JPH10194975A - Hepatic disease medicine - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject medicine capable of improving the syndromes of a hepatic disease patient, especially hyperammonemia, and little in side effects by including highly pure raffinose as an active ingredient. SOLUTION: This hepatic disease medicine contains highly pure raffinose as an active ingredient. The raffinose is preferably administered at a daily dose of 0.3-0.9g per kg of a body. The raffinose is produced e.g. from beet roots by a known method. The medicine may further contain a known vehicle, a known shape-retaining agent, a known filler, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for liver disease containing high-purity raffinose as an active ingredient, which is administered mainly for the purpose of improving hyperammonemia associated with liver disease.

[0002]

2. Description of the Related Art The liver collects blood flow from the gastrointestinal tract via the portal vein, and 3/4 of the hepatic blood flow is occupied by the portal blood flow.
Although the liver performs various important functions, a main function particularly associated with the digestive tract is a function of the liver retaining nutrients absorbed from the digestive tract and metabolizing the nutrients into substances necessary for living organisms. Unnecessary and harmful substances absorbed from the digestive tract are ingested by hepatocytes, metabolized into hydrophilic substances, and excreted from the biliary tract or kidneys.
Furthermore, the liver also has a detoxification function such as processing ammonia in a urea cycle to form urea.

[0003] Cirrhosis is the terminal picture of all liver diseases, and abnormalities in portal venous blood flow occur together with hepatocellular dysfunction. Particularly in the decompensated period of cirrhosis, toxic substances originating in the gastrointestinal tract (ammonia, lower fatty acids, mercaptan). , Indole compounds, etc.)
Detoxification function is significantly inhibited, and various symptoms such as hepatic encephalopathy develop based on this.

[0004] On the other hand, the intestinal flora is involved in digestion and absorption of meals and in the production and absorption of various harmful substances, and it is known that the relationship between the role of the liver and the intestinal flora is extremely large. (Intestinal flora and metabolism, edited by Japan Bifidobacterium Center, pages 38-56, 7th training textbook, 1991).

[0005] For example, a comparative experiment of the intestinal flora of a liver disease patient and a healthy person showed that the number of anaerobic bacteria in the feces of the cirrhosis patient was significantly increased as compared with the healthy person, and the main bacterial species was Bacteroides. (Journal of the Japanese Society of Gastroenterology, Vol. 77, No. 10, pp. 28-36, 1980)
Year). Further, among anaerobic bacteria, there are many bacteria having urease activity involved in ammonia production in the intestinal tract, and the bacteroides are known as the main bacteria (Clinical Physiology of Gastroenterology, Vol. 5, pp. 71-90). Page, 1981
Year).

[0006] In view of these findings, various methods have been tried for a long time for the purpose of treating severe liver disease, and remedies for hepatic encephalopathy observed in decompensated cirrhosis have been remarkably advanced, and liver function has been improved. Can also be expected. For example, administration of hyperbranched amino acid preparations for liver failure mainly due to decompensated cirrhosis is known (Meiji Watanabe and Misako Okita, "Liver Disease and Therapeutic Nutrition", Daiichi Shuppan, 199).
2 years). In addition, lactulose preparations are widely used for the treatment of hepatic encephalopathy and hyperammonemia with the intention of improving the intestinal flora (Japan Pharmaceutical Information Center, edited by “Nippon Pharmaceutical Information Center”, Pharmaceutical Industry Co., 1996) Year).

[0007] As described above, a drug for the purpose of improving the symptom in the treatment of liver disease and the relationship between liver disease and intestinal microflora and the like have been hitherto known, but lactulose has a wide range of utilization properties to bifidobacteria. Non-digestible oligosaccharides, and various types of galactooligosaccharides, various fructooligosaccharides, raffinose, and the like are known as similar oligosaccharides. These oligosaccharides have the effects of improving the intestinal environment, improving convenience, suppressing the production of intestinal spoilage products as bifidobacteria growth factors, and are widely used mainly in the field of food (Monthly Food Chemical, 10, Vol. 79-84, 1986).

[0008] However, among these oligosaccharides, the only drugs that have been used for the treatment of liver diseases are lactulose and lactitol for improving hepatic encephalopathy and hyperammonemia alone (Japanese clinical study, vol. 52, No. 1, No. 1
10-118, 1994).

