JP2000297042A - Active oxygen scavenger - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an active oxygen scavenger originated from a neutral product, high in safety and useful as a component of a health food having an active oxygen scavenging activity by adding an extract of matico. SOLUTION: This scavenger contains an extract of matico. Matico is a plant of Piperaceae Piper, and the plant body or its crushed product (either raw or dried material) can be used for the extraction. A method for obtaining the extract comprises, for example, suitably crushing a dried product of leaves, stems, buds or the like of matico as the raw material, and subsequently extracting the crushed product with water, alcohol, a hydrated alcohol or the like. The active oxygen scavenger contains the obtained extract of matico, for example, in an amount of 0.001-100 (w/w)%, preferably 0.01-30 (w/w)% in terms of a dried matter. Commonly speaking, the daily dosage is, for example, about 1-5,000, preferably about 100-3,000 mg/adult of the extract of matico (dried product).

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 more particularly to a novel active oxygen scavenger derived from natural products and having high safety, and a health food having an active oxygen scavenging action. Things.

[0002]

2. Description of the Related Art Superoxide anion radicals, hydrogen peroxide, hydroxy radicals, singlet oxygen, and peroxy radicals and alkoxy radicals generated by peroxidation of lipids are known as active oxygens generated in vivo. Reactive oxygen causes damage to tissues due to crosslinking of collagen fibers, DNA damage, and generation of chain radicals in the living body, causing aging of skin and living bodies, inflammation,
It is known that the myocardial infarction causes damage to smooth muscle, exacerbation of liver damage, and dementia due to destruction of brain tissue.

[0003] Therefore, elimination of active oxygen generated in a living body is considered to be important for prevention or treatment of these diseases. From such a viewpoint, a search for a substance that eliminates active oxygen has been widely performed. For example, substances that eliminate active oxygen include those derived from natural substances such as tocopherol and ascorbic acid, BHT (3,5-di-t-butyl-4-hydroxytoluene), and BHA (2
(3) -t-butyl-4-hydroxyanisole). However, these substances have insufficient active oxygen scavenging activity or have problems in safety and the like.

[0004] Piper aduncum
Is commonly called "Matico" (hereinafter referred to as "Matico"), a plant mainly produced in South America and the like, and the plant is usually used as a hemostatic agent.
The decoction is said to be effective for diseases of the chest and pharynx, ie, pneumonia and pharyngitis. Furthermore, the obesity prevention effect is also known (Japanese Patent Application Laid-Open No. 62-81).
No. 324). However, its active oxygen scavenging action is not known.

[0005]

DISCLOSURE OF THE INVENTION The present invention has an activity of sufficiently eliminating active oxygen generated in a living body, is contained in a large amount in plants, is easy to manufacture and process, and has high safety. It is an object of the present invention to provide an active oxygen scavenger containing the substance as an active ingredient, and a health food having an active oxygen scavenging action.

[0006]

Means for Solving the Problems In order to solve the above-mentioned problems, the present inventors have conducted intensive studies on the biological functions of various edible plant extracts. The present inventors have shown that they exhibit oxygen scavenging activity and have high safety, and have completed the present invention based on these findings.

[0007] That is, the present invention is an active oxygen scavenger comprising an extract of Matico as an active ingredient,
The extract is an active oxygen scavenger, an extract of Matico extracted with either water, alcohol or aqueous alcohol. Further, the present invention is a health food having an active oxygen scavenging action, which contains an extract of Matico.

[0008]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.

The raw material used in the present invention, Matico, is a plant belonging to the genus Pepper of the family Pepper, and its plant body or its crushed material (they may be raw or dried) can be used.

[0010] The active ingredient is obtained from matico by extracting its leaves, stems, buds and the like with water or an organic solvent or a mixture thereof. Examples of the organic solvent include methanol, ethanol, isopropanol, N, N-dimethylformamide, acetone, ethyl acetate, chloroform, dichloromethane, pentane,
Hexane and heptane can be mentioned. Among them, hot water, alcohols such as methanol and ethanol, and hydrated alcohols thereof are particularly preferable in terms of extraction rate and safety.

In order to obtain an extract using the above-mentioned solvent,
Although any method may be used, for example, a dried material such as leaves, stems, and buds of matico is used as a raw material, and after appropriately crushing the crushed material, the crushed material is subjected to a known method using the solvent described above. To process. Specifically, the solvent is used in an amount of 1 to 100 times (weight ratio), preferably 3 to 20 times (weight ratio) the amount of the raw material, at a temperature of 0 ° C. or higher, preferably 10 ° C. to the boiling point of the solvent or lower. 1 minute to 8 weeks with stirring under conditions
Preferably, it can be obtained by performing the extraction treatment for 10 minutes to 1 week. If necessary, a method of extracting while heating and refluxing may be used.

