JP2001017122A - Food for preventing inflammation of digestive system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject food effective for preventing inflammation of a digestive system caused by bacteria belonging to the genus Helicobacter by bringing the food to contain an aggregating factor against bacteria of the genus helicobacter, a bactericidal factor against the bacteria of the genus Helicobacter and the like. SOLUTION: This food for preventing inflammation of a digestive system contains (A) preferably a polyclonal antibody originated from avian eggs which is an aggregating factor against bacteria belonging to the genus Helicobacter, (B) preferably a polyphenol compound [e.g. (+)-catechin, etc.], which is a germicidal factor against bacteria of the genus Helicobacter and (C) a milk protein or an egg protein (e.g. casein, etc.). The aforesaid bacteria of the genus Helicobacter is preferably one or more selected from helicobacter pylori, Helicobacter cinaedi, Helicobacter fenneliae and the like. The oral dose of the ingredient A is preferably, e.g. 0.1 mg-0.1 g/kg body weight in case of a polyclonal antibody originated from hen eggs having >=256 aggregated antibody titer.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a food for preventing inflammation of the digestive system, which comprises using an aggregating factor against Helicobacter bacteria, a bactericidal factor against Helicobacter bacteria, and milk protein or egg protein in combination.

[0002]

BACKGROUND ART The gastrointestinal system has gastritis, stomach and duodenal ulcer,
Gastric cancer, hyperacidity, acute appendicitis, intestinal tuberculosis, imported infectious disease,
There are many diseases such as ischemic bowel disease. Until now, the cause of many of these diseases is unknown, and various factors such as stress, diet, genetic predisposition, personal constitution, drinking, and smoking have been mentioned. In addition, the stomach is considered to be sterile for a long time because the stomach is in a strongly acidic state due to secretion of stomach acid. However, in 1983 Mershall and Warren showed gastritis and Helicobacter pylori was detected in gastric biopsies from gastric ulcer patients at a high rate (Warren JR, Maskall B).
J: Lancet, 1273-1275 (1983))
Since it was reported that diseases that were thought to be caused by eating habits and stress, such as gastritis, stomach or duodenal ulcer, have been gradually revealed to be actually related to Helicobacter bacteria. At present, eradication of Helicobacter bacteria as a treatment for gastric ulcer is considered to be a complete treatment for preventing recurrence of ulcer. Various methods such as antibiotics, herbal medicines, and food ingredients have been considered as eradication therapy for Helicobacter bacteria. Helicobacter bacteria are susceptible to antibiotics such as penicillin, cephalosporin, macrolide, and nitroimidazole in vitro. However, in vivo, even if the drug is administered alone, a sufficient eradication effect cannot be obtained. As a reason,
It is considered that the antibacterial activity of the administered antibiotic is attenuated by stomach acid, that the effective concentration of antibiotics does not reach the cells present in the mucus layer, that the cells acquire drug resistance, etc. . Therefore, treatments using a combination of several antibiotics have been performed. When three agents were used in combination, a high eradication rate could be obtained, but the incidence of side effects such as diarrhea was high, which led to a decrease in drug compliance, and the incidence of resistant bacteria was high. Can not.

