JP4402897B2 - Inflammatory bowel disorder preventive or therapeutic agent - Google Patents

Description

[0001]
[Technical field to which the invention belongs]
The present invention relates to a preparation used for prevention or treatment of inflammatory bowel disorder.
[0002]
[Prior art]
Among non-specific enteritis, ulcerative colitis and Crohn's disease are on the rise year by year worldwide, and are increasing at a rate of about 4,000 people in Japan in the year. It is called inflammatory bowel disease (IBD; Inflammatory bowel disease). Treatment of IBD depends on symptomatic treatment, but IBD is extremely refractory. Treatment methods for IBD include diet therapy, drug therapy (such as salazopyrine, prednin), immunosuppressive therapy (such as azathioprine and cyclosporine), and surgical therapy. In recent years, leukocyte removal therapy, granulocyte removal therapy, and active oxygen elimination New therapies such as therapies have been developed, and the treatment results have been dramatically improved. These new therapies are deeply involved in the control of inflammatory cells and immunocompetent cells and require treatment at specific medical institutions. In addition, the treatment of IBD other than diet therapy has problems such as restrictions on the medical system. Furthermore, in recent years, probiotic therapy such as improvement of stool properties with lactic acid bacteria preparations has attracted attention, and expectations for live bacteria preparations have increased, and its effectiveness in the prevention and treatment effects on human clinical and experimental animal models has been recognized. (Karen M, Anthony C, Paul S, Conor M, Humberto J, Christine Y, Jason D, Lawrence J, Claudio de S. Probiotic bacteria enhance murine and human intestinal epithelial barrier function. Gastroenterology 2001; 121 : 580-91. (Non-Patent Document 1), Malin M, Suomalainen H, Saxelin M and Isolauri E. Promotion of IgA immune response in patients with Crohn's disease by oral bacteriotherapy with Lactobacillus GG. Ann Nutr Metabol 40: 137-145, 1996 (Non-Patent Document 2), Oren S, Fanny K, Rami E, Erwin S, Patrizia B, Paulo G, Massimo C, Sara M, Daniel R. Variable response to probiotics in two models of experimental colitis in rats. Inflammatory Bowel Diseases 2002; 8: 390-406 (Non-Patent Document 3))
[0003]
However, conventional probiotic therapy has not always produced satisfactory results in experimental animal models (Kenney RJ, Hoper M, Deodhar K, Kirk SJ, Gardiner KR. Probiotic therapy fails to improve gut permeability in a hapten model of colitis. Scand J Gastroenterol 2000; 35: 1266-71. In addition, there is often no substantial improvement in the number of diarrhea, stool characteristics, fatigue, and inflammatory response, which are very important in human clinical findings and patient quality of life (Prantera C, Scribano). ML, Falasco G, Andreoli A, Luzi C. Ineffectiveness of probiotics in preventing recurrence after curative resection for Crohn's disease: a randomized controlled trial with Lactobacillus GG. Gut 2002; 51: 405-9 (non-patent document 4)). In particular, improvement of diarrhea, which is regarded as clinically important, suppression of inflammatory reactions, and maintenance of blood essential metals (hemoglobin deficiency, zinc / selenium deficiency, etc.) are actually serious problems that are required. .
[0004]
JP-A-2001-48796 (Patent Document 1) describes a food material capable of preparing immunity against Crohn's disease, ulcerative colitis and the like mainly composed of a pure culture of Enterococcus faecalis strain. ing. However, in the examples of this publication, there is only a description that the specific Enterococcus faecalis heat-treated cells were administered into the abdominal cavity of mice to improve the resistance to intravenous infection of Escherichia coli. There is no description about the effectiveness of such. In fact, when the present inventors tested in a TNB-induced IBD model by rats, administration of lactic acid bacteria alone improved the effects on indicators such as weight gain, large intestine wet weight, degree of damage, and radical damage-causing enzyme (MPO) in colon tissue. Was not observed (see Example 1).
Japanese Patent Application Laid-Open No. 7-126177 (Patent Document 2) describes a therapeutic agent for ulcerative colitis containing Bifidobacteria cells as an active ingredient. However, according to the test by the present inventors, improvement of the number of diarrhea and the reactive protein (CRP) content of serum, which is an index of acute inflammatory reaction, was not observed with bifidobacteria alone (see Examples 5 to 8). ).
[0005]
[Non-Patent Document 1]
Gastroenterology 2001; 121: 580-91
[Non-Patent Document 2]
Ann Nutr Metabol 40: 137-145, 1996
[Non-Patent Document 3]
Inflammatory Bowel Diseases 2002; 8: 390-406
[Non-Patent Document 4]
Scand J Gastroenterol 2000; 35: 1266-71
[Non-Patent Document 5]
Gut 2002; 51: 405-9
[Patent Document 1]
JP 2001-48796 [Patent Document 2]
JP-A-7-126177 [0006]
[Problems to be solved by the invention]
The purpose of the present invention is not limited to a specific medical institution or medical system, and is not only limited to clinical findings such as fecal improvement but also simple and aimed at improving quality of life by improving systemic conditions. It is an object of the present invention to provide an orally ingestible preventive or therapeutic agent for inflammatory bowel disorder capable of safe home therapy.
