JP2003012526A - Pharmaceutical preparation including useful live bacterium - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide pharmaceutical preparation capable of preventing killing of useful live bacteria included in an enteric coating with a bile after the enteric coating is dissolved in the small intestine and proliferating the useful live bacteria after arrival at the large intestine. SOLUTION: This pharmaceutical preparation is obtained by carrying out the enteric coating with a composition composed of a useful live bacterial powder, a soybean protein and a gelling agent.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a preparation which allows useful live bacteria to reach the large intestine. Specifically, useful live bacterial powder,
Capsules containing a composition containing soybean protein and a gelling agent, or a tableted product obtained by tableting the composition, which has an enteric coating, passed through the stomach without dissolving. The present invention relates to a large intestine-delivering preparation, which protects useful live bacteria from bile even after the enteric coating film is dissolved in the small intestine and proliferates after reaching the large intestine.

[0002]

BACKGROUND OF THE INVENTION It is known that useful live bacteria such as bifidobacteria and lactic acid bacteria have an intestinal regulating action such as suppressing the growth of bad bacteria in the intestine, improving the intestinal environment, and eventually improving fecal properties. ing. Therefore, attempts have been made to obtain health effects such as intestinal regulation by ingesting foods containing these useful live bacteria in various forms. However, in order for these useful live bacteria to reach the lower part of the digestive tract, especially the large intestine, it is necessary to pass through gastric acid and bile, which have a strong bactericidal action, and allow the useful live bacteria to reach the large intestine, where they are active, alive Is said to be difficult.

As a means for solving this problem,
A method of encapsulating useful live bacteria in an enteric-coated capsule or coating useful live bacteria powder with fats and oils is known. For example, the method disclosed in JP-A No. 11-302158 provides an enteric seam capsule that dissolves in the intestinal tract without dissolving with gastric acid by using gelatin containing sodium alginate or zein. In addition, Japanese Patent Publication No. 5-68446 provides a soft capsule containing Bifidobacterium that does not dissolve in gastric acid, which forms a film with gelatin and sodium alginate or pectin on hardened oil in which Bifidobacterium is suspended. That is, the problem that useful live bacteria are killed by gastric acid can be solved by a method of coating useful live bacteria with a component that does not dissolve in acid but dissolves in neutral as in these inventions.

However, these methods do not solve the problem that the useful live bacteria are exposed to the bile having a high bactericidal action and die when the film dissolves after reaching the small intestine. As a method of solving this, Japanese Patent Laid-Open No. 10-324
As disclosed in Japanese Laid-Open Patent Publication No. 642, useful viable bacteria are contained in a structure composed of an inner layer film of chitosan which is decomposed by intestinal bacteria in the large intestine and an outer layer film of a gastric acid resistant material which is neutral but not acidic and soluble. There is a method of encapsulation, but since the action of intestinal bacteria is used when chitosan is decomposed in the large intestine, the degradability of the film varies greatly with intestinal bacteria with large individual differences, and depending on the person, The film may not decompose. Further, if the intestinal bacteria are out of balance, the chitosan film may not be decomposed and the structure may be excreted as it is. Furthermore, when useful bacteria are required, the balance of intestinal bacteria such as constipation or diarrhea is often unbalanced, and when useful live bacteria are required, there is a contradiction that the structure does not work effectively. There is a fear.

On the other hand, various methods have been provided so far for the sustained release of drugs. For example, Japanese Patent Laid-Open No. 02-8
In 3316, a formulation in which a gelling agent and a drug are mixed provides a means for suppressing the release of the drug by forming a gel layer on the surface when contacted with water or a digestive fluid, and providing a sustained release. There is. By applying this, if the formulation is a gelling agent mixed with useful live bacteria, the gel layer formed can prevent bile from contacting the useful live bacteria and prevent the useful live bacteria from being killed. Is supposed to be. However,
Live cells cannot be expected to diffuse or be released from the formed gel layer, and the proliferative ability of the cells after reaching the large intestine may deteriorate. To solve this problem, by mixing useful live bacteria with a gelling agent in addition to a substance that promotes disintegration, the gel layer is passed through the region of the small intestine where the bile concentration is high and the bacteria are released in the large intestine. It is thought that the disintegration time should be controlled. However, such a method has not been provided so far. For example, JP 20
In Japanese Patent Publication No. 00-63289, a gelling agent and caseins are contained together with useful living bacteria, but it is described that they rapidly disintegrate in artificial intestinal fluid, and it is difficult to control the disintegration in the small intestine. Shows.

