JP2001064253A - Substance for inducing activity of flutathione-s- transferase and food containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an activity-inducing substance which can activate or increase glutathione-S-transferase(GST) as one of the second phase foreign matter- metabolizing enzymes in biological foreign matter metabolism, and exhibits a strong activity-inducting action, and to obtain a solid, semi-fluidized or fluidized food containing the activity-inducing substance. SOLUTION: This GST activity-inducing substance comprises an ω- methylsulfinylalkylisothiocyanate (the alkyl group has five to eight carbon atoms), especially 6-methylsulfinylhexylisothiocyanate. The food containing the GST activity-inducing substance is obtained by adding the substance to a solid, semi-fluidized or fluidized food, or the like. The food includes health foods.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to glutathione-S-
The present invention relates to an activity inducer that activates or increases the amount of transerase (hereinafter, referred to as “GST”) and a food containing the same, and particularly to a GST activity inducer that exhibits strong activity induction and a food containing the same.

[0002]

2. Description of the Related Art Today, the environment in which we live is flooded with foreign substances such as chemical substances having harmful effects on living organisms. Some of these are carcinogens, and we are forced to live such carcinogens on a daily basis.

On the other hand, our human body has a detoxification and metabolism function consisting mainly of two phases, a first phase and a second phase, as a protective function for the metabolism of xenobiotics including these carcinogens. It is found in organs such as the liver, stomach, intestines, and lungs where direct contact is common.

[0004] In the first phase, the polarity of the xenobiotic is increased by an oxidation reaction, a reduction reaction, a hydrolysis reaction, etc., and the water solubility is increased. Furthermore, in the above-mentioned second phase, the water solubility of the xenobiotics (including the xenobiotic metabolized in the first phase) is further enhanced by various conjugation reactions and the like, so that the xenobiotics are detoxified and easily excreted outside the body.

GST is an enzyme that conjugates reduced glutathione with various electrophilic compounds, specifically, an enzyme that catalyzes the conjugation reaction between a xenobiotic and glutathione, and is one of the second-phase xenobiotic metabolizing enzymes. A multifunctional enzyme consisting of a group of molecular species having different functions. That is, it has a function as a detoxifying enzyme, for example, a function of conjugating various exogenous organic compounds to glutathione, and also has a function as a binding protein that binds bile acids, bilirubin, steroid hormones, and the like. .

[0006] Lipids constituting the body or lipids ingested as food are automatically oxidized in the body into peroxides, and the peroxides are further oxidized to produce oxidized products such as 4-hydroxynonenal. Things. These lipid peroxides and oxidation products, particularly oxidation products, are substances that damage DNA and living tissues, induce various diseases such as arteriosclerosis and Alzheimer, and promote aging of living bodies. I know that. This is what is called oxidative stress. GST also catalyzes the conjugation of these lipid peroxides and oxidation products, detoxifies them, and facilitates their excretion outside the body.

[0007] If the activity of such GST is further improved or increased, the above-described second phase conjugation reaction is further activated, and as a result, the detoxification and metabolic function of xenobiotics such as carcinogens is improved. Thus, the risk of carcinogenesis is reduced.

Further, by improving the activity and increasing the amount of GST,
Oxidative stress due to lipid oxidation products can be further reduced, and as a result, arteriosclerosis, Alzheimer's, etc. caused by lipid oxidation products can be effectively prevented.

As a substance for improving or increasing the activation of GST, sulforaphane (4-methylsulfinylbutyl isothiocyanate) conventionally contained in broccoli is particularly strong GS as compared with other isothiocyanates.
It has been confirmed to exhibit T activity induction. Furthermore, it has been confirmed that carcinogenesis in rats by a chemical carcinogen is effectively suppressed by the action of sulforaphane.

[0010]

However, although the above-mentioned sulforaphanes exhibit considerably strong GST activity induction,
In recent years, which are still inadequate, harmful chemicals and xenobiotics are increasingly flooded, and oxidative stress due to lipid oxidation products is further increased, these days, stronger GST activity inducers or foods containing them The development of is strongly desired.

