KR20090039838A - Antioxidant Composition Containing Acacia Bark Derivatives - Google Patents

Abstract

The present invention provides a composition having excellent antioxidant activity in vivo. The present invention relates to an antioxidant composition characterized by containing acacia bark derivatives.

Description

ANTIOXIDANT COMPOSITION CONTAINING COMPONENT ORIGINATING IN THE BARK OF TREE BELONGING TO THE GENUS ACACIA}

The present invention relates to an antioxidant composition derived from a tree belonging to the genus Acacia, and to the use of the antioxidant composition as a food, animal feed, medicine, and quasi-drug.

In the atmosphere, in general, oxygen is a stable substance called triplet oxygen. When some of the oxygen is changed into a highly reactive substance called active oxygen, such as superoxide, hydroxy radicals, singlet oxygen, hydrogen peroxide, etc. in the body, some of it is produced by neutrophils or macrophages. While acting as a bio-defense against foreign bodies, if excess oxygen is present in the living body, it attacks lipids, proteins, enzymes, nucleic acids and the like to damage biological membranes and tissues, and atherosclerosis, cerebrovascular disorders, emphysema, It has also been found to be a cause of the development of rheumatoid arthritis, cataracts, hypertension, senile dementia, Alzheimer's disease, or blemishes, freckles or aging.

Conventionally, in order to prevent various diseases as described above caused by oxidative stress caused by these excess active oxygen, it is possible to catalyze the reaction for generating hydrogen peroxide and oxygen molecules by disproportionating superoxide (0 ₂ ⁻ ), which is a starting material. SOD-like substances that function similar to the enzyme superoxide dismutase (SOD), and antioxidants that add active oxygen-capturing substances (active oxygen scavenging active substances) to food have been widely searched.

For example, such substances include fat-soluble α-tocopherol (vitamin E) and water-soluble ascorbic acid (vitamin C) as substances derived from natural products, and BHT (butylhydroxytoluene) as a synthetic substance, Phenolic substances such as BHA (butylhydroxyanisole) and TBHQ (tertiary butylhydroquinone).

However, these substances are generally used as antioxidants in foods, but the active oxygen scavenging action to a living body is not sufficient, and in particular, problems such as suspicion of carcinogenicity occur in BHA of synthetic substances.

Therefore, in recent years, research and development which searched for highly stable active oxygen scavenging active substance from natural products, such as animals and plants, are actively performed, for example, patent documents 1, 2, 3, 4, 5, etc. are mentioned. In addition, in fact, polyphenol-based compounds such as tea leaves, catechins extracted from cacao beans extracts, anthocyanin compounds contained in blueberry peels, and isoflavones obtained from fermented soybeans have very high activity. Attention has been drawn in that it has an oxygen scavenging effect.

Among the polyphenol compounds found to have high active oxygen scavenging effects, (-) epigallocatechin, (-) epigallocatechin-3-gallate, (-) epicatechin, and (-) contained in tea leaves. It has been found that catechins, which are flavanols based on flavan-3-ol, such as epicatechin-3-gallate and (+) catechin, have a higher active oxygen scavenging effect than other polyphenols.

On the other hand, tannins extracted from the bark of the Acacia genus Acacia mearnsii De Wild. ), It is mainly used very cheaply as leather tanning agent and wood adhesive. In addition, the scientific name: Acacia mangium Willd. Is growing rapidly, and recently, forests have been actively grown in subtropical or tropical regions including Indonesia and Malaysia. This wattle tannin is classified as a condensed tannin.

Recently, the extract of the genus Acacia has a selective inhibitory effect of COX-2 (Patent Document 6), or the whitening effect by the active oxygen scavenging effect (Patent Document 7) or the tyrosinase activity inhibitory effect on the bark of the acacia genus (Patent Document 8). Is disclosed.

However, it is not known that acacia bark or bark-derived polyphenols can be expressed in a specific dose in vivo to exhibit excellent antioxidant activity.