[0006]

Therefore, the problem to be solved by the present invention is to prevent the useful live bacteria contained in the coating from being killed by bile after the enteric coating is dissolved in the small intestine. It is to provide a means for viable bacteria to grow after reaching the large intestine.
More specifically, by blending a mixture of useful viable bacteria and a gelling agent as the contents of the enteric preparation, the contents form a gel layer in the region of the small intestine where the bile concentration is high to protect the useful viable bacteria from the bile. In the large intestine, it is to provide means for releasing and growing the bacteria from the mixture of the gelling agent and the useful live bacteria.

[0007]

[Means for Solving the Problems] Therefore, the present inventors have prepared a composition in which various compositions that can be ingested as food are mixed with useful live bacteria into capsules or tablets, and an enteric-coated coating is prepared. did. As a result, it was found that when soybean protein was mixed with a gelling agent, the contents formed a gel layer after the capsule film was dissolved, and the gel layer slowly disintegrated with time. Furthermore, when the formulation was put into a bile model and the survival rate of useful bacteria was confirmed, it was found that useful live bacteria survived in bile for a long time. Furthermore, when the formulation was added to a liquid medium and cultivated, it was found that useful viable bacteria grow, and the present invention was completed.

That is, according to the present invention, by applying an enteric coating to a composition comprising useful live bacterial powder, soybean protein, and a gelling agent, the useful live bacteria pass through gastric acid and bile in the small intestine alive. , A colonic reachable preparation that proliferates in the large intestine is provided.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The viable bacteria useful in the present invention are not particularly limited as long as they have a health effect for improving the intestinal environment. For example, lactic acid bacteria of the genus Lactobacillus can be used. Lactobacillus acidophilus, L. amylophilus
s), Lactobacillus amylovorus (L. amylo)
volus), Lactobacillus crispatus (L. cr)
ispatus), Lactobacillus plantarum (L.
plantarum), Lactobacillus brevis (L.
brevis), Lactobacillus helveticus (L.
helveticus), Lactobacillus gallinarum
(L. gallinarum), Lactobacillus gasseri
(L. gasseri), Lactobacillus johnsoni
(L. johnsonii), Lactobacillus casei
(L. casei), Lactobacillus rhamnosus (L. casei)
Rhamnosus), Lactobacillus zeae (L.z.
eae), Lactobacillus delbruecki subsp
・ Bulgaricus (L. delbrueckii subs
p. bulgaricus), Lactobacillus lactis, Lactobacillus reuteri
(L. reuteri), Lactobacillus fermentum (L. fermentum), Lactobacillus marinus (L. murinus), Streptococcus (Str)
Streptococcus salivarius subsp. thermophilus (S. sali of the genus Eptococcus)
varius subsp. thermophilu
s), Streptococcus lactis (S. lacti)
s), Enterococcus
Enterococcus faecium (E. faeciu)
m), Enterococcus faecalis (E. fecali
s).

Also, Bifidobacterium longum (B. longum), Bifidobacterium bifidum, Bifidobacterium breve (B) belonging to the genus Bifidobacterium, which is Bifidobacterium. .Breve), Bifidobacterium adrescentis (B. adoles)
centis), Bifidobacterium infantis (B. infantis), Bifidobacterium
B. animalis, Bifidobacterium pseudolongum (B. pseudolongu)
m),

In addition, Bacillus
Genus Bacillus coagulans
s), Bacillus natto (B. natto), and Clostridium butyricum (C. butyricum) belonging to the genus Clostridium. Of these, lactic acid bacteria and bifidobacteria are preferably used, and in particular, Bifidobacterium (Bifidobac)
terium) Bifidobacterium longum
(B. longum), Bifidobacterium bifidum (B. bifidum), Bifidobacterium breve (B. breve), Lactobacillus (Lactob)
acillus) Lactobacillus acidophilus
(L. acidophilus), Lactobacillus gasseri (L. gasseri), Lactobacillus casei
(L. casei), Lactobacillus delbrukki s
ubsp. bulgaricus (L. delbrueckii
subsp. bulgaricus, Enterococcus faecium of the genus Enterococcus, and E. fecalis are preferred, and B. longum and Bifidobacterium bifidum. bifidum) and Bifidobacterium breve are most preferred.

