JPS62201823A - Material containing beneficial enterobacterium and production thereof - Google Patents

Abstract

PURPOSE:To provide the titled material, by dispersing beneficial enterobacterium or powder supporting beneficial enterobacterium in an oil and filling the dispersion in a specific capsule. CONSTITUTION:An oil containing dispersed beneficial enterobacterium or dispersed powder supporting said bacterium is poured out through an inner nozzle of a vibrating multiple nozzle and, at the same time, a liquid coating material is poured out through the outermost nozzle of the vibrating nozzle to form a multi-layered droplet having a diameter of <=3mm. The coating material constituting the outermost layer of the multi-layered droplet is hardened to obtain a material containing beneficial enterobacterium. The product may be further coated with an enteric coating layer. The material produced by this process has excellent storage stability and high implanting ratio. It can be easily taken to enable easy administration of a desired amount of the beneficial enterobacterium. The product is extremely suitable as a drug or food. The capsule is preferably a seamless capsule and the coating material is e.g. gelatin, alginic acid, etc.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to a stable intestinal beneficial bacterial content.

[Conventional technology]

A large number of bacteria reside in the human intestine, and their activities are involved in the chemical reactions of substances in the human intestinal tract, and have a major impact on the function of the intestine and, ultimately, on the state of human health. Yo (known)

Among these bacteria, B. bifidus and lactic acid bacteria do not themselves have any harmful effects, but when they multiply, they act to suppress the proliferation of other bacteria that have harmful effects.

Therefore, it is desirable to allow B. bifidus and lactic acid bacteria to proliferate in the intestines, since this can prevent the generation of harmful products due to abnormal fermentation in the intestines caused by these harmful bacteria.

In general, during infancy, humans mainly consume amino-tJ! It is supplied with a component called the so-called bifidus factor. Therefore, it is said that bifidus bacteria and lactic acid bacteria proliferate in the intestines, and more than 90% of the intestinal bacteria are occupied by these bacteria, forming an extremely good bacterial flora.

However, as children grow older and stop ingesting breast milk, the supply of bifidus factors decreases, resulting in a decrease in these beneficial intestinal bacteria and a deterioration of the bacterial flora.

Therefore, attempts have been made to improve the bacterial flora by orally administering useful intestinal bacteria, and lactic acid bacteria preparations and foods and drinks containing Bifidus bacteria are commercially available for this purpose.

[Problem that the invention seeks to solve]

However, when ingested only from conventional foods and drinks, there is a limit to the amount that can be contained in the food and drink itself, and a considerable portion is killed by gastric juices, so it is generally difficult to secure an effective amount. In particular, for patients whose intestinal function deteriorates due to deterioration of the bacterial flora and exhibit symptoms such as abnormal fermentation and diarrhea, the effective bacteria should be administered to ensure that an effective amount of the above-mentioned useful bacteria reaches the intestines. It is necessary to. However, formulations of the above-mentioned useful bacteria have problems such as poor storage stability because they are anaerobic bacteria, and difficulty in increasing the concentration of bacteria necessary to administer an effective amount. At high concentrations, they are killed by the action of self-produced lactic acid, making it impossible to maintain this high concentration.Currently, no satisfactory preparations have been obtained.

An object of the present invention is to provide a technique that allows a desired amount of useful bacteria to reach the intestine reliably and enables effective utilization of the useful bacteria.

[Means for solving problems]

The outline of the present invention is such that a capsule having a diameter of 3 s or less is filled with oil in which intestinal useful bacteria or powder carrying the bacteria is dispersed.

In addition, from the inner nozzle of the vibrating multi-nozzle, oil in which beneficial intestinal bacteria or powder carrying them is dispersed,
In addition, the coating material liquid is simultaneously flowed out from the outermost nozzle to form multilayer droplets with a diameter of 3 mm or less, and then the outermost coating material of the multilayer droplets is hardened to remove the intestinal beneficial bacteria-containing material. It is manufactured.

Further, after the step of curing the coating material, an enteric coating is applied on the coating material to produce the above-mentioned material containing beneficial bacteria for the intestine.

