JP2008156270A - Hyaluronidase activity inhibitor, antibacterial agent against helicobacter pylori, and bamboo extract composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a hyaluronidase activity inhibitor containing as a component a safe and easily ingestable bamboo extract composition using the skin of Phyllostachys pubescens of a natural product as a raw material, to provide an antibacterial agent against Helicobacter pylori, and to provide a safe and easily ingestable bamboo extract composition using the skin of the Phyllostachys pubescens of the natural product. <P>SOLUTION: The hyaluronidase activity inhibitor contains the bamboo extract composition obtained by adding an organic solvent or the organic solvent and water to a treated material obtained by treating the bamboo skin by a hydrolysis method to carry out the extraction as an active ingredient. The inhibitor can inhibit the development of allergy affected by the hyaluronidase activity and gastritis or the like caused by the Helicobacter pylori, because the bamboo extract composition has the hyaluronidase activity-inhibiting activity and the germicidal activity effective to the Helicobacter pylori. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

When bamboo is cut and left untreated, mold will grow on the cut surface, but it will not grow on the surface and will not rot even if left as it is for a long time. Also, when insects enter, they do not enter from the surface, but always enter the inside. From such a phenomenon, it has been known for a long time that the part called bamboo epidermis and cuticle has antibacterial activity. For this reason, bamboo extract extracted from Somune bamboo using water and organic solvents, etc. is used as an antibacterial agent, shelf life improver, and designated as a food additive. It is added and demonstrates its effect. For example, an application method (Patent Document 1) in which an extract obtained by extracting bamboo pulverized material with an organic solvent such as ethanol to dryness is used as a deodorant, and an ultrafinely pulverized bamboo epidermis and cuticle part are applied. Method of use for fabric products with excellent antibacterial and deodorizing properties (Patent Document 2), Deodorizing and disinfecting agent for external use (Patent Document 3) using an extract from organic solvent after dry-distilling partial skin particles in a high-pressure kettle In addition, a nutritional drink (patent document 4) in which bamboo skin and vinegar are mixed has been developed. It is also known that Kumassasa has an anti-inflammatory effect (Non-patent Document 1).
The present invention relates to a hyaluronidase activity inhibitor, a Helicobacter pylori antibacterial agent, and a bamboo extract composition extracted from the bamboo skin, which contain as an ingredient a bamboo extract composition extracted from the bamboo skin.

Currently, there are people with various allergies, and drugs with antiallergic effects have been developed. Even with bamboo extracts, it is known that the organic solvent extract of the sect of Buddhism bamboo or bee leaves has an antiallergic effect (Patent Documents 5 and 6).
However, among the components contained in the bamboo extract, no component that contributes to the antiallergic effect has been identified, which allergy the bamboo extract is effective against, and which component of the bamboo extract is allergic It is not completely understood whether it has an effect on.

  It may be possible to determine which allergy an anti-allergic agent has an effect on depending on the components contained in the anti-allergic agent, but it is usually determined based on the results of various tests that measure the anti-allergic effect. Has been. This is because the mechanism of action of allergy is complicated, and a test method that can legitimately evaluate all antiallergic effects has not yet been established.

As a general method for measuring an antiallergic effect, a histamine release inhibition test, a leukotriene secretion inhibition test and the like are known.
The histamine release inhibition test is a test method that measures the antiallergic effect based on the fact that many active substances such as histamine are released from mast cells and basophils have an effect on allergic symptoms. It is.
The leukotriene secretion inhibition test is a test method for measuring the antiallergic effect based on the fact that leukotriene is one of chemical mediators released from mast cells and basophils during an immediate allergic reaction.

In recent years, a method for testing hyaluronidase inhibitory activity has been developed as another test method for measuring the antiallergic effect.
Hyaluronidase is a hyaluronic acid hydrolase widely distributed in animal connective tissues. This hyaluronidase is said to increase its activity prior to histamine release in vivo. Moreover, there is a positive correlation between hyaluronidase inhibitory activity and histamine release inhibition, and antiallergic activity is expected for hyaluronidase inhibitory activity (Non-patent Document 2). The effect is measured.

  Currently, hyaluronidase activity inhibitors include hyaluronidase activity inhibitors containing theaflavins as active ingredients (Patent Document 7) and hyaluronidase activity inhibitors using acetone-insoluble components derived from cereal-derived organic solvents (Patent Document 8). Are known.

  However, an antiallergic agent containing bamboo extract as a component and having an antiallergic effect based on hyaluronidase inhibitory activity has not been reported.

