JPH07223940A - Active oxygen eliminating agent and composition containing the same - Google Patents

Abstract

PURPOSE:To obtain an active oxygen eliminating agent having sufficiently removing action on active oxygen occurring in an organism and high safety, containing 1,2-dihydroxy-4-(2-hydroxyethyl) benzene. CONSTITUTION:This active oxygen eliminating agent contains a compound of the formula and/or its salt as an active ingredient. The agent is effective for treating various diseases related to active oxygen, such as inflammation, senile dementia, ischemic disease, e.g. myocardial infarction, allergic disease, liver disorder and rheumatism and for improving aging of organism such as skin. A proper dose is 10-1,000mg calculated as the compound of the formula per adult daily by oral administration and 5-500mg by injection. A food such as cake, drink or bread is mixed with 0.1-10wt.% of the compound of the formula and can be used. The compound of the formula is extracted from Stachys riederi of labiate plant.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an active oxygen scavenger and a composition containing the same, and more specifically to 1,2-dihydroxy-4- (2-hydroxyethyl) benzene and / or
Alternatively, it relates to an active oxygen scavenger consisting of a salt thereof and a composition containing the same.

[0002]

2. Description of the Related Art Generally, the effects of active oxygen on the living body include cross-linking of collagen fibers, partial cleavage of DNA helix, and tissue damage due to generation of chained radicals, resulting in wrinkles and elasticity. Aging of the skin and living body such as disappearance, induction of allergic reaction such as bronchial asthma and inflammation due to histamine release, damage of smooth muscle in ischemic myocardial infarction, deterioration of diseases such as liver injury, and brain tissue It is known that destruction causes induction of dementia and the like. Furthermore, although the detailed cause is unknown, it is said that active oxygen is involved in the onset of rheumatism.

Therefore, suppressing the generation of active oxygen in the living body is very important in treating or preventing these diseases, and therefore, the active oxygen generated in the living body is eliminated from the conventional case. The search for substances that have the action to do so has been widely conducted.

[0004] For example, as a drug having such an action, conventionally used are those derived from natural products such as fat-soluble tocopherol (vitamin E),
Examples include water-soluble ascorbic acid (vitamin C), and synthetic compounds include BHT (butylhydroxytoluene),
BHA (butylhydroxyanisole) etc. are mentioned. However, these agents have insufficient active oxygen scavenging action, and in synthetic compounds, both BHT and BHA are suspected to be carcinogenic, and it was difficult to say that any of them was practical as an active oxygen scavenger.

In recent years, many attempts have been made to obtain a substance having an active oxygen scavenging action from a crude drug extract in search of sufficient medicinal effect and safety, for example, JP-A-60-224629. , JP-A-61-24522,
JP-A-2-193930, JP-A-2-243632
No. 2, JP-A-2-264727, JP-A-3-15362
No. 9, JP-A-3-221587, JP-A-4-6934.
3, JP-A-4-202138, and JP-A-4-2470.
The inventions described in the respective publications of No. 10 each utilize a substance having an active oxygen scavenging action derived from a crude drug. However, although these crude drug extracts have no safety problems, they have not been sufficiently obtained in terms of active oxygen scavenging action.

Further, attempts have been made to eliminate active oxygen generated in the living body by administering superoxide desmutase (SOD), one of the enzymes in the living body, but since SOD is a protein, Not only is it difficult to obtain, but it is digested, so oral administration is not possible, and even when administered by injection,
The blood half-life was short and it was not satisfactory.

On the other hand, 1,2-dihydroxy-4- (2-
(Hydroxyethyl) benzene is a compound known to be contained in large amounts in the extract of sesame, a plant of the Labiatae family, but it was not known that this compound has a function of eliminating active oxygen. . Furthermore, there has been no attempt to use this compound for the prevention or treatment of various diseases such as those mentioned above, prevention of aging, or improvement by incorporating it into a drug or food.

[0008]

The present invention has been made from the above point of view, and has an action of sufficiently eliminating active oxygen generated in the living body and a highly safe active oxygen eliminating agent, and It is an object to provide a composition containing this.