[Effect]

By configuring the intestinal useful bacteria content as described above, the beneficial intestinal bacteria can be protected with a capsule, so the useful bacteria can be stored for a long time, and when ingested orally, the number of viable bacteria at the time of adjustment can be reduced. The above-mentioned useful bacteria can be delivered to the intestines without being reduced, and at the same time, the oil containing the useful bacteria can be easily emulsified into the digestive juices in the intestines.
The above objective is achieved. In this case, the achievement of the above purpose is further ensured by coating the capsule coating material with an enteric substance that is insoluble or poorly soluble until it reaches the intestine after taking it, but quickly dissolves when it reaches the intestine. It is something that can be done.

By the way, research has been carried out on the stabilization of these anaerobic intestinal useful bacteria, especially Bifidobacterium, and Japanese Patent Application Laid-open No. 57-33543 describes a method of coating with oils and fats that become almost solid at temperatures below 30°C. is proposed. However, this alone has the disadvantage that a considerable amount of the oil is discharged while remaining contained in the oil and fat, and is not effectively utilized.

The present inventors previously reported in JP-A-57-42616 and JP-A-57-70815 that when a drug dispersed or dissolved in oil is filled into capsules with a diameter of 3 mm or less, oil droplets form from the beginning. Because it is fragmented, it is easily dispersed by stirring in the gastrointestinal tract or by emulsification by digestive juices, and it has a large surface area from the beginning, so it has high bioavailability. However, the above-mentioned Japanese Patent Application Laid-open No. 1987-
No. 42616 and No. 57-70815 are concerned with the absorption of drugs from the intestinal wall, whereas the present invention is concerned with colonization of bacteria in the intestines, and their purpose is completely different.

Therefore, the present inventor conducted intensive research to apply this technology to capsules containing oil dispersed with intestinally beneficial latin, and as a result, a formulation with not only high storage stability but also a high implant rate was obtained. This is what I discovered.

The beneficial intestinal bacteria-containing material according to the present invention has the advantage that it is easy to take, and even if the dosage varies depending on the age, the desired dose can be easily taken. No. 57-70815, and it is suitable both as a preparation and as a food.

Intestinal useful bacteria that can be used in the present invention include B.
bHidum, B, longum, , B, ado
lescentis.

Btfido-bacterium such as B, breve, B, 1nfantis, L
, acidophHus, L. 1actis,
L.

Lactobacillus casei, L. bulgaricus, etc. and 5tre
ptococcus 1act4s and the like. These orchids can be used as they are or as a powder supported on skim milk powder, starch, etc.

In addition, oils used in the present invention include vegetable oils such as soybean oil, rapeseed oil, cottonseed oil, corn oil, sesame oil, linseed oil, olive oil, palm oil, safflower oil, safflower oil, rice bran oil, and castor oil, as well as phospholipids, Intermediate to higher fatty acid esters of glycerin and polyglycerin, unsaturated higher fatty acids such as linoleic acid, lylunic acid, and eicosapencitric acid, higher hydrocarbons such as squalene and squalane, higher alcohols, mineral oils, and non-containing substances such as silicone oil. Alternatively, poorly water-soluble edible or oily substances that are harmless to the human body are exemplified. The concentration of bacteria dispersed in the oil needs to be at an effective concentration when administered, and is not limited to this, but is preferably 10 6 cells/g or more, preferably 10 9 cells/g or more.

In the present invention, it is essential to fill capsules with a diameter of 3 ss or less with oil in which bacteria are dispersed, but seamless capsules are usually suitable for such capsules, and the covering material is gelatin, alginic acid, etc. , carrageenan, pectin, etc. are used.

It goes without saying that it is possible to fill the above-mentioned capsule with the liquid content and produce a substance containing bacteria useful for the intestines by using ordinary soft capsule manufacturing techniques, but by employing the method according to the present invention, , which can be easily manufactured.

