JP4847712B2 - Antioxidant composition containing acacia bark - Google Patents

Description

  The present invention relates to an antioxidant composition derived from a tree belonging to the genus Acacia, and uses of the antioxidant composition as foods, animal feeds, medicines, and quasi drugs.

  In general, in the atmosphere, oxygen is a stable substance called triplet oxygen. When some of the oxygen changes into highly reactive substances called active oxygen such as superoxide, hydroxy radicals, singlet oxygen, and hydrogen peroxide in vivo, some of the oxygen is produced by neutrophils and macrophages. While it plays a role in protecting the body against foreign substances, if excessive active oxygen is present in the body, it attacks lipids, proteins, enzymes, nucleic acids, etc., and damages biological membranes, tissues, etc. It has also been shown to cause onset such as disability, emphysema, rheumatoid arthritis, cataracts, hypertension, senile dementia, Alzheimer's disease, or blemishes or aging.

Conventionally, in order to prevent the above-mentioned various diseases caused by oxidative stress caused by these excessive active oxygens, a reaction that generates hydrogen peroxide and oxygen molecules by disproportionating superoxide (0 ₂ ⁻ ) as a starting material. There is a widespread search for antioxidants that add to foods SOD-like substances that act similarly to superoxide dismutase (SOD), an enzyme that catalyzes oxidative activity, and substances that trap active oxygen (active oxygen scavenging active substances). I came.

Examples of such substances include fat-soluble α-tocopherol (vitamin E) and water-soluble ascorbic acid (vitamin C) for substances derived from natural products, and BHT (butylhydroxytoluene) and BHA for synthetic substances. There are phenolic substances such as (butylhydroxyanisole) and TBHQ (tertiary butylhydroquinone).
However, although these substances are generally used as food antioxidants, they do not have a sufficient active oxygen scavenging action on the living body, and in particular, the synthetic substance BHA also causes problems such as suspected carcinogenicity. Is coming out.

  Thus, in recent years, research and development for searching for active oxygen scavenging active substances having high safety from natural products such as animals and plants has been actively conducted, for example, Patent Documents 1, 2, 3, 4, and 5. In addition, the products that have actually been commercialized include polyphenolic compounds such as tea leaves, catechins extracted from cacao bean extract, anthocyanin compounds contained in blueberry peels, isoflavones obtained from fermented soybeans, etc. Has attracted attention due to its extremely high active oxygen scavenging effect.

  Among these polyphenol compounds that have been shown to have a high active oxygen scavenging effect, (−) epigallocatechin, (−) epigallocatechin-3-gallate, (−) epicatechin, 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, tannin extracted from the bark of Acacia mearnsii De Wild., Which is a kind of Acacia genus, has been produced in large quantities in South Africa and Brazil since ancient times. It is used at a very low price as a tanning agent and an adhesive for wood. Also, the scientific name: Acacia mangium Willd. (Acacia mangium) grows very quickly, and in recent years, plantations have been flourishing in the subtropical to tropical regions including Indonesia and Malaysia. This walt tannin is classified as condensed tannin.

Recently, an extract of the genus Acacia has a selective inhibitory effect on COX-2 (Patent Document 6), and a whitening effect by an active oxygen scavenging effect (Patent Document 7) and an inhibitory effect on tyrosinase activity on an Acacia bark (Patent Document) 8) is disclosed.
However, it has not been known that an excellent antioxidant action can be expressed in vivo by ingesting acacia bark or polyphenol derived from bark at a specific dose.
Japanese Unexamined Patent Publication No. 01-25726 JP-A-6-65074 JP 7-300422 A Japanese Patent Laid-Open No. 11-5975 JP 2001-98264 A JP-T-2004-532811 JP 2004-352639 A Japanese Patent Laid-Open No. 10-025238

  The present invention provides a highly safe composition that exhibits excellent antioxidant action in vivo.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that an acacia bark-derived product exhibits an excellent antioxidant effect in vivo when taken at a specific dose. The present invention has been completed.
That is, this invention relates to the antioxidant composition characterized by including the substance derived from an Acacia bark at a specific dose.

ADVANTAGE OF THE INVENTION According to this invention, the composition which can exhibit the outstanding antioxidant effect by ingesting by a specific dose can be obtained.
In addition, the composition of the present invention is less likely to cause side effects even when taken for a long time, and has high safety.