  In addition, Helicobacter pylori (H. pylori) infection rate in Japan is about 50% of the total population, and Helicobacter pylori is said to be involved in more than 80% of gastritis, the National Institute of Health in the United States It is recommended that patients with gastric and duodenal ulcers should receive H. pylori eradication therapy in addition to acid secretion inhibitors. Regarding the relationship with gastric cancer, it was recognized as a group 1 of the WHO carcinogen classification in 1994 by WHO as "H. pylori causes pathological conditions leading to the development of gastric cancer."

  For the treatment of Helicobacter pylori, sterilization treatment is considered to be the most effective, but the fact that easy and insufficient sterilization treatment is performed increases the emergence of resistant bacteria and new reflux esophagitis after eradication About 10% of cases have been reported in Japan, and caution must be exercised in sterilization treatment.

  At present, it is known that bamboo extract mainly composed of carbohydrate arabinoxylan has antibacterial activity against Helicobacter pylori (Patent Document 9). Arabinoxylan is a generic name for arabinose and xylose, is the main component of hemicellulose, which is a major component of the cell wall, and is contained in hemicelluloses such as grass family plants and olive leaves.

  However, the technique of Patent Document 9 is a bamboo extract extracted with water or ethanol, and there is no example in which lipids such as glycolipids contained in the bamboo extract have reported on the antibacterial activity of Helicobacter pylori.

Japanese Patent Laid-Open No. 9-094290 JP-A-6-073665 JP-A-5-306232 JP-A-5-095769 JP-A-9-278662 JP 2003-212786 A Japanese Patent No. 3242997 Japanese Patent Laid-Open No. 2003-252778 Japanese Patent Application Laid-Open No. 2002-322079 Showa Medical Society Vol. 48 No. 5 595-600 (1988) Toshio Kakegawa et al .: Inflammation, 4,437 (1984)

  In view of the above circumstances, the present invention uses as a raw material a hyaluronidase activity inhibitor, a Helicobacter pylori antibacterial agent, and a natural product, Munetake bamboo epidermis, which contains a safe and easy-to-take bamboo extract composition made from the natural product, Munetake bamboo epidermis. An object of the present invention is to provide a bamboo extract composition that is safe and easy to ingest.

The hyaluronidase activity inhibitor of the first invention is obtained by extracting an organic solvent or an organic solvent and water from a substance to be treated obtained by treating bamboo skin by a hydrolysis method. It contains a component contained in the aqueous phase as an active ingredient.
In the present specification, the aqueous phase means a phase having polarity, and the aqueous phase is not only a phase composed of water but also a phase composed of water and a relatively strong organic solvent such as alcohol. Moreover, the phase which consists only of organic solvents with comparatively strong polarity, such as alcohol, is also contained.
The hyaluronidase activity inhibitor of the second invention is characterized in that, in the first invention, the hydrolysis method is an acid hydrolysis method using a mixed solution in which an acid, an organic solvent and water are mixed.
The hyaluronidase activity inhibitor of the third invention is characterized in that, in the first invention, the hydrolysis method is an alkali hydrolysis method using a mixed solution in which an alkali, an organic solvent and water are mixed.
The Helicobacter pylori antibacterial agent of the fourth invention is an extract of a bamboo extract composition extracted by adding an organic solvent or an organic solvent and water to a treated substance obtained by treating bamboo skin by a hydrolysis method, It contains a component contained in the organic solvent phase as an active ingredient.
The Helicobacter pylori antibacterial agent of the fifth invention is characterized in that, in the fourth invention, the hydrolysis method is an acid hydrolysis method using a mixed solution in which an acid, an organic solvent and water are mixed.
The Helicobacter pylori antibacterial agent of the sixth invention is characterized in that, in the fourth invention, the hydrolysis method is an alkali hydrolysis method using a mixed liquid in which an alkali, an organic solvent and water are mixed.
The bamboo extract composition of the seventh invention is an extract obtained by adding an organic solvent or an organic solvent and water to a material to be treated obtained by treating bamboo skin by a hydrolysis method. It is characterized by an acid hydrolysis method using a mixed solution of water, an organic solvent and water.
The bamboo extraction composition of the eighth invention is an extract obtained by adding an organic solvent or an organic solvent and water to a substance to be treated obtained by treating bamboo skin by a hydrolysis method. It is characterized by being an alkali hydrolysis method using a mixed liquid in which water, an organic solvent and water are mixed.
The bamboo extract composition of the ninth invention is the seventh or eighth invention, wherein the organic solvent is one of toluene, ethyl acetate, methanol, ethanol, or a combination of two or more of them. Features.