[0009]

In order to solve the above-mentioned problems, the present inventors extensively screened compounds derived from various medicinal plants using active oxygen scavenging activity as an index, and as a result, dog sesame (Stakis reederi) which is a Labiatae family plant. (Stachys ri
ederi)) contained in 1,2-dihydroxy-4- (2
The inventors have found that -hydroxyethyl) benzene and its salts have an excellent active oxygen scavenging action, and completed the present invention.

That is, the present invention is an active oxygen scavenger consisting of 1,2-dihydroxy-4- (2-hydroxyethyl) benzene represented by the following structural formula (I) and / or a salt thereof, and a composition containing the same. It is a thing.

[0011]

[Chemical 2]

The present invention will be described in detail below.

<1> Active Oxygen Scavenger of the Present Invention The active oxygen scavenger of the present invention is 1,2-dihydroxy-4.
-(2-hydroxyethyl) benzene and / or a salt thereof.

The above-mentioned 1,2-dihydroxy-4- (2-hydroxyethyl) benzene is a component which is present in the extract from almost any part of sesame seeds, and the sesame seeds are widely distributed throughout Japan. As a plant, it is easy to obtain the above compound.

1,2-dihydroxy-4-from dog sesame
A method for obtaining (2-hydroxyethyl) benzene is, for example, extracting raw or dried sesame seeds with a solvent, and removing an insoluble portion from the obtained extract by filtration or the like,
Examples of the method include removing the solvent from the filtrate and then purifying the filtrate using a conventional purifying means such as column chromatography.

In this case, the extraction may be carried out batchwise or continuously. Examples of a preferable extraction method include a method in which whole grass of sesame seeds or a dried product of whole grass is immersed in a solvent for several days at room temperature, and a method in which it is immersed at a temperature near the boiling point for several hours.

The solvent used for the extraction is preferably a polar solvent, and examples thereof include alcohols such as methanol, ethanol and propanol, ketones such as acetone and methyl ethyl ketone, and water. These solvents may be used alone or in combination of two or more.

As the active oxygen scavenger of the present invention, in addition to 1,2-dihydroxy-4- (2-hydroxyethyl) benzene obtained as described above, a salt of this compound can be used. In this case 1,2-dihydroxy-4
One kind of-(2-hydroxyethyl) benzene and its salt may be used alone, or two or more kinds may be mixed and used.

Examples of such salts include salts obtained by reacting 1,2-dihydroxy-4- (2-hydroxyethyl) benzene with an inorganic or organic base in the same amount, for example, alkali metals such as sodium and potassium. With salts, salts with alkaline earth metals such as calcium and magnesium, ammonia,
Examples thereof include salts with amines such as triethylamine and triethanolamine, salts with basic amino acids such as arginine and lysine, and the like.

<2> Composition Containing Active Oxygen Scavenger of the Present Invention The composition of the present invention is a mixture of the above active oxygen scavengers according to a conventional method, and examples thereof include pharmaceuticals and foods.

When the active oxygen scavenger of the present invention is formulated as a medicine, the dosage form is not particularly limited, but for example, injections, powders, granules, tablets, capsules, liquids and the like, which are commonly used, The formulation can be formulated according to a conventional method. In addition, in forming a dosage form, in addition to the active oxygen digestive agent, an excipient, a binder, a disintegrating agent, a lubricant,
Arbitrary components usually used in pharmaceuticals such as a flavoring agent, a bulking agent and a coating agent may be used in any amount.

Regarding the dose of the above-mentioned medicine, although it depends on the type of disease, symptoms, age of patient, body weight, etc., adult 1
The amount of 1,2-dihydroxy-4- (2-hydroxyethyl) benzene per person per day is 10 mg to 1000
Orally administer mg once or several times or 5m
It is suitable to administer g-500 mg by injection. Examples of the administration method of the injection include intravenous administration, intraarterial administration, portal vein administration, intraperitoneal administration, intramuscular administration, subcutaneous administration and the like.

When the active oxygen scavenger of the present invention is added to foods, it can be added to various foods together with optional components usually used in foods. Examples thereof include confectionery such as candy, gummy candy, jelly, drinks such as juice, and staple food such as bread. The blending amount varies depending on the type of food, but is preferably 0.1 to 10% by weight, which can expect a sufficient effect of eliminating active oxygen without impairing the taste of the food.