The Acacia bark-derived product that can be used in the present invention is not particularly limited as long as it is obtained from the bark of a tree belonging to the genus Acacia (hereinafter referred to as “Acacia” or “Acacia genus”), For example, acacia bark debris, powder and suspensions thereof, extracts such as acacia bark extract, concentrated extract, and extract powder, and purified products obtained by purifying the extract include Can be mentioned. An extract of acacia bark, particularly an acacia bark polyphenol, is preferable from the viewpoint of obtaining excellent antioxidant activity.
In the present invention, only one kind of these Acacia bark-derived materials may be used, or two or more kinds thereof may be used in combination.

The acacia that can be used in the present invention is not particularly limited as long as it is a tree belonging to the genus Acacia, but the scientific name: Acacia mearnsii De Wild. (Bartle), scientific name: Acacia mangium Willd. (Generic name Acacia mangium), scientific name: Acacia dealbata Link., Scientific name: Acacia decurrens Willd., And scientific name: Acacia pycnantha Benth. Acacia mearnsii De Wild. And Acacia mangium Willd. Are particularly preferable.
In the present invention, only one kind of these Acacia barks may be used, or two or more kinds thereof may be used in combination.

The bark of the genus Acacia is usually obtained by cutting only a bark after harvesting it as a tree, and then collecting and drying it. Preferably, the bark is further dried in the sun.
Acacia bark consists of an outer skin and a slightly fibrous endothelium, and when dried to a moisture content of about 20% or less, it is easily pulverized with a pulverizer such as a hammer mill. In the present invention, as the acacia bark, both the inner skin and the outer skin of the acacia may be used together, or only one of them may be used.

The acacia bark strips can be obtained by pulverizing the acacia bark to an appropriate size according to a conventional method.
The powder of the acacia bark can be obtained by pulverizing the acacia bark by a conventional method. In particular, a powder having a particle size of 100 μm or less, particularly 50 to 70 μm is preferable. In the powder fractionation, bark dried to a moisture content of 20% or less is pulverized to an appropriate size, for example, a particle size of 1.6 mm or less, and the obtained powder is classified with a vibration sieve or the like to obtain the required powder. Obtainable.

The acacia bark extract can be obtained by extracting the acacia bark according to a conventional method. In order to obtain an extract of acacia bark having an excellent antioxidant action, it is preferable to extract the acacia bark with alcohol or a polar solvent.
Ethanol can be used as such an alcohol, and water can be used as a polar solvent, and these solvents can be used alone or in combination of two or more as required. In particular, in order to obtain an excellent antioxidant action, a mixed solvent of water and an alcohol such as ethyl alcohol is preferable.
Furthermore, the extraction operation may be performed multiple times with the same or different solvents.
In order to obtain an extract having an excellent antioxidant action, an extract obtained by further extracting an extract from water or hot water from acacia bark with ethanol may be used.

Extraction is carried out by adding a solvent to acacia bark strips or powder and stirring as necessary, but the temperature, time, or solid-liquid ratio is not particularly limited. When water is used as the solvent, it may be extracted 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 in various forms such as an extract, a solution, a powder, a concentrated solution, and a paste, and can be widely used as needed.
Further, the acacia bark extract of the present invention obtained in these forms can be used as it is as an antioxidant composition, and further purified as necessary, and the purified product can be used as an antioxidant active ingredient. it can.

In the present invention, components contained in the bark of the genus Acacia are also exemplified as the acacia bark-derived material. Examples of such components include acacia bark polyphenols. In particular, acacia bark polyphenol is a preferred component because it exhibits an excellent antioxidant action.
The acacia bark polyphenol of the present invention means a kind of condensed tannin which is a polymer of flavanols having flavan-3-ol as a basic skeleton. Here, such a condensed tannin preferably has a molecular weight of 500 to 3,000. The acacia bark polyphenol used in the present invention can be obtained by hot water extraction of the acacia bark powder and the like.
Acacia bark polyphenol products include registered trademarks of MIMOSA Central Co-operative Ltd. Examples include MIMOSA AL POWDER, MIMOSA CR POWDER, and GOLDEN MIMOSA POWDER.

