JPH05186769A - Antioxidant which eliminates active oxygen - Google Patents

Abstract

PURPOSE:To provide an antioxidant which has a high capability of eliminating active oxygen and a high safety and hence can be widely used in industries as the material of food and drink, medicine, and cosmetics. CONSTITUTION:The title antioxidant contains a hydrolyzable tannin, e.g. 1,2,6-tri- O-galloyl-beta-D-glucose and/or 2,3-di-O-galloyl-4,6-(O-4,4',5,5',6,6'-hexahydroxydiphenoyl)-D-glucose, as the main component.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antioxidant having a hydrolyzable tannin as an active ingredient and having an active oxygen scavenging action. More specifically, the present invention is 1,2,6-tri-O, which is particularly effective among hydrolyzed tannins.
-Galloyl-β-D-glucose and / or 2,3
-Di-O-galloyl-4,6- (O-4,4 ', 5,
5 ', 6,6'-hexahydroxydiphenoyl) -D
-It relates to an antioxidant containing glucose as a main component, and is applied to the food industry, the pharmaceutical industry, the cosmetic industry, and the like.

[0002]

BACKGROUND OF THE INVENTION Tannin generally refers to polyphenols of plant origin and is widely distributed as a natural organic compound. This tannin has various uses as a tanning material, dye, paper, cloth, thread, etc. reinforcement and waterproofing agent. In addition, it is also used as a medicine such as an anti-astringent agent and an intestinal regulating agent which utilize the astringent property of tannin.

Recently, it has been assumed that tannin prevents lipid peroxidation in vivo from the action of reducing and antioxidant tannin. Metabolism of arachidonic acid, which is a peroxidation reaction in vivo, and liver microsomes and mitochondria Studies were conducted focusing on the lipid peroxide production reaction, and each of the extracts contained in ginkgo biloba and geraniin, which is the main tannin of ginkgo biloba, was administered to rats treated with lipid peroxide to increase serum GOT, GPT and lipid peroxide. Suppressed,
It was reported that the hydrolyzed tannin was more effective than the condensed tannin (Chem. Pharm.
l. ) 31, 1625, (1983); 32, 1866.
(1984)).

Reactive oxygen species including superoxide anion radicals generated in the xanthine (XA) -xanthine oxidase (XOD) system in vivo are inflammation, cataract, arteriosclerosis, myocardial ischemia, aging, convulsion, stroke, It is known to be involved in diabetes and the like. In addition, superoxide dismutase (SOD), which is an enzyme that exists in the body and has an active oxygen scavenging action, inactivates superoxide anion radicals and plays a role in preventing various diseases caused by radical reactions occurring in the body. Acting.

For the prevention and treatment of various diseases such as inflammation, carcinogenesis, radiation damage, immunity, cataract, arteriosclerosis, myocardial ischemia, aging, convulsion, stroke, diabetes caused by active oxygen generated by metabolism in the living body. A prophylactic and therapeutic agent for a disorder derived from in vivo active oxygen using superoxide dismutase (SOD) as an effective agent is also known (JP-A-56-32422).

Vitamin E and vitamin C are used as substances for eliminating active oxygen in the prevention and treatment of the above-mentioned diseases.

[0007]

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention In the above-mentioned prior art, the superoxide dismutase (SO
Since D) is an enzyme protein, it has drawbacks such that the production yield cannot be increased to a high degree of purity and it is difficult to maintain the blood concentration when administered in vivo. On the other hand, vitamin E and vitamin C have drawbacks such as insufficient action of scavenging active oxygen in the living body, and further have strong action and availability of low molecular weight active oxygen scavenger (low SOD-like action). In addition to superoxide, a highly versatile active oxygen scavenger that can be widely used to eliminate not only superoxide but also various active oxygen such as hydrogen peroxide, hydroxy radicals, and singlet oxygen Is requested.

[0008]

[Means for Solving the Problems] As a result of the research conducted by the present inventors for the purpose of obtaining a substance that strongly eliminates active oxygen,
1, which is a hydrolyzable tannin isolated from eucalyptus
2,6-tri-O-galloyl-β-D-glucose and 2,3-di-O-galloyl-4,6- (O-4,4 ′,
5,5 ', 6,6'-hexahydroxydiphenoyl)
-D-Glucose strongly eliminates the active oxygen generated in the body, and superoxide anion, which is considered to be a bad body in the body, was not sufficiently eliminated by the conventional antioxidant substances Vitamin E and Vitamin C. The inventors have found that radicals are strongly eliminated, and have completed the present invention.
The present invention relates to an antioxidant having an active oxygen scavenging action, which has hydrolyzed tannin as an active ingredient, completed by such research.

In the present invention, any hydrolyzable tannin can be used. For example, 1, 2,
6-tri-O-galloyl-β-D-glucose and 2,
3-di-O-galloyl-4,6- (O-4,4 ', 5
5 ', 6,6'-hexahydroxydiphenoyl) -D
-Glucose is preferred. These substances are known {Chemical and Pharmaceuticals.
Bulletin (Chem. Pharm. Bull.) 2
9, 2862 (1981), Fight Chemistry (P
hytochemistry 15, 211 (197)
6)}. However, the physiological action of eliminating active oxygen is the first in the present invention.

Since the active ingredient compound of the present invention is derived from a natural product, its toxicity is low.
No death was observed and no change in body weight was observed even when the rats were administered with 50 mg / kg for a long period of 100 days. Moreover, since the active ingredient compound has a strong active oxygen scavenging action and can be used in various dosage forms, it can be freely used in foods and drinks, pharmaceuticals, cosmetics and the like.

