JP3249778B2 - Anti-Helicobacter pylori agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an anti-Helicobacter pylori agent.

[0002]

2. Description of the Related Art Helicobacter pylori
Bacter pylori was introduced in 1983 by Warren and Marshall.
l) is a gram-negative spiral bacillus specifically found in the antrum of the stomach pylorus discovered by [Lancet (La).
ncet), i, 1273-1275 (1983)].

[0003] Helicobacter pylori is considered to be strongly involved in the onset of gastritis, peptic ulcer, etc. since it is detected at high rates in the gastric mucosa of patients. Recently, involvement in gastric cancer has been suggested, and social interest has been increasing. Helicobacter pylori eradication therapy has been actively pursued in Europe and the United States, and the results indicate that eradication of Helicobacter pylori is extremely effective in preventing recurrence of gastric ulcer and duodenal ulcer.

[0004]

To eradicate Helicobacter pylori, β-lactam antibiotics, tetracycline antibiotics, macrolide antibiotics, and new quinolone antibacterial agents are used. The effect was not enough and a large amount of drug had to be used for a long time to completely eradicate the bacteria. In addition, taking a large amount of antibiotics for a long period of time is not preferable because side effects may occur.

[0005] Therefore, there has been a demand for a substance having no or few side effects and showing a strong anti-Helicobacter pylori action. The present invention addresses this need.

[0006]

BACKGROUND OF THE INVENTION The present inventors have focused on garlic having a strong antibacterial property and searched for a substance having an anti-Helicobacter pylori action. As a result, garlic extracted in fats and oils by mixing ground garlic with fats and oils Found that the oil-soluble fraction significantly inhibited the growth of Helicobacter pylori, thereby completing the present invention.

Incidentally, Cellini et al.
The anti-Helicobacter pylori action of a freeze-dried product of squeezed juice obtained by pulverizing garlic and removing the fiber content has been reported [FEMS Immunology and Medical.
Microbiology (FEMS Immuno. Md.
Microbiol. ), 13, 273-277 (19)
96)]. However, raw garlic and its squeezed juice have an antibacterial effect, but are strongly irritating, and if consumed too much, may cause ulcers in the digestive organs such as the esophagus and stomach, and thus cannot be used directly.

[0008]

The anti-Helicobacter pylori agent according to claim 1 for solving the above-mentioned problems comprises an oil-soluble fraction of garlic as an active ingredient. The anti-Helicobacter pylori agent according to claim 2 is the claim 1.
The anti-Helicobacter pylori agent, wherein the garlic is
Oil-soluble fraction of garlic or its juice
Fat and oil-soluble fraction extracted into the fat and oil.
And features.

The anti-Helicobacter pylori agent according to claim 3 is the anti-Helicobacter pylori agent according to claim 1 , wherein the oil-soluble fraction of garlic is obtained by mixing crushed garlic or its squeezed oil with fat or oil. After extraction into fats and oils
It is an oil-soluble fraction separated from fats and oils .

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The anti-Helicobacter pylori agent of the present invention is characterized in that an oil-soluble fraction of garlic is used as an active ingredient, and garlic (Allium sativum) is used.
L. Any kind of) is acceptable.

Similarly, the method of extracting the oil-soluble fraction is not limited, but the method of processing garlic and the method of producing ajoene-containing fats and oils disclosed in, for example, Japanese Patent No. 2608252 can be applied. This patent No. 2608252
The method of No. 1 is a method in which crushed garlic or its squeezed juice is mixed with fats and oils to produce and accumulate ajoene at a high concentration in the fats and oils. The present invention can be said to give a novel use to ajoene-containing fats and oils produced by the method of Japanese Patent No. 2608252, but the active ingredient of the anti-Helicobacter pylori agent of the present invention is not limited to ajoene.

The oils and fats used for extracting the oil-soluble fraction of garlic by the method of Japanese Patent No. 2608252 or a method similar thereto are not particularly limited, but are considered harmless to the human body when taken orally. Are preferred. The garlic crushed (cut or crushed) may be directly mixed with fats and oils, but in order to efficiently obtain the active ingredient, that is, the oil-soluble fraction, an appropriate amount (for example, equal to or less than garlic) of garlic is used. It is effective to use squeezed juice obtained by pressing after crushing by adding water. When the squeezed juice is mixed with the oil and fat and allowed to stand, it is separated into an oil layer and an aqueous layer, but the oil-soluble fraction is dissolved in the oil layer. The oil-soluble fraction can be efficiently extracted by leaving it for 3 hours or more, though it depends on the treatment temperature and pH value at that time. The oil layer can be separated from the aqueous layer by a known method such as decantation.