Patent Document 1: Japanese Patent Application Laid-Open No. 01-25726

Patent Document 2: Japanese Patent Application Laid-Open No. 6-65074

Patent Document 3: Japanese Patent Application Laid-Open No. 7-300422

Patent Document 4: Japanese Patent Application Laid-Open No. 11-5975

Patent Document 5: Japanese Patent Application Laid-Open No. 2001-98264

Patent Document 6: Japanese Patent Publication No. 2004-532811

Patent Document 7: Japanese Patent Application Laid-Open No. 2004-352639

Patent Document 8: Japanese Patent Application Laid-Open No. 10-025238

<Start of invention>

Problems to be Solved by the Invention

The present invention provides a composition having high stability and exhibiting excellent antioxidant activity in vivo.

Means for solving the problem

As a result of intensive studies to solve the above problems, the present inventors have found that the acacia bark derivative is ingested at a specific dose to exert an excellent antioxidant effect in vivo, thus completing the present invention.

That is, the present invention relates to an antioxidant composition characterized by containing the acacia bark derivative at a particular dose.

The invention also relates to an anti-oxidation method using acacia bark derivatives.

The invention also relates to a method of using acacia bark derivatives for the production of antioxidant compositions.

Effect of the Invention

According to the present invention, it is possible to obtain a composition capable of exhibiting an excellent antioxidant activity by being taken in a specific dose.

In addition, even if the composition of the present invention is taken for a long time, there is little concern about side effects and the like, and safety is high.

Best Mode for Carrying Out the Invention

The acacia bark derivative which can be used in the present invention is not particularly limited as long as it is obtained by using a bark of a tree belonging to the acacia genus (hereinafter referred to as "acacia" or "acacia") as a raw material. And extracts such as acacia bark fragments, powders and suspensions thereof, acacia bark extracts, concentrated extracts, and extract powders and purified products obtained by refining the extracts. In order to obtain excellent antioxidant activity, extracts of bark of the genus Acacia, in particular acacia bark polyphenols, are preferred.

In the present invention, only one kind of these acacia bark derivatives may be used, or two or more kinds thereof may be used in combination.

The acacia which can be used in the present invention is not particularly limited as long as it is a tree belonging to the genus Acacia, but in view of obtaining an acacia bark derivative having excellent antioxidant action, the scientific name: Acacia mearnsii De Wild., Common name Black Wattle, Scientific name: Acacia mangium Willd, Common name Acacia mandium, Scientific name: Acacia dealbata Link, Scientific name: Acacia decurrens Willd. And scientific name: Acacia bark of the genus Acacia is selected from the group consisting of Acacia pycnantha Benth. Particularly preferred are Acacia Myrnaid Wild and Acacia Mani Wild.

In the present invention, one kind of these acacia bark may be used, or two or more kinds thereof may be used in combination.

The bark of the genus Acacia is usually obtained by logging as a tree, then peeling off only the bark, harvesting and drying. Preferably, the bark is further sun-dried bark.

The bark of the genus Acacia is composed of the outer skin and the fibrous inner skin, and when it is dried at a water content of about 20% or less, it is easily micronized with a grinder such as a hammer mill. In the present invention, as the bark of the acacia genus, both the inner and outer skins of the acacia genus may be used together, or only one of them may be used.

The three pieces of the bark of the acacia genus can be obtained by crushing the bark of the acacia genus according to the usual method.

The acacia bark powder may be obtained by pulverizing the bark of the acacia tree by a conventional method, and particularly preferably a powder having a particle diameter of 100 μm or less, particularly 50 to 70 μm. The powder fraction can be crushed to a bark dried at a water content of 20% or less to an appropriate size, for example, a particle diameter of 1.6 mm or less, and the powder obtained can be classified by vibrating sieve or the like to obtain the required powder.

The extract of the bark of the Acacia genus can be obtained by extracting the bark of the Acacia genus according to the conventional method. In order to obtain the extract of the bark of the acacia which has excellent antioxidant action, it is preferable to extract the bark of the acacia genus with an alcohol or a polar solvent.

Ethanol can be used as such an alcohol, water etc. can be used as a polar solvent, and these solvent can also be used individually or in combination of 2 or more types as needed. In particular, a mixed solvent of water and an alcohol such as ethyl alcohol is preferable in order to obtain excellent antioxidant action.