[0003] Subsequently, a new acid secretion inhibitor, proton pump inhibitor (PPI), was found to have an antibacterial effect against Helicobacter bacteria, and was used as an eradication therapy. However, since the antibacterial activity of PPI is lower than that of antibiotics, a method of removing bacteria by adding antibiotics and antiprotozoal agents has been adopted. As antibiotics, β-lactam agents (penicillin, ampicillin, etc.), macrolide agents (erythromycin, clarithromycin, etc.), aminoglycoside agents (streptomycin), tetracycline agents, bismuth agents have been used. At present, three new
The combination of PPI and three antibiotics, which is a combination therapy, is predominant, has a considerably higher eradication effect, and has a lower incidence of side effects than the previous triple therapy. However, this method still has problems of producing antibiotic-resistant bacteria and problems of side effects, so that long-term administration cannot be performed, and it cannot be said that it is an absolutely safe treatment. A high method is eagerly awaited. Therefore, as a result of studying the eradication effect of Helicobacter against foods and herbal medicines, tea, sialyl lactose, propolis, lactoferrin, vitamin C, E, garlic, algae, lactic acid bacteria, garcinia, pepper, senna, aloe, lemon Many foods, such as rosemary, mugwort, sagebrush, and life-prolonging herbs, and the herbicidal effect of Chinese herbs have been reported. For example, in tea, a polyphenol compound of a green tea component suppresses the growth of Helicobacter sp., Suppresses the adhesion of Helicobacter sp. To gastric mucosa and duodenal mucosa (Japanese Patent Laid-Open No. 5-139977), and has a bactericidal effect against Helicobacter sp. (Yoshito Komatsu, Daizo Saito, Atsushi Hirayama, etc .: Progress of
Digestive Endoscopy, 47, 1
36-137 (1995)) (Katsu Kato, Takeshi Noda, Daizo Saito, et al .: Effects of tea polyphenols on Helicobacter pylori, 53rd Annual Meeting of the Cancer Society of Japan, p117 (1994)). On the other hand, it has been reported that a mammalian antibody or an avian-derived antibody immunized with a specific bacterium is effective for prevention of infection and eradication of the bacterium (H. Hatta, M. Kim and T. Yamada).
moto: Japanese Journal ofD
airy and Food Science, 41
(16), 217-221 (1992)) Also, it has been argued that mammalian antibodies or bird-derived antibodies are safe and effective against the present Helicobacter bacteria (Japanese Patent Laid-Open No. 4-275232). ). However, there is room for practical improvement in the above-mentioned methods, and there is a need for a more effective and safe method for removing bacteria.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide an effective and safe food for preventing digestive inflammation caused by Helicobacter bacteria.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION The present inventors have conducted intensive studies in order to solve the above-mentioned problems. As a result, a factor having an agglutinating activity against Helicobacter bacteria, a factor having a bactericidal effect against Helicobacter bacteria, and milk protein Alternatively, it has been found that by using egg proteins in combination, high-efficiency eradication can be achieved while ensuring high safety, and the present invention has been completed. That is, the present invention provides an aggregating factor for Helicobacter bacteria,
A food for preventing digestive system inflammation, comprising a bactericidal factor against Helicobacter bacteria and milk protein or egg protein.

The genus Helicobacter according to the present invention is Helicobacter pylori.
r pylori), Helicobacter sinaedi (H
elobacter cinaedi, Helicobacter fenneliae
fennelliae), Helicobacter heilmanni
i), Helicobacter rapini
cter rappini) and at least one member selected from the group consisting of Helicobacter felis. The aggregating activity in the present invention refers to the aggregating force generated by specifically binding to Helicobacter bacteria. Specifically, it refers to the cohesive force caused by the antigen-antibody reaction. The agglutinating activity factor in the present invention is not particularly limited, and examples thereof include avian eggs, mammalian colostrum, normal milk, and polyclonal antibodies produced by organisms contained in blood. In particular, polyclonal antibodies obtained from chicken eggs are highly safe and can be mass-produced, so that they can be suitably used for the purpose. Furthermore, polyclonal antibodies obtained from chicken eggs, whole eggs or egg yolk liquid of chickens immunized with an inactive cell of Helicobacter bacteria as an antigen, or powder or egg yolk liquid dried and powdered by ordinary methods such as spray drying Egg yolk water-soluble protein powder obtained by removing egg yolk lipoprotein using carrageenan or the like, or yolk water-soluble protein as ion-exchange chromatography, gel filtration, sodium sulfate salting-out, ammonium sulfate salting-out, etc. A variety of forms and purity can be used, such as a purified chicken egg antibody purified by a known protein purification method. The purity of the hen's egg antibody of the various prepared samples obtained in this way is calculated as the amount of the hen's egg antibody with respect to the weight of the powder. 8 to 30%, and 95% or more in the form of a purified chicken egg antibody. In general, the titer of polyclonal antibody is determined by boosting the target animal with a booster.
Can be increased. In addition, polyclonal antibodies obtained from bird eggs using a diet supplemented with Miyai bacteria (Japanese Patent Laid-Open No. Hei 5-227899), lactic acid bacteria, chitin, chitosan, dietary fiber, sodium alginate, yoke peptide, etc., which promote immunological activity, It is known that high antibody titers are easily obtained, and in practicing the present invention, it is possible to obtain a polyclonal antibody using the above-mentioned method without any problem.

