JPH08188535A - Viable bacterium preparation - Google Patents

Abstract

PURPOSE: To obtain a viable bacterium preparation containing a viable bacteri um and chlorella powder as an excipient, useful for medicines, foods, etc., and capable of enhancing the survival rate of bacterial cells after prepared. CONSTITUTION: The viable bacterium preparation contains a viable bacterium, preferably lactobacillus, as a main ingredient and 60-90wt.% of chlorella powder as an excipient. The chlorella powder has following properties; a particle degree distribution: particle sizes of <=149μm: 80-98%, particle sizes of 150-250μm: 2-20%; a bulk density of 0.37±0.05; a tap density: 0.46±0.05g/cm<3> ; and an angle of repose: 42±5 degree. The addition of a conventional binder such as crystalline cellulose, calcium carboxymethylcellulose or α-starch to the chlorella in an amount of 3-20wt.% gives tablets high in hardness without killing the viable lactobacillus. If necessary, a lubricant, a perfume, a pigment, a pharmacodynamic ingredient, etc., may be added to prepare a formulation. The tabletting enables to prevent abrasion and defects generated on storage or carriage.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a probiotic preparation useful for medicines, foods and the like.

[0002]

2. Description of the Related Art Conventionally, freeze-dried preparations of lactic acid bacteria have been used as intestinal medicines and health foods. Lactic acid bacteria used for such purposes need to be commercialized as they are (live cells), and there are many concerns regarding the effects of freeze-drying conditions on the survival of bacteria and the effects of storage conditions on dry cells. Is being researched. (Michi Honma, Chimitsu Mitsuoka, "Bifidobacterium", Yakult Co., Ltd., Tokyo, 197
In addition, when processing lactic acid bacteria into tablets, granules, etc., the decrease in the viable cell count during processing and storage is also a major problem. In particular,
In the case of tablets, bacteria are killed when high pressure is applied during tableting (Koichiro Sonoike and Tamotsu Seto, "Effects of temperature and pressure on the killing rate of microorganisms", Genetics, 48 (1).
, 36-41 (1994)). Therefore, it is necessary to select stable conditions that do not kill the bacteria.

[0003]

At the present time, when tablets are produced using live bacteria such as lactic acid bacteria, soft tablets are produced by lowering the compression pressure during tableting in order to prevent the killing of the bacteria. There is. However, when tableting is carried out at such a low pressure, fluctuations in operating conditions at the time of tableting are reflected as they are in the number of surviving bacteria, resulting in unstable product quality.

Tablets compressed under low pressure are soft and fragile, and are liable to cause problems such as scratches, abrasion and chipping during storage and transportation. Therefore, it is an object of the present invention to provide a viable cell preparation containing a living fungus (live cell) as a main component, with less killing of the cell. Another object of the present invention is to provide a live bacterial tablet having a high hardness and a low friability in addition to the low killing of bacteria.

[0005]

In view of the above circumstances, the present inventors have extensively studied the influence of excipients on the survival of viable bacteria during processing of pharmaceutical preparations. As a result, they have found that the above problems can be solved by using chlorella powder, which has been conventionally used as a health food, as an excipient, and completed the present invention. That is, the viable cell preparation according to the present invention is characterized by containing viable cells as a main component and chlorella powder as an excipient.

The present invention can be applied to any fungi to be formulated, for example, lactic acid bacteria, natto bacteria, yeasts, etc., and lactic acid bacteria are particularly preferable because of wide application range. Examples of lactic acid bacteria that can be used for the live bacterial preparation according to the present invention include Streptococcus-faecali.
s, Streptococcus-faecium, Lactobacillus-acidophil
Examples include us, Lactobacillus-bulgaricus, and Bifidobacterium. The viable cell preparation according to the present invention usually contains a dry powder obtained by subjecting viable cells of these fungi to a drying treatment. The drying treatment of live cells can be carried out by a conventional method, for example, freeze-drying.

In the present invention, chlorella powder obtained by subjecting a single-cell green alga Chlorella to a drying treatment is used as an excipient. As the chlorella used here, chlorella which has been subjected to photosynthetic culture in an outdoor pool or tank culture can be used, and its drying treatment can also be carried out by an ordinary method, for example, spray drying. The blending amount of the chlorella powder is preferably 15% by weight or more, more preferably 60 to 90% by weight, based on the whole preparation. If the blending amount is less than 15% by weight, the number of viable bacteria to be killed tends to increase.

