JP2003104880A - Antimutagenic agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new antimutagenic agent by using a readily available substance having a specified quality as an active ingredient. SOLUTION: This antimutagenic agent comprises at least one kind of an organic acid selected from a group consisting of citric acid, malic acid, lactic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, acetic acid, α- ketoglutaric acid, isocitric acid, oxalacetic acid, cis-aconitic acid and salts thereof as an active ingredient.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antimutagenic agent.

[0002]

2. Description of the Related Art In recent years, development of foods for specified health use and health foods has been rapidly increasing due to high interest in health. Also,
The development of new materials having various functionalities is also increasing, and antimutagenicity has been confirmed in extracts such as bark and roots of various plants including mushrooms.

[0003] On the other hand, conventionally, fruits are considered to be good for health, and interest in components contained in fruits is increasing from the expectation of prevention of lifestyle-related diseases. For example, an extract of kiwifruit is antimutant. It has been reported to have an original effect and an antiviral effect (JP-A-3-254340). It has also been reported that juices and extracts of fruits of Rosaceae have antioxidant effect, blood pressure lowering effect, antimutagenicity, etc. (JP-A-7-285876).

However, since these substances are extracts or squeezed products of natural products, it takes time and labor to produce them, and their contents and components vary depending on the place of production and the time of harvesting, and the stability is poor. There is a problem that the price is high and it is difficult to use.

[0005]

The main object of the present invention is to:
It is intended to provide a novel antimutagenic agent using a substance having a constant quality and easily available as an active ingredient.

[0006]

Means for Solving the Problems As a result of intensive studies to achieve the above-mentioned object, the present inventor has found that a specific organic acid or a salt thereof, which is easily available and relatively inexpensive, has an excellent mutation. It has been found that it has an original inhibitory action, and has completed the present invention.

That is, the present invention provides the following antimutagenic agents. 1. Citric acid, malic acid, lactic acid,
Succinic acid, maleic acid, fumaric acid, tartaric acid, acetic acid, α-
Ketoglutaric acid, isocitric acid, oxaloacetic acid, cis-
An antimutagenic agent comprising at least one organic acid selected from the group consisting of aconitic acid and salts thereof as an active ingredient. 2. Item 1. The antimutagenic agent according to Item 1, which is used as a food additive.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The antimutagenic agent of the present invention is citric acid, malic acid, lactic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, acetic acid, α-ketoglutaric acid, isocitric acid, oxaloacetic acid, cis. -It contains at least one organic acid selected from the group consisting of aconitic acid and salts thereof as an active ingredient.

All of these components are substances that can be easily obtained, and by using these components as active ingredients, antimutagenic substances of constant quality and low cost and having an excellent antimutagenic action are obtained. A sex agent can be obtained.

Of the above-mentioned organic acids, citric acid may be either anhydrous or monohydrate. As for malic acid, not only the DL-form but also the D-form and L-form of optical isomers can be used. Also, as tartaric acid, L-form, D-
Body or DL-body may be used, and as lactic acid,
Any of the L-form, D-form and DL-form may be used.

Examples of the salts of the above organic acids include sodium salt, potassium salt, calcium salt and the like.

In the present invention, the above organic acids may be used alone or in combination of two or more.

In the antimutagenic agent of the present invention, the above-mentioned organic acids may be used as a powder as they are, or may be dissolved in an appropriate solvent such as water and used as a liquid. When used as a liquid, the concentration is not particularly limited, and may be appropriately dissolved and used within a soluble range. In addition, if necessary, it may be used in combination with other acidulants such as ascorbic acid and acetic acid, seasonings such as amino acids and nucleic acids such as glycine, nutritional components such as vitamins and minerals, and mixed in advance to prepare a preparation. You may use.

The method of ingesting the antimutagenic agent of the present invention is not particularly limited, and either oral or parenteral is possible, but it is preferable to ingest orally, for example, as a food additive. It is preferably added to food and ingested.