The bactericidal factors in the present invention are not particularly limited, but include tea, sialyl lactose, propolis, lactoferrin, vitamins C and E, garlic, algae, lactic acid bacteria, garcinia, pepper, senna, aloe, lemon, and rose. Extracts such as marie, mugwort, gajutsu, and life-extinging grasses, and active ingredients such as powders are included. In particular, tea-derived polyphenol compounds (+)-catechin, (-)-gallocatechin, (-)-gallocatechin gallate, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin , (-)-Epigallocatechin gallate, free theaflavin, theaflavin monogallate A, theaflavin monogallate B, and one or more compounds selected from the group consisting of theaflavin gallate can be added to tea for daily drinking. It has been reported that it is contained and has a very strong bactericidal effect against Helicobacter bacteria (Yoshito Komatsu, Daizo Saito, Atsushi Hirayama, et al .: Progress of Diges)
five Endoscopy, 47, 136-137
(1995)) (Katsu Kato, Takeshi Noda, Daizo Saito, others: He
Effect of tea polyphenol compounds on L. pylori, Article of the 53rd Annual Meeting of the Japanese Cancer Society,
p117 (1994)), it is desirable to use a tea-derived polyphenol compound. The tea-derived polyphenol compound is preferably a hot water extract of tea leaves, but can also be obtained from a water or alcohol extract, or may be of another origin or a chemically synthesized product. Examples of the raw tea leaves include fresh tea leaves, unfermented tea, semi-fermented tea, fermented tea, sencha, instant green tea, and the like. Such tea-derived polyphenol compounds are contained in large amounts as components of tea, and are effective in preventing hyperlipidemia and cancer. This security is very high because it is used. An example of the method for preparing the polyphenol compound of the present invention is disclosed in detail in patents (JP-A-2-6499, 63-214183). The milk protein or egg protein of the present invention is not particularly limited, but ovalbumin, ovotransferrin, ovomucoid, ovomucin, lysozyme and the like in egg protein, casein, α-lactoglobulin, β-lactoglobulin and the like in milk protein. Is mentioned.

In the present invention, the concentration of the agglutinating activator is not particularly limited as long as it is higher than the concentration that specifically agglutinates Helicobacter bacteria, and may be appropriately selected according to the dosage according to the dosage form. For example, an aggregate antibody titer of 256
In the case of the above-mentioned chicken egg-derived polyclonal antibody, when 0.1 mg to 0.1 g / kg of body weight per day is orally administered as a chicken egg antibody, it exerts a specific aggregating effect on Helicobacter bacteria and has no side effects. The concentration of the bactericidal factor is
There is no particular limitation as long as the concentration is higher than the concentration that can kill Helicobacter bacteria in the digestive tract. In the case of tea-derived polyphenols, oral administration of 0.001 to 0.03 g / kg of body weight per day produces a specific and strong therapeutic and preventive effect on inflammation of the upper gastrointestinal tract such as gastritis and duodenal ulcer, And there are no side effects. Further, the dose of milk protein or egg protein is not particularly limited as long as it can maintain the pH of the digestive tract in an acidic environment at 3 or more. The use forms of the present invention include tablets, granules, capsules, syrups, drinks or various foods (ice cream, yogurt, shortcake, gum and other frozen desserts, confectionery, prepared milk powder, milk, cocoa, coffee, etc. Beverages, margarine, butter, cheese, baby food, etc.) forms are possible. Hereinafter, the present invention will be described in detail with reference to examples. However, these do not limit the invention.

[0009]

[Embodiment 1] Preparation of Helicobacter pylori Helicobacter pylori ATCC4
Using 3504 cells, select medium (Blood Agar)
Base No. 2 (manufactured by OXOID) and 7% horse defibrinated serum (manufactured by NBL), 37 ° C., 10% CO ₂
The cells were cultured under the conditions for 5 days. Embodiment 2. FIG. Preparation of Anti-H. Pylori Egg Yolk Antibody Helicobacter pylori described in Example 1 was prepared so as to contain about 108 cells per ml, heated at 60 ° C. for 30 minutes, and used as an antigen solution. One laying hen was injected intramuscularly with 1 ml of the antigen solution, and then 8 weeks later, the antigen solution was administered again (booster). From the 4th week after the booster, chicken eggs were collected for 3 months and the yolk was separated. The yolk was homogenized with a homomixer, and this solution was used as an egg yolk solution. 1 kg of water was added to 1 kg of the obtained egg yolk solution to homogenize, and 4 kg of a 0.15% λ-carrageenan aqueous solution was added thereto, followed by stirring and left to stand for 2 hours. 8000r after standing still
pm 20 minutes centrifugation, about 5 kg from the supernatant
Of yolk water-soluble protein was obtained. 150 g of sodium sulfate was added little by little to 1 L of the obtained egg yolk water-soluble protein solution to dissolve it, and then allowed to stand for 30 minutes and centrifuged (normal temperature, 8000 rpm).
× 15 minutes). Discard the supernatant and add 10 mM to the precipitate.
200 ml of Na ₂ HPO ₄ buffer was added and dissolved.
Using the obtained solution, salting out was performed again in the same manner as described above. This solution was dialyzed overnight using a 10 mM Na ₂ HPO ₄ buffer. The obtained solution was freeze-dried to obtain an anti-H. Pylori egg yolk antibody. The obtained powder was confirmed by gel filtration ("Protein I", edited by The Japanese Biochemical Society, Chapter 11, Tokyo Kagaku Dojin (1990)) to have a ratio of antibody to total protein of 90% or more.