These useful live bacterial powders are contained in the formulation of the present invention in an amount of 5 to 50% by weight, particularly 10 to 4% by weight based on the total amount of the composition.
It is preferable to add 0% by weight. If the blending amount is less than 5% by weight, the intestinal regulating effect due to useful live bacteria cannot be obtained, and if it exceeds 50% by weight, the protective effect according to the present invention becomes difficult to obtain.

The useful live bacterium of the present invention is cultivated by a predetermined method,
Used as freeze-dried live bacterial powder. In order to prevent the live bacteria from being damaged when these live powders are freeze-dried,
Known protective agents such as amino acids, sugars, sugar alcohols, milk components and the like can be added. The viable cell powder preferably contains 1 × 10 ⁶ cells / g or more, particularly 1 × 10 ⁸ cells / g or more of viable cells.

The soybean protein used in the present invention is not particularly limited as long as it is provided as a food, but raw soybean flour, kinako, commercially available purified soybean protein and the like are preferable examples. Among them, purified soybean protein is particularly preferable because various types of soybean protein having uniform properties are available. These soy proteins are 10 to 70% by weight in the total amount of the composition contained in the capsule of the present invention,
In particular, 20 to 65% by weight can be blended, and 30 to 50
Weight percent is more preferred. If the blending amount is less than 10% by weight, the protective effect from bile cannot be obtained, and the gel layer is prevented from collapsing to prevent the contents from diffusing, and if it exceeds 70% by weight, the gel layer is collapsing. Premature bile protection is not fully exerted.

The gelling agent used in the present invention is not particularly limited as long as it reacts with water to form a gel, but xanthan gum, sodium alginate, pectin, carrageenan, locust bean gum, gum arabic, etc. Among these, xanthan gum, sodium alginate, pectin and carrageenan are preferable, and xanthan gum is most preferable. These gelling agents can be used alone or in combination of two or more.

These gelling agents are contained in the capsule of the present invention in an amount of 1 to 15% by weight, particularly 3 to 1% by weight based on the total amount of the composition.
2 wt% can be compounded, and 5 to 10 wt% is particularly preferable. If the blending amount is less than 1% by weight, the gel layer is not formed and the bile protection effect cannot be obtained.
Above, the gel layer becomes too stiff and prevents the contents from diffusing.

Further, in order to assist the gelling of the gelling agent, a salt of divalent cation such as calcium or magnesium may be contained. The composition encapsulated in the capsule of the present invention may contain, in addition to the above-mentioned useful live bacteria, soybean protein, gelling agent, additives usually used in foods, food materials, or flavors.

The enteric preparation of the present invention can be provided in the form of a capsule preparation such as a hard capsule or a soft capsule, or a tablet preparation. The capsule preparation is constituted as a structure composed of a composition to be encapsulated, a capsule film enclosing the composition, and an enteric coating film outside the capsule film. In the case of a hard capsule, the volume ratio of the capsule film to the composition to be encapsulated is approximately 1: 2-1: 1.
5 and about 1: 1 for soft capsules
It is 1: 3. The material of the capsule is not particularly limited, but gelatin, whey protein, or cellulose derivative is preferably used. Capsules can be manufactured by a conventional method. The tablet formulation is constituted as a structure consisting of a tableted product obtained by tableting the encapsulated composition by a conventional method, and an enteric coating film on the outside of the tableted product.

The coating agent used in the enteric preparation of the present invention can be exemplified by zein, shellac and the like. The enteric coating can be carried out by a conventional method, and fatty acid, glycerin fatty acid ester, glycerin, pigment and the like can be added to the coating agent. In the case of capsules, the coating amount is preferably 0.1 to 7%, particularly 0.5 to 5% by weight based on the capsule weight, and in the case of tablets, 0.05% to 5%, particularly It is preferably 0.1 to 3%.