The properties of the chlorella powder preferably used as an excipient in the present invention are as follows. Particle size distribution: 149 μm or less 80-98% 150-250 μm 2-20% Bulk density: 0.37 ± 0.05 g / cm ³ Tap density: 0.46 ± 0.05 g / cm ³ Angle of repose: 42 ± 5 degrees In addition to the above chlorella powder, it is possible to add a binder such as crystalline cellulose, carboxymethyl cellulose calcium, pregelatinized starch, etc., which are usually used in the preparation, and particularly 3 to 20% by weight of these binders relative to the chlorella powder. % Added,
A tablet with higher hardness can be obtained without killing the viable bacteria, which is preferable. If necessary, additives such as lubricants such as magnesium stearate, fragrances, pigments and the like which are usually used in preparations can be added.

Further, the viable cell preparation according to the present invention is, in addition to the above-mentioned viable cells, provided that it does not adversely affect the viable cells.
It is also possible to contain medicinal components such as antacids, stomachic agents, digestive agents and vitamins.

The dosage form of the probiotic preparation according to the present invention is a tablet,
Although any substance such as granules may be used as long as it requires an excipient, the effect of the present invention is remarkably exhibited in tablets to which high pressure is applied during formulation.

The viable cell preparation according to the present invention can be produced by a conventional method. That is, when the preparation is a tablet, it can be produced by thoroughly mixing a predetermined mixture containing viable bacteria and chlorella powder and compression-molding the mixture by a compression-molding means such as a tableting machine. Also,
When the preparation is a granule, the above-mentioned predetermined compound may be well kneaded and granulated by a granulator.

[0012]

The mechanism by which the killing of viable bacteria is suppressed in the viable cell preparation according to the present invention has not been completely elucidated and is not beyond the presumption, but chlorella powder functions as an impact buffer, It is considered to protect live bacteria from the shearing force during stirring and the pressure during compression molding.

[0013]

【Example】

Example 1 I.D. Preparation of tablets Lactic acid bacterium Streptococcus faecalis, corn starch, lactose, Avicel (crystalline cellulose manufactured by Asahi Chemical Industry Co., Ltd.), anhydrous calcium hydrogen phosphate, magnesium stetaphosphate and chlorella powder (spray dried product) with four compositions shown in Table 1 below. Several kinds of tablets each having different hardness were prepared by blending.

Table 1 Composition of tablets (% by weight) ------------------------------------------- ABCD D ---- −−−−−−−−−−−−−−−−−−−−−−−−−−−− Streptococcus faecalis 0.7 0.7 0.7 0.7 Lactose-68.8 68.8-Anhydrous calcium hydrogen phosphate--68.8 Corn starch- 29.5 − − Avicel 9.9 − 29.5 29.5 Chlorella powder 89.4 − − − Magnesium stearate − 1.0 1.0 1.0 −−−−−−−−−−−−−−−−−−−−−−−−−−−−− As is clear from Table 1, composition A is an example of the present invention, in which chlorella powder was mainly used as an excipient, to which a small amount of Avicel was added. Tableting was possible without adding a lubricant. Compositions B to D are comparative examples. Composition B
In C and C, lactose was the main excipient and cornstarch or Avicel was added thereto. In addition, magnesium stearate was added as a lubricant. Composition D
Was composed mainly of anhydrous calcium hydrogen phosphate as an inorganic excipient, Avicel was added thereto, and magnesium stearate was further added as a lubricant.

Tablets were tableted using a rotary tableting machine HT-P22 manufactured by Hata Tekko Co., Ltd. using a die and a punch having a diameter of 8 mm (R10). The hardness of the tablet was changed by weighing the formulation so that the weight of the tablet was 190 to 230 mg, and adjusting the tablet thickness adjusting scale of the tableting machine.

As a result, all of the four powders having the above compositions A to D were excellent in moldability, and tablets having various hardnesses of 0.5 to 11.2 kg could be obtained as measured by a Monsanto hardness meter. II. Measurement of the number of viable cells of lactic acid bacteria The number of viable cells of lactic acid bacteria was measured by the plate dilution method. That is, each produced tablet was dissolved in water, diluted to an appropriate ratio, inoculated into a medium for lactic acid bacteria, and then at 37 ° C.
Incubate for 2 days. Then, the number of colonies was counted, and the number of viable cells was calculated from the obtained number of colonies and the dilution ratio. The measurement was repeated twice for each composition. The result is shown in FIG.