[0013] The oil layer thus obtained has anti-Helicobacter pylori activity, which is
It can be used as an active ingredient of a pylori agent. In addition,
By removing trace amounts of water and solids contained in the oily layer, garlic odor can be reduced and storage stability is improved.

Further, by a technique such as liquid distribution or liquid chromatography generally used for substance purification, all or one of the oil-soluble fraction of the garlic is more than the oil-soluble fraction containing the oil-soluble fraction (anti-Helicobacter pylori component). Part can be extracted. The anti-Helicobacter pylori agent of the present invention may be obtained by using an oil-soluble fraction of garlic as an active ingredient, separating and purifying the oil-soluble fraction (all or a part of the components) from fats and oils, or using an oil-soluble fraction. A fat or oil in which the components are dissolved may be used, or both may be used in combination.

The anti-Helicobacter pylori agent of the present invention comprises:
Allows addition and mixing of excipients, binders, diluents, etc. Also,
It may be compounded with other drugs as needed. The administration form is not limited, and it can be administered orally or parenterally, for example, in the form of powder, granules, tablets, capsules, syrups, injections and the like.

The present preparation can also be prepared as general foods. In that case, the oil-soluble fraction or the fat or oil in which the oil-soluble fraction is dissolved can be directly added to any food, or can be processed as powder, granules, bulk solid food or beverage food, Things or fragrances may be added. Alternatively, an excipient, a binder,
Diluents and the like can be mixed and processed into health foods in the form of, for example, powders, granules, tablets, capsules, and beverages.

Next, embodiments of the present invention will be described in more detail with reference to examples.

[0018]

Example 1 (Preparation) An oil or fat containing an oil-soluble fraction of garlic was prepared according to the method of Japanese Patent No. 2608252.

First, 300 g of water was added to 1 kg of raw garlic (6 pieces of white), and a food processor (Cuisin) was added.
The mixture was pulverized using a DLC-X PLUS type (manufactured by Art Co., Ltd.) and squeezed by hand using a nylon filter cloth to obtain 800 g of juice. From this, take 5 samples of 80g each, 80g each
Medium-chain fatty acid triglyceride (MCT) (manufactured by NOF CORPORATION, trade name: Panasate 810), refined rapeseed oil (manufactured by Nisshin Oil Co., Ltd.), refined corn oil (manufactured by Ajinomoto Co., Ltd.), refined safflower oil (manufactured by Ajinomoto Co., Ltd.) ) And salad oil (soybean oil: rapeseed oil = 4: 6, manufactured by Nisshin Oil Co., Ltd.) and a homomixer (M type manufactured by Tokushu Kika Kogyo Co., Ltd.)
After mixing, the mixture was kept at 37 ° C. for 24 hours to separate an upper oil layer. The water and solids contained in the oil layer were adsorbed on Celite (manufactured by Wako Pure Chemical Industries, Ltd.) and removed with a filter paper. As a result, 70 g of each of clear oils and fats containing a component having an anti-Helicobacter pylori action was obtained. (Measurement of anti-Helicobacter pylori activity) The anti-Helicobacter pylori activity of the oil and fat obtained above was measured by calculating the minimum growth inhibitory concentration.

That is, 0.25% of yeast extract (manufactured by Difico), 10% of fetal bovine serum (manufactured by Wako Pure Chemical Industries, Ltd.), and agar (manufactured by Wako Pure Chemical Industries, Ltd.) were used in 1.%.
An agar medium was prepared by adding anti-Helicobacter pylori components at various concentrations to a Brain Heart Infusion medium (manufactured by Difico) added to 5%, and a culture solution of Helicobacter pylori (10 ⁸⁾ was prepared. Cells / ml) and appropriately spread 1000 Helicobacter pylori on 1 agar medium, and at 37 ° C under microaerobic conditions.
After culturing for 3 days, the number of colonies on the agar medium is counted, and the concentration at which no colony grows is defined as the growth inhibitory concentration.
This minimum was determined.

The results are as shown in Table 1 below.
From these results, it was found that the oil or fat containing the oil-soluble fraction of garlic inhibited the growth of Helicobacter pylori and inhibited the growth of 25 to 35 mg / g.
It is found to be effective at a concentration of ml.

[0022]

[Table 1]

[0023]

Example 2 To 1 kg of fresh garlic, 300 g of water was added, and crushed and squeezed in the same manner as in Example 1 to obtain 800 g of squeezed juice. To this was added 800 g of MCT, and after mixing as in Example 1,
The mixture was kept at 37 ° C. for 24 hours to separate an upper oil layer. In the same manner as in Example 1, water and solids contained in the oil layer were adsorbed on celite and removed with filter paper. As a result, 750 g of a clear fat or oil containing a component having an anti-Helicobacter pylori action was obtained.