According to the first invention, since the bamboo extract composition has a hyaluronidase activity inhibitory action, allergies affected by the hyaluronidase activity can be suppressed.
According to the second invention, the bamboo fiber is effectively hydrolyzed by the acid, so that an effective ingredient effective for inhibiting hyaluronidase activity can be easily extracted.
According to the third invention, the bamboo fiber is effectively hydrolyzed by the alkali, so that an effective component effective for inhibiting hyaluronidase activity is easily extracted.
According to the fourth invention, since the bamboo extract composition has an effective bactericidal action against Helicobacter pylori, the occurrence of gastritis caused by Helicobacter pylori can be suppressed.
According to the fifth invention, the bamboo fiber is effectively hydrolyzed by the acid, so that an effective component effective for sterilization of Helicobacter pylori is easily extracted.
According to the sixth invention, the bamboo fiber is effectively hydrolyzed by the alkali, so that an effective component effective for sterilization of Helicobacter pylori is easily extracted.
According to the seventh invention, the bamboo extract composition contains various glycolipids effective for the living body. Therefore, if the bamboo extract composition is used as a functional food material, it is effective for the living body. Various glycolipids can be supplied to the living body. In addition, the bamboo fiber is effectively hydrolyzed by the acid, and the active ingredient is easily extracted.
According to the eighth invention, since the bamboo extract composition contains various glycolipids effective for the living body, if the bamboo extract composition is used as a functional food material, it is effective for the living body. Various glycolipids can be supplied to the living body. Moreover, the bamboo fiber is effectively hydrolyzed by the alkali, and the active ingredient is easily extracted.
According to the ninth aspect, the bamboo fiber is effectively hydrolyzed and the active ingredient is easily extracted.

  The present invention relates to a composition extracted from bamboo skin and having various functions (hereinafter referred to as a bamboo extract composition), a Helicobacter pylori antibacterial agent and a hyaluronidase activity inhibitor containing the bamboo extract composition as components. About.

Bamboo used as a raw material for the bamboo extract composition of the present invention includes various plants belonging to bamboos such as, for example, Soso bamboo and true bamboo. Bamboo is widely distributed from so-called subtropical regions to temperate regions and cold regions, and there are many varieties. Bamboos that live in any region and bamboos belonging to any varieties can be used as raw materials. It is.
Bamboo skin means the part of the blue skin mainly on the trunk of the bamboo. For example, what is cut out from the surface of the bamboo trunk with a thickness of about 0.1 to 2.0 mm can be used as the bamboo skin. At this time, even if a portion other than the epidermis in the bamboo trunk described above is included, although the difference in the yield of the bamboo extract composition is seen, the same effect as when extracted from only the bamboo trunk epidermis A bamboo extract composition that yields is obtained.

  In addition, although the bamboo skin used as an extraction raw material may be used for extraction as it is, it may be subjected to pretreatment such as drying, pulverization, and dry distillation. For example, bamboo skin powder obtained by pulverizing bamboo skin to about 50 to 400 mesh or bamboo skin chip obtained by making bamboo skin fine without pulverization can be used as an extraction raw material.

  In the present invention, the bamboo extract composition is produced from the bamboo skin through several processes. The process will be briefly described as follows.

First, a bamboo skin as an extraction raw material, for example, a crushed bamboo skin or a material obtained by cutting bamboo into an appropriate length is treated by a hydrolysis method using a hydrolysis solvent (hydrolysis step). .
During the hydrolysis, it is preferable to carry out the hydrolysis while stirring the solvent for hydrolysis in order to decompose the bamboo skin quickly and uniformly. Hydrolysis may be performed in the atmosphere at atmospheric pressure, but may be performed under an inert gas atmosphere such as nitrogen, or under a pressure pressurized to atmospheric pressure or higher.

Next, an extraction solvent is put into the material to be treated obtained by the hydrolysis treatment, and the soluble components are extracted with occasional stirring as necessary. Thereafter, the extraction solvent is filtered to remove the residue to obtain an extract, and a crude extract is obtained by removing the hydrolysis solvent and extraction solvent from this extract (extraction step).
In addition, as a substance to be treated, a hydrolysis solvent containing a solid content of bamboo skin may be used as it is, but vacuum filtration, suction filtration, filter press, cylinder press, decanter, centrifuge, filtration centrifuge, etc. It may be used after removing the residue and put into the extraction solvent.

  In addition, even if it is a crude extract, it can be used as the bamboo extract composition of the present invention, but if the crude extract is purified, impurities can be removed and the purity of the bamboo extract composition can be improved. The method for purifying the crude extract is not particularly limited. For example, purification using water washing, acetone washing, hexane washing, silica gel column, resin column, reverse phase column, etc., partitioning with solvents of different polarity, recrystallization, etc. The method can be adopted.

  Next, each step will be described in detail.

(Hydrolysis step)
As a method for performing hydrolysis, there are an acid hydrolysis method and an alkali hydrolysis method.