Therefore, an object of the present invention is to provide a more powerful GST.
GST activity-inducing substance which exhibits activity induction, thereby exhibiting a carcinogenesis-suppressing action by improving the detoxification metabolic function, and also exhibiting an oxidative stress-reducing action, and a solid form containing the GST activity-inducing substance, which has improved the disadvantages of the above-mentioned known techniques, It is to provide a semi-liquid or liquid food.

[0012]

In order to achieve the above object, according to the GST activity inducer of the present invention, ω-methylsulfinylalkyl isothiocyanate (alkyl group having 5 to 8 carbon atoms), preferably It is characterized by comprising 6-methylsulfinylhexyl isothiocyanate.

Further, in order to achieve the above-mentioned object, according to the solid, semi-liquid or fluid food containing the GST activity-inducing substance of the present invention, the GST activity-inducing substance is ω-.
It is characterized by including methylsulfinylalkylisothiocyanate (alkyl group having 5 to 8 carbon atoms), preferably 6-methylsulfinylhexylisothiocyanate.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

The above-mentioned GST activity inducer ω-methylsulfinylalkylisothiocyanate according to the present invention may be a chemically synthesized substance.
It may be an extract from cruciferous plants. Specific examples of these substances include 5-methylsulfinylpentyl isothiocyanate, 6-methylsulfinylhexyl isothiocyanate, 7-methylsulfinylheptyl isothiocyanate and 8-methylsulfinyloctyl isothiocyanate. Of these, 6-methylsulfinylhexylisothiocyanate is particularly preferred.

The method for synthesizing the above-mentioned GST activity inducer will be described as follows.

In principle, it follows the method of Kiaer et al. (Kiaer
etal. Acta chem. Scand, 11, 1298, 1957). Using ω-chloroalkenol as a starting material, CH ₃ -SN
The mixture was refluxed with a to obtain ω-methylthioalkenol, which was then treated with SOCl ₂ to obtain ω-chloroalkenol methyl sulfide.

Next, an amino group was introduced using the Gabriel method to produce N- (ω-methylthioalkyl) -phthalimide, to which hydrazine hydrate was added and refluxed to obtain ω-
A methylthioalkylamine is obtained. Further, according to the method of Li et al. (Lietal. J. Org. Chem., 62, 4539, 1997), the methylthio group of ω-methylthioalkylisothiocyanate obtained via thiuram disulfide is oxidized with mCPBA, and ω -To obtain methylsulfinylalkylisothiocyanate.

Examples of the cruciferous plants include various vegetables and fruits, and especially, wasabi (Eutrema wasabi or Wasabia japonica) and watercress are preferable because they exhibit strong GST activity induction.

In extracting ω-methylsulfinylalkylisothiocyanate from wasabi and watercress, the wasabi and watercress may be extracted with water or an organic solvent, or by a distillation method such as steam distillation or molecular distillation. Is preferred, but the method is not particularly limited to these methods.

For example, a specific method for extracting wasabi with an organic solvent is as follows. After the grated horseradish is extracted, it is extracted with an ethyl acetate solvent, and the extract is dehydrated with anhydrous sodium sulfate and concentrated with an evaporator. Then, ω-methylsulfinylalkyl isothiocyanate is obtained. (See Example 1 below.) This method is particularly suitable for the extraction of 6-methylsulfinylhexylisothiocyanate.

When ω-methylsulfinylalkylisothiocyanate is extracted from watercress, it is extracted in the same manner as wasabi. For example, watercress is ground, extracted with an ethyl acetate solvent, and the extract is dehydrated with anhydrous sodium sulfate and concentrated by an evaporator to obtain ω-methylsulfinylalkylisothiocyanate. This method is particularly suitable for extracting 7-methylsulfinylheptyl isothiocyanate and 8-methylsulfinyloctyl isothiocyanate.

The above-mentioned extract is extracted and concentrated, and then purified by any method such as a liquid-liquid distribution method, a chromatographic method, molecular distillation, and rectification.