[0024]

The active oxygen scavenger and the composition containing the same of the present invention are 1,2-dihydroxy-4 which is the active ingredient thereof.
-Ischemic diseases such as inflammation, senile dementia, myocardial infarction and the like, which are said to involve active oxygen, due to the excellent active oxygen scavenging action of-(2-hydroxyethyl) benzene and / or its salt. Alternatively, it works effectively for treatment of various diseases such as allergic diseases, liver disorders, rheumatism, and improvement of biological aging such as skin.

The active oxygen scavenger of the present invention and the composition containing the same can be effectively used for the prevention of the above-mentioned diseases and biological aging. This is because the active oxygen generated in the living body can be quickly eliminated and detoxified by allowing the component having the active oxygen scavenging effect to exist in the living body in advance.

[0026]

EXAMPLES Examples of the present invention will be described below. First, examples of the active oxygen scavenger of the present invention will be described.

[0027]

Example 1 10 kg of dried sesame seeds were added to 5-10 mm
The mixture was cut into small pieces, to which 5 l of ethanol and 5 l of purified water were added, and the mixture was heated under reflux for 3 hours with stirring. Then, the extract was cooled, filtered to remove insoluble matter, and the filtrate was concentrated under reduced pressure to remove the solvent. The obtained extract was dissolved in purified water and poured into a column filled with Diaion HP-20 (manufactured by Mitsubishi Kasei Co., Ltd.), and further 2 l of purified water was flowed to wash it.

After that, the column adsorbate was eluted with 2 l of ethanol, which was subjected to preparative high performance liquid chromatography (manufactured by Tosoh Corp., HLC837, ODS column, elution solvent; 5% → 50% acetonitrile aqueous solution, ultraviolet 22
0 nm) to obtain 1,2-dihydroxy-4- (2-hydroxyethyl) which is the active oxygen scavenger of the present invention.
263 mg of benzene was obtained.

<Evaluation of Active Oxygen Scavenger of the Present Invention> The active oxygen scavenger obtained in Example 1 above was evaluated for safety and active oxygen scavenging action.

(1) Acute toxicity test (intraperitoneal administration) The active oxygen scavenger obtained in Example 1 was dissolved in physiological saline to 5 ICR male mice (body weight 25 to 30 g) to prepare 1
It was intraperitoneally administered at a rate of 000 mg / kg. 14 after administration
Life or death was judged on the day, but no death was observed. From this, LD by intraperitoneal administration of the active oxygen scavenger of the present invention
_{The 50} value is greater than 1000 mg / kg, which shows that it is excellent in safety.

(2) Acute toxicity test (oral administration) Six male ICR mice (weight: 25 to 30 g) were dissolved in physiological saline to dissolve the active oxygen scavenger obtained in Example 1
It was orally administered at a rate of 000 mg / kg. Although life or death was determined 14 days after administration, no death was observed. From this, it is understood that the LD ₅₀ value by oral administration of the active oxygen scavenger of the present invention is larger than 1000 mg / kg, and the safety is excellent.

[0032] (3) based on the reaction formula shown in the measurement of the active oxygen scavenging action (in vitro) of 3, xanthine - generate ^- superoxide anions which is one of active oxygen by xanthine oxidase (XOD) system (O ₂₎ Then, the production rate of generated O ₂ ⁻ was measured by the nitrous acid method, and this value was corrected by the xanthine oxidase inhibition rate value to obtain the active oxygen elimination action value.

[0033]

[Chemical 3]

0.1 ml of an active oxygen scavenger aqueous solution containing the active oxygen scavenger obtained in Example 1 at a rate of 20 μg / ml was added to 65 mM potassium dihydrogen phosphate, 35 m.
M sodium borate, 0.5 mM EDTA disodium aqueous solution (hereinafter referred to as buffer A) 0.2 ml, 0.5 m
It was added to a mixed solution of 0.2 ml of M xanthine solution, 0.1 ml of 10 mM aqueous solution of hydroxylamine hydrochloride and 0.2 ml of pure water, and well stirred to give a test solution. Similarly, a control solution using 0.1 ml of pure water instead of the active oxygen scavenger was prepared.