  The composition of the present invention may be an acacia bark-derived material such as an acacia bark, an extract thereof, a purified product thereof, or an acacia bark polyphenol itself, but other substances having an antioxidant action, such as Vitamin C, Vitamin E, Coenzyme Q10, α-tocopherol, proanthocyanin, isoflavone, quercetin, green tea, or wheat germ. In particular, it is preferable to contain coenzyme Q10, green tea, or wheat germ in terms of obtaining an excellent antioxidant effect due to a synergistic effect.

  The composition of the present invention may be an acacia bark-derived material, for example, an acacia bark, an extract thereof, a purified product thereof, or an acacia bark polyphenol itself, but as long as the effects of the present invention are not impaired, It may contain additives, ingredients generally used in pharmaceuticals and foods, and other preparations, sweeteners, acidulants, thickeners, fragrances, pigments, emulsifiers, and the like.

  The composition of the present invention can be used for the prevention or treatment of diseases involving active oxygen. In the present invention, active oxygen-related diseases are cell or tissue disorders caused by active oxygen such as superoxide, hydroxy radical, singlet oxygen, or hydrogen peroxide, or lipid peroxide oxidized by these. If there is no particular limitation, arteriosclerosis, cerebrovascular disorder, pulmonary emphysema, rheumatoid arthritis, cataract, hypertension, senile dementia, Alzheimer's disease, stain / sobacas or aging are exemplified. Arteriosclerosis is preferred.

The intake amount of the composition according to the present invention is not particularly limited, but can be appropriately selected according to the dosage form and the age, weight and symptoms of the intake person or intake animal such as the user or patient. For example, the amount of the active ingredient is 0.005 to 1 g, preferably 0.005 to 0.5 g, more preferably 0.005 to 0.1 g in terms of the amount of acacia bark polyphenol per 1 kg of the body weight of the ingestor or ingestion animal per day. Ingestion, particularly by oral intake, provides an excellent antioxidant effect.
The intake period can be arbitrarily determined according to the age and symptoms of the intake person or animal.

  The composition according to the present invention can be used as a food or animal feed, for example, as a health food, a functional food, a health supplement, a specific health food, a beauty food, and a nutritional supplement (supplement). These foods and animal feeds may be in the form of, for example, drinking water such as tea and juice; and ice cream, jelly, candy, chocolate, chewing gum and the like. Moreover, the form of a liquid agent, a powder agent, a granule, a capsule, or a tablet may be sufficient. Here, animals for animal feed include all animals that require prevention or treatment of the above-mentioned diseases involving active oxygen, including pet animals, livestock animals, and animals raised in zoos, etc. It is.

  The composition according to the present invention is orally administered in the form of, for example, a powder, a tablet, a coated tablet, a sugar-coated tablet, a hard or soft gelatin capsule, a liquid, an emulsion, or a suspension as a medicine or quasi drug. It can also be administered rectally, for example in the form of suppositories; it can also be administered parenterally, eg locally or transdermally in the form of ointments, creams, gels or solutions.

Hereinafter, the present invention will be described in more detail with reference to Production Examples, Test Examples, and Formulation Examples, but the present invention is not limited thereto. In particular, the embodiment shows the acacia bark of the present invention without dividing the bark into the outer skin and the endothelium, but the outer skin can be used separately from the inner skin.
In the following production examples and test examples, each acacia of the present invention is indicated by a number shown in parentheses after each scientific name. For example, an acacia with the scientific name: Acacia mearnsii De Wild. 1 is written.
Scientific name: Acacia mearnsii De Wild. (No. 1), Scientific name: Acacia mangium Willd. (No. 2), Scientific name: Acacia dealbata Link. (No. 3), Scientific name: Acacia decurrens Willd. (No. 4), Scientific name : Acacia pycnantha Benth. (No. 5).

Production example 1 of Acacia bark powder
Acacia No. 1 bark was dried to a moisture content of 20% or less, and the dried bark was pulverized to a powder of 1.6 mm or less (10 mesh sieve (Tyler) passed) with a hammer mill (made by Raymond), and then further sieved. The fine powder of 63 micrometers or less (under 250 mesh sieve) was obtained.
Similarly, the remaining four Acacia No. 2 to 5 bark were pulverized to obtain a fine powder of 63 μm or less. Although there were some differences in the yield of the fine powder that passed through the 250 mesh sieve depending on the type, the desired fine powder was obtained.