In addition, the extraction operation may be performed multiple times with the same or different solvents.

In obtaining an extract having excellent antioxidant activity, an extract obtained by further extracting with water or hot water from the bark of the acacia genus with ethanol may be used.

The extraction is carried out by adding a solvent to the pieces or powder of bark of the acacia genus and stirring them as necessary. The extraction is not particularly limited in terms of temperature, time or solid-liquid ratio. When water is used for a solvent, it can also extract with hot water. The obtained extract may be freeze dried or spray dried as it is, or may be freeze dried or spray dried after concentration under reduced pressure. The obtained extract can be made into various forms, such as an extract, a solution, a powder, a concentrate, and a paste, and can be used widely as needed.

In addition, the bark extract of the genus Acacia of the present invention obtained in these forms can be used as an antioxidant composition as it is, and further purified as needed, and the purified product can also be used as an antioxidant active ingredient.

In this invention, the component contained in the bark of the acacia genus is also illustrated as an acacia bark derivative. As such a component, an acacia bark polyphenol and the like are exemplified. In particular, acacia bark polyphenols are preferred components because they exhibit good antioxidant activity.

The acacia bark polyphenol of the present invention is based on flavan-3-ols such as (-)-picetinidol, (-)-rovinitynidol, (+)-catechin, and (+)-gallocatechin. It means a kind of condensed tannin which is a polymer in which flavanols made into a skeleton are C4-C8 and C4-C6 bonded. Here, as such a condensation type tannin, the thing of molecular weight 300-3000, especially 500-3000 are preferable. The acacia bark polyphenol used in the present invention can be obtained by hot water extraction of the powder of the bark of the acacia genus.

In addition, acacia bark polyphenol products include Mimosa Central Co., Ltd. Mimosa ME POWDER, Mimosa MS Powder, Mimosa GS Powder, Mimosa FS Powder, Mimosa WS Powder, Mimosa RG Powder, Mimosa RN Powder, Mimosa DK Powder, Mimosa AL Powder, Mimosa CR Powder, Golden GOLDEN MIMOSA POWDER, etc. are illustrated.

The composition of the present invention may be an acacia bark derivative, for example an acacia bark, an extract thereof, a purified product thereof, or an acacia bark polyphenol itself, but a substance having another antioxidant activity, for example , Vitamin C, vitamin E, coenzyme Q10, α-tocopherol, proanthocyanin, isoflavones, quercetin, green tea, or wheat germ. In particular, it is preferable to contain coenzyme Q10, green tea, or wheat germ from the point that the excellent antioxidant effect by a synergistic effect is obtained.

The composition of the present invention may be an acacia bark derivative, for example an acacia bark, an extract thereof, a purified product thereof, or an acacia bark polyphenol itself, so long as it does not impair the effect of the present invention, It may contain excipients, sweeteners, acidulants, thickeners, flavors, colorants, emulsifiers, and other additives and materials generally used in medicines and foods.

The composition of the present invention can be used for the prevention or treatment of diseases related to free radicals. In the present invention, the disease associated with active oxygen is not particularly limited as long as it is a cell or tissue disorder caused by active oxygen such as superoxide, hydroxy radical, singlet oxygen, hydrogen peroxide, or lipid peroxide oxidized by them. , Atherosclerosis, cerebrovascular disorders, emphysema, rheumatoid arthritis, cataracts, hypertension, senile dementia, Alzheimer's disease, or blemishes, freckles or aging. Preferably atherosclerosis.

The intake amount of the composition according to the present invention is not particularly limited, but may be appropriately selected depending on the dosage form and the age, weight and symptoms of the intake or ingested animal such as a user or a patient. For example, 0.001 to 1 g, preferably 0.001 to 0.5 g, more preferably 0.005 to 0.1 g of the amount of acacia bark polyphenols per 1 kg of body weight of the intaker or ingested animal per day as the active ingredient amount, By ingestion, especially orally, good antioxidant action is obtained.