The thus obtained G according to the present invention
The ST activity inducer ω-methylsulfinylalkyl isothiocyanate has a strong GST activity inducing effect, and is detoxified by administering it to a human or animal body as a pharmaceutical or as a food as described below. Metabolic function is strongly improved, carcinogenesis is suppressed, and oxidative stress is reduced.

When the above-mentioned GST activity-inducing substance is administered to humans or animals as foods, the detoxification metabolic function is strongly improved, carcinogenesis is suppressed, and oxidative stress is reduced, as described above.

Examples of the above-mentioned food according to the present invention include solid, semi-fluid, and fluid foods which are generally recognized as foods in the society. Among these foods, of course, the solid state, which is recognized as a health food by social wisdom,
It also includes semi-liquid and fluid foods.

Examples of solid foods include general foods and health foods in the form of biscuits, sheets, tablets, granules and the like. Examples of the semi-liquid food include general foods and health foods in the form of paste, paste, jelly, gel and the like. Examples of the fluid food include general foods and health foods in the form of juice, tea, soft drinks, drinks, and the like. In addition,
These health foods include special health foods licensed by the Ministry of Health and Welfare, specifically, intestinal control, cholesterol control, blood pressure control, high absorbable minerals, maintenance of dental health (prevention of dental caries), blood sugar control, postprandial blood levels. Foods in which components having various functions such as suppression of essential fats are also included.

Further, it may be a simple food consisting of the above-mentioned GST activity-inducing substance itself or a food derived from cruciferous plants, or may be appropriately mixed with excipients, seasonings, flavors and the like to improve edibility. It may have improved palatability.

Further, the above-mentioned GST activity-inducing substance or a cruciferous plant containing the same can be prepared by adding spice seasonings such as powdered wasabi, kneaded wasabi and kneaded mustard, animal meat products, fish meat products,
Processed foods such as canned foods, freeze-dried foods, retort pouch foods, frozen foods, noodles, side dishes, seasonings, mayonnaise, dressings, broths, sauces, breads, Japanese and Western sweets, pickles, etc. May be added.

[0030]

The present invention will be described below with reference to examples, but the present invention is not limited to these examples.

Example 1 After wasabi waster was ground, double weight ethyl acetate solvent was added thereto, and the mixture was allowed to stand overnight. Next, the ethyl acetate layer was separated, dehydrated with anhydrous sodium sulfate, and concentrated with an evaporator to obtain an extract.

Next, the components of the obtained wasabi extract were separated and identified as follows.

First, the Sawasabi washi extract was subjected to silica gel column chromatography, and a mixed solvent of hexane / ethyl acetate (ethyl acetate content: 0, 25, 50, 75, 100
%), And eluted with acetone. The obtained acetone-eluted fraction was concentrated by an evaporator to obtain a fraction. This fraction was further purified using high performance liquid chromatography (column: Develosil ODS-HG-5, elution solvent: methanol / water = 40/60, flow rate: 2.0 ml / min), and 6-methylsulfur Finylhexyl isothiocyanate was obtained. The obtained 6-methylsulfinylhexyl isothiocyanate was confirmed to be the same as the result of the instrumental analysis by NMR, IR and MS.

Next, 6-methylsulfinylhexylisothiocyanate was administered to mice, and the GST activity-inducing action was measured. The administration method and the measurement of GST activity were performed by the methods described below.

Administration Experiment The experimental animals were 4-week-old ICR mice (females) (manufactured by Slc Japan) and were fed a normal diet (20% casein diet) for 4 weeks. Then, using an oral sonde (NIS flexible sonde, manufactured by Natsume Seisakusho), 100 μl of sesame oil
6-Methylsulfinylhexylisothiocyanate and 4-methylsulfinylbutylisothiocyanate (sulforaphane) were separately dissolved and prepared at a concentration of 15 μmol / mouse / day. These were administered to different groups of mice by intragastric gavage at 10 am each morning.
Went on for consecutive days. As a control, a group to which only sesame oil was administered and a group to which nothing was administered were prepared. In addition, 5 mice were used for each group.