Test Example 1 Aggregation activity of anti-H. Pylori egg yolk antibody The agglutination antibody titer of the anti-H. Pylori egg yolk antibody obtained in Example 2 was measured using the cells obtained in Example 1. S. as control. Similarly, the agglutinating antibody titer of the anti-H. pylori egg yolk antibody obtained in Example 2 was measured using mutans bacteria (JP-A-4-71465), and the results are shown in Table 1.

[0011]

[Table 1]

Embodiment 3 FIG. Preparation of Polyphenol Compound To 1 kg of commercially available green tea, about 15 L of water was added, stirred, and extracted at 80 ° C. for 3 hours. The extract obtained by filtration was concentrated to dryness to obtain 350 g of a hot water extract of green tea (purity 38% as a mixture of polyphenol compounds). This hot water extract 3
8 L of water was added to 50 g to dissolve, and the mixture was sequentially partitioned with hexane and chloroform. 10 L of ethyl acetate was added to the aqueous layer after distribution, and the mixture was vigorously stirred and allowed to stand. The ethyl acetate layer was separated, and the ethyl acetate was distilled off, followed by drying to obtain 70 g of an ethyl acetate-soluble fraction (as a mixture of polyphenol compounds). Purity 74.
5%). The proportion of each polyphenol compound in the present ethyl acetate-soluble fraction was (+)-catechin 3.5%, (+)-gallocatechin 14.8%, (-)-gallocatechin gallate 1
1.6%, (-)-epicatechin 7%, (-)-epicatechin gallate 4.6%, (-)-epigallocatechin 1
5% and (-)-epigallocatechin gallate 18%. 10 g of the obtained ethyl acetate-soluble fraction was subjected to silica gel column chromatography (solvent, chloroform: methyl alcohol, 20: 1, 10: 1, v / v) Sephadex LH-20 column chromatography (solvent,
Methyl alcohol) and recycle HPLC (LC-908, GS-320 column, solvent methyl alcohol, manufactured by Nippon Kagaku Kogyo Co., Ltd.) are sequentially used to obtain (+)-
0.3 g of catechin, 1.22 g of (+)-gallocatechin,
0.9 g of (-)-gallocatechin gallate, 0.5 g of (-)-epicatechin gallate, 0.1 g of (-)-epicatechin gallate.
38 g, (−)-epigallocatechin gallate 1.2 g, and (−)-epigallocatechin gallate 1.5 g polyphenol compound were obtained.

Embodiment 4 FIG. Preparation of Polyphenol Compound 1 kg of commercially available green tea was extracted with 20 L of hot water at 85 ° C. while stirring for 30 minutes, and the tea leaves were removed by filtration to obtain a 17 L extract. This solution is passed through an ultrafiltration device (DDS, membrane type GR
-81PP, fractionation molecule 6000) using a flow-through of 15 L
I got 5 L of water was added to the concentrated residue, and the same operation was carried out to obtain 6 L of the passing solution. The two solutions were combined and concentrated with a reverse osmosis membrane (manufactured by DDS, membrane type HC-50) to 1 L to obtain 233 g of a polyphenol compound having a purity of 35%. The obtained concentrated product is allowed to flow through a column filled with an adsorption resin (Duolite S-876, manufactured by Sumitomo Chemical Co., Ltd.), adsorbed, washed with deionized water, eluted with 50% ethanol, and concentrated under reduced pressure to remove ethanol. The resulting solution was distilled off to form a concentrated aqueous solution, and then freeze-dried by a conventional method to obtain 70 g of a polyphenol compound having a purity of 74.5%. The components of the obtained polyphenol compound were (+)-catechin 3.5%, (+)-gallocatechin 14.8%, and (-)-gallocatechin gallate.
6%, (-)-epicatechin 7%, (-)-epicatechin gallate 4.6%, (-)-epigallocatechin 15%
And (-)-epigallocatechin gallate 18%.