Lactulose has assimilation properties for a wide range of bifidobacteria, and grows bifidobacteria and is metabolized into organic acids to lower the pH in the intestinal tract.
It is known that it inhibits the absorption of ammonia produced by putrefactive bacteria from the intestinal tract, promotes intestinal peristalsis, shortens the residence time of intestinal contents in the intestinal tract, and promotes defecation (Japanese clinical, 46th volume extra number, pages 1077-1083, 1988). It has been reported that these actions suppress the absorption of various toxic substances, including ammonia, from the intestinal tract, and recently suppress the production of ammonia by the intestinal tract [Hepatolo.
gy), Vol. 6, p. 1133, 1986].

[0010] However, lactulose is also utilized by Escherichia coli, Clostridium and the like, and there are cases where there is a slight difficulty in the selective utilization of useful bacteria such as Bifidobacterium.
In addition, depending on the dose, diarrhea is induced and abdominal bloating is easily caused. Furthermore, lactulose has a problem that it has a strong sweetness and it may be difficult for some people to take a large amount.

On the other hand, lactitol is a substance belonging to sugar alcohol, and its properties are similar to those of oligosaccharides.
Like lactulose, it is also used in the treatment of hepatic encephalopathy and hyperammonemia (Clinical Medicine, Vol. 11, No. 7).
No. 1439-1473, 1995). However, lactitol has the drawback that it is not widely available to various human-derived bifidobacteria (Bifidus, Vol. 9, No. 1, pp. 19-26,
1995), the improvement of the intestinal flora becomes insufficient, and the effect may not be obtained depending on the patient.

As described above, lactulose and lactitol, which are conventionally known as therapeutic agents for liver diseases, still have room for improvement in the intestinal environment and the like for the purpose of improving the symptoms of liver diseases. The development of a more desirable therapeutic agent for liver disease from a practical point of view has been expected.

Next, besides the conventionally known therapeutic agents for liver diseases, raffinose is a saccharide that has been studied in recent years from the viewpoint of improving the intestinal environment. Raffinose does not have as wide assimilation properties of bifidobacteria as lactulose, but is hardly assimilated by putrefactive bacteria and ammonia-producing bacteria, and may be an excellent oligosaccharide as a selective sugar source for the growth of bifidobacteria. It is known, and the effect of suppressing the production of intestinal putrefaction products and improving the convenience has been clarified [Bifidobacteria and microflora (Bifidobacteria and
Microflora), Volume 6, Issue 2, pages 34-40,
1993].

However, the conventional use of raffinose has been mainly used for foods utilizing processing characteristics, edible dehydrating agents utilizing hygroscopicity, stabilizers for transport liquids of transplanted organs, stabilization of viable bacterial agents and the like. (Japan Food Science,
9, No. 34-40, 1993).

In addition, as a composition for improving the intestinal environment,
It is known that a composition comprising a combination of galactooligosaccharides such as raffinose and stachyose and dietary fiber is effective for treatment and prevention of hepatic encephalopathy and colorectal cancer (Japanese Unexamined Patent Publication (Kokai) 3-1).
No. 51854), however, the effect of the composition is limited to the reduction of the amount of intestinal putrefaction products, and the effect on liver disease is not clear.
It was not intended to be used as a drug for liver disease in earnest. In addition, the publication states that administration of oligosaccharide alone does not show any effect of improving intestinal flora. It has also been known that a liquid food containing an indigestible oligosaccharide such as raffinose and polydextrose has an effect of improving bowel movement (Japanese Patent Application Laid-Open No. 62-220169). Was limited to bowel movements. Further, it has been known that a food containing a saccharide mixture of oligosaccharides containing raffinose and edible dietary fiber as a main component has an intestinal action and is effective in promoting health (Japanese Patent Laid-Open No. 63-63366). However, the food was not known at all for its effect on liver disease.

In addition to the above, there are many known studies in which various known oligosaccharides contribute to physical health by improving the intestinal environment and suppressing the production of harmful substances by putrefactive bacteria. (Bifidus, Vol. 7, No. 1, pp. 1-12, 1993), regarding the effects on the functions of organs other than the intestinal tract, studies on oligosaccharides other than the above-mentioned lactulose and lactitol have not been carried out, and are clear. Was not evaluated.