The intake period can be arbitrarily determined according to the age and symptoms of the intaker or ingested animal.

The composition according to the invention can be used as a food or animal feed, for example as a health food, a functional food, a dietary supplement, a specific health food, a beauty food, and a nutritional supplement (supplement). have. These food and animal feeds include beverages, such as tea and juice; And ice cream, jelly, candy, chocolate, chewing gum and the like. It may also be in the form of a liquid, powder, granule, capsule, or tablet. Here, the animal of the animal feed includes all animals which need the prevention or treatment of the disease related to the active oxygen, including pets, livestock animals, or animals raised in zoos.

In addition, the composition according to the present invention can be orally administered as a medicament or quasi-drug, for example, in the form of powder, tablet, coated tablet, dragee, hard or soft gelatin capsule, liquid, emulsion, or suspension. Into the rectum, for example in the form of suppositories; For example, it may be administered parenterally or topically, in the form of ointment, cream, gel, or liquid.

Hereinafter, although a manufacture example, a test example, and a compounding example are given and this invention is demonstrated further in detail and concretely, this invention is not limited to this. In particular, although the embodiment of the present invention is described without dividing the bark of the genus Acacia of the present invention into the shell and the endothelium, the shell may be used separately from the endothelium.

In the following manufacture examples, test examples, etc., each acacia of this invention is shown by the number shown in the following parenthesis of each scientific name. For example, the scientific name: Acacia Miarci de Wilde describes the acacia as Acacia No.1.

Scientific name: Acacia Miarci de Wilde (No. 1), Scientific name: Acacia Diploma Wild (No. 2), Scientific name: Acacia Dilbatta Link (No. 3), Scientific name: Acacia Deckerence Wilde (No. 4), Scientific name: Acacia Pikkanta Benz (No. 5).

In addition,% means weight% unless there is particular notice.

Preparation Example 1 of Acacia Bark Powder

The bark of Acacia No. 1 was dried to a water content of 20% or less, and the dried bark was pulverized with a powder of 1.6 mm or less (passed by 10 mesh sieves (tyler) Tyler) with a hammer mill, and then again classified into a vibrating sieve. Fine powders of 63 μm or less (under a 250 mesh body) were obtained.

In the same manner, the remaining four types of bark of Acacia Nos. 2 to 5 were ground to obtain fine powders of 63 μm or less. Although there exist some differences in the yield of the fine powder which passed the 250 mesh sieve according to the kind, the target fine powder was obtained.

Preparation Example 2 of Acacia Bark Extract

Each of the bark of each Acacia No. 1 to 5 of the present invention was dried up to a moisture content of 20% or less, and the dried bark was pulverized into a powder of 1.6 mm or less with a hammer mill, and then, to 100 g of this dry crushed bark. Multiple volumes of hot water were added, boiled, extracted for 15 minutes and filtered using a 10-20 μm filter. The obtained filtrate was spray dried with a spray drier to obtain 40 g of each bark extract.

Hereinafter, each bark extract is described as acacia No. 1 to 5 hot water extract.

Preparation Example 3 of Acacia Bark Extract

The bark of the acacia of the present invention was dried to a moisture content of 20% or less, and the dried bark was pulverized into a powder of 1.6 mm or less with a hammer mill, and then 5 times of ethanol was added to 100 g of the dry crushed bark, Extracted for 15 minutes while boiling to reflux and filtered using a filter of 10 to 20 μm. After evaporating ethanol from the obtained filtrate, the concentrated liquid was spray dried with a closed spray dryer to obtain 40 g of bark extract (hereinafter described as Acacia No. 1 ethanol extract).

In the same manner, ethanol extracts of Acacia Nos. 1 to 5 were obtained.

Preparation Example 4 of Acacia Bark Extract

Three times the amount of ethanol was added to 10 g of the acacia hydrothermal extract obtained in Preparation Example 2, the mixture was boiled and extracted under reflux for 15 minutes, and filtered using a filter of 10 to 20 µm. Ethanol was evaporated from the filtrate obtained, water was added thereto, followed by freeze drying to obtain 9 g of extract (hereinafter described as Acacia No. 1 hydrothermal extract ethanol fraction).