[0020]

EXAMPLES The present invention will be described below with reference to Examples and Test Examples.
As will be described in more detail, the present invention is not limited to this. Unless otherwise specified, [%] is [% by weight].
Represents

First, the compositions shown in Table 1 (Nos. 1 to 1)
0) was produced and the total amount was 2 kg. The obtained composition was encapsulated in a gelatin hard capsule by a conventional method using a capsule filling machine. The obtained capsules were coated with a spray coating device. The coating solution is 2% zein, 6% shellac, 67.7 ethanol.
%, Distilled water 24.3%, and fatty acid 1.2% (each weight%) were used. Capsule charge 2kg,
Coating was performed at an air supply temperature of 45 ° C., a spray speed of 50 g / min, a spray time of 10 minutes, and a pan rotation number of 10 rpm, and the coating amount was 2% of the capsule weight ratio. At this time, the viable cell count per gram of the composition in the capsule was 1.2 × 10 ¹⁰ .

[0022]

[Table 1]

Test Example 1 (gastric acid resistance) An enteric test was performed on the capsules 1 to 10 according to the disintegration test of the Japanese Pharmacopoeia. As a result, for all capsules, the first liquid (p
It was confirmed that the product did not disintegrate in H1.2 hydrochloric acid) for 120 minutes and the contents did not elute. In addition, when the capsules that had not been disintegrated were taken out and the number of Bifidobacterium contained therein was measured, the same amount of Bifidobacteria as the charged amount was recovered.

Test Example 2 (bile resistance) As artificial intestinal juice, 0.02 in a pH 6.8 phosphate buffer solution was used.
What added the bile powder of the weight% concentration was used. 100 ml of artificial intestinal juice was put into a screw cap bottle, a new capsule was put therein, and the mixture was stirred with a magnetic stirrer at a speed of 800 rpm. After 3 hours, the entire test solution was homogenized and the number of Bifidobacteria was measured. The results are shown in Table 2.

[0025]

[Table 2]

From this result, N containing no gelling agent was found.
o. 1, No. In No. 6, the bifidobacteria were markedly killed by the bile, but in other test examples, it was found that the killing of the bifidobacteria by the bile was suppressed regardless of the type of gelling agent. No. 2, No. 7, No.
8, and No. It can be seen from the comparison of 9 that xanthan gum has the greatest protective effect. In addition, No. From the results of No. 10, it was found that when casein was used instead of soybean protein, the contents of the capsule rapidly disintegrated even when forming a gel, and as a result, the mortality by bile was markedly deteriorated. Thus, it is speculated that the disintegration time of the formed gel layer depends on the type of protein because the isoelectric point and hydrophobicity of the protein differ depending on the protein and the interaction with the gelling agent. It From the above, it was found that the combination of soybean protein and gelling agent, especially xanthan gum, improves the bile resistance of useful live bacteria.

Test Example 3 (Proliferation in colon) New capsules were placed in sterilized MRS medium and 37 ° C.
The cells were statically cultured at. After 16 hours, the culture broth was sampled, and the bacterial growth was evaluated by measuring the acidity and the number of Bifidobacterium in 1 mL of the culture broth. The results are shown in Table 3.

[0028]

[Table 3]

No. 3, which has a high gelling agent content. 4 and N
o. In the prototype of No. 5, the contents of the capsule were solidified into a gel. Also, from the results of acidity measurement, it can be seen that the acidity is less likely to increase in the prototype having a high gelling agent content.
From this result, it can be seen that if the content of the gelling agent is too high, the growth is inhibited. That is, it is speculated that when the content of the gelling agent is high, the bacteria are not released into the medium and the growth is inhibited. From the above-mentioned tests, it was shown that the capsule of the present invention is characterized by protecting useful living bacteria from gastric acid and bile and disintegrating in the large intestine to grow useful living bacteria.