Production example 2 of Acacia bark extract
Each Acacia No. of the present invention. Each bark of 1 to 5 was dried to a moisture content of 20% or less, and the dried bark was pulverized to a powder of 1.6 mm or less with a hammer mill, and then 5 times the amount of hot water was added to 100 g of the dry crushed bark. After boiling, the mixture was extracted for 15 minutes and filtered using a 10 to 20 μm filter. The obtained filtrate was spray-dried with a spray dryer to obtain 40 g of bark extract.
Hereinafter, each bark extract is acacia no. 1-5 hot water extract.

Production Example 3 of Acacia Bark Extract
The acacia bark of the present invention is dried to a moisture content of 20% or less, the dried bark is pulverized to a powder of 1.6 mm or less with a hammer mill, and then 5 times the amount of ethanol is added to 100 g of the dried crushed bark. Extraction was performed for 15 minutes while boiling and refluxing, followed by filtration using a 10-20 μm filter. After evaporating ethanol from the obtained filtrate, the concentrate was spray-dried with a closed spray dryer to obtain 40 g of a bark extract (hereinafter referred to as Acacia No. 1 ethanol extract).
Acacia No. 1 to 5 ethanol extracts were obtained.

Production Example 4 of Acacia Bark Extract
Three times the amount of ethanol was added to 10 g of the acacia hot water extract obtained in Production Example 2, and the mixture was boiled and refluxed for 15 minutes, and filtered using a 10 to 20 μm filter. Ethanol was evaporated from the obtained filtrate, and water was added thereto, followed by freeze-drying to obtain 9 g of an extract (hereinafter referred to as an Acacia No. 1 hot water extract ethanol fraction).
Acacia No. 1 to 5 hot water extract ethanol fractions were obtained.

Test Example 1 Antioxidation test (1) Preparation of test sample Add water for injection (Otsuka Pharmaceutical Factory Co., Ltd.) to 1.50 g of sodium carboxymethylcellulose (CMC-Na, Nacalai Tesque Co., Ltd.) and dissolve to dissolve the total amount. The solution was adjusted to 500 mL, and a 0.3% (W / V) CMC-Na solution was prepared.
Acacia No. 1 described in Production Example 2 6.0 g of 1 hot water extract was weighed, finely ground in a mortar, suspended in the above 0.3% (W / V) CMC-Na solution to make a total volume of 15 mL, and a test sample was prepared.

(2) Test animals Rats (Slc: SD, SPF, male, 6 weeks old) were purchased from Japan SLC Co., Ltd., and were subjected to a test after a quarantine / acclimation period of 8 days.
During the quarantine / acclimation period, the general condition was observed once a day, and the body weight was measured the day after the arrival of the animals and the end of the quarantine / acclimation period.
Based on the body weight at the end of quarantine and habituation, 8 mice were divided into a control group and a test group so that the average body weight of each group became equal by a completely random extraction method using a computer.
The animals were set to a temperature of 20 to 26 ° C., a relative humidity of 40 to 70%, a ventilation rate of 10 to 20 times / hour, and a lighting time of 12 hours (7: 0 to 19:00) throughout the quarantine / acclimation period and the test period. In a clean breeding room, they were raised in a stainless steel cage installed on a stainless steel hanger rack. The number of animals per cage was 2-3.
In addition, the feed was freely ingested commercially available solid feed (F-2, manufactured by Funabashi Farm Co., Ltd.) throughout the quarantine / acclimation period and test period. As drinking water, tap water was freely consumed by a water bottle.

(3) Test method 0.3% (W / V) CMC-Na solution for the control group and the test sample prepared above for the test group at a dose of 5 mL / kg each day It was orally administered once for 14 days.

(4) Observation and examination items (i) Measurement of serum 8-OHdG (8-hydroxy-2'-deoxyguanosine) On the 14th day from the start of administration, about 1 mL of blood was collected from the jugular vein without anesthesia, The collected blood was centrifuged at 3000 rpm for 15 minutes to obtain serum. The 8-OHdG concentration in the obtained serum was measured using a highly sensitive 8-OHdG Check (manufactured by Nikken Zeil Co., Ltd.).
(Ii) Measurement of urinary 8-OHdG (8-hydroxy-2'-deoxyguanosine) Rats were metabolically caged for about 16 hours from 5:00 pm on the day of blood collection to 9:00 am on the following day. And urine was collected. The collected urine was centrifuged at 1500 rpm for 5 minutes to obtain a supernatant. The 8-OHdG concentration in the obtained supernatant was measured using New 8-OHdG Check (manufactured by Nikken Zeil Co., Ltd.).