In the same manner, the hydrothermal extract ethanol fractions of Acacia Nos. 1 to 5 were obtained.

Test Example 1 Antioxidant Test (1)

(1) test method

The acacia No. 1 hydrothermal extract described in Preparation Example 2 was suspended in 0.3% (W / V) carboxymethylcellulose sodium solution (CMC-Na solution), and rats (Slc: SDF, male) at a daily dose of 2 g / kg. , 7 weeks old) was orally administered for 14 days. The control group was administered with CMC-Na solution.

On day 14 of dosing, 8-OHdG (8-hydroxy-2'-dioxyguanosine) concentrations in the serum and urine of rats were determined using a high sensitivity 8-OHdG check and a new 8-OHdG check (Nikken Xyl). Measured by Kaisha Corporation).

Each measurement was expressed as mean ± standard error. Significant difference test with the control group was Student's t test. Significance levels are expressed as 5%.

(2) test score

The results are shown in Table 1. In all cases, no mortality and general abnormalities were seen.

Test Example 2 Antioxidant Test (2)

(1) Preparation of Test Sample Liquid

Acacia No. 1 hydrothermal extract described in Preparation Example 2 prepared in the same manner as in Test Example 1 was orally administered to rats (Slc: SD, male, 7 weeks old) for 14 days at a daily dose of 0.1 to 2.0 g / kg. . The control group was administered with CMC-Na solution.

At 14 days after the administration, 8-OHdG concentration in urine was measured in the same manner as in Test Example 1.

Measurements are expressed as mean ± standard deviation. Significant differences between the control group and each dose group were tested by Dunnett's multiple comparison test. Significance levels are expressed as 5% and 1%.

(2) test score

The results are shown in Table 2 below. In all cases, no mortality and general abnormalities were seen.

From the results of Test Examples 1 and 2 above, it became clear that the acacia bark polyphenol had an antioxidant action.

Test Example 3 Mutation Organic Test

Mutation organic tests were carried out in accordance with Japanese Labour Department Notice No. 77 (Sm. 1, 63). As a result of testing with a sample of a dose of 156 to 5,000 μg / plate (Acacia No. 1 to 5 hydrothermal extract of Preparation Example 2), the number of revertant colonies did not increase for any strain.

Test Example 4 Micronucleus Test

According to the conventional method, the presence or absence of the micronucleus inducing ability in vivo was investigated. Acacia No. 1 hydrothermal extract was orally administered to male ICR mice twice daily at 24-hour intervals at 2,000, 1,000, and 500 mg / kg doses, and micronucleus specimens were prepared 24 hours after the final administration.

Neither dose of Acacia No. 1 hydrothermal extract was positive. In addition, there was no constant change in the ratio of total polycytic erythrocytes to total erythrocytes observed at the same time, and there was no inhibition of proliferation of erythrocytes even when compared to the negative control group.

Test Example 5 Mouse Acute Toxicity Test (Oral)

In accordance with OECD (Guidelines for the Testing of Chemicals 401, 1987), an acute oral toxicity test was performed using ICR male and female mice. As a result, the LD 50 value of the Acacia No. 1 hydrothermal extract was 4,468 mg / kg in males and 3,594 mg / kg in females.

The test was carried out on the acacia No. 2 to 5 hydrothermal extract of Preparation Example 2 to obtain the same result.

Test Example 6 Rat Repeat Dose Toxicity Test (Oral)

According to the conventional method, the rat was subjected to repeated dose toxicity test for 13 weeks.

Mixed feeds containing 0.5, 1.5, and 5.0% of Acacia No. 1 hydrothermal extracts were fed to the male and female of Slc: SD rats.

As a result, there were no deaths of rats. There was no abnormality in the inspection including the general condition.

Test Example 7 Human Single Dose Test

Five healthy adult males aged 32 to 43 years were fed 1500 mg of Acacia No. 1 hot water extract (12 tablets of Formulation Example 4 below) before ingestion, 3 hours after ingestion, 8 hours after ingestion, and then ingested. After 24 hours and one week after ingestion, general examination, hematologic examination, blood biochemical examination, and urine examination were performed, but there were no changes in the clinically problematic test values. There were no adverse events related to purification.