The effects of various oligosaccharides in the feed on the onset of galactosamine liver injury were examined using rats, and the effects of galactose-containing lactulose, raffinose, two types of galactooligosaccharides, galactose, lactose and galactosamine on liver injury were investigated. Although an onset-suppressing effect has been reported (Journal of the Japan Nutrition and Food Science, Vol. 48, No. 3, pp. 181-187, 1995), the results of this experiment indicate that galactosamine-induced liver injury occurs specifically in rats. It only revealed that galactose was deeply involved as an oligosaccharide or monosaccharide in the suppression, and did not suggest any effect of the oligosaccharide on the effect of improving human hyperammonemia.

The present inventors have found that raffinose improves the intestinal environment, invented a food composition for liver disease patients containing raffinose, and filed a patent application (filed on December 26, 1996). Reference number P000000102
9. Hereinafter, it is described as a prior application. ). However, the invention of the prior application relates to food or the like consumed by a liver disease patient who is being treated with a medicine, and assists in the treatment with the medicine from the aspect. Furthermore, in the invention of the prior application, raffinose has a low solubility, and has a disadvantage that it cannot be administered in an effective amount for directly treating liver disease due to the balance between the raffinose and other components of the food. there were.

As described above, there is no known known therapeutic agent for liver disease which has a main effect of improving the intestinal environment and which has the effect of improving the symptoms of liver disease. As an oligosaccharide used for improvement of bloodemia, there has been a long-awaited demand for a drug having an oligosaccharide as a main component which is effective with little side effects and is easy to drink and ingest.

[0020]

DISCLOSURE OF THE INVENTION In view of the above prior art, the present inventors conducted a detailed study on the effect of administration of a large amount of raffinose on liver function in patients with liver disease after filing the earlier application. went. As a result, a drug containing high-purity raffinose as an active ingredient is effective for ameliorating symptoms of liver disease patients, particularly for improving hyperammonemia, and a drug containing high-purity raffinose as an active ingredient is useful for treating liver disease. The inventors have found that the intestinal environment of the patient has been remarkably improved and that there is little concern about side effects, and the present invention has been completed.

An object of the present invention is to provide a therapeutic agent for liver disease which can improve the symptoms of liver disease patients, particularly hyperammonemia, and has few side effects.

[0022]

The present invention for solving the above-mentioned problems is a therapeutic agent for liver disease containing high-purity raffinose as an active ingredient, wherein raffinose is weighed 1 kg per day.
In a preferred embodiment, the dose is 0.3 to 0.9 g per administration.

Next, the present invention will be described in detail.

[0024]

BEST MODE FOR CARRYING OUT THE INVENTION Raffinose used in the present invention may be a commercially available product, and may be a known method (for example,
Raffinose produced from sugar beet root by the method described in JP-A-54-49345, and raffinose produced from soybean whey by a known method (eg, the method described in JP-A-59-179064). You may. Further, from the oligosaccharide mixture produced as soybean oligosaccharide, a raffinose content increased or a purely separated raffinose content can be used by a known method such as chromatography, but as high as possible. It is desirable to use one of purity.

The therapeutic agent for liver disease of the present invention may be the raffinose powder itself, but in addition to raffinose, which is an essential component, a well-known excipient for the purpose of enhancing storage stability, adjusting the dosage form, and the like. It can be processed into a liquid, powder, granule, or the like by a conventional method by adding a shaping agent, a shape retaining agent, a bulking agent, an emulsifier, a flavor, a coloring agent, a preservative, and the like.

The effective dose of these raffinoses to patients with liver disease varies depending on the patient's symptoms, age, etc., but as will be apparent from the test examples described below, the body weight per day is 1k.
It is desirable to administer in the range of 0.3 to 0.9 g per g.

The therapeutic agent for liver disease of the present invention can be used in combination with a known therapeutic agent for liver disease. Known drugs for liver disease include, for example, various amino acids, various essential fatty acids, lactulose, lactitol and other various oligosaccharides, vitamins (eg, vitamins A, D, K, etc.), and minerals (eg, zinc preparations, etc.). Examples can be given. In particular, when used in combination with lactulose, the ratio of lactulose 1/6 to 2 parts to 1 part of raffinose (weight; hereinafter the same unless otherwise specified) is most desirable.
Raffinose has an advantage that it has a sweetness of about 1/10 or less of sucrose and is easy to drink and ingest.

