JPH0840913A - Secondary bile acid lowering agent - Google Patents

Abstract

(57) [Summary] [Structure] Represented by the general formula Gal- (Gal) n-Glc (wherein Gal represents a galactose residue, Glc represents a glucose residue, and n represents an integer of 1 to 4). A secondary bile acid lowering agent containing galactooligosaccharide as an active ingredient. [Effect] It is effective in preventing colon cancer by preventing the production of secondary bile acids, which are said to be involved in carcinogenesis, from bile acids in the intestine.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a secondary bile acid lowering agent which suppresses the production of secondary bile acid in the intestine.

[0002]

2. Description of the Related Art In recent years, the number of patients with colorectal cancer has been increasing in Japan, and the proportion of deaths from colorectal cancer among the deaths from various cancers is also increasing. It is considered that the cause is that the amount of substances having a carcinogenic effect produced in the intestine is increased due to westernization of diet.

One of the substances thought to be involved in carcinogenesis in the intestine is secondary bile acid (SB Reddy).
Cancer Res., Vol.37, p.3238-3242 (1977)).
The secondary bile acid is a primary bile acid, that is, a bile acid secreted from the liver to the intestinal tract, which is changed by the action of some intestinal bacteria. Deoxycholic acid and lithocholic acid which are not present in the primary bile acid. , Ursodeoxycholic acid, etc. Therefore, if the production of this secondary bile acid can be suppressed by a substance that can be taken orally on a daily basis, it is considered to be very effective in the prevention of colon cancer. However, such substances are not yet known.

[0004]

An object of the present invention is to provide a secondary bile acid lowering agent which is highly safe, can be taken orally on a daily basis, and is effective in suppressing the production of secondary bile acids in the intestine. It is in.

[0005]

The secondary bile acid lowering agent successfully provided by the present invention is represented by the general formula Gal- (Gal) n-G.
A galactooligosaccharide represented by lc (wherein Gal represents a galactose residue, Glc represents a glucose residue, and n represents an integer of 1 to 4) is used as an active ingredient. (Hereinafter, galactooligosaccharide means the oligosaccharide represented by the above general formula.)

The galactooligosaccharide constituting the secondary bile acid lowering agent of the present invention is an oligosaccharide known from, for example, Japanese Patent Application Laid-Open No. 2-299582 and Japanese Patent Publication No. 58-20266, wherein lactose is β-galactosidase. It can be obtained by treating with a.

When this galacto-oligosaccharide is produced from lactose, β-galactosidase used in the transfer reaction is Streptococcus thermophilus, Lactobacillus bulgaricus, Kleveromyces fragilis, Kleveromyces lactis, Aspergillus oryzae, Bacillus circulans, Β-galactosidase and the like produced by Brera singularis, Cryptococcus lorenti, Lipomyces lipofa, Sterigmatomyces elivieae, Rhodotorula minuta, Syrobasidium magnum and the like can be used. Among them, β-galactosidase derived from Bacillus circulans is preferable because it has a characteristic of giving galactooligosaccharide in a high yield. The β-galactosidase may be used as it is in a microbial cell.

As the raw material lactose, those commercially available for food can be used as they are. In addition to lactose, milk containing lactose, milk powder, cheese whey, or the like may be used.

The β-galactosidase treatment of lactose is preferably carried out by adding an enzyme to a lactose solution having a concentration of about 4.5-80% w / v and a temperature of about 5-80 ° C. At this time, from lactose, in addition to galactooligosaccharide, Gal-β (1-2) -Glc,
Transfer disaccharides such as Gal-β (1-3) -Glc and Gal-β (1-6) -Glc, and glucose and galactose due to hydrolysis are also produced, but the changes over time in the amounts produced differ. By selecting the time, a reaction product mainly composed of galactooligosaccharide can be obtained.

The reaction product can be used as it is in the secondary bile acid lowering agent of the present invention. For example, it is fractionated by, for example, activated carbon column chromatography, chromatographic separation using an ion exchange resin or a gel filtration agent, and the like. It is desirable to separate monosaccharides and unreacted lactose produced by hydrolysis. The transfer disaccharide is considered to be effective for lowering secondary bile acid, though not as much as galactooligosaccharide, and therefore it is advantageous to add it to a secondary bile acid lowering agent together with galactooligosaccharide without separation.

[0011] Conventionally, galactooligosaccharides have been known to promote the growth of Bifidobacterium in the intestine, but they have not been known to be effective in suppressing the production of secondary bile acids.

If the galactooligosaccharide or the product of the above-mentioned rearrangement reaction containing the galactooligosaccharide is formulated into a dosage form suitable for oral ingestion, such as a powder, granules, syrups and tablets, by a conventional method,
The secondary bile acid lowering agent of the present invention can be obtained. For formulation, use of commonly used excipients (eg glucose, lactose), disintegrants (eg starch, sodium alginate), binders (eg gelatin), pigments, diluents, etc., depending on the dosage form. You can

Since the secondary bile acid lowering agent of the present invention contains lactose-derived galacto-oligosaccharide as a main component and is not toxic, it has been confirmed that when it is administered orally for the purpose of preventing colorectal cancer. There is no strict limit to the dose of
Generally, a suitable dose is 0.1 to 10 g per dose, 1
The total daily dose is 0.1 to 20 g.