Test Example 8 Human 4-week continuous administration test

To 25 healthy adult males aged 23 to 44 years, 750 mg / day (six tablets of Formulation Example 4) and 1000 mg / day (tablets of Formulation Example 4) were added to the Acacia No. 1 hydrothermal extract of Formulation Example 4 below. 8 tablets) for 4 weeks each.

Before the ingestion of each group, after 2 weeks of ingestion, after 4 weeks of ingestion, and after 2 weeks of ingestion, general tests, blood tests, and urine tests were performed. There was no change in the problematic test values. There were no adverse events.

Formulation Example 1 Preparation of Internal Reproduction

Using the acacia bark hydrothermal extract ethanol fraction of Preparation Example 4, an internal preparation was prepared with the composition shown below.

Extract fraction 1.0 of the preparation example 4 (wt%)

Lactose 30.0

Corn Starch 60.0

Crystalline Cellulose 8.0

Polyvinylpyrrolidone 1.0

Total 100.0

Formulation Example 2 Preparation of Pet Food

The pet food was manufactured using the acacia bark hydrothermal extract of manufacture example 2 with the composition shown below.

1.0 extract (weight%) of Preparation Example 2

Oatmeal 88.0

Starch 5.0

Saline 2.5

Whole War 3.0

Seasoning 0.5

Total 100.0

Formulation Example 3 Preparation of Tablets

Using the acacia bark hydrothermal extract ethanol fraction of Preparation Example 4, a tablet (sweet) was prepared with the composition shown below.

Extract fraction 1.0 of the preparation example 4 (wt%)

Citric Acid 1.0

Skim milk powder 15.0

Sucrose Ester 1.0

Flavor 0.5

20.0 powdered sugar

Lactose 61.5

Total 100.0

Formulation Example 4 Preparation of Tablet

The tablet was manufactured with the composition shown below using the acacia bark No. 1 hydrothermal extract of manufacture example 2.

Acacia bark No. 1 hydrothermal extract 125 of manufacture example 2 (mg)

Sucrose ester 9

Lactose 166

Total 300

According to the present invention, a composition for preventing and / or treating a disease related to active oxygen can be obtained.

More specifically, the compositions of the present invention are useful for the prevention and / or treatment of atherosclerosis.

These compositions can be used for food or animal feed such as pharmaceuticals, quasi-drugs, cosmetics, or health foods, dietary supplements, certain health foods, or nutritional supplements.

Claims (14)

An antioxidant composition comprising acacia bark derivatives as acacia bark polyphenols at a dose of 0.005 to 1 g per 1 kg of body weight of the intake per day. The acacia bark of claim 1, wherein the acacia bark is acacia mearnsii de wild., Acacia mangium willd. (Acacia decurrens Willd.), And Acacia pycnantha Benth. The composition is a bark selected from the group consisting of. The composition of claim 1 wherein the acacia bark is the bark of the acacia myarnsid de wild or the acacia materia. The composition of claim 1, wherein the Acacia bark derivative is an extract of the Acacia bark. The composition according to claim 1, wherein the acacia bark derivative is acacia bark polyphenols. 6. The composition of claim 5 wherein the acacia bark polyphenol is a condensed tannin. The composition of claim 6 wherein the condensed tannin has a molecular weight of 500 to 3000. 8. The composition according to claim 6 or 7, wherein the condensed tannin is a polymer of flavanols based on flavan-3-ol. The composition according to any one of claims 1 to 8, wherein 0.001 to 1 g of acacia bark polyphenol is orally ingested with respect to 1 kg of body weight of the intaker or ingested animal per day as an active ingredient amount. The composition according to any one of claims 1 to 9, which is a food. The composition according to any one of claims 1 to 9, which is an animal feed. The composition of claim 1, which is a medicament. The composition according to any one of claims 1 to 9, which is a quasi drug. The composition according to any one of claims 1 to 9, which is a cosmetic.