The therapeutic agent for liver disease of the present invention produced as described above is effective in improving various symptoms of liver disease, particularly hyperammonemia, as described below.

Next, the present invention will be described in detail with reference to test examples. Conventionally, the problems of the tests that have been performed to develop oligosaccharides that are the main components of therapeutic agents for liver diseases are not sufficiently understood and reflected on the intestinal environment of patients with liver diseases. As described above, the effect of improving the various symptoms of liver disease by the conventional oligosaccharide preparation was insufficient in this respect.

The present inventors have selected the oligosaccharides in the preparation on the assumption that the anaerobic ammonia-producing bacteria are present in the intestine in a large amount as described above and the intestinal tract of a liver disease patient whose intestinal environment has deteriorated. In this case, the main constituent bifidobacteria in the intestine are widely assimilated, have a high growth rate, and the added oligosaccharides are rarely used by other spoilage bacteria. The acid production in the intestinal tract is high as a result of the added oligosaccharide being sufficiently utilized by useful bacteria such as bifidobacteria. The pH value in the intestinal tract is reduced as much as possible as a result of the added oligosaccharide being sufficiently utilized by useful bacteria such as bifidobacteria. However, it was considered to be an important factor to quickly improve the deteriorated intestinal environment and enhance the effect of improving various symptoms of liver disease.

When the conditions (1) to (4) are satisfied, the production of toxic substances such as ammonia, indole and skatole by putrefactive bacteria in the intestinal tract can be suppressed early, reliably and effectively. By reducing the burden of liver function,
It is considered that the present invention realizes a certain improvement effect on various symptoms of liver disease. Therefore, the present inventors conducted a preliminary test under conditions as close as possible to these three points.

As an experimental method, in order to properly test a desired oligosaccharide preparation under conditions most suitable for a patient with liver disease, the assimilation and growth rate of major bifidobacterium mixed bacteria in the intestine by various oligosaccharides, A method of comprehensively evaluating each index of acid generation and pH value was adopted. Preliminary test This test is intended to improve the symptoms of liver disease patients by determining the growth promoting effect of added oligosaccharides on a mixed culture system of various Bifidobacteria frequently detected in the human intestinal tract. This was performed to select the oligosaccharides in the formulation to be administered. This test was carried out by the same method as the preliminary test 1 of the prior application, except that the strain used was changed. The outline of the test method is as follows.

Bifidobacterium adolescentis ATC is one of the most frequently detected Bifidobacterium strains in adults and the elderly in Japan.
C15703, Bifidobacterium bifidum (Bi
fidobacterium bifidum) ATCC29521, Bifidobacterium longum
Three strains of ATCC 15707 were selected and a mixed system of these three strains was used assuming intestinal flora.

Basal medium in the test method [yeast extract 0.5% (weight; hereinafter the same unless otherwise specified), meat extract 0.5%, glucose 2%, monopotassium phosphate 0.1%.
1%, dipotassium phosphate 0.1%, anhydrous sodium acetate 0.2%, cystine 0.05%, Tween 80 (Tween
en80) 0.05%, purified water (ion-exchanged water)
5%], and a test medium prepared by adding 2% of various oligosaccharides shown in Table 1 in place of the glucose added to the preculture solution and the basal medium obtained by culturing the mixed system of the three strains using the basal medium was prepared. . The mixed system of the above three strains was cultured in this test medium and tested.

In order to comprehensively evaluate the effects of various oligosaccharides on a mixed culture system of bifidobacteria in a test medium, the growth rate, the acidity of the medium and the pH of the medium were determined from the following three points.
The growth activity value was calculated according to the following formula, and the evaluation was examined.

Growth activity value = (medium turbidity increase) × (medium acidity) / (medium pH) In this equation, “medium turbidity increase” refers to the degree of rapid growth of the main bifidobacteria. The “acidity of the medium” means the acid generating power for the added oligosaccharide, and the “pH of the medium” means the pH lowering power due to the use of the added oligosaccharide, and the growth activity value is determined by the above equation. Are summarized in order to comprehensively determine the effects of

The test results are as shown in Table 1.
From Table 1, it was found that raffinose has the highest growth activity value, lactulose follows, and lactitol and galactooligosaccharide have low growth activity values.

The values of any of the other known oligosaccharides such as fructooligosaccharides, xylo-oligosaccharides, lactosucrose, and isomaltooligosaccharides,
Neither effect reached raffinose or lactulose.