The extracted product obtained as described above may be used in the present invention as it is. Preferably, the organic solvent used for the extraction is removed by a usual method, for example, using a rotary evaporator under reduced pressure. Is good. The organic solvent-removed product may be subjected to a conventional drying treatment such as freeze-drying and heat-drying, if necessary.

The above-mentioned extract of Matico has a strong elimination effect on superoxide anion radicals and hydroxy radicals as shown in Experimental Examples 1 and 2 described later. And, as shown in Experimental Examples 3 and 4 described later, this extract of Matico is extremely high in safety.

Therefore, the extract of Matico as it is,
Alternatively, a product containing the extract thereof can be used as an active oxygen scavenger or a health food which can be expected to have an effect of eliminating active oxygen in a living body or an effect of suppressing generation of active oxygen species.

[0015] The active oxygen scavenger of the present invention can be prepared by converting the extract of matico into a dry matter, for example, from 0.001 to 100%
(W / w), preferably 0.01 to 30% (w / w)
It is used by containing. Alternatively, an appropriate emulsifier, stabilizer, extender, or the like may be mixed with the extract of Matico alone or as a mixture with another drug.
If necessary, an appropriate excipient, such as lactose, starch, or fat, may be added to the extract of Matico. Further, the active oxygen scavenger can be administered by any of oral, external use and injection methods. As the oral preparation, any dosage form such as a powder, a tablet, a granule, a capsule, a syrup and the like can be adopted. External preparations include emulsions,
Ointments and the like are preferred. As an injection, a physiological saline solution of an extract of Matico is preferably used.

The dose of the active oxygen scavenger of the present invention varies depending on the condition to be treated, age, body weight, etc., for example, usually, 1 to 5000 mg of Matico extract (dry matter) per adult per day. Preferably 100-3000mg
It is about.

When the extract of Matico is used as a health food, for example, sweeteners such as sugars, acidulants such as citric acid and lactic acid, etc., as well as proteins, lipids, highly unsaturated fatty acids, vitamins, minerals, etc. In addition, the content may be the same as the content of the active oxygen scavenger described above.
The shape at that time is not particularly limited, tablet, pill, powder, powder, granule, capsule, aqueous solution,
It can be in the form of a suspension, syrup or the like.

Further, as shown in Experimental Example 5 described below, the extract of Matico has an effect of preventing gastric ulcer induced by administration of indomethacin beforehand, and is useful as a therapeutic agent for gastric ulcer.

The active oxygen scavenger containing the extract of Matico and the health food containing the extract of Matico have an active oxygen scavenging effect and a gastric ulcer treatment effect at the same time.

[0020]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference examples, experimental examples, and examples. The technical scope of the present invention is:
They are not limiting in any way. Reference Example 1 (Production of Matico Extract) 16 L of water was added to 2 kg of dried material such as leaves, stems and buds of Matico, heated, and extracted at 90 ° C. for 1 hour with stirring. The residue obtained by removing the residue by suction filtration using a glass filter was concentrated under reduced pressure by a rotary evaporator, and then freeze-dried to obtain 440 g of Matico extract.
I got The obtained Matico extract was subjected to the following experiments and implementations. Experimental Example 1 (Measurement of Superoxide Anion Radical Scavenging Activity) Using the extract (dry product) of Matico obtained in Reference Example 1, the superoxide anion radical scavenging activity was measured as follows (K. Mitsuya , et al., Bull.Chem.S
oc. Jpn., 63, 187-191, 1990).

As a substrate solution, hypoxanthine (Sig
ma) (manufactured by Ma Corporation) to 100 mM HEPE
S (N-2-hydroxyethylpiperazine-N'-2-
Ethanesulfonic acid) buffer solution (pH 7.4) was used, and the enzyme solution was xanthine oxidase (Sig
(made by Ma Co., Ltd.) dissolved in a 100 mM HEPES buffer solution (pH 7.4) to a concentration of 0.4 U / ml. Superoxide anion radical generated by hypoxanthine-xanthine oxidase system is DMP
O (5,5-dimethyl-1-pyrroline-N-oxide)
And quantified by a JER-FR30 type electron spin resonance apparatus (manufactured by JEOL Ltd.). In this system, the extract of Matico dissolved in water was added at various concentrations (0.1, 0. 1).
(0.01, 0.001, 0.0001%). The superoxide anion radical scavenging activity was defined as the reduction rate of the superoxide anion radical generation amount at this time.