(Acid hydrolysis method)
First, acid hydrolysis is a hydrolysis method using an acid adjusted solvent as a solvent for hydrolysis, and an acid such as sulfuric acid, hydrochloric acid or other mineral acid is appropriately added to a mixture of water and an organic solvent. An acid hydrolysis solvent is used. If the acid hydrolysis solvent is a mixture of acid in an amount of 5% by weight or less, preferably 0.01% to 2% by weight, based on the total weight of the acid hydrolysis solvent, acid hydrolysis of the bamboo skin is performed. be able to.

In the case of acid hydrolysis, after the hydrolysis, the material to be treated is neutralized, and then extraction with an organic solvent or the like described later is performed. Any neutralization may be used as long as it is an alkali. For example, sodium hydrogen carbonate can be used.
Furthermore, any acid may be used as long as it shows acidity, and for example, mineral acids such as sulfuric acid and hydrochloric acid can be used.

(Alkaline hydrolysis method)
Alkaline hydrolysis is a hydrolysis method using a solvent adjusted to be alkaline as a solvent for hydrolysis. Alkaline hydrolysis by appropriately adding sodium hydroxide or the like to a mixed solution of water and an organic solvent. Use a solvent. If the alkali hydrolysis solvent used is a mixture of sodium hydroxide in a proportion of 10% by weight or less, preferably 0.01% to 2% by weight, based on the total weight, the alkali hydrolysis of the bamboo skin It can be performed.

In the case of alkaline hydrolysis, after the hydrolysis, the material to be treated is neutralized, and then extraction with an organic solvent or the like described later is performed. For neutralization, any acid can be used. For example, citric acid can be used.
Furthermore, any alkali may be used as long as it shows alkalinity, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium hydrogen carbonate, potassium hydrogen carbonate can be used.

An organic solvent is used for the hydrolysis solvent used in the above hydrolysis method, but the organic solvent to be used is not particularly limited. For example, alcohol such as methanol, ethanol, acetone, 1-propanol, 2-propanol, 1-butanol, 2-butanol, tert-butanol, chloroform, dichloromethane, acetonitrile, ethyl acetate, hexane, pentane, diethyl ether, petroleum, etc. Can be mentioned. Of these, ethanol is particularly preferable.
In addition, although these organic solvents may be used independently, you may use them in combination of 2 or more types.

  The weight of the solvent for hydrolysis used for hydrolysis is usually 2 to 10 times, preferably 2 to 5 times the weight of the bamboo skin as the raw material. If the amount of the solvent for hydrolysis is less than this range, there is a possibility that the solvent for hydrolysis does not reach the entire raw material and the extraction efficiency may decrease. If this amount is exceeded, the solvent for hydrolysis used for hydrolysis later This increases the burden of performing removal.

The temperature for the hydrolysis is 50 to 240 ° C, preferably 130 to 180 ° C. For example, the temperature when using a 30 wt% aqueous ethanol solution as a solvent is usually 50 to 240 ° C., preferably 130 to 180 ° C.
The time for the hydrolysis is usually 30 minutes to 24 hours, preferably 1 to 6 hours, and can be appropriately adjusted depending on conditions such as the type of solvent and temperature.
Even when the hydrolysis solvent includes a plurality of organic solvents, the same hydrolysis temperature and hydrolysis time are preferable.

  In the hydrolysis solvent, the mixing ratio of the organic solvent and water is not particularly limited. For example, when the organic solvent is ethanol, the ethanol concentration is 0.001 to 99.9% by weight, preferably 10 to 40% by weight, based on the total weight of the mixed solution before adding the acid and alkali. Can be used.

  And you may hydrolyze using the solvent for hydrolysis which consists only of the organic solvent mentioned above and water, without adding an acid and an alkali. Even in this case, the mixing ratio of the organic solvent and water is not particularly limited.

(Extraction process)
After the hydrolysis, an extraction operation is performed to extract the active ingredient from the substance to be treated using an extraction solvent such as an organic solvent.
The substance to be treated includes both a hydrolysis solvent containing a solid content of bamboo skin and a hydrolysis solvent from which a residual is removed.

In the extraction operation, the substance to be processed is put into an extraction processing tank filled with an organic solvent, and a soluble component is extracted with occasional stirring as necessary. Thereafter, the residue is removed by filtration to obtain an extract, and a crude extract is obtained by removing (evaporating) water and an organic solvent from the extract.
In addition, the removal of a residue can be performed using suction filtration, a filter press, a cylinder press, a decanter, a centrifuge, a filtration centrifuge, etc., for example.
Concentration of the extract (removal of water and organic solvent) can be performed using, for example, a vacuum concentrator such as an evaporator.

In addition, it is not necessary to employ | adopt a special extraction method also in the extraction process after acid hydrolysis or alkali hydrolysis, and arbitrary apparatuses can be used at room temperature or under heating. For example, a separatory funnel used for general extraction work can be used.
Furthermore, when extracting water and an organic solvent above the boiling point, a pressure vessel may be used.