When used as a food additive, the amount added is not particularly limited, and may be appropriately determined depending on the type of food, taking into consideration the degree of sourness and the like. For example, when used by being added to liquid foods such as soft drinks and carbonated drinks, as the concentration of the above organic acids, usually,
The amount may be about 0.005% by weight or more, but it is preferably in the range of about 0.005 to 3% by weight in consideration of sourness, antimutagenic action and the like. Also, when used by adding to solid foods such as confectionery and other various foods, similarly, it may be appropriately determined according to the type of food, but generally,
As the content rate of the above-mentioned organic acids, 0.01 to 5% by weight
It is preferable to set the degree.

An example of a preferable addition amount depending on the type of food is, for example, in the case of soft drinks in liquid drinks, the concentration of the above-mentioned organic acids is about 0.05 to 0.5% by weight. In the case of carbonated drinks, the concentration of the above-mentioned organic acids may be about 0.1 to 0.5% by weight. Among confectionery, in the case of gummy, the addition amount may be about 2% by weight, and in the case of jelly,
The amount added may be about 0.5 to 0.8% by weight.

In addition, an example of the administration method and dose when the antimutagenic agent of the present invention is administered to the human body is as follows.

The administration can be carried out by various methods, for example, oral administration such as tablets, capsules, granules and syrups. Regarding oral administration, in the case of oral administration, an appropriate amount of the active ingredient in an adult is usually about 0.01 to 1000 mg / kg, which may be administered once to several times a day. . The orally-administered agent can be produced according to a usual production method.
For example, excipients such as starch, lactose, mannitol, binders such as sodium carboxymethyl cellulose, hydroxypropyl cellulose, disintegrants such as crystalline cellulose and carboxymethyl cellulose calcium, lubricants such as talc and magnesium stearate, light anhydrous By appropriately combining and formulating a fluidity improver such as silicic acid, it can be produced as a tablet, capsule, granule or the like.

The antimutagenic agent of the present invention can be used as a food additive as described above, or can be taken or administered by other administration methods to prevent various diseases based on mutation, such as cancer, etc., It can be effectively used for treatment and the like. In addition to this, it can be used widely in the field of biochemistry when it is necessary to suppress the mutation of bacteria, for example, in the case of culture, biochemical analysis and the like.

[0020]

INDUSTRIAL APPLICABILITY The antimutagenic agent of the present invention contains, as an active ingredient, organic acids which are relatively easily available,
It is inexpensive, has good quality stability, and has an excellent antimutagenic effect.

[0021]

EXAMPLES The present invention will be described in more detail with reference to test examples and formulation examples.

Test Example 1 A method for detecting a mutagen using the mutagen suppressing effect of the antimutagenic agent of the present invention as an index for the induction of SOS reaction (UMU test;
Science and Industry, Vol. 62, No. 4, p. 142, 1988
Year). Here, the "UMU test" is a mutagen detection test that utilizes the SOS reaction that is observed when DNA is damaged in Escherichia coli, and has many advantages such as results in a short time.

Trp-P-1 (manufactured by Wako Pure Chemical Industries, 0.3 μg / ml) and furylflamid were used as mutagens, and Salmonella typhimurium T was used as a strain.
A1535 / pSK1002) was used. The outline of the test method will be described below.

That is, a test bacterial solution cultured overnight at 37 ° C. in an LB medium (1% tryptone, 0.5% yeast extract, 0.5% sodium chloride) was added to a TGA medium (1% tryptone, 0.1% sodium chloride).
5%, glucose 0.2% and ampicillin 50 μg /
1/50 amount was inoculated into the mixture (added in the amount of ml) and cultured at 37 ° C. with shaking.