(5) Statistical method Each of the above measured values was expressed as an average value ± standard error. Student's t-test was performed as a significant difference test from the control group. Significance level is expressed as 5%.

(6) Test results There were no deaths or abnormal general conditions in all cases.
Serum and urine 8-OHdG concentrations were decreased in the test group compared to the control group, and in particular, the urinary 8-OHdG concentration was significantly decreased in the test group compared to the control group. (Table 1).

  From the above results, it was confirmed that acacia bark polyphenol has an effect of lowering 8-OHdG in serum and urine, and it was confirmed that it exhibited an antioxidant effect in vivo by oral administration.

Test Example 2 Acute toxicity test (oral administration)
An oral acute toxicity test using mice was performed according to OECD (Guidelines for the Testing of Chemicals 401, 1987).
For ICR male and female mice, specimens of 9 doses each with a tolerance of 500 mg / kg, up to 7,000 mg / kg for males and 6,500 mg / kg for females (Acacia No. 1 hot water extract of Production Example 2 above) Was orally administered to male and female mice once. As the specimen, the product of the present invention was suspended in pure water. As a result, the LD50 value was 4,468 mg / kg for males and 3,594 mg / kg for females, and no death was observed at a dose of 3,000 mg / kg.
The above test was conducted using the Acacia No. Similar results were obtained with 2-5 hot water extracts. Thereby, it turned out that the composition of this invention is excellent in safety | security.

Test Example 3 Mutagenicity Test A mutagenicity test was conducted according to Ministry of Labor Notification No. 77 (September 1, 1988).
Using the Escherichia coli WP2 uvrA strain and the Salmonella typhimurium TA strain 4 strains, a reverse mutation test including metabolic activation was conducted at a dose of 156 to 5,000 μg / plate (Acacia No. 1 to 5 hot water of Production Example 2). Extract). As a result, no increase in the number of revertant colonies was observed for any of the strains. It was concluded that mutagenicity of 1-5 hot water extracts was negative, and it was found to be excellent in safety.

Formulation Example 1 Preparation of internal use Using the acacia bark hot water extract ethanol fraction of Production Example 4, an internal preparation was prepared with the composition shown below.
Extract fraction of Production Example 4 1.0 (% by weight)
Lactose 30.0
Cornstarch 60.0
Crystalline cellulose 8.0
Polyvinylpyrrolidone 1.0
Total 100.0

Formulation Example 2 Preparation of Pet Food Using the acacia bark hot water extract of Production Example 2, a pet food was prepared with the composition shown below.
Extract of Production Example 2 1.0 (% by weight)
Oatmeal 88.0
Starch 5.0
Salt 2.5
Whole egg 3.0
Seasoning 0.5
Total 100.0

Formulation Example 3 Preparation of Tablet (Confectionery) Using the ethanol fraction of Acacia bark hot water extract of Production Example 4, tablets (confectionery) were prepared with the composition shown below.
Extract fraction of Production Example 4 1.0 (% by weight)
Citric acid 1.0
Nonfat dry milk 15.0
Sucrose ester 1.0
Flavor 0.5
Powdered sugar 20.0
Lactose 61.5
Total 100.0

According to the present invention, a composition for preventing and / or treating a disease associated with active oxygen can be obtained.
More specifically, the composition of the present invention is useful for the prevention and / or treatment of arteriosclerosis.
These compositions can be used for pharmaceuticals or quasi drugs, foods such as health foods, health supplements, foods for specified health, or dietary supplements, or animal feed.

Claims (3)

Scientific name:. Acacia mearnsii De Wild, scientific name:. Acacia mangium Willd, scientific name: Acacia dealbata Link, scientific name:.. Acacia decurrens Willd, and scientific name: the Acacia pycnantha Benth bark of genus Acacia selected from the group consisting of. An antioxidant for oral consumption , characterized in that it contains 0.005 to 1 g of acacia bark polyphenol per day per kg of the intaker's body weight and does not contain free B-ring flavonoids as active ingredients . The antioxidant of Claim 1 whose acacia genus bark origin is an extract of the acacia bark. The antioxidant of Claim 1 whose acacia bark origin thing is an acacia bark polyphenol.