[0007]
[Means for Solving the Problems]
Looking at the current state of IBD therapy that meets the above challenges, probiotic therapy is becoming a major trend. Today's probiotic therapy is based on lactic acid bacteria. However, according to epidemiological studies, the incidence of IBD in Japan is much lower than in the West. Italy's morbidity is more than twice as high as that of Japan, the UK is more than three times, and Norway and Sweden are more than five times. These countries are considered to have a much higher intake of lactic acid bacteria dairy products than Japan, contradicting the onset of IBD. Therefore, it is speculated that there is a fundamental defect in the current probiotic therapy with lactic acid bacteria. Japan has been taking miso and soy sauce on a daily basis since ancient times. The source of miso and soy sauce is germ rice cake. The inventors of the present invention focused on this ancient gonococcus and tried to improve the IBD pathological condition systematically by supplementing the part lacking in lactic acid bacteria with culm culture.
[0008]
Inventors of the present invention, in actual clinical settings, after fully providing informed consent, seeks treatment of inflammatory bowel disorder by various oral administration, and is effective in improving patient QOL Have been tested for formulations and ingredients. As a result, the inventors have found that a composition comprising a koji mold powder and lactic acid bacteria brewed by koji mold fermentation is extremely useful and completed the present invention.
The present invention is a preventive or therapeutic agent for inflammatory bowel disorder comprising a koji mold powder and a lactic acid bacteria culture powder.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
As the koji mold of the koji mold used in the present invention, it is not particularly limited, but Aspergillus oryzae, Aspergillus sojae, Asp. Niger, Aspergillus awamori (Asp. ), Aspergillus saitoi, Monascus, Rhizopus, and Penicillium, particularly preferably Aspergillus oryzae. In addition, molds and lactic acid bacteria having fermentation effects similar to those of gonococcal mutants and molds are also available.
The lactic acid bacteria used in the present invention are not particularly limited, but Enterococcus faecalis, Enterococcus faecium, Lactobacillus, and Bifidobacterium are preferred. Particularly preferred are Enterococcus hecheum and Enterococcus faecalis.
Yeast can be added to the preventive or therapeutic agent of the present invention as long as the effects of the present invention are not impaired. The yeast used in this case is not particularly limited, but Saccharomyces cerevisiae, Sacch. Boulardii, Sacch. Sacch. Sake) is preferable, and Saccharomyces cerevisia and Saccharomyces bowlaride are particularly preferable.
[0010]
The preventive or therapeutic agent of the present invention preferably contains 40-70 w / v% of koji mold powder and 10-50 w / v% of lactic acid bacteria powder. Moreover, when yeast is contained in the preventive or therapeutic agent of this invention, it is preferable to make it contain 5-40 w / v%. The gonococcal culture powder used in the present invention is preferably obtained by sterilizing a defatted germ after steam-pressure sterilization, mixing a small amount of gonococcal seeds, culturing at a constant temperature in a cocoon room, drying by ventilation, and milling. The lactic acid bacteria culture powder is sterilized by steam heat sterilization and then cooled, and after culturing lactic acid bacteria in a lactic acid bacteria culture solution mainly composed of glucose, the mixture is stirred and mixed well, then placed in a culture room and incubated at a constant temperature. After that, it is preferably air-dried into a powder.
Moreover, the preventive or therapeutic agent of the present invention may contain a pharmaceutically acceptable excipient as necessary.
The preventive or therapeutic agent of the present invention can be in the form of tablets, dragees, capsules and pills, for example. When the prophylactic or therapeutic agent of the present invention is administered orally, the preferred daily dose is 4 to 6.75 g.
The preventive or therapeutic agent of the present invention is obtained by sieving the above koji mold powder, lactic acid bacteria culture powder, yeast and excipients, mixing them uniformly, and then adding water to 7% to 12%, followed by mixing and mixing. It can be manufactured by tableting using a granulator.
【Example】
[0011]
[Formulation example] Tablets Strong Wakamoto (registered trademark) Unit is% by weight
Aspergillus oryzae NK culture powder 50%
Lactic acid bacteria culture powder (Enterococcus faecalis) 10%
Yeast 37%
3% excipient
[0012]
The above Aspergillus oryzae NK culture powder, lactic acid bacteria culture powder, yeast and excipients are sifted and mixed uniformly, and then mixed with water and mixed to a moisture content of 7% to 12%. And tableted.