[0039]

[Table 1]

Test Example 1 Raffinose, an active ingredient of the present invention, was subjected to an acute oral toxicity test using rats.

(1) Test method Five 4-week-old Wistar rats (purchased from Japan SLC, Inc.) each of 5 males and 5 females were preliminarily bred for 1 week on normal rat chow to confirm that there was no abnormality in health. After confirming, the animals were fasted for about 13 hours and weighed. A raffinose powder (manufactured by Nippon Sugar Beet Sugar Co., Ltd.) was mixed with purified water to give a 400 mg / ml solution to each rat. Observe frequently on the day, and 1 day a day for 13 days
Observations were made. In addition, body weight was measured every week after administration, and at the end of the observation, all rats were dissected, and the presence or absence of abnormality was visually observed.

(2) Test Results In all the rats used in the test, no deaths were observed during the observation period, and no abnormalities were observed in all the rats clinically.

Table 2 shows the results of body weight measurement one week and two weeks after administration, but no inhibition of weight gain was observed in both males and females. In addition, no abnormalities were found in the main organs of all rats at the autopsy after the observation.

Therefore, it was found that the lethal dose of raffinose, which is an active ingredient of the present invention, to rats after a single oral administration was 6,000 mg / kg or more for both males and females.

[0045]

[Table 2]

Test Example 2 The mutagenicity of raffinose, which is an active ingredient of the present invention, was examined for Escherichia coli WP2 uvrA strain and Sa.
Using 4 lmonella typhimurium TA strains, a reversion test including the metabolic activation method (Ames test) was performed.

(1) Test method 1) Test strain Escherichia coli WP2 uvrA strain, Salmonella typhimuri
Five strains of um TA100, TA1535, TA98 and TA1537 were used.

In the test, Nutrient broth No. 2
(OXOID) Inoculate each strain cryopreserved at -80 ° C into an L-shaped test tube into which 10ml has been dispensed, shake culture at 37 ° C for about 7 hours, and test the resulting bacterial suspension. It was used for.

2) Preparation of Test Solution Raffinose (manufactured by Nippon Sugar Beet Sugar Co., Ltd.) was precisely weighed and subjected to high-pressure steam sterilization (121 ° C., 20 minutes) in pure water (hereinafter referred to as “sterilized water”).
It is described. ) Was added and stirred by a test tube mixer to prepare a 50 mg / ml solution. This was used as a stock test solution, diluted appropriately with sterile water, and used for the test.

3) Test Concentration The concentration of raffinose was set at 5,000 μg / plate, and the test was carried out at 2,500, 1,250, 625, 313 and 156 μg / plate in six steps.

4) Method of test operation The following tests were carried out by the preincubation method (both conditions without the metabolic activation method and with the metabolic activation method).

0.1 ml of a test solution prepared to a predetermined concentration is weighed into a sterilized small test tube, and 0.5 ml of an S9 Mix or 0.1 M Na-phosphate buffer (pH 7.4) having the following composition is prepared.
And 0.1 ml of each bacterial suspension were sequentially added. Each small test tube was shaken (pre-incubation) for 20 minutes in a thermostat at 37 ° C., then 2 ml of the next top agar was added and mixed, and a minimum glucose agar plate medium (manufactured by Nisshin Seifun KK).
(Diameter 100 mm, 30 ml) was spread evenly on a Petri dish and solidified. This petri dish was cultured in a thermostat at 37 ° C. for 48 hours, and the number of colonies generated by reversion was counted. In addition, it was confirmed that the test liquid and S9Mix did not contain various bacteria.

Further, AF-2 [2- (2-furyl) -3- (5-nitro)
-2-furyl) acrylamide], ENNG [N-ethyl-N'-nitro
-N-nitrosoguanidine], 9-AA [9-aminoacridine
] (Above, no S9Mix), 2-AA [2-aminoac
ridine] (presence of S9Mix).

[Composition of S9Mix] S9 (manufactured by Kikkoman) 0.1 ml (/ ml) MgCl ₂ 8 μmol KCl 33 μmol glucose-6-phosphate 5 μmol NADPH 4 μmol NADH 4 μmol Na-phosphate buffer (pH 7.4) 100 μmol [ Top agar] Bacto agar (manufactured by DIFCO) 0.6
% And NaCl 0.5% after autoclaving, the S. typhimurium TA 4 strains were separately sterilized 0.5 mM histidine-biotin solution in 1/10 volume, and the E. coli WP2 uvrA strain was , 0.5 mM tryptophan solution was added to 1/10 volume and mixed.