To the above test solution and control solution, 0.2 ml of buffer solution A containing xanthine oxidase at a concentration of 1 μl / ml was added and stirred, and then incubated at 37 ° C. for 30 minutes. As a blank, 0.2 ml of xanthine oxidase-free buffer solution A was added to the test solution and control solution prepared in the same manner as above, and a solution treated in the same manner as above was prepared.

Each of the solutions thus obtained was mixed with 30 ml of N-1-naphthylethylenediamine hydrochloride, 3 mM of sulfanilic acid and 2 ml of 25% glacial acetic acid.
And leave it for 30 minutes at room temperature.
The generation amount of active oxygen was measured at the absorbance of 50 nm, and the generation amount of uric acid was measured at the absorbance of 295 nm.

Using the obtained values, based on the following equation,
The active oxygen elimination activity value was calculated. Active oxygen generation rate = [(A _550-3 -A _550-4 ) / (A _550-1
-A _550-2 )] x 100 Urea production rate = [(A _295-3 -A _295-4 ) / (A _295-1 -A
_295-2 )] × 100 active oxygen scavenging activity = 100− (active oxygen generation rate / uric acid production rate) × 100 where the symbols in the formula are the absorbance values of the solutions prepared under the conditions shown in Table 1. To do.

[0038]

[Table 1]

The active oxygen scavenging activity value of the active oxygen scavenger of Example 1 obtained by the above method was 74.7%.
From this result, it is clear that the active oxygen scavenger of the present invention has an excellent active oxygen scavenging action.

(4) Measurement of active oxygen scavenging action (in vivo) Blood was collected from 43-week-old ICR MCH mice (weight: 34 g), and 50 mg of the active oxygen scavenger obtained in Example 1 was added to the mice. It was orally administered at a rate of / kg. Blood was collected again 30 minutes after the administration and 4 hours after the administration. Serum was taken out from the blood obtained under each of the above conditions by centrifugation, diluted 10-fold with buffer A, and the active oxygen scavenging activity was measured by the same method as in the above (3). The results are shown in Table 2.

[0041]

[Table 2]

From these results, it is clear that the orally administered active oxygen scavenger of the present invention is rapidly absorbed and spreads in the body to exhibit an active oxygen scavenging action. It is also clear that the action is maintained for a long time.

Next, examples of the composition of foods, pharmaceuticals and the like containing the active oxygen scavenger obtained in Example 1 will be described. The amounts used below are all parts by weight unless otherwise specified.

[0044]

Example 2 Candy The component A in Table 3 was melted by heating at 150 ° C., cooled to 120 ° C., then the component B was added and stirred to homogenize. After molding this, it was cooled to obtain a candy.

[0045]

[Table 3]

[0046]

Example 3 Gummi A component of Table 4 was heated and dissolved at 110 ° C., separately swelled B component was added, and further C component was added and poured into a mold,
After leaving it for a whole day and night, I removed it from the mold and got a gummy.

[0047]

[Table 4]

[0048]

Example 4 Juice The components in Table 5 were well solubilized by stirring, sterilized, aseptically filled, and sealed to produce juice.

[0049]

[Table 5]

[0050]

Example 5 Hot Cake A hot cake was prepared by thoroughly mixing the ingredients shown in Table 6 and baking them in an oiled frying pan.

[0051]

[Table 6]

[0052]

Example 6 Granules A component of Table 7 was mixed well, and 100 ml of 20%
The B component dissolved in an ethanol aqueous solution was gradually added while kneading and granulated. This was blow-dried at 40 ° C. for 2 days, sieved and sized to obtain granules.

[0053]

[Table 7]

[0054]

Example 7 Injectable Solution The components in Table 8 were dissolved, filtered, sterilized, and aseptically filled into ampoules and sealed to obtain an injectable agent.

[0055]

[Table 8]

[0056]

The active oxygen scavenger of the present invention is excellent in active oxygen scavenging action and highly safe. In addition, the composition of the present invention, such as the drug or food, containing the same can be used safely and effectively for a long period of time for the prevention and treatment of diseases involving active oxygen.

Claims (2)

[Claims] 1. An active oxygen scavenger composed of 1,2-dihydroxy-4- (2-hydroxyethyl) benzene represented by the following structural formula (I) and / or a salt thereof. [Chemical 1] 2. A composition containing the active oxygen scavenger according to claim 1.