[0013]
[Example 1]
Sprague-Dawley male rats (7-week-old: Charles River Japan, Inc.) that arrived at 3-week-olds were raised in a breeding room with a temperature of 23 ± 3 ° C and humidity of 50 ± 20%. Lot: 1Y08) Powdered feed mixed with 1 to 5% and powdered feed mixed with 1% frozen raw powder (Lot: 17B7) of lactic acid strain Lactobacillis Salivarius WB21 were allowed to be freely ingested in the same manner as water for one month. Then, after fasting for 2 days under free water intake, pentobarbital sodium (Nembutal (registered trademark), Dainippon Pharmaceutical) was administered intraperitoneally (40 mg / kg) to give anesthesia, and using silicon sonde, anal 8 cm from the rectum, 2,4,6-trinitrobenzenesulfonic acid (TNBS, Tokyo Chemical Industry Co., Ltd.) 30 mg-50% ethanol / 0.25 mL / rat was administered in the intestine and the lumbar region was raised until anesthesia arousal. Created. After creation of the IBD model, the mice were bred with a powerful Wakamoto (registered trademark) mixed feed or a frozen bulk powder mixed diet of Lactobacillis Salivarius WB21 as before TNBS induction. The control group was two groups in which distilled water and TNBS were enemaerated in rats fed in the same manner with a normal powdered feed (CE-2, Claire Japan). The test group and the control group consisted of 10 animals per group. Seven days after TNBS induction, each animal was killed by anesthesia with ether (Wako Pure Chemical Industries, Ltd.). The colon from 2 cm to 10 cm was removed from the anus, and the incision was made longitudinally and washed with ice-cold physiological saline. After observation, the wet weight was weighed using a balance (SHINKO DENSHI, Co Ltd). Samples were stored at −80 ° C. until the activity of MPO in colon tissue as an indicator of the degree of inflammation was measured. Morris GP et al. Hapten-induced model of chronic inflammation and ulceration in the rat colon.Observed and examined edema, thickening, bleeding and ulcers in the isolated colon. Gastroenterology 96: 795-803, 1989) and scored. In addition, the measurement of MPO activity, an enzyme that causes radical damage in colon tissue, is based on the method of Krawisz et al. (Krawisz JE et al. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Gastroenterology 87: 1344-50, 1984). went.
As a result, one week after TNBS enema, the TNBS-induced group showed significant changes in post-enema weight loss, increased colon wet weight, and increased MPO activity compared to the normal group (distilled water enema). . Compared with the TNBS-induced control group, the group administered before and after “Strong Wakamoto” showed an improvement trend in body weight gain, large intestine wet weight, and degree of injury. Moreover, a significant reduction was observed in the evaluation index of the degree of disability by damage score (p <0.05). However, no change was observed in the MPO activity in the large intestine tissue. On the other hand, the improvement effect was not recognized in all the indices in the groups administered before and after Lactobacillis Salivarius WB21, which is a lactic acid bacterium alone. In addition, it was similarly invalid in the group administered before or after “Strong Wakamoto”.
[0014]
[Example 2]
Four patients with Crohn's disease (male, 20 to 60 years old) who have diarrheal stool 8 to 16 times a day, "Strong Wakamoto" 1 day after breakfast and dinner twice a day, 6 to 10 tablets once I took it for months. As a result, the number of diarrheal stools decreased to 5 to 14 and a significant difference was observed (p <0.05). It was not recognized. On the other hand, acute inflammatory reaction CRP (C-reactive protein) was positive in strong positive patients, and positive patients (5 mg / dL serum) were improved and suppressed to mild patients (0.9 mg / dL serum). It was. The patient also had zinc deficiency (30 μg / dL serum) and selenium deficiency (undetectable), but one month after taking “Strong Wakamoto”, serum zinc improved to 63 μg / dL serum (normal value) 120 ± 20 μg / dL serum). Selenium rose to detectable 12 μg / dL serum, but not about 23 μg / dL serum in normal individuals. Serum selenium concentration is an essential trace element for the biosynthesis of glutathione peroxidase that plays an important role in redox regulation using selenocysteine as a constituent amino acid. The above improvement of CRP, recovery of essential trace metals, etc. are improvement of clinical factors not found in other lactic acid bacteria preparations.
[0015]
[Example 3] Effect of GWT on patients with ineffective yeast preparations 40-year-old male with Crohn's disease, after onset, clinical indices returned to normal after drug therapy and nutrition therapy, but beer was recovered to recover physical strength The yeast preparation was taken at 3 g per day for 3 months. However, there was a strong decrease in energy and fatigue, and no improvement was observed. The brewer's yeast preparation was discontinued, 6 tablets (1.5 g) per day of GWT, and 3 doses for 3 months. As a result, there was almost no fatigue and a marked improvement in physical strength was recognized.