[0030]

The above formulations were mixed so that the total amount was 1.5 kg. The obtained mixture was encapsulated in gelatin hard capsules using a capsule filling machine according to a standard method. The obtained capsules were subjected to enteric coating with a spray coating machine according to a standard method. The coating solution is 7% zein, 68.7% ethanol, 24.3% distilled water,
A fatty acid containing 1.2% (each weight%) was used. Coating was performed at a capsule charging amount of 2 kg, an air supply temperature of 45 ° C., a spraying speed of 50 g / min, a spraying time of 10 minutes, and a pan rotation number of 10 rpm, and the coating amount was 2% of the capsule weight ratio.

[0032]

The above formulations were mixed so that the total amount was 1.5 kg. The obtained mixture was encapsulated in gelatin hard capsules using a capsule filling machine according to a standard method. The obtained capsules were subjected to enteric coating with a spray coating machine according to a standard method. The coating solution used contained zein 2%, shellac 6%, ethanol 67.7%, distilled water 24.3%, and fatty acid 1.2% (each weight%). Coating was performed at a capsule charging amount of 2 kg, an air supply temperature of 45 ° C., a spraying speed of 50 g / min, a spraying time of 10 minutes, and a pan rotation number of 10 rpm, and the coating amount was 2% of the capsule weight ratio.

[0034]

The above formulation was mixed so that the total amount became 1 kg, and then mixed with 4 kg of hardened oil heated to 40 ° C. to obtain a core liquid of a soft capsule. The obtained core liquid was encapsulated with a gelatin film in a soft capsule manufacturing machine. The obtained capsules were dried for one day and then subjected to enteric coating with a spray coating machine according to a conventional method.
The coating solution is 2% zein, 9% shellac, 65.7% ethanol, 24.3% distilled water, 1.2% fatty acid.
Those containing (each weight%) were used. Capsule charge 2 kg, air supply temperature 45 ° C, spray speed 50 g /
Coating was carried out at min, spraying time 20 minutes, and pan rotation number 10 rpm, and the coating amount was 4% of the capsule weight ratio.

[0036]

The above formulations were mixed so that the total amount would be 1.5 kg and filled into gelatin hard capsules. The obtained capsules were subjected to enteric coating with a spray coating machine according to a conventional method. The coating liquid is shellac 8%, ethanol 67.7%, distilled water 24.3%,
A fatty acid containing 1.2% (each weight%) was used. Coating was performed at a charged amount of capsules of 2 kg, an air supply temperature of 45 ° C., a spray rate of 50 g / min, a spray time of 6 minutes, and a pan rotation number of 10 rpm, and the coating amount was 1% of the capsule weight ratio.

[0038]

The above formulations were mixed so that the total amount was 10.0 kg, and the mixture was compressed according to a conventional method to give a compressed product. The obtained tableted product was subjected to enteric coating by a spray coating machine according to a conventional method. The coating solution is shellac 5%, tween 3%, ethanol 67.2%,
Distilled water 24.2%, fatty acid 0.6% (each weight%)
Was used. The amount of the tableting material charged was 10 kg, the air supply temperature was 45 ° C., the spraying speed was 50 g / min, the spraying time was 18 minutes, and the pan rotation speed was 10 rpm. The coating amount was 3% based on the weight of the tableting material.

[0040]

Industrial Applicability The enteric preparation of the present invention containing a composition containing useful soybean bacteria together with soybean protein and a gelling agent prevents the useful live bacteria from being killed by gastric acid, and after disintegrating in the small intestine. Also, since the useful bacteria are prevented from being killed by the bile, and the useful viable bacteria proliferate after reaching the large intestine, more useful viable bacteria reach the intestine, and further health effects can be expected.

Claims (4)

[Claims] 1. A preparation coated with enteric coating which essentially contains useful live bacteria powder, soybean protein, and gelling agent. 2. The preparation according to claim 1, wherein the useful live bacteria are one or more selected from lactic acid bacteria and bifidobacteria. 3. The preparation according to claim 1, wherein the gelling agent is xanthan gum. 4. The amount of gelling agent is 1 with respect to the total amount of the composition.
The formulation according to any one of claims 1 to 3, which is -15% by weight.