When the bacterial concentration reaches the logarithmic growth phase (A ₆₀₀ is 0.25 to 0.30), 2.0 ml of bacterial solution is added.
Take each in a test tube and add it to the mutagen (Trp-P-1 (0.
3 μg / ml) or 0.1 ml of furyl flamid and 0.
0.3 ml of 1M phosphate buffer (pH 7.0) was added, and the test substances were added at the concentrations shown in the graphs of FIGS. 1 and 2 below, followed by incubation at 37 ° C. for 2 hours.

As test substances, citric acid (anhydrous), citric acid (crystal), DL-malic acid, D-malic acid, L-
Malic acid, DL-lactic acid, L-lactic acid, fermented lactic acid, succinic acid, maleic acid, fumaric acid, L-tartaric acid or D-lactic acid was used.

After culturing, the bacterial solution was spun down to collect the cells, and then the cells were resuspended in physiological saline. The amount of bacterial cells was measured in a part of this bacterial solution, and β-galactosidase activity was measured in another part. It was measured.
Here, the logarithmic growth phase is the time when the number of bacteria and cells increases logarithmically, and is also called the exponential growth phase.

The β-galactosidase activity was measured by Mill
er's method (Miller, JH: Experiments in molecular gen
etics, Cold spring Harbor Laboratory, New York, P3
52-355 (1972)). That is, Z buffer 2.2
After adding 0.25 ml of the above test bacterial solution to 5 ml,
% Sodium dodecyl sulfate aqueous solution (50 μl) and chloroform (10 μl) were added, and the mixture was vigorously stirred. Substrate (o
-nitrophenyl-β-D-galactopyranoside 4 mg / m
l) 0.25 ml was added and reacted at 28 ° C. After 15 minutes, 1.25 ml of 1 M Na ₂ CO ₃ was added to stop the reaction, and A ₄₂₀ , A ₅₅₀ and A ₆₀₀ (absorbance) were measured by a spectrophotometer.

Here, the β-galactosidase activity value is
It was calculated by the following formula.

Β-galactosidase activity value (unit) = 1
000 (A ₄₂₀ -1.75 x A ₅₅₀ ) /1.5 x A ₆₀₀ The SOS reaction inhibition rate was calculated by the following formula.

SOS reaction inhibition rate (%) = [1- (A-
C) / (B−C)] × 100 where A is the β-galactosidase activity value when the test substance is added to the mutagen, and B is the β-galactosidase activity induced only by the mutagen. The value C indicates the control β-galactosidase activity value. The same amount of pure water was used for control. In addition, each test was conducted as one set of trials, and the average was taken.

The above test results are shown as graphs showing the SOS reaction inhibition rate in FIGS.

Formulation Example 1 A powdered health drink having the following formulation was prepared.

(Formulation) Citric acid (anhydrous) 35% by weight DL-malic acid 3% by weight Na citrate 3% by weight Glucose 45% by weight Vitamin 5% by weight Minerals 4% by weight Baking soda 5% by weight Powdered beverage of the above composition 5 g is dissolved in 100 to 200 ml of water and taken as a drink.

[Brief description of drawings]

FIG. 1 is a graph showing the inhibitory effect on furylflamid in Test Example 1.

FIG. 2 is a graph showing the inhibitory effect on Trp-P-1 in Test Example 1.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 43/00 105 A61P 43/00 105 (72) Inventor Takaho Jiyo 4-26 Tajima, Saitama City, Saitama Prefecture No. 11 F term (reference) 4B018 MD09 ME14 4B035 LC06 LG04 LG06 LK19 LP21 4C206 AA01 DA01 DA07 DA09 DA34 DA36 MA01 MA04 NA05 ZB21 ZB26

Claims (2)

[Claims] 1. A method selected from the group consisting of citric acid, malic acid, lactic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, acetic acid, α-ketoglutaric acid, isocitric acid, oxaloacetic acid, cis-aconitic acid and salts thereof. An anti-mutagenic agent comprising at least one organic acid as an active ingredient. 2. The antimutagenic agent according to claim 1, which is used as a food additive.