The organic solvent used in the extraction step is not particularly limited. For example, alcohols such as methanol, ethanol, acetone, 1-propanol, 2-propanol, 1-butanol, 2-butanol, and tert-butanol, chloroform , Dichloromethane, acetonitrile, ethyl acetate, hexane, pentane, diethyl ether, petroleum and the like. Of these, ethyl acetate is particularly preferable.
In addition, although these organic solvents may be used independently, you may use 2 or more types as an extraction solvent in combination.

The number of times of performing the extraction operation on the substance to be treated is not particularly limited, and may be one time. After the first extraction, a fresh extraction solvent is added again, and the second and subsequent extraction operations are performed. You may give it.
Further, the extraction solvent used once for extraction may be reused, and the extraction operation of the substance to be treated may be performed a plurality of times.

  The bamboo extract composition extracted from the bamboo skin by the method as described above can be used as a hyaluronidase activity inhibitor or a Helicobacter pylori antibacterial agent.

Hyaluronidase activity inhibitor containing bamboo extract composition as an active ingredient is not only bamboo extract composition used alone, but also mixed with appropriate excipients such as gelatin, sodium alginate, etc., solvent such as water, alcohols, etc. In combination with a diluent such as carboxymethylcellulose.
And about the usage-amount of the hyaluronidase activity inhibitor of this invention, when it administers to a human body as a chemical | medical agent, if it is orally taken so that a daily dose may be about 0.01-30g normally, Preferably it is about 0.1-3.5g. Good. The dosage form is arbitrarily used as a powder, tablet, capsule or the like. Moreover, when mix | blending with cosmetics etc., what is necessary is just to add so that a final concentration may be set to 1-1000 ppm.

In addition, the Helicobacter pylori antibacterial agent containing the bamboo extract composition as an active ingredient is used in combination with an appropriate excipient such as gelatin, sodium alginate, water, alcohol, etc. Or a solvent such as carboxymethyl cellulose can be used in combination.
And about the usage-amount of the Helicobacter pylori antibacterial agent of this invention, when administering to a human body as a chemical | medical agent, it should just be taken orally so that a daily dose may be 0.01-500g normally, Preferably it is about 1-30g. . The dosage form is arbitrarily used as a powder, tablet, capsule or the like.

Moreover, although the aqueous phase and the organic solvent phase exist in the extract obtained in the extraction step, the aqueous phase and the organic solvent phase each contain an active ingredient extracted from the bamboo skin. The active ingredient contained in each solvent phase has a hyaluronidase activity inhibitory effect, while the active ingredient contained in a large amount in the aqueous phase has a Helicobacter pylori antibacterial effect.
For example, 孟宗竹 powder is subjected to acid hydrolysis using water, ethanol and sulfuric acid, neutralized, and then ethyl acetate is added to extract the active ingredient and filtered. Then, after filtration, among the active ingredients of the bamboo extract composition contained in the extract, the ethanol aqueous solution (aqueous phase), which is a polar solvent, contains a lot of hyaluronidase activity-inhibiting components and acetic acid, which is an organic solvent phase. The ethyl layer contains many Helicobacter pylori antibacterial components.

Therefore, if only the active ingredient contained in the aqueous phase is used, the concentration of the active ingredient having a hyaluronidase activity inhibitory effect is increased, and a hyaluronidase activity inhibitor having a higher hyaluronidase activity inhibitory effect can be produced.
Moreover, if only the active ingredient contained in the organic solvent phase is used, the concentration of the active ingredient having a high Helicobacter pylori antibacterial effect increases, and a Helicobacter pylori antibacterial agent having a higher Helicobacter pylori antibacterial effect can be produced.
It should be noted that some of the active ingredients having an inhibitory effect on hyaluronidase activity are also contained in the organic solvent phase, and some of the active ingredients having a high Helicobacter pylori antibacterial effect are also contained in the aqueous phase. Absent.

Bamboo extract compositions usually contain “lipids”, which are roughly classified into simple lipids, complex lipids, and derived lipids, of which “complex lipids” include “glycolipids”. It is.
“Glycolipid” is a general term for substances containing both water-soluble sugar chains and fat-soluble groups in the molecule, and is broadly classified into sphingoglycolipids and glyceroglycolipids by fat-soluble groups. In addition, glycosides having fat-soluble groups such as hydroxy fatty acids (for example, steryl glycosides, steroid glycosides, rhamnolipids, etc.) are also included in the glycolipids in a broad sense. In addition, what are called acidic glycolipids that are distinguished from neutral glycolipids include glycolipids having sialic acid, uronic acid, sulfuric acid, phosphoric acid, etc. (eg, ganglioside, sulfatide, sulfolipid, etc.) Included in glycolipids.
The bamboo extract composition of the present invention is presumed to contain “lipid”, and “glycolipid” is also presumed to be contained. The “glycolipid” is assumed to include any type of glycolipid other than the glycolipids described above.