At the end of the administration on the 5th day, 24 hours later, blood was collected and sacrificed. The liver was excised and rapidly frozen under liquid nitrogen for -8 days.
Stored at 0 ° C. After that, the obtained liver was 4 times the weight of T
An ED buffer was added and homogenized while cooling on ice. After pre-centrifugation of the homogenate solution, the supernatant
Ultracentrifuged at 000 G for 1 hour. GST activity was measured by the following measurement method using a 100-fold dilution of each of the obtained supernatants as a sample.

GST activity is measured in principle by WHHabig (J. Biol. Chem. 249, 713, 1974).
Year). That is, 0.1 ml of the sample obtained above was added to a mixture of 0.5 ml of a 0.2 M phosphate buffer (pH 6.5), 0.1 ml of an aqueous 10 mM reduced glutathione solution, and 0.2 ml of purified water, followed by stirring. Followed by 10 mM CDN
B (1-chloro-2,4-dinitrobenzene) 100 μ
1 was added to start the reaction. Then, the absorbance at 340 nm was established every 15 seconds for 3 minutes, and the activity was measured from the calibration curve.

[0038] Measurement of protein content of the protein mass measurement the supernatant using a BCA Protein Assay Li Age entry kit (PIERCE Co. type), the measurement mixed reagent 200μl added to the supernatant 10 [mu] l, incubated for 30 minutes, absorbance at 562nm Was measured to determine the amount of protein.

The GST activity value was determined by the following arithmetic expression. GST activity value (unit / mg protein) = (34
Absorbance at 0 nm / 3 × 10 × 100) / (9.6 × protein amount)

FIG. 1 shows the measurement results. From FIG. 1, it can be seen that the GST activity of the mouse administered with 6-methylsulfinylhexylisothiocyanate according to the present invention is considerably higher than that of the mouse administered with conventional sulfolphane.

[0041]

As described above, the ω-methylsulfinylalkylisothiocyanate of the present invention, in particular,
Methylsulfinylhexyl isothiocyanate, or an extract of cruciferous plants, especially saw Wasabi or watercress (extracted extract), exhibits high GST activity induction, and detoxification function by administering these as a pharmaceutical or food in vivo. Thus, the present invention exhibits a carcinogenesis-suppressing action and also has an oxidative stress-reducing action, and is a practically useful invention.

[Brief description of the drawings]

FIG. 1 is a graph showing GST activity in a mouse administration experiment.

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Yasujiro Morimitsu 805 (B-201) F-term (Reference) 4B047 LB03 LB08 LG48 LG49 LP01 4C088 AB15 AC11 BA08 BA09 CA01 CA03 CA05 CA11 CA14 MA16 MA27 MA28 MA34 MA35 MA41 MA52 NA05 NA14 ZA18 ZB26 ZC19 ZC21 ZC37 4H006 AA01 AA03 AB10 AB21 AB23 AB28

Claims (8)

[Claims] 1. A glutathione-S-transferase activity-inducing substance comprising ω-methylsulfinylalkyl isothiocyanate. However, the alkyl group has 5 to 8 carbon atoms. 2. The glutathione-S-transferase activity inducer according to claim 1, wherein the ω-methylsulfinylalkyl isothiocyanate of claim 1 is 6-methylsulfinylhexylisothiocyanate. 3. The glutathione-S-transferase activity inducer according to claim 1, wherein the ω-methylsulfinylalkylisothiocyanate of claim 1 is an extract from a cruciferous plant. 4. The glutathione-S according to claim 3, wherein the cruciferous plant of claim 3 is a wasabi or watercress.
-An inducer of the activity of transfelase. 5. A solid, comprising a glutathione-S-transferase activity inducer comprising ω-methylsulfinylalkylisothiocyanate.
Semi-liquid or liquid food. However, the alkyl group has 5 to 8 carbon atoms. 6. The food according to claim 5, wherein the ω-methylsulfinylalkyl isothiocyanate of claim 5 is 6-methylsulfinylhexyl isothiocyanate. 7. The food according to claim 5, wherein the ω-methylsulfinylalkyl isothiocyanate according to claim 5 is a cruciferous plant or an extract thereof. 8. The food according to claim 7, wherein the cruciferous plant according to claim 7 is a wasabi or watercress.