750 g of this fat and oil and 75 g of silica gel (Wako Gel C-10, manufactured by Wako Pure Chemical Industries, Ltd.)
0) was mixed to adsorb the active ingredient onto silica gel, and the silica gel separated from the fat was washed with 300 ml of hexane to remove the fat. Next, the silica gel was added to ethanol 30
After washing with 0 ml, the adsorbed components were eluted. Further, ethanol was removed under reduced pressure to obtain 18.8 g of the active ingredient.

The minimum growth inhibitory concentration of this active ingredient was measured in the same manner as in Example 1. The results are as shown in Table 2 below. The results show that the garlic oil-soluble fraction was extremely effective in inhibiting the growth of Helicobacter pylori, and the minimum inhibitory concentration for the three strains tested was 750 μg /
ml. This was 2.5 to 6.5 times higher than the antibacterial activity against the Helicobacter pylori of the freeze-dried garlic juice by Cellini et al.

[0026]

[Table 2]

[0027]

Example 3 Further, the active ingredient was subdivided and the minimum growth inhibitory concentration was measured. The active ingredient was adsorbed on silica gel in the same procedure as in Example 2 and washed with hexane.
The silica gel was washed successively with 200 ml of hexane containing 200%, 10%, 20%, and 50% of ethanol and 200 ml of 100% ethanol to elute the components. After removal of the solvent, the recovered components were weighed to find 12.3 g for the 5% ethanol fraction and 4.8 for the 10% ethanol fraction.
g, 1.3 g in the 20% ethanol fraction, 0.3 g in the 50% ethanol fraction, and 0.1 g in the 100% ethanol fraction. These minimum growth inhibitory concentrations were measured in the same manner as in Example 1.

The results are shown in Table 3 below. 400-450 μg / ml for 5% ethanol fraction, 200-250 μg / ml for 10% ethanol fraction, 110-120 μg / ml for 20% ethanol fraction, 50 μg / ml for 100% ethanol fraction, 100% ethanol fraction Per minute, the growth of Helicobacter pylori was inhibited at a concentration of 50-60 μg / ml. Based on these results, all fractions were obtained from Helicobacter
It was found to be effective against H. pylori.

[0029]

[Table 3]

Although the embodiments of the present invention have been described with reference to the embodiments, the present invention is not limited to such embodiments, and it is possible to implement variously without departing from the gist of the present invention. Needless to say.

[0031]

As described in detail above, the anti-Helicobacter pylori agent according to claim 1 can inhibit the growth of Helicobacter pylori. Garlic as claimed in claim 2
Crushed garlic or its juice from oil-soluble fraction
Fats and oils, and collect as oil-soluble fraction extracted in the fats and oils.
It does not require special processing operations and can handle large amounts of processing.
Large amount of anti-Helicobacter pylori drug
It can be provided and is suitable for industrialization.

The oil-soluble fraction of garlic is mixed with crushed garlic or its squeezed oil and extracted with the oil and fat, and then separated from the oil and fat to extract the oil-soluble fraction. No special processing operations are required,
In addition, since it can be processed in a large amount, it is possible to provide a large amount of an anti-Helicobacter pylori agent, which is suitable for industrialization.

────────────────────────────────────────────────── ─── Continuation of the front page (56) References JP-A-8-84570 (JP, A) Applied and Environmental Microbiology, 1996, Vol. 62, No. 11, p. 4238-4242 Journal of the Japanese Society of Gastroenterology, 1996, Vol. 93, no. 9, p. 688 (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 35/78 A01N 65/00 BIOSIS (STN) CA (STN) MEDLINE (STN)

Claims (3)

(57) [Claims] 1. An anti-Helicobacter pylori agent comprising an oil-soluble fraction of garlic as an active ingredient. 2. The anti-Helicobacter piro according to claim 1.
In the repellent, the oil-soluble fraction of the garlic was pulverized.
Mix the garlic or its juice with the fat and extract into the fat and oil
An anti-Helicobacter pylori agent, which is the oil-soluble fraction thus obtained . 3. The anti-Helicobacter pylori agent according to claim 1 , wherein the oil-soluble fraction of garlic is obtained by mixing ground garlic or its squeezed juice with oil and fat, extracting the oil into the oil and fat, and separating the garlic from the oil and fat. An anti-Helicobacter pylori agent, which is an oil-soluble fraction.