Test Example 2. Helicobacter pylori eradication effect test using Helicobacter pylori-infected gerbils Mongolian gerbil (Me
riones unguiculatus) MGS / S
ea (SPF)), a 7-week-old male (manufactured by CLEA Japan, Inc.), and the H. cerevisiae obtained by the method described in Example 1. p
yoriri's method (Journal of G)
astroenterology, 31 (5), 755
(1996)). pylori infection. One month after administration, the above gerbils infected with H. pylori were divided into five groups (1). Physiological saline, (2). milk,
(3). Milk + anti-H. Pylori IgY solution obtained in Example 3 (20 μg / 10 g BW), (4). Milk + polyphenol obtained in Example 4 (40 μg / 10 g
B. W. ), (5). Milk + anti-H. Pylori IgY obtained in Example 3 (20 μg / 10 g BW) + Example 4
(40 μg / 10 g B.I.)
W. )) Were administered twice a day for 20 days. After sacrifice, the stomach is opened, gross findings, CLO test of gastric surface tissue (manufactured by Kokusai Reagent Co., Ltd.), and H. pylori culture method in gastric tissue (Skirow medium, 37 ° C., 48 hours, microaerobic culture) The number of positive animals was counted and the eradication effect (the number of positive animals / the total number of animals (12)) was determined. The results are shown in Table 2.

[0015]

[Table 2]

From the results shown in Table 2, it was not possible to eradicate H. pylori bacteria infected with the stomach by IgY alone, but it was possible to effectively eliminate H. pylori by using a food and a polyphenol compound in combination. .

The embodiments of the present invention and the desired products are as follows. (1) A food for preventing gastrointestinal inflammation, comprising a combination of an aggregating factor against Helicobacter bacteria, a bactericidal factor against Helicobacter bacteria and milk protein or egg protein. (2) The food for preventing gastrointestinal inflammation according to (1), wherein the agglutinating factor is a polyclonal antibody derived from avian eggs. (3) The food for preventing digestive inflammation according to any one of (1) and (2), wherein the bactericidal factor is a tea-derived polyphenol compound. (4) The bacterium of the genus Helicobacter is at least one selected from the group consisting of Helicobacter pylori, Helicobacter sinaedi, Helicobacter fenneliae, Helicobacter helmannii, Helicobacter rapini, and Helicobacter ferris.
(3) The food for preventing digestive system inflammation according to any of the above.

[0018]

According to the present invention, it has been shown for the first time that a strong eradication effect against Helicobacter bacteria is exerted by using an agglutinating factor, a bactericidal factor and a milk protein or egg protein in combination with Helicobacter bacteria. The invention leads to a safe and effective method for eradication of Helicobacter bacteria.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A61K 35/78 A61K 35/78 X 38/00 39/395 D 39/395 45/06 45/06 A61P 1/04 A61P 1/04 31/04 31/04 A61K 37/02 F term (reference) 4B018 LB08 MD08 MD20 MD49 MD72 ME09 ME11 4C084 AA02 AA23 BA44 CA41 DA36 DA38 DA41 DC01 DC50 MA02 MA52 NA06 NA07 ZA661 ZA681 ZB352 4C085 AA13 AA17 BA20 CC04 CC04 EE03 GG08 4C088 AB45 AC05 CA05 MA02 MA52 ZA66 ZA68 ZB35 4C206 AA01 AA02 CA19 MA03 MA72 ZA66 ZA68 ZB35

Claims (5)

[Claims] 1. A food for preventing gastrointestinal inflammation, comprising a coagulation factor for Helicobacter bacteria, a bactericidal factor for Helicobacter bacteria, and milk protein or egg protein. 2. The food for preventing digestive system inflammation according to claim 1, wherein the agglutinating factor is a polyclonal antibody derived from avian eggs. 3. The food for preventing gastrointestinal inflammation according to claim 1, wherein the bactericidal factor is a polyphenol compound. 4. The food for preventing digestive system inflammation according to claim 1, wherein the bactericidal factor is a tea-derived polyphenol compound. 5. The bacterium of the genus Helicobacter is at least one selected from the group consisting of Helicobacter pylori, Helicobacter sinaedi, Helicobacter fenneliae, Helicobacter helmannii, Helicobacter rapini, and Helicobacter ferris. The food for preventing digestive system inflammation according to any one of the above.