That is, a manufacturing vt comprising multiple nozzles, a coating liquid supply means connected to the outermost nozzle of the multiple nozzles, a filler liquid supply means connected to the inner nozzle thereof, and a means for curing the coating material. can be easily produced by the method according to the present invention using

The above-mentioned equipment includes, for example, a Dutch-made capsule covering machine, GLOBE X INTERNATIONA (product name, GLOBE x INTERNATIONA, sold by Mucheal Trading Co., Ltd., Tenroku Hankyu Building, 7-1-10 Tenjinbashi, Oyodo-ku, Osaka).
There is LIM [manufactured by TED].

Furthermore, in the manufacturing apparatus having the basic configuration described above, it is also possible to use an apparatus in which a vibrating means is connected to multiple nozzles, and in this case, even better results can be obtained.

Here, an example of such a manufacturing apparatus will be described with reference to the drawings.

The figure is a schematic configuration diagram of the above manufacturing apparatus.

In this manufacturing equipment, 1 is a double nozzle (multiple nozzles)
A vibrator 3 connected to a vibration generator 2 is connected to the rear end of the double nozzle.

In addition, the double nozzle 1 has a first nozzle (inner nozzle>
la and the second nozzle located outside it (outermost nozzle)
It is composed of lb.

A pipe 5 extending from a tank 4 in which water is stored is communicated with the first nozzle 1a. A pump 6 and a valve 7 are interposed in the middle of the pipe 5. Similarly, a pipe 5a extending from the tank 4a is communicated with the second nozzle 1b, and a pump 6a and a valve 7a are interposed in the middle of the pipe 5a.

Further, below the double nozzle 1, a curing tank 8 is provided.
is located.

A case will be described in which a substance containing beneficial intestinal bacteria is produced using the above-mentioned production apparatus.

First, filler liquid 9, which is oil in which bacteria or powder carrying bacteria is dispersed, is placed in one tank 4, and the other tank 4 is filled with filler liquid 9.
A coating material liquid 10 such as a hot gelatin aqueous solution is stored in each a, and a hardening liquid 11 made of cold oil or the like is stored in a hardening tank 8.Next, the pulps 7 and 7a are adjusted and vibration is applied. A two-layer liquid stream 12 consisting of the filler liquid 9 and coating material liquid IO flows downward from the double nozzle 1, forming two-layer droplets (multilayer droplets) 13 in the air. The formed two-layer droplet 13 falls into the curing tank 8,
The coating material liquid, which is the outermost layer of the droplets, is cured, and the production of a material containing beneficial bacteria for the intestine consisting of seamless minicapsules 13a is achieved.

It goes without saying that the apparatus equipped with multiple nozzles to which vibration means is added that can be applied to the production of the intestinal useful bacteria-containing material according to the present invention is not limited to the one shown in the drawings. It has the above basic configuration! Any device may be used as long as it can be placed in a position and achieve the intended purpose. For example, instead of arranging multiple nozzles in the air as in the above device, they may be placed in a flowing curing liquid and a multilayered liquid flow may flow into the curing liquid. Various combinations of variations are possible. Needless to say.

In addition, as a method for curing the outermost layer of the droplets made of the coating material liquid, a method was shown in which a hot gelatin aqueous solution, which is a thermoreversible sol-gel conversion substance, is cooled; however, as the conversion substance,
The coating material is not limited to a hot gelatin aqueous solution, and carrageenan, alginic acid, or pectin can also be used as a coating material. Appropriate means can be adopted depending on the capsule material, such as a curing method using a chemical reaction.

Since the bacteria contained in the intestine useful bacteria of the present invention are dispersed in the oil, a considerable portion of the bacteria does not come into direct contact with the gastric juices, but passes through the stomach while remaining contained in oil droplets, and is transported from the duodenum to the small intestine. After moving to the large intestine, the oil is digested by pancreatic juice, bile, etc., so the useful bacteria contained therein can be brought into sufficient contact with the digestive juices.

The above-mentioned beneficial intestinal bacteria are sufficiently stable and effective as they are; It is desirable to apply a coating of a soluble substance. The enteric substances include, but are not limited to, zein, shellac, carboxymethyl cellulose, cellulose acetate phthalate, hydroxypropyl methyl cellulose phthalate, hydroxypropyl methyl cellulose acetate succinate, polyvinyl alcohol phthalate, etc. Among them, zein, shellac, carboxymethylethyl cellulose, etc. are particularly suitable.