Among the active ingredients of the bamboo extract composition of the present invention, “glycolipid” is mainly contained in the aqueous phase, and “lipid” that is more hydrophobic than “glycolipid” etc. is contained in the organic solvent phase. It is thought that there is.
Then, as described above, the active ingredient contained in the aqueous phase has a hyaluronidase activity inhibitory effect, and the active ingredient contained in the organic solvent phase has a Helicobacter pylori antibacterial effect. It is presumed that “glycolipid” has an inhibitory effect on hyaluronidase activity, and “lipid” that is more hydrophobic than glycolipid and the like has an antibacterial effect on Helicobacter pylori.

(Use of bamboo extract composition)
Moreover, the bamboo extract composition obtained by the method described above is useful not only as an active ingredient of a hyaluronidase activity inhibitor or a Helicobacter pylori antibacterial agent but also as a material for functional foods.
In addition, the bamboo extract composition used as a material for functional food includes not only a bamboo extract composition obtained in the form of an extract obtained from bamboo epidermis as an extraction raw material, but also a diluted or concentrated extract, or an extract. A dried product obtained by drying the product, or a crude product or a purified product thereof.

  In addition, the function of the functional food is not particularly limited as long as it is a function that can be exhibited by lipids and glycolipids contained in the bamboo extract composition. For example, moisturizing skin, improving rough skin, beautiful skin, atopic Improvement of dermatitis, improvement of allergic dermatitis, improvement of breakout, hair restoration, hair growth, anti-cancer and the like.

Furthermore, as functional food forms, soft drinks, carbonated drinks, nutrition drinks, fruit drinks, lactic acid drinks and other drinks (including concentrated concentrates and adjustment powders of these drinks); ice cream, ice sherbet, shaved ice, etc. Noodles such as buckwheat, udon, harusame, gyoza skin, sushi mai, Chinese noodles, instant noodles, etc. Confectionery such as confectionery; processed fishery and livestock products such as kamaboko, ham and sausage; dairy products such as processed milk and fermented milk; fats and oils such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream and dressing Foods; seasonings such as sauces and sauces; soups, stews, salads, prepared dishes, pickles . Moreover, the feed given to livestock is also included in the form of functional food.
In addition, the compounding quantity of the bamboo extraction composition to a functional food can be suitably adjusted according to the form, function, etc. of a functional food.

(Example)
Hereinafter, examples will be shown to verify that the active ingredient of the bamboo extract composition is effective as a hyaluronidase activity inhibitor or a Helicobacter pylori antibacterial agent.
(Manufacturing method of Somune bamboo skin powder)
First, in this example, the bamboo skin used as the raw material for the bamboo extract composition was crushed to a thickness of about 0.1 to 2.0 mm from the surface of the bamboo scorpion and pulverized to about 50 to 400 mesh to obtain a bamboo skin powder. It was.

(Hydrolysis step)
The bamboo skin powder 40 g, 30 wt% aqueous ethanol solution 160 g, placed in a pressure reactor to sulfate 1g, after nitrogen substitution, was then sealed and pressurized to 2 kgf / cm ² initial pressure with nitrogen.
Subsequently, it is heated to 150 ° C. and the internal pressure is adjusted to 10 kgf / cm ² . While maintaining this state, hydrolysis was carried out with stirring for 3 hours.
After cooling, the extraction device was opened to return to normal pressure, and the solution was neutralized with sodium bicarbonate.

(Extraction process)
Next, 100 ml of ethyl acetate was added, followed by extraction with stirring for 1 hour, followed by vacuum filtration to remove bamboo skin residue.

(Extract analysis)
A crude extract was obtained by concentrating the extraction solution obtained in the above extraction step without dispensing it into an organic solvent phase and an aqueous phase. The crude extract (4.0 g) is separated and purified by silica gel chromatography (column length: 20 cm, inner diameter: 2.6 cm, packing material: Wako Pure Chemical Industries, Ltd., Wako Gel C-300). As a moving layer, ethyl acetate, acetone, and methanol (Me-OH) were developed in order to obtain fractions 1 to 36 (FIG. 1).

  It is generally said that this ethyl acetate extract fraction is a lipid (simple lipid), and the acetone extract fraction and the methanol extract fraction are glycolipids (including phospholipids).