(2) Test Results Table 3 shows the results when the metabolic activation method was not used (S9Mix was not present), and Table 4 shows the results when the metabolic activation method was used (S9Mix was present). .

In each case, the compound used as a positive control showed a remarkable increase in the number of revertant colonies as compared with the solvent control, but raffinose showed no increase in the solvent at all six concentrations. No increase in the number of revertant colonies was observed as compared with the control.

From the above results, it is apparent that raffinose, which is an active ingredient of the present invention, has a negative mutagenicity under the test conditions.

[0058]

[Table 3]

[0059]

[Table 4] Test Example 3 This test was performed to examine the effective dose of the therapeutic agent for liver disease of the present invention.

(1) Test Method Thirty-nine male and female SD rats aged 7 to 9 weeks were preliminarily reared for one week to acclimate to the rearing environment. . During preliminary breeding and the test period, tap water and commercially available solid feed were freely ingested.

A total of 39 animals were randomly selected into two groups of 18 animals (A
Group and group B) and the control group were divided into three groups, and the rats in the group A and group B were further randomly divided into three groups of six groups.

The group A rats were treated with the therapeutic agent for liver disease described in Example 1 as raffinose at 0.5, 1.0, 1.5, 2.0, 3.0 and 6 per kg of body weight per day. .0
g was orally administered twice a day in equal doses for 7 days.

On the other hand, rats in group B were given lactulose syrup (Morinaga Milk Industry Co., Ltd., lactulose content 50%).
As lactulose in an amount of 0.1 kg / kg body weight per day.
5, 1.0, 1.5, 2.0, 3.0 and 6.0 g,
Oral administration was repeated twice a day for 7 days.

The control group was reared under the same environment as the preliminary rearing for 7 days without treatment. Several hours after the end of the final administration on day 7, portal blood was collected from all rats under anesthesia, plasma was separated by centrifugation, and the ammonia concentration in the portal blood was measured (Wako Pure Chemical Industries, Ltd.) (Manufactured by the company).

(2) Test Results The test results are shown in Table 5 as the average value of three animals in each group. From Table 5, it can be seen that in group A, the dose of raffinose was 0.5.
And 1.0 g / kg / day, there was no significant change in ammonia concentration in portal vein compared to the control group, but at doses of 1.5 g / kg / day or more, As a result, the ammonia concentration decreased. On the other hand, in group B, 1.
No effect was observed at a dose of 5 g / kg / day or less,
At doses of 2.0 g / kg / day and above, the concentration of ammonia decreased in a dose-dependent manner. In addition, in both the group A and the group B, the tested maximum dose of 6.0 g / kg / day did not
No side effects such as loose feces and reduced food consumption were observed.

From the above results, it is clear that the preparation containing raffinose as an active ingredient significantly reduces the amount of ammonia absorbed into the body, and is useful for improving hyperammonemia in patients with liver disease. . The effective dose of raffinose is about 3 times the lactulose dose.
Since the same effect is shown at / 4 times, the clinical use amount of lactulose is 20 to 60 g [65% syrup,
30-90 ml (bile-hepatic pancreas, Vol. 26, No. 2, 297
~ P.303, 1993)], the effective dose of raffinose is calculated to be 0.3 to 0.9 g per kg of body weight per day as a standard body weight of 50 kg.

The effective dose was calculated by changing the type of raffinose and tested, and almost the same results were obtained.

[0068]

[Table 5]

Test Example 2 This test was conducted to examine the effect of the therapeutic agent for liver disease of the present invention on the improvement of hyperammonemia in a chronic liver failure model animal.

(1) Test Method Twelve 9-week-old SD male rats were preliminarily reared for one week to acclimate them to the rearing environment, and after confirming that there was no problem with their health condition, the inferior portal vena cava was used. An anastomosis operation (PCS plastic operation) was performed to obtain a chronic liver failure model rat. During preliminary breeding and the test period, tap water and commercially available solid feed were freely ingested.