The intestinal beneficial bacteria-containing material according to the present invention includes substances that have a propagating effect on intestinal bacteria such as fructooligosaccharides, lactylose, and N-acetylglucosamine, which are dispersed in oil or in the outer layer of a capsule, along with useful bacteria. Coating with a certain dressing material or as an intermediate layer between the dressing material and an Ill-soluble coating is preferable because it can further improve the implantation rate in the intestines.

Next, examples of the present invention will be described along with various tests.

[Example 1] Lactic acid bacteria (Lacfobacillus actdphi)
lus) Supported starch powder (Number of countries of birth 6.5xlO@/g) 2
00g was uniformly dispersed in 800g of safflower oil.

Separately, add 1 kg of gelatin and 300 g of gum arabic to 101
The mixture was added to 00O purified water and left in the apparatus overnight to swell and dissolve by heating.

The above two types of liquids were respectively charged into predetermined portions of the Glowpex Mark ■ capsule coating material, and spherical non-coated minicapsules with a particle size of 2 Tsuru were obtained using the same machine.

This non-coated minicapsule 1k+4 was prepared using a centrifugal granulation coating device ficF-360 (trade name, manufactured by Freund Sangyo Co., Ltd.), containing 8% by weight of zein (enteric substance), 1% of glycerin, 78% of ethanol,
A mixture containing 13% water was coated until the capsule weight increased by 10% to obtain zein cord minicapsules.

Through the above operations, two types of intestinal useful bacteria-containing products were obtained, consisting of non-coated minicapsules and zein-coated minicapsules filled with lactic acid bacteria-containing oil.

(Storage test) Wrap the minicapsules obtained in this example in aluminum foil.
It was stored at ℃ for 31 months, and the number of viable bacteria was checked every month.

The test was carried out by culturing the sample on Cystine's commercially available BL agar medium supplemented with 5% hydrochloric acid fiO.

As a control standard, the raw material powder used in this example was diluted five times with dry starch.

The results are shown in Table I.

Table 1 - (Dissolution test using artificial gastric fluid) The lactic acid zein coated minicapsules obtained in this example were
Based on the Japanese Pharmacopoeia disintegration test method, the first liquid (
1 hour and 2 hour dissolution tests were conducted in artificial gastric fluid).

Next, the cells were similarly cultured on the aforementioned BL agar medium (containing 0.05% cysteine hydrochloride) to determine the number of viable bacteria, and the results are shown in Table 2.

-1 knee (temperature: 37°C) As mentioned above, no death was observed in the artificial gastric fluid. Therefore, the second liquid (artificial gastric juice) of the Japanese Pharmacopoeia disintegration test
When similarly tested, the capsules disintegrated rapidly within 3 to 12 minutes.

That is, the corchid minicapsules obtained in this example can be delivered to the intestines in a viable state without being contaminated by gastric juice.

As explained above, the non-coated capsules and zein-coated capsules produced in Example 1 were useful as health foods and medicines.

In particular, those coated with zein were suitable as medicines.

[Example 2] Bifidobacterium lon
gum) supported on skim milk powder (viable bacteria count 1.2
10”/g) 400g medium chain fatty acid triglyceride; Miglyol (trade name, manufactured by Dynamite Nobel)
A filler solution was prepared by uniformly dispersing the filler in 1600 g.

Separately, a 20% aqueous gelatin solution was prepared as a capsule coating material liquid.

The above-mentioned filler liquid is stored in the tank 4 and the coating material liquid is stored in the tank 4a of the manufacturing apparatus schematically shown in the figure, and the above-mentioned operations are performed to store the cold miglyol (curing liquid) 11 in the curing tank 8. Two-layer droplets 13 are dropped into the interior at a rate of 300 droplets/second.

In this way, uncoated minicapsules with a particle size of about 1 liter were obtained.

Non-coated mini male busel 100 obtained by the above operation
0 g was coated with enteric material in the same manner as in Example 1.