  For fractions 1 to 36, 0.5 μL each was placed on a TLC plate (TLC plate silica gel, Merck) using a microcapillary syringe, and a developing solution in which chloroform, methanol, acetic acid, and calcium chloride were mixed in advance (chloroform: acetone). : Methanol: Acetic acid: Water = 30: 12: 9: 6: 3) Place the TLC plate in the developing tank, and develop until the developing solution is about 1 cm from the top of the plate. And sprayed with anthrone reagent, heated on a hot plate and analyzed (thin layer chromatography method).

  The analysis of the crude product and the purified product can be performed by, for example, not only thin layer chromatography (TLC) but also high performance thin layer chromatography (HPTLC).

  As a result (FIG. 2), it was classified into 6 fractions from the spot shape to fractions A to F (FIG. 3). Fractions having the same shape were combined and concentrated under reduced pressure using an evaporator to remove the developing solvent.

Next, the hyaluronidase inhibition test was done about the fraction classified into six.
1) Preparation of enzyme solution Bovine testis hyaluronidase is dissolved in 0.1 M acetate buffer (pH = 4.0) to adjust the final enzyme concentration to 400 units / ml.
2) Preparation of enzyme activation solution
Compound 48/80 is dissolved in 0.1 M acetate buffer (pH = 4.0) to a final concentration of 0.1 mg / ml.
3) Preparation of substrate solution Dissolve potassium hyaluronate in 0.1 M acetate buffer (pH = 4.0) to adjust the final concentration to 0.4 mg / ml.
4) Preparation of boric acid solution Add 50 ml of water to 4.95 g of boric acid, adjust the pH to 9.1 with 1N sodium hydroxide solution, and add distilled water to make 100 ml.
5) Preparation of p-dimethylaminobenzaldehyde (p-DAB) reagent
Dissolve 10 g of p-DAB in a mixture of 12.5 ml of 10N hydrochloric acid and 87.5 ml of acetic acid and store in a refrigerator. Use 10-fold diluted with acetic acid just before use.
6) Preparation of measurement sample The upper limit of the extract belonging to each fraction is diluted with an aqueous ethanol or DMSO solution to a concentration of 1 mg / ml. To do.
7) Measurement procedure Add 0.05 ml of the enzyme solution to 0.1 ml of this sample, and heat at 37 ° C. for 20 minutes.
Next, 0.1 ml of the enzyme activation solution is added and heated at 37 ° C. for 20 minutes.
Further, add 0.25 ml of the substrate solution and react at 37 ° C. for 40 minutes.
The reaction is stopped by adding 0.1 ml of 0.4N aqueous sodium hydroxide and cooling with ice.
Add 0.1 ml of boric acid solution and heat in a boiling bath at 100-120 ° C for 5 minutes, then cool on ice.
Add 3 ml of p-DAB reagent and heat at 37 ° C. for 20 minutes to develop color, and measure the absorbance at 585 nm using distilled water as a control.
The results are shown in FIGS. 4 and 5 in which the extraction solvent was used instead of the sample for the control, and 0.1 M acetate buffer (pH = 4.0) was added instead of the enzyme solution for the blank and the same operation was performed.
As shown in FIGS. 4 and 5, when compared at the same concentration, a strong hyaluronidase inhibitory effect was observed in fraction E.

Next, a Helicobacter pylori antibacterial test was conducted.
1) (Preparation of plate medium)
Dissolve 4.3 g Burucella Agar in 83 ml distilled water and sterilize. After sterilization, incubate in a constant temperature water bath at 58-60 ° C. Separately, keep the horse serum warm.

2) (Test fraction solution)
In the hydrolysis step, a part of the solution obtained by neutralizing the solution with sodium hydrogencarbonate was concentrated under reduced pressure using an evaporator to obtain fraction Z. Fraction Z and Fraction A were suspended in water to prepare 250, 500, and 1000 μg / mL fraction solutions for testing. Next, Fraction Z was suspended in ethanol, and 250, 500, 1000 μg / mL sample solutions were prepared and allowed to stand for 12 hours. The supernatant was used as a fraction solution for Fraction Z ethanol soluble fraction test. .

3) (Burucella plate medium)
Place 1.0 ml of the test fraction solution prepared in 2 above in the petri dish, add 9.0 ml of the medium in which 83 ml of Burucella agar medium and 7 ml of horse serum that had been kept warm are added immediately, mix quickly, and cool to room temperature. And flat plate.

4) (Preparation of Helicobacter solution)
The above culture medium pre-cultured for 1 week is diluted with Burcella liquid medium (no serum added). Empirically, the number of bacteria is expected to be 3-4 × 10 ³ cells / ml at 10 ⁵ times dilution. When diluted 2 × 10 ⁵ times, the number of bacteria is 1.5-2 × 10 ³ cells / ml, and the number of colonies to be counted is 200-300.

5) (Inoculation of bacteria)
3. above. On the flat plate prepared in 4 above. Inoculate 100 μl of the bacterial solution and smear with a large stick.