On the 7th day after the operation, 12 animals were randomly divided into 4 groups of 3 animals each, and the therapeutic agent for liver disease described in Example 1 was administered.
As raffinose, 0 (control group), 2.0, 3.0 and 4.0 g per kg of body weight per day were repeatedly orally administered twice a day in equal amounts for 7 days.

Several hours after the end of the final administration on day 7, blood was collected from all rats under anesthesia from the abdominal vena cava.
After separating the plasma by centrifugation, the ammonia concentration in the blood was measured using a measurement kit (manufactured by Wako Pure Chemical Industries, Ltd.),
The average of each group was calculated and tested.

(2) Test Results As a result of this test, the ammonia concentration of venous blood (μg / d
l) means that the mean of the control group is 162, whereas 2.
The average of the 0 g / kg / day administration group was 132, 3.0 g / kg
The average for the / day administration group was 122 and the average for the 4.0 g / kg / day administration group was 118. Therefore, in a rat model of chronic liver failure, 2.0 to 4.0 g / raffinose was used.
By administering the therapeutic agent for liver disease of the present invention at a dose of kg / day, the concentration of ammonia in the blood is remarkably reduced, and the therapeutic agent for liver disease of the present invention can be used in a patient with chronic hepatic failure. It is clear that it is useful for improving the quality.

A similar test was carried out using the therapeutic agent for liver disease described in Example 2 in place of the therapeutic agent for liver disease described in Example 1, and almost the same effect was confirmed. Furthermore, similar effects have been confirmed in preliminary clinical studies in humans.

[0075]

Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples.

Example 1 20 kg of raffinose (manufactured by Nippon Sugar Beet Co., Ltd.) and 80 kg of purified water (ion-exchanged water) were mixed, sterilized, and then mixed.
The mixture was dispensed in a sterilized polyethylene container at 0 g each to obtain 490 syrup-like agents for treating liver diseases.

Example 2 16 kg of raffinose (manufactured by Nippon Beet Sugar Co., Ltd.), 32 kg of lactulose syrup (manufactured by Morinaga Milk Industry Co., Ltd., 50% lactulose content) and 52 purified water (ion-exchanged water)
of syrup-shaped liver disease treatment agent 490.
Got a piece.

Example 3 40 kg of raffinose (manufactured by Nippon Sugar Beet Sugar Co., Ltd.) and 60 kg of dextrin as an excipient were uniformly mixed, 20 g of each was filled in an aluminum foil bag, sealed, and powdered therapeutic agent 4 for liver disease was prepared. , 800 were obtained.

[0079]

As described above in detail, the present invention relates to a therapeutic agent for liver disease containing raffinose as an active ingredient. The effects of the present invention are as follows. 1) The therapeutic agent for liver disease of the present invention can reduce the burden on liver function by improving the intestinal environment of a liver disease patient early and effectively. 2) The therapeutic agent for liver disease of the present invention suppresses the generation of toxic substances such as ammonia in the intestinal tract and the absorption of toxic substances from the intestinal tract into the body by proliferating bifidobacteria and suppressing ammonia-producing bacteria. It is effective in preventing and treating hepatic encephalopathy and hyperammonemia. 3) The therapeutic agent for liver disease of the present invention is less likely to cause side effects such as diarrhea, loose stool, and abdominal distension. 4) The therapeutic agent for liver disease of the present invention is a preparation that is low in sweetness and easy to drink and ingest.

Continuing from the front page (72) Inventor Toshio Ohashi 5-1-83 Higashihara, Zama City, Kanagawa Prefecture Morinaga Dairy Co., Ltd. Nutrition Science Research Institute, Inc. (72) Inventor Kozo Kawase 5-1-83 Higashihara, Zama City, Kanagawa Prefecture Morinaga Milk Company Stock (72) Inventor Hirohiko Nakamura 5-1-83 Higashihara, Zama City, Kanagawa Prefecture Morinaga Milk Industry Co., Ltd. (72) Inventor Koji Sayama Nisshu 13 Nishi-ku, Inadacho, Obihiro City, Hokkaido Japan Sugar Beet Sugar Co., Ltd.

Claims (2)

[Claims] 1. A therapeutic agent for liver disease, comprising high-purity raffinose as an active ingredient. 2. The therapeutic agent for liver disease according to claim 1, wherein the raffinose is administered at a rate of 0.3 to 0.9 g per kg of body weight per day.