In FIG. 1, (a) shows composition A, (b) shows composition B, (c) shows composition C and (d) shows composition D, respectively. In each figure, the vertical axis represents the number of viable cells of lactic acid bacteria, and the horizontal axis represents the hardness of tablets.

As shown in FIG. 1, in all compositions,
It was recognized that the number of viable cells tended to decrease as the hardness of the tablet increased. However, the rate of decrease in the number of viable cells is greatly different, and the rate of decrease is greatest in the composition D composed mainly of anhydrous calcium hydrogen phosphate, which is an inorganic excipient, and the composition C in which lactose and Avicel are excipients is the largest. It is the next largest. Like composition C, composition B (lactose + corn starch) is mainly composed of lactose, but the rate of decrease in the number of viable cells is smaller than that of composition C. Composition A according to the present invention, which is mainly composed of chlorella powder which is a spray-dried product of unicellular green alga Chlorella
Is the most stable of lactic acid bacteria.

Further, the above results were tested for statistical significance. That is, the relationship between the hardness of tablets and the number of viable lactic acid bacteria was approximated by a curve by the method of least squares, and the obtained regression curves were compared with each other. The formula of the calculated approximate curve is shown below.

A: Y = −0.0206X + 1.09 (R = 0.84) B: Y = −0.0415X + 0.97 (R = 0.91) C: Y = −0.0714X + 0.98 (R = 0.98) D: Y = -0.0910X + 1.14 (R = 0.97) As described above, the relationship between the hardness of the tablet and the viable lactic acid bacteria count can be approximated by a straight line for all compositions ( Correlation coefficient R: 0.84 to 0.98), the slope of which is composition A, composition B, composition C
And composition D increased in that order. The slope of the regression line obtained here represents the ease with which the lactic acid bacteria die when the hardness of the tablet is increased. That is, the larger the slope of the regression line, the easier the lactic acid bacteria die when the hardness of the tablet is increased. The slope of this straight line includes composition A and composition B, C or D.
There was a significant difference between the
that's all). Example 2 Using lactic acid bacterium Bifidobacterium longum as a viable bacterium, a tablet having the composition shown in Table 2 below was prepared in the same manner as in Example 1, and the viable cell count was measured. As a result, the survival rate of lactic acid bacteria was as high as 81% even in a tablet having a very hard hardness of 9 kg.

Table 2 Composition of tablets --------------- Bifidobacterium longum 0.7 Avicel 9.9 Chlorella powder 89.4 ------------------ Example 3 Streptococcus faecalis was used as the lactic acid bacterium, and a mixture of chlorella powder and Avicel was used as the excipient (9: 1).
A tablet using the above and a tablet using only the chlorella powder were prepared, and their hardness and the survival rate of lactic acid bacteria were examined. The results are shown in Table 3 below. As is clear from Table 3, the tablets to which Avicel was added showed the same survival rate of lactic acid bacteria, although the tablets had higher hardness than the tablets containing Chlorella alone.

Table 3 Effect of addition of Avicel on the survival rate of lactic acid bacteria ------------------ Excipients Tablet hardness Lactic acid bacteria Survival rate (kg) (%) −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−− Chlorella powder + Avicel (9: 1) 5.5 75 Chlorella powder alone 3.0 72 −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−

[0023]

INDUSTRIAL APPLICABILITY As described above, according to the present invention, it is possible to obtain a viable cell preparation in which the killing of the bacteria after the preparation is small (the survival rate of the bacteria is high), and particularly when the preparation is a tablet, The hardness can be increased while maintaining the survival rate of viable bacteria. Therefore, it is possible to prevent wear and loss that occur during storage and transportation. Further, by adding an appropriate amount of a binder to chlorella powder and using it as an excipient, the hardness of the tablet can be further increased while maintaining the survival rate of viable bacteria.

[Brief description of drawings]

FIG. 1 is a graph showing the relationship between tablet hardness and the number of viable lactic acid bacteria for each tablet prepared by changing the excipient in the examples of the present invention. (A) is an embodiment of the present invention in which a mixture of chlorella powder and Avicel is used as an excipient. (B) to (d) are comparative examples using conventional excipients, (b) is a mixture of lactose and corn starch, (c) is a mixture of lactose and Avicel, and (d) is anhydrous hydrogen phosphate. A mixture of calcium and Avicel was used as an excipient.

Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display area // (C12N 1/20 C12R 1:46 1:89) (C12N 1/20 C12R 1:01 1:89)

Claims (1)

[Claims] 1. A viable cell preparation containing viable cells and chlorella powder as an excipient.