6) (Culture and measurement)
After culturing at 37 ° C. under microaerobic conditions using aneropack microaerobic, the number of colonies formed is counted (3-4 days, 4 days later are easy to count). The growth inhibitory activity was evaluated from the difference from the blank (the test group using distilled water instead of the sample solution) (FIGS. 6, 7, and 8).

  As shown in FIGS. 6, 7, and 8, a strong Helicobacter pylori inhibitory action is observed in fraction A. And even if a density | concentration is about 250 microgram / ml, since the high inhibition rate is shown, it is guessed that it is suitable as an active ingredient of Helicobacter pylori inhibition.

The extraction solution obtained in the above extraction step is divided into an aqueous phase (ethanol and water) and an organic solvent phase (ethyl acetate), and each is concentrated. The aqueous phase is extracted X and the organic solvent phase is extracted Y. did.
And the hyaluronidase activity inhibition test (FIG. 1) was done about the extract X and Y obtained at the extraction process, and the Helicobacter pylori inhibition test was done about the extract Y. FIG.
The method of Example 1 was used for the hyaluronidase activity inhibition test.

  As shown in FIG. 9, in the hyaluronidase activity inhibition test, even when the concentration is about 125 μg / ml, the inhibition rate is 40% or more. Moreover, the higher the concentration, the higher the inhibition rate. From this, it can confirm that the substance contained in an aqueous phase has a hyaluronidase activity inhibitory effect.

  The bamboo extract composition of the present invention is used not only as a raw material for hyaluronidase activity inhibitors and Helicobacter pylori antibacterial agents, but also as a raw material for antiallergic agents that suppress allergic symptoms such as allergic rhinitis and atopic dermatitis Is possible. Moreover, since the activity of hyaluronidase is inhibited by anti-inflammatory agents and anti-allergic agents, it can be used as a raw material for drugs and quasi drugs that reduce inflammation and the like.

It is a fraction list. It is the figure which showed the TLC analysis result. It is the table | surface which showed the fraction classification | category from a TLC analysis result. It is a table | surface of the hyaluronidase inhibition test in each extraction fraction. It is the graph which showed the hyaluronidase inhibition test in each extraction fraction. It is a table | surface of the Helicobacter pylori inhibition rate in each extraction fraction. It is the table | surface which showed the sulfuric acid 0.5 weight% extract ethyl acetate fraction bacteria inhibitory activity (fraction A-4,5,6). It is a photograph (Fraction A-4,5,6) of a Helicobacter pylori antibacterial test. It is the table | surface which showed the hyaluronidase inhibition test result in the extract X and Y.

Claims (9)

The substance to be treated obtained by treating the bamboo skin by the hydrolysis method is extracted by adding an organic solvent or an organic solvent and water. Among the extracted bamboo extract compositions, the components contained in the aqueous phase are used as active ingredients. Hyaluronidase activity inhibitor characterized by containing. The hydrolysis method comprises
The hyaluronidase activity inhibitor according to claim 1, which is an acid hydrolysis method using a mixed solution obtained by mixing an acid, an organic solvent and water. The hydrolysis method comprises
The hyaluronidase activity inhibitor according to claim 1, which is an alkali hydrolysis method using a mixed solution obtained by mixing an alkali, an organic solvent and water. The substance to be treated obtained by treating the bamboo skin by the hydrolysis method is extracted by adding an organic solvent or organic solvent and water, and the components contained in the organic solvent phase of the extracted bamboo extract composition are the active ingredients. Helicobacter pylori antibacterial agent characterized by containing as. The hydrolysis method comprises
The Helicobacter pylori antibacterial agent according to claim 4, which is an acid hydrolysis method using a mixed solution obtained by mixing an acid, an organic solvent and water. The hydrolysis method comprises
5. The Helicobacter pylori antibacterial agent according to claim 4, wherein the antibacterial agent is an alkali hydrolysis method using a mixed liquid obtained by mixing an alkali, an organic solvent and water. The substance to be treated obtained by treating the bamboo skin by the hydrolysis method is extracted by adding an organic solvent or an organic solvent and water,
The hydrolysis method comprises
Bamboo extraction composition characterized by being an acid hydrolysis method using a mixed liquid in which an acid, an organic solvent and water are mixed. The substance to be treated obtained by treating the bamboo skin by the hydrolysis method is extracted by adding an organic solvent or an organic solvent and water,
The hydrolysis method comprises
A bamboo extraction composition characterized by being an alkali hydrolysis method using a mixed liquid in which an alkali, an organic solvent and water are mixed. The bamboo extract composition according to claim 7 or 8, wherein the organic solvent is one or a combination of two or more of toluene, ethyl acetate, methanol, and ethanol.