The coating liquid was 8% by weight of carboxymethylethyl cellulose (enteric-coated material) (manufactured by Freund Sangyo Co., Ltd.), 1% of glycerin triacetate,
Coating treatment was performed using a mixed solution of 46% methylene dichloride and 45% ethanol until the weight of the capsule increased by 15%.

Through the above operations, two types of minicapsules were obtained: non-coated minicapsules filled with Bifidobacterium-containing oil and carboxymethylethyl cellulose (hereinafter abbreviated as CMEC) coated minicapsules.

(Storage Test) A storage test was conducted in the same manner as in Example 1 above. The results are shown in Table (2).The reference standard was also obtained by diluting the raw material powder five times with dry starch, and the numerical values are used in the same sense.

-1 knee (temperature: 37°C) (Dissolution test using artificial gastric juice) A dissolution test using artificial gastric juice was conducted on the 0MEC coated minicapsules obtained in Example 2.

This test was also conducted in the same manner and under the same conditions as in Example 1 above. The results shown in the following table (■) were obtained.

-Human IV (Temperature: 37°C) According to the results in Tables ■ and ■ above, the intestinal useful bacteria-containing material of Example 2 also has a good preservability as in Example 1.
It was revealed that it has extremely excellent solubility resistance in artificial gastric fluid. Furthermore, when the 0MEC-coated minicapsules were subjected to a dissolution test using artificial intestinal fluid, they were rapidly dissolved as in Example 1.

As explained above, the intestinal beneficial bacteria-containing material of Example 2 was useful as a health food and medicine, as in Example 1, and the 0MEC-coated minicapsules were particularly suitable as medicines.

[Example 3] In Example 3, a 0.8% sodium alginate aqueous solution was used instead of the gelatin aqueous solution as the coating material liquid in the above-mentioned Example 2, and a 20% calcium chloride aqueous solution was used as the curing liquid. It is something.

Droplets were formed at a rate of 200 droplets/second to obtain a material containing beneficial bacteria for the intestines consisting of minicapsules with a particle size of about 1 m. In the above capsule, the filler liquid was the same as that of Example 2, and the coating material was made of calcium alginate.

A storage test was conducted on the intestinal useful bacteria-containing material of Example 3, and the results shown in the following Table 3 were obtained.

Note that the test method and conditions were the same as in the previous example.

-Jjijin- (Temperature change: 37°C) The intestinal useful bacteria-containing material of Example 3 was effective as a health food and medicine as in Examples 1 and 2 above.

Although the present invention has been specifically explained based on Examples above, it goes without saying that the present invention is not limited to what was shown in the Examples.

For example, in the examples, only two-layer capsules have been described, but they may have three or more layers, and the coating material and filler liquid for the capsules are not limited to those in the examples above, and various types may be used. Modifications are available.

In addition, although Lactobacillus lactis and Bacteria bifidus have been described as useful intestinal bacteria, it goes without saying that the present invention is also effective when applied to other useful bacteria. Furthermore, although the above-mentioned useful bacteria are supported on starch powder or skim milk powder, they may be directly dispersed in oil.

In addition, the manufacturing equipment is not limited to the Glowpex mark and shown in the diagram, and is equipped with double or more nozzles.
Any device may be used as long as it can be used for the intended purpose.

〔Effect of the invention〕

The beneficial bacteria in the intestine is contained in a capsule with a diameter of 3 dragons or less, which is filled with oil in which beneficial bacteria in the intestine or a powder supporting the bacteria is dispersed. Since it can be protected, the useful bacteria can be stored for a long time, and when ingested orally, the useful bacteria can reach the intestines without reducing the number of viable bacteria at the time of preparation. Oil containing useful bacteria can be easily emulsified and dispersed in digestive juices in the intestines.

Therefore, a desired amount of useful bacteria can be reliably delivered to the intestine, and thus accurate intestinal regulation can be performed.

The above effects can be further improved by coating the capsule with an enteric substance.