[0016]
[Example 4] Improvement of pathological condition by GWT in a patient ineffective in yeast preparation A 42-year-old male with Crohn's disease was introduced with remission after colostomy. In order to maintain remission, the brewer's yeast preparation was taken at 3 g / day for 3 months, but CRP was 0.7 μg / mL (index 0.7), the amount of defecation was large, and improvement in QOL was not observed. When the yeast preparation was discontinued and GWT was switched to 6 tablets (1.5 g) 3 times a day for 3 months, the normal value of CRP 0.2 μg / mL (index 0.2) returned to normal. Was reduced to 1/2 to 1/3 when taking the yeast preparation, and the feces were maintained in shape and markedly improved QOL.
[0017]
[Example 5] Effect of GWT on patients with ineffective bifidobacterial preparation (Bifidobacterium longum, infantise) pathological improvement effect by GWT A 38-year-old male with Crohn's disease was introduced with remission by drug therapy and nutrition therapy. The maintenance treatment was conducted for 3 months by taking 3 g of bifidobacteria preparation daily, but the frequency of diarrhea lasted 8-9 times a day, CRP 0.6-0.5 μg / mL (index 0.5), and progress in improvement was observed. There wasn't. Discontinuation of bifidobacterial preparation, GWT 6 tablets (1.5 g) daily, 3 doses continued for 1.5 months, resulting in a dramatic decrease in diarrhea 3-4 times a day, CRP 0.2 μg / mL (index 0.2) The feeling of fatigue was reduced and the QOL was significantly improved.
[0018]
[Example 6] Effect of GWT on patients with ineffective bifidobacterial preparation (Bifidobacterium longum, infantise), 43-year-old male with Crohn's disease, after onset / treatment, the number of diarrhea severely 12-13 times a day It was accompanied by hypokalemia and hypoproteinemia. Bifidobacteria-day formulation, 3 tablets (2~8x10 ^8/1 tablet) three times took a month, but improvement was observed. As a result of discontinuing bifidobacterial preparations and taking GWT 6 tablets (1.5 g) daily for 3 months, the number of diarrhea decreased to 5-6 times a day, and serum potassium levels from 0.3 mEq / L A marked improvement was observed with an increase to 2.0 mEq / L. Normal serum potassium concentration; 3.2-4.5 mEq / L
[0019]
[Example 7] Pathophysiological improvement effect by GWT of bifidobacteria preparation (Bifidobacterium longum, infantise) ineffective patient 54-year-old woman with Crohn's disease, bifidobacteria preparation 3 g daily for more than 1 year to maintain remission However, CRP was about 5.0 μg / mL (index 5.0) and hemoglobin was 8.4 g / dL, which was not improved from the abnormal value. Discontinuation of bifidobacterial preparation, GWT 6 tablets a day (1.5 g), 3 doses continued for 1 month. As a result, CRP decreased to 2.1 μg / mL (index 2.1) and hemoglobin was 9.4 g. The patient recovered to / dL, and significant progress was made in improving the condition.
[0020]
[Example 8] Pathophysiological improvement effect by GWT of bifidobacteria preparation (Bifidobacterium longum, infantise) ineffective patient A 29-year-old male with Crohn's disease, 3 g of bifidobacteria preparation daily for 3 months or more to maintain remission However, there was a great feeling of general fatigue and a significant decrease in QOL. As a result of discontinuing the bifidobacterial preparation and taking 3 tablets of GWT (1.5 g) daily for 3 months for 3 months, the feeling of fatigue was significantly reduced and systemic vitality returned.
[0021]
[Example 9] Improvement of pathological condition by GWT in patients with ineffective butyric acid preparations 35-year-old Crohn's disease male, colostomy, 3 g of butyric acid preparation 3 times a day for 3 months from October 1, 2002 to maintain remission The dose was improved and CRP 0.3 μg / mL (index 0.3) and serum albumin 4.0 g / dL were improved, but hemoglobin 10.4 g / dL was not improved. Further, fecal excretion was large and there was a problem with QOL. As of January 2003, the administration of butyric acid was discontinued and 6 tablets (1.5 g) of GWT per day, followed by 3 doses for 3 months showed no significant change in CRP. A slight increase in g / dL and a significant increase in hemoglobin (13.4 g / dL) were recovered. In addition, fecal excretion was significantly reduced, stool eased and QOL was greatly improved.
【The invention's effect】
The purpose of the present invention is not to be restricted by a specific medical institution or medical system due to the preventive or therapeutic agent of the present invention, but also to improve the quality of life by improving the general pathological condition as well as clinical findings such as improvement of feces. Simple and safe home therapy is possible, and it has become possible to provide a preventive or therapeutic agent for inflammatory bowel disorder that can be taken orally.

Claims (1)

A prophylactic or therapeutic agent for inflammatory bowel disorder comprising a koji mold culture powder and a lactic acid bacteria culture powder.