use BS agar, BL agar, EG agar medium with Eubacter
ium is ES agar, BL agar, EG agar medium, Bacteroidace
ae is NBGT agar, BL agar, EG agar medium, Peptococca
ceae was BL agar, EG agar medium, Veillonella and
Megasphaera is VS agar, BL agar, EG agar medium, Cl
The ostridium was separated using BL agar and EG agar media, respectively. These isolates were subjected to a glucomannan assimilation test using the following medium. The determination was carried out by measuring the pH of the acid produced after the culture was completed. GAM Glycolysis semi-fluid medium 52.5g Glucomannan or glucose 5g Purified water 1000ml

Furthermore, two strains of butyric acid bacteria Clostridium
Butyricum Miyairi 588, C, butyricum NIP 100
6, C. pasteurianum ATCC6 which is another standard strain of the same genus
013, C. beijerinckii ATCC 25752, C. acetobut
For ylicum IAM19012, the following medium was used for the glucomannan utilization test. Kashiton 15 g Yeast extract 1 g Sodium chloride 2.5 g L-cystine hydrochloride 0.05 g Sodium thioglycolate 0.5 g Glucomannan or glucose 5 g Purified water 1000 ml

The assimilation capacity of the bacterium is 37
After culturing at 3 ° C for 3 days, it was examined by adding BCP or BTB.

As a result, as shown in Table 1, in the glucose-containing medium, good growth was observed in all the test bacteria. However, in the glucomannan-added medium, good growth was observed in Clostridium
Buchirikum Miyairi 588 and C.I. butyricum NIP 10
There were only two strains of 06.

[0026]

[Table 1]

Example 3 Feces of 0.5 g of a healthy person were collected and immediately suspended in 30 ml of PBS containing 0.03% sodium thioglycolate in an anaerobic glove box to prepare a fecal suspension. Separately, a solution was prepared by adding 0.3 g of starch or 0.3 g of glucomannan to 9 ml of the same PBS, and mixed with the fecal suspension in the same amount. Butyric acid bacterium Clostridium butyricum Miyairi 588 was added to these mixed solutions at a concentration of 5 × 1.
The cells were inoculated at a rate of 0 ⁴ / ml, and statically cultured at 37 ° C for 24 hours. As a control, the same operation was performed for the system to which the saccharide was not added. The proliferation was judged by using the pH, foaming and speculum of the test solution as indexes. As a result, as shown in Table 2, it was confirmed that the glucomannan-added system markedly lowered pH and foamed as compared with the control starch-added system, and that butyric acid bacteria were major under the microscope.

[0028]

[Table 2]

Example 4 Clostridium butyricum miyairi 588 was added to an ICR mouse (about 30 g) in which 5% of glucomannan was mixed in the water supply and which was freely ingested for 3 days to obtain 3.1 × per head.
10 ⁶ were orally administered. The animals were sacrificed 3 hours and 6 hours after administration, the entire digestive tract was excised, and the viable count of Clostridium butyricum Miyairi 588 in the contents was measured using a GAM plate medium in an anaerobic glove box. . As a control, the same operation was performed for a system in which glucomannan was not ingested. As shown in Table 3, the number of bacteria ingested glucomannan was higher than that of the control, particularly 3 hours after the administration.

[0030]

[Table 3]

[0031]

EFFECTS OF THE INVENTION According to the butyric acid bacterium composition of the present invention, it is excellent in terms of formulation and also from the viewpoint of bioavailability.
It is possible to provide a composition having excellent characteristics.

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Claims (2)

[Claims] 1. A butyric acid bacterium composition comprising a viable intestinal bacterium represented by butyric acid bacterium and glucomannan. 2. The butyric acid bacterium composition according to claim 1, wherein the butyric acid bacterium is Clostridium butyricum Miyairi 588.