The results are as shown in FIG. 1. The extract of Matico eliminated the superoxide anion radical generated by the hypoxanthine-xanthine oxidase system in a concentration-dependent manner, and showed a strong active oxygen scavenging effect. . Experimental Example 2 (Measurement of hydroxy radical scavenging activity) The hydroxy radical scavenging activity was measured as follows using the extract (dried product) of Matico obtained in Reference Example 1 (N. Ogawa, et al. , Brain Research, 657,207-21
3,1994).

To a 100 mM HEPES buffer solution (pH 7.4) containing 2 μM ferrous sulfate and 1.1 μM diethylenetriaminepentaacetic acid, 0.01% hydrogen peroxide was added, and the hydroxyl radical generated by the Fenton reaction was removed by DM.
It was trapped by PO and quantified in the same manner as in Experimental Example 1. In this system, the extract of Matico dissolved in water was added at various concentrations (0.1, 0.01, 0.001, 0.0001).
%), And the reduction rate of the amount of hydroxy radical generated at this time was defined as hydroxy radical scavenging activity.
The results are as shown in FIG. 2. The extract of Matico eliminated hydroxy radicals generated by the Fenton reaction in a concentration-dependent manner, and showed a strong active oxygen scavenging effect. Experimental Example 3 (Single Dose Toxicity Test) A single dose toxicity test was performed according to the research report guidelines of the Pharmaceutical Toxicity Laboratory. That is, F344 / DuCrj of 5 weeks old
Rats (5 males and 5 females) were orally administered with Matico extract (dried product) at a rate of 4 g / kg and observed for 16 days.

As a result, none of the animals died, no side effects were observed, and no microscopic abnormalities in tissues and organs were observed at necropsy on the 16th day. The extract of Matico was found to be very low in toxicity. Experimental example 4 (mutagenicity test) According to the guidelines of the Industrial Safety and Health Law, Salmonella typhi
A mutagenicity test was performed by the Ames method using murium TA100 strain and TA98 strain. That is, various weights of extract of matico (dried, 5000, 2500, 1250, 625, 3
33 μg), a phosphate buffer solution or S9Mix was added, the pre-cultured cells were added, the mixture was incubated at 37 ° C. for 20 minutes, and then soft agar was added to spread on a minimal glucose agar plate. After culturing at 37 ° C. for 48 hours, the number of revertant colonies that appeared was counted.

As a result, the remarkable increase in the number of revertant colonies was not observed in the TA100 strain and the TA98 strain irrespective of the presence or absence of metabolic activation by S9Mix. The product was found to be very low mutagenic. Experimental Example 5 (Effect on Indomethacin-Induced Gastric Ulcer) In the test group, the extract (dried product) of Matico obtained in Reference Example 1 was used as a comparative group, and commercially available cetraxate and cimetidine, which are therapeutic agents for gastric ulcer, were used for 24 hours. Fasted 6-week-old Wistar male rats weighing 170-190 g
Each animal was orally administered 200 mg each. Untreated rats served as control groups. After 30 minutes, 1% sodium carboxymethylcellulose and 0.1% Tween 2
50 mg of indomethacin suspended in
/ Kg subcutaneously. Seven hours later, the animals were exsanguinated from the abdominal aorta under Nembutal anesthesia. The stomach is excised and the esophagus and duodenum are ligated with arterial klemme.
10 ml of phosphate buffered saline containing 0% formalin was injected. Thirty minutes later, the stomach was incised along the great bay, and the length of the major axis of the bleeding spot due to the ulcer in the glandular stomach was measured, which was used as the damage factor. The results are as shown in FIG. 3. The extract of Matico suppressed the increase in the area of ulcer occurrence induced by indomethacin administration. Also, its anti-gastric ulcer activity,
The strength was equivalent to that of the commercially available gastric ulcer treatment cetraxate or cimetidine. Example 1 (Production of powder) The extract (dried product) of matico obtained in Reference Example 1, lactose and crystalline cellulose were mixed in the following composition, kneaded with purified water, and then kneaded by a conventional method. 1 kg of powdered or finely divided powder was produced. <Composition of Powder> Matico Extract 200 g Lactose 600 g Crystalline Cellulose 200 g This powder is an active oxygen scavenger containing 200 mg of Matico extract in one packet per 1 g. Example 2 (Production of Syrup) The extract (dried product) of Matico obtained in Reference Example 1 was made into a fine powder in a mortar, suspended in sterile purified water, and cherry syrup was gently added to the suspension. Add with stirring 1
A syrup having the following composition was prepared to be 200 ml. <Composition of syrup> Matico extract 48 g Cherry syrup 1000 ml Sterile purified water Residue This syrup can be taken 1 to 3 times per day, 5 ml per dose and 200 mg of Matico extract containing active oxygen elimination Agent. Example 3 (Production of powdered health food) 1 kg of a powdered health food having an active oxygen scavenging action having the following composition was produced by containing the extract (dried product) of Matico obtained in Reference Example 1.