[0014]

【Example】

Example 1 25 kg of lactose was dissolved in 20 liters of hot water, and this solution was added with 500 ml of 1M potassium phosphate buffer having pH 7.0 and β-.
150,000 units of galactosidase violacta (trade name, product of Daiwa Kasei Co., Ltd.) were added and reacted at 60 ° C. for 12 hours. After that, the reaction solution is heated to deactivate the enzyme, and activated carbon is added to decolorize the cation exchange resin (Diaion P
K218) and an anion exchange resin (PA316) were passed through a resin column mixed and packed, followed by filtration and concentration to obtain a viscous sugar mixture. The sugar composition of this sugar mixture was as follows: galactooligosaccharide 38%, transfer disaccharide 20%, unreacted lactose 20.
% And monosaccharides 22%. The main component of galactooligosaccharide was Gal-β (1-4) -Gal-β (1-4) -Glc.

Example 2 The secondary bile acid lowering action of galactooligosaccharide was confirmed by the following method. As the galactooligosaccharide, the galactooligosaccharide-containing sugar mixture produced in Example 1 was used as it was.

Test method: 8 healthy boys (36.1 ± 9.8)
A group of 9 years old and 9 healthy males (39.7 years old)
A group B subject consisting of ± 10.8 years old) was prepared, 10 g of galacto-oligosaccharides for group A subjects and 2.5 g of galacto-oligosaccharides for group B subjects were dissolved in 10 ml of lemon water, and after 3 weeks, after each breakfast. Was administered. All subjects were kept under observation for 3 weeks before the start of galactooligosaccharide administration, 3 weeks during the administration, and 3 weeks after the termination of the administration, and fecal samples were collected once a week for a total of 9 times. In addition,
Meanwhile, the intake of foods and drinks containing dairy products, fermented foods and oligosaccharides, and the taking of antibiotics were prohibited. The dosing study was conducted in compliance with the Helsinki Convention. The stool sample was diluted with 10 times the amount of phosphate buffer (0.05M, pH7),
Homogenized, filtered through gauze and lyophilized. After extracting the dried feces with ethanol at 70 ° C. for 2 hours, the centrifugal supernatant was dried to dryness under a nitrogen stream, the obtained dried product was dissolved in methanol, and the bile acid analysis system manufactured by JASCO Corporation (separation column) was used. : Bilepak-11, immobilized enzyme color: Enzymepak-HSD)
Was used to quantify bile acids. Cholic acid and chenodeoxycholic acid were quantified as primary bile acids, and deoxycholic acid, lithocholic acid, and ursodeoxycholic acid were quantified as secondary bile acids. The total amount of primary bile acid and secondary bile acid were calculated and averaged for each period.

The results are shown in FIGS. In both groups A and B, administration of galactooligosaccharides significantly reduced the concentration of secondary bile acids in feces (FIG. 1,
2). No effect of galactooligosaccharide administration was observed on the concentration of primary bile acids during the entire study period (Fig. 3,
4). For the significance test of the results, nonparametric multiple comparison was performed, and it was determined that there was a significant difference when p <0.01.

From the above results, it was found that the administration of galacto-oligosaccharide reduces the concentration of secondary bile acids in human feces. This is probably because galactooligosaccharides suppress the production of secondary bile acids in the human intestine.

Example 3 Twenty healthy males were used as subjects, and after overnight fasting Example 1
15 g of galactooligosaccharide-containing sugar mixture according to 70 ml of water
A test was conducted in which the product dissolved in was drunk. After administration,
When the condition of the abdomen of the subject was observed for 8 hours,
There were no symptoms such as abdominal pain or diarrhea.

[0020]

INDUSTRIAL APPLICABILITY As described above, the secondary bile acid lowering agent of the present invention can extremely effectively reduce the intestinal concentration of secondary bile acid which is deeply involved in the development of colon cancer. The secondary bile acid lowering agent of the present invention has a good taste because it contains a galacto-oligosaccharide obtained from lactose as a main component, and it is considered that there is almost no adverse effect of continuous administration. It is also an advantage that it can be used on a daily basis to prevent colorectal cancer.

[Brief description of drawings]

FIG. 1 is a graph showing the quantitative values of secondary bile acids in feces of group A subjects in Example 2.

FIG. 2 is a graph showing quantitative values of secondary bile acids in feces of group B subjects in Example 2.

FIG. 3 is a graph showing the quantitative values of primary bile acids of feces of group A subjects in Example 2.

FIG. 4 is a graph showing primary bile acid quantitative values of feces of group B subjects in Example 2.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Innovator Norie Onodera 1-1-19 Higashishimbashi, Minato-ku, Tokyo Yakult Honsha Co., Ltd. (72) Inventor Masaaki Watanuki 1-1-19 Higashishimbashi, Minato-ku, Tokyo Stocks Company Yakult Head Office

Claims (1)

[Claims] 1. A galactooligosaccharide represented by the general formula Gal- (Gal) n-Glc (wherein Gal represents a galactose residue, Glc represents a glucose residue, and n represents an integer of 1 to 4). A secondary bile acid lowering agent as an active ingredient.