In addition, from the inner nozzle of the multi-layered nozzle, oil in which intestinal bacteria or powder carrying the same is dispersed is simultaneously flowed out, and from the outermost nozzle, the coating material liquid is simultaneously flowed out into multilayer droplets with a diameter of 3 or less. and then its multiplicity N? &, The coating material of the outermost layer of the droplet is cured, and the above-mentioned substance containing beneficial intestinal bacteria can be produced.

Furthermore, after the step of curing the coating material, an enteric coating is applied on the coating material, thereby making it possible to produce the above-mentioned intestinal beneficial bacteria-containing material.

[Brief explanation of drawings]

The figure is a schematic configuration diagram of a manufacturing apparatus according to an embodiment of the present invention. 1...Double nozzle (multiple nozzle), 1a...
1st nozzle (inner nozzle), 1b...2nd nozzle (
outermost nozzle), 2... Vibration generator, 3... Vibrator, 4.4a... Tank, 5.5a... Pipe, 6.6a... Pump, 7.7a... Pulp, 8...Curing tank, 9...Filler liquid, 10...Coating material liquid, 11...Curing liquid, 12...Two layers Liquid flow, 13...Two-layer droplet (multilayer droplet), 13a...
Seamless mini capsule. Patent applicant Freund Sangyo Co., Ltd. agent Patent attorney Dai Tsutsui Sodo Patent attorney Hidetoshi Matsukura w Rooster engraving (no change in content) Figure 1 Procedural amendment (1 button May 22, 1986 Commissioner of the Patent Office Uga Michibe Beating 1, Incident Display K
” 1985 Patent No. 43651 2 Name of the invention Intestinal bacteria-containing material and method for producing the same 3 Relationship with the case of the person making the amendment Patent applicant name Freund Sangyo Co., Ltd. 4 Agent Address: 160 Address: 7-21-21 Nishi-Shinjuku, Shinjuku-ku, Tokyo Name (8000) Yamato Tsutsui, Patent Attorney, 1.゛、.・11-' 5. Date of amendment order (voluntary) 7. Contents of amendment (1), r on line 11 of line 14 of the specification (Lacfob
acillu=acidphi 1us) J to r(L
actobacillus acidphilus)
Correct to. (2), page 15, lines 16 to 18, “The test...
····went. ” is corrected as follows. "The test was carried out by culturing the sample on a commercially available BL agar medium supplemented with 0.05% cysteine hydrochloride." I let it happen. ”. Procedural formalities (methods) 1. Indication of the case Patent Application No. 43651 of 1985 2. Name of the invention Intestinal useful bacteria-containing material and its manufacturing method 3. Person making the amendment Relationship to the case Name of patent applicant Title Freund Sangyo Co., Ltd. 4, Agent 160 Address 7-21-21-5 Nishi-Shinjuku, Shinjuku-ku, Tokyo
Date of amendment order: May 7, 1986 (Shipping date: 1986)
amended on May 27, 2013). (2), “The figure is...” on page 25, line 20 of the same page is replaced with “1st
It is corrected to "It's the drawing number...". (3) Correct the figure number of Figure 1 to "Figure 1" as shown in the attached sheet.

Claims (5)

[Claims] (1) A product containing beneficial intestinal bacteria, which is obtained by filling a capsule with a diameter of 3 mm or less with a capsule having a diameter of 3 mm or less and filled with oil in which beneficial intestinal bacteria or powder carrying the beneficial intestinal bacteria is dispersed. (2) The enteric bacteria-containing material according to claim 1, wherein the capsule is coated with an enteric substance. (3) The oil in which intestinal beneficial bacteria or powder carrying the intestinal beneficial bacteria is dispersed is poured out from the inner nozzle of the multi-nozzle, and the coating material liquid is flowed out from the outermost nozzle to form a diameter of 3 mm.
A method for producing a substance containing beneficial intestinal bacteria, which comprises forming the following multilayer droplets and curing the outermost coating material of the multilayer droplets. (4) The method for producing a substance containing beneficial intestinal bacteria according to claim 3, characterized in that the multiple nozzles are vibrated. (5) The method for producing a substance containing beneficial intestinal bacteria according to claim 3, characterized in that the outermost coating material of the cured multilayer droplet is coated with an enteric substance.