As the soybean separated protein in the composition, "New Fuji Pro 1200" manufactured by Fuji Purina Protein Co., Ltd. was used. <Composition of health food> Matico extract 8.0 g Soybean isolated protein 946.446 g Hydrous crystalline glucose 30.3 g Dextrin 3.0 g Vitamin B ₁ 0.05 g Vitamin B ₂ 0.06 g Vitamin B ₆ 0.15 g Vitamin B ₁₂ 0.00018 g Folic acid 0.024 g Vitamin A 0.22 g Ascorbic acid 2.0 g α-dl-tocopherol 0.75 g Calcium lactate 9.0 g By eating 25 g of this health food a day, 200 mg of Matico extract is taken. be able to. Example 4 (Production of Soft-Capsulated Health Food) A soft capsule-shaped health food having the following composition and having an active oxygen-eliminating action was produced by containing the extract (dried product) of Matico obtained in Reference Example 1. <Composition of health food (content per capsule)> Matico extract 10.0 mg β-carotene 20.0 mg α-dl-tocopherol 0.75 mg DHA 30.0 mg olive oil 939.25 mg By eating 1 to 30 a day, 10 to 300 mg of Matico extract
Can be taken.

[0027]

Industrial Applicability The active oxygen scavenger of the present invention containing the extract of Matico as an active ingredient has remarkable superoxide anion radical scavenging activity and hydroxy radical scavenging activity. In addition, the Matico extract is highly safe in use. Therefore, the active oxygen scavenger of the present invention is extremely useful for the prevention and improvement of various diseases involving active oxygen. Furthermore, the Matico extract of the present invention can be used for pharmaceuticals, health foods and the like, and the present invention is extremely useful industrially.

[Brief description of the drawings]

FIG. 1 is a diagram showing the relationship between the concentration (%) of an extract (dried product) of Matico and the superoxide anion radical scavenging activity (%) in Experimental Example 1.

FIG. 2 is a diagram showing the relationship between the concentration (%) of an extract (dried matter) of Matico and the hydroxyl radical scavenging activity (%) in Experimental Example 2.

FIG. 3 is a graph showing the damage factor (mm) of the glandular stomach induced by administration of indomethacin in the group administered with the extract of Matico (dry matter), the group administered with cimetidine, the group administered with cetraxate, and the control group in Experimental Example 5.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Minoru Saito 339 Noda, Noda-shi, Chiba Prefecture Kikkoman Corporation (72) Inventor Hiroharu Ishikawa 339 Noda, Noda-shi, Chiba Prefecture Kikkoman Corporation (72) Inventor Nagasaki Katsuko 339 Noda, Noda City, Chiba Prefecture Kikkoman Corporation (72) Inventor Shigeru Nagano 339 Noda City, Noda City, Chiba Prefecture Kikkoman Corporation (72) Inventor Toshiaki Ariga 339 Noda, Noda City, Chiba Prefecture Kikkoman Corporation (72 ) Inventor Koichiro Tobe 339 Noda, Noda-shi, Chiba F-term (reference) 4B018 MD61 ME06 MF01 MF05 MF06 4C088 AB36 AC05 BA08 CA05 CA06 CA07 CA08 CA09 MA52 NA14 ZA68 ZC21

Claims (4)

[Claims] 1. An active oxygen scavenger comprising a matico extract as an active ingredient. 2. The active oxygen scavenger according to claim 1, wherein the extract of Matico is an extract of Matico extracted with any of water, alcohol and hydrous alcohol. 3. A health food having an active oxygen scavenging effect, comprising a extract of matico. 4. The health food according to claim 3, wherein the extract of Matico is a extract of Matico extracted with any of water, alcohol and hydrous alcohol.