When the compound of the present active ingredient is used in a form other than food and drink, it is used by oral administration, coating, or other parenteral administration. For example, as a preparation for oral administration, tablets, pills, Granules, soft / hard capsules, powders, fine granules, powders, emulsions, suspensions, syrups, pellets, elixirs and the like can be mentioned. Formulations for parenteral administration include injections, drops, infusions, ointments, lotions,
Examples include tonics, sprays, suspensions, oils, emulsions and suppositories. To formulate the active ingredient of the present invention, a conventional method may be followed, including a surfactant, an excipient, a coloring agent, a flavoring agent,
Preservatives, stabilizers, buffers, suspensions, isotonic agents and other commonly used suppositories are used as appropriate. The same applies to external preparations.

The dose of the antioxidant according to the present invention varies depending on the type, the type of disease to be treated or prevented, the administration method, the age of the patient, the symptoms of the patient, the treatment time, etc. Is 0.01 active ingredient per adult per day
~ 2000 mg / kg, 0.0 for intramuscular administration
1-3000 mg / kg, and the same for oral administration.
It is preferably administered within the range of 5-4000 mg / kg. When used as a cosmetic or external preparation for skin,
Apply an appropriate amount to the affected area according to a conventional method.

Next, examples showing the effect of hydrolyzable tannin as an active ingredient of the present invention on the elimination of active oxygen and the effect of suppressing lipid peroxidation will be shown below as examples.

[0014]

Example 1

1) Method In a test tube, 1.0 ml of a mixed solution of 0.4 mM xanthine and 0.24 mM nitroblue tetrazolium was added to 1, 2, 6
-Tri-O-galloyl-β-D-glucose and / or 2,3-di-O-galloyl-4,6- (O-4,
4 ', 5,5', 6,6'-Hexahydroxydiphenoyl) -D-glucose was added to 0.1 ml of a dimethylsulfoxide aqueous solution to prepare a test solution, and xanthine oxidase (0.049 unit / ml) was prepared. After adding 1.0 ml, the mixture is reacted at 37 degrees Celsius for 20 minutes. After the reaction
The reaction was stopped by adding 2.0 ml of 69 mM sodium dodecyl sulfate, stirred sufficiently, and the absorbance at a wavelength of 560 nm was measured.

2) Test results Superoxide dismutase (C) purified from bovine erythrocytes instead of tannin in the same method as above
u, Zn-SOD) was added and a calibration curve was prepared from the absorbance. From the calibration curve obtained, 1,2,6-tri-O
-Galloyl-β-D-glucose and 2,3-di-O-
Galloyl-4,6- (O-4,4 ', 5,5', 6,6
The activity of 6'-hexahydroxydiphenoyl) -D-glucose was calculated. Table 1 below shows the superoxide dismutase (SOD) -like activity of these tannins.

[0017]

[Table 1]

As is clear from the above results, these hydrolyzable tannins have a scavenging action on superoxide anion radicals and are put to practical use as low molecular weight substances.

[0019]

Example 2

1) Method A rabbit red blood cell ghost membrane (2.5 mg protein / m 2) was placed in a test tube.
l) 0.9 ml and 1,2,6-tri-O-galloyl-β
-D-glucose and 2,3-di-O-galloyl-4,
6- (O-4,4 ′, 5,5 ′, 6,6′-hexahydroxydiphenoyl) -D-glucose in 0.1 ml of dimethyl sulfoxide was added, and tert-butyl hydroper was dissolved in dimethyl sulfoxide. The reaction was initiated by the addition of 9 μl of oxide (10 mg / ml). The reaction was carried out at 37 degrees Celsius for 30 minutes, and the reaction was stopped by adding 1.0 ml of 2M trichloroacetic acid. 0.6 for this
By adding 2.0 ml of 7% thiobarbituric acid and reacting for 15 minutes in a boiling water bath, the amount of thiobarbituric acid reaction positive substances such as malondialdehyde produced by this reaction was measured by absorbance at a wavelength of 535 nm. I asked. Based on this value, the concentration at which lipid peroxidation was suppressed by 50% was determined, and the antioxidant activity against butylhydroxyanisole, which is a commercially available synthetic antioxidant, was calculated.

2) Test Results 1,2,6-tri-O-galloyl-β-D-glucose and 2,3-di-O-, using the concentration at which the peroxidation of rabbit erythrocyte membrane lipids is suppressed by 50% as an index. Galloyl-4,6- (O-
The antioxidant activity of 4,4 ′, 5,5 ′, 6,6′-hexahydroxydiphenoyl) -D-glucose is shown in Table 2 below.
Shown in.

[0022]

[Table 2]

As is clear from the above results, the antioxidative activity of hydrolyzed tannin is about 3 times as potent as the molar activity of butylhydroxyanisole which is a commercially available synthetic antioxidant.

[0024]

The hydrolyzable tannin, which is the active ingredient of the present invention, is generated by metabolism in the body and causes inflammation, carcinogenesis, radioactivity disorder, immunity, cataract, arteriosclerosis, myocardial ischemia, aging, convulsion, stroke, It is a useful antioxidant for directly scavenging active oxygen, especially superoxide anion radicals, which causes various diseases such as diabetes, and is widely industrially used as a material for foods, pharmaceuticals, cosmetics and the like.

Claims (2)

[Claims] 1. An antioxidant having an active oxygen scavenging action, which comprises hydrolyzable tannin as an active ingredient. 2. The hydrolyzable tannin is 1,2,6-tri-O-galloyl-β-D-glucose and / or 2,3-di-O-galloyl-4,6- (O-4, 4 ',
5,5 ', 6,6'-hexahydroxydiphenoyl)
The antioxidant having an active oxygen scavenging action according